Recognition of murine integrin beta 1 by a rat anti-stromal cell monoclonal antibody.
Previous studies with the rat monoclonal antibody KMI6 had localized its antigen in vivo to a discrete subpopulation of marrow stromal cells. The KMI6 antigen has now been identified as the murine homolog of integrin beta 1 by amino acid sequence analysis and by cross-reactivity with antiserum to the avian integrin beta 1. The relative tissue abundance of murine integrin beta 1 was determined by Western blot. Although immunoperoxidase staining of fixed murine hematopoietic tissues demonstrated an abundance of intracellular beta 1, few primary-derived cells of lymphohematopoietic origin were surface positive as assessed by immunofluorescence and flow cytometry. Fetal erythroblasts provided the only exception. In contrast, the antigen was readily detected on the surface of several cultured cell lines in association with a variety of alpha chains. The biochemical properties of the surface labeled murine integrin beta 1 were similar to those of its human counterpart, exhibiting an altered electrophoretic migration under reduced conditions or following N-glycanase treatment. The antibody recognition of the protein was insensitive to glycosylation state, presence of divalent cations, detergents, or transfer to a nitrocellulose membrane. However, on Western blot, the epitope was lost on reduction of the protein, suggesting that it is conformation dependent. These data indicate that although KMI6 epitope is widely distributed, its surface expression in vivo may be restricted within lymphohemopoietic tissues.


Perioperative outcomes for laparoscopic and robotic compared with open prostatectomy using the National Surgical Quality Improvement Program (NSQIP) database.
To examine contemporary outcomes of minimally invasive radical prostatectomy (MIRP) compared with open prostatectomy, using a national, prospective perioperative database reflecting diverse practice settings. The National Surgical Quality Improvement Program database was queried from 2005 to 2010 for laparoscopic or robotic prostatectomy (Current Procedural Terminology code 55866) and open retropubic prostatectomy (Current Procedural Terminology codes 55840, 55842, 55845). Perioperative outcomes examined were surgical and total operation duration, transfusion rates, length of stay, major morbidity (cardiovascular, pulmonary, renal, and infectious), and mortality. The study identified 5319 radical prostatectomies: 4036 MIRP and 1283 open. Although operative time was significantly longer in the MIRP group, there were significantly fewer perioperative blood transfusions and shorter mean length of stay. Major postoperative morbidity and mortality were 5% in the MIRP group and 9% in the open group (P <.001). Age, body mass index, presence of medical comorbidities, and open surgical technique were all independently predictive of major complications and mortality on multivariate analysis. In a nationwide database of diverse medical centers, MIRP was associated with longer operative time, but a significantly decreased rate of blood transfusions, length of stay, perioperative complication rate, and mortality compared with open prostatectomy. The minimally invasive surgical approach was independently associated with significantly fewer complications and deaths on multivariate analysis. Compared with other administrative databases that capture only inpatient events, the National Surgical Quality Improvement Program identifies complications up to 30 days postoperatively, providing more detailed characterization of complications after prostatectomy. These data reflect contemporary practice patterns and suggest that MIRP can be performed with low perioperative morbidity.


Effects of severe mental illness on survival of people with diabetes.
People with mental health problems are more likely to die prematurely than the general population but no study has examined this in individuals with diabetes. To compare survival rates in people with diabetes with and without schizophrenia or bipolar disorder. A total of 43,992 people with diabetes were drawn from the QRESEARCH database population of over 9 million patients. Survival rates during the study period, between 1 April 2000 and 1 April 2005, and hazard ratios for deaths associated with schizophrenia and bipolar disorder were adjusted by age and gender and additionally for socioeconomic status, obesity, smoking and use of statins. Among the participants, we identified 257 people diagnosed with schizophrenia, 159 with bipolar disorder and 14 with both conditions. Although crude survival rates did not show significant differences between the groups during the study period, people with schizophrenia or bipolar disorder and diabetes, compared with those with diabetes alone, had a significantly increased risk of death after adjusting for age and gender, with hazard ratios for schizophrenia of 1.84 (95% CI 1.42-2.40) and for bipolar disorder of 1.51 (95% CI 1.10-2.07). After adjusting for the other factors, hazard ratios were 1.52 (95 CI 1.17-1.97) for schizophrenia and 1.47 (95% CI 1.07-2.02) for bipolar disorder. People with schizophrenia or bipolar disorder in addition to diabetes have a relatively higher mortality rate. This suggests that diabetes either progresses more rapidly or is more poorly controlled in these individuals, or that they have higher levels of comorbidity and so are more likely to die of other causes.


Assessment of fitness in patients with cystic fibrosis and mild lung disease.
Maximal exercise testing is used in patients with cystic fibrosis to assess functional status and prognosis. The lactate threshold is an index of aerobic fitness with significant advantages over maximal exercise tests. This study was undertaken to determine if the lactate threshold might be identified, non-invasively, in adult patients with cystic fibrosis and mild lung disease by measurement of ventilatory and gas exchange parameters. Ten subjects with mild cystic fibrosis (forced vital capacity (FVC) > 70% predicted) and 10 healthy controls undertook an incremental exercise test on a bicycle ergometer. Ventilation and gas exchange parameters were measured continually and arterialised venous blood pH, carbon dioxide tension (PCO2), and lactate concentrations were measured at intervals throughout the tests. In subjects with cystic fibrosis there was no significant difference between the mean gas exchange and lactate thresholds (mean difference 1.0 (95% confidence interval (CI) of the mean -1.5 to 3.44) ml/kg/min). In contrast, there was a significant difference between the mean ventilatory and lactate thresholds (3.8 (95% CI 0.9 to 6.7) ml/kg/min). Arterialised venous PCO2 increased significantly during the exercise tests. In healthy subjects the mean differences between these thresholds were not significantly different from zero and PCO2 fell significantly during the tests. The ventilatory threshold significantly overestimates the lactate threshold in subjects with cystic fibrosis induced lung disease because of impaired carbon dioxide excretion during exercise. However, the gas exchange threshold may be used to determine the lactate threshold in this patient group.


The biology of epidermal growth factor receptor and its value as a prognostic/predictive factor.
Receptor tyrosine kinases play a major role in human carcinogenesis. Overexpression of the epidermal growth factor receptor (EGFR) has been associated with poor clinical outcome in several types of cancer. In principle, as with HER2, the EGFR status of a tumor should predict the likelihood of response to EGFR-targeted therapy. However, clinical data have failed to demonstrate a relationship between EGFR expression and response to the EGFR-targeted compounds cetuximab, gefitinib and erlotinib. Recently, patients reported to be EGFR negative have been shown to respond to cetuximab. Possible explanations include methodological failures or most likely heterogeneity and complexity of the mechanisms of EGFR-mediated molecular carcinogenesis. Immunohistochemistry is the most widely used method for measuring EGFR expression; however, its value is limited by lack of methodological standardization. Other approaches to measuring EGFR such as amplification assays are currently being introduced but need further testing before they can enter clinical practice. Mutational analysis seems also fruitful in predicting response to anti-EGFR small molecules. Further work is needed to identify how EGFR contributes to the carcinogenic and metastatic processes.


Mentoring programs for medical students--a review of the PubMed literature 2000-2008.
Although mentoring is acknowledged as a key to successful and satisfying careers in medicine, formal mentoring programs for medical students are lacking in most countries. Within the framework of planning a mentoring program for medical students at Zurich University, an investigation was carried out into what types of programs exist, what the objectives pursued by such programs are, and what effects are reported. A PubMed literature search was conducted for 2000 - 2008 using the following keywords or their combinations: mentoring, mentoring program, medical student, mentor, mentee, protégé, mentorship. Although a total of 438 publications were identified, only 25 papers met the selection criteria for structured programs and student mentoring surveys. The mentoring programs reported in 14 papers aim to provide career counseling, develop professionalism, increase students' interest in research, and support them in their personal growth. There are both one-to-one and group mentorships, established in the first two years of medical school and continuing through graduation. The personal student-faculty relationship is important in that it helps students to feel that they are benefiting from individual advice and encourages them to give more thought to their career choices. Other benefits are an increase in research productivity and improved medical school performance in general. Mentored students also rate their overall well-being as higher. - The 11 surveys address the requirements for being an effective mentor as well as a successful mentee. A mentor should empower and encourage the mentee, be a role model, build a professional network, and assist in the mentee's personal development. A mentee should set agendas, follow through, accept criticism, and be able to assess performance and the benefits derived from the mentoring relationship. Mentoring is obviously an important career advancement tool for medical students. In Europe, more mentoring programs should be developed, but would need to be rigorously assessed based on evidence of their value in terms of both their impact on the career paths of juniors and their benefit for the mentors. Medical schools could then be monitored with respect to the provision of mentorships as a quality characteristic.


Mixer/Bon and FoxH1/Sur have overlapping and divergent roles in Nodal signaling and mesendoderm induction.
Transcription factors belonging to the FoxH1 and Mixer families are required for facets of Nodal signaling during vertebrate mesendoderm induction. Here, we analyze whether zebrafish proteins related to FoxH1 [Schmalspur (Sur)] and Mixer [Bonnie and clyde (Bon)] act within or downstream of the Nodal signaling pathway, test whether these two factors have additive or overlapping activities, and determine whether FoxH1/Sur and Mixer/Bon can account for all Nodal signaling during embryogenesis. We find that sur expression is independent of Nodal signaling and that bon is expressed in the absence of Nodal signaling but requires Nodal signaling and Sur for enhanced, maintained expression. These results and the association of FoxH1 and Mixer/Bon with phosphorylated Smad2 support a role for these factors as components of the Nodal signaling pathway. In contrast to the relatively mild defects observed in single mutants, loss of both bon and sur results in a severe phenotype characterized by absence of prechordal plate, cardiac mesoderm, endoderm and ventral neuroectoderm. Analysis of Nodal-regulated proteins reveals that Bon and Sur have both distinct and overlapping regulatory roles. Some genes are regulated by both Bon and Sur, and others by either Bon or Sur. Complete loss of Nodal signaling results in a more severe phenotype than loss of both Bon and Sur, indicating that additional Smad-associated transcription factors remain to be identified that act as components of the Nodal signaling pathway.


Activation of gene expression by herpes simplex virus type 1 ICP0 occurs at the level of mRNA synthesis.
ICP0 is a nuclear phosphoprotein involved in the activation of herpes simplex virus type 1 (HSV-1) gene expression during lytic infection and reactivation from viral latency. Although available evidence suggests that ICP0 acts at the level of transcription, definitive studies specifically addressing this issue have not been reported. In the present study we measured the ability of ICP0 to activate gene expression (i) from promoters representing the major kinetic classes of viral genes in transient expression assays and (ii) from the same promoters during viral infection at multiplicities of infection ranging from 0.1 to 5.0 PFU/cell. The levels of synthesis and steady-state accumulation of mRNA, mRNA stability, and levels of protein synthesis were compared in cells transfected with a reporter plasmid in the presence and absence of ICP0 and in cells infected with wild-type HSV-1 or an ICP0 null mutant, n212. In transient expression assays and during viral infection at all multiplicities tested, the levels of steady-state mRNA and protein were significantly lower in the absence of ICP0, indicating that ICP0 activates gene expression at the level of mRNA accumulation. In transient expression assays and during infection at low multiplicities (< 1 PFU/cell) in the presence or absence of ICP0, marked increases in the levels of viral mRNAs accompanied by proportional increases in the levels of protein synthesis were observed with increasing multiplicity. At a high multiplicity (5 PFU/cell) in the presence or absence of ICP0, mRNA levels did not increase as a function of multiplicity and changes in the levels of protein were no longer related to changes in the levels of mRNA. Collectively, these tests indicate that transcription of viral genes is rate limiting at low multiplicities and that translation is rate limiting at high multiplicities, independent of ICP0. Consistent with the lower levels of mRNA detected in the absence of ICP0, the rates of transcription initiation measured by nuclear run-on assays were uniformly lower in cells infected with the ICP0 null mutant at all multiplicities tested, implying that ICP0 enhances transcription at or before initiation or both. No evidence was found of posttranscriptional effects of ICP0 (i.e., effects on the stability of mRNA, nuclear-cytoplasmic distribution, polyribosomal mRNA distribution, or rates of protein synthesis). Taken together, these results suggest that ICP0 activates gene expression prior to or at the level of initiation of mRNA synthesis in transient expression assays and during viral infection. Based on these findings; we hypothesize that the exaggerated multiplicity-dependent growth phenotype characteristic of ICP0 null mutants reflects the requirement for ICP0 under conditions where the steady-state level of mRNA is rate limiting, such as during low-multiplicity infection and reactivation from latency.


A first phylogenetic analysis of Giant Pill-Millipedes (Diplopoda: Sphaerotheriida), a new model Gondwanan taxon, with special emphasis on island gigantism.
The phylogeny of the Giant Pill-Millipedes, order Sphaerotheriida, is investigated using a new morphological character matrix comprising 89 characters. The majority of these characters are employed for the first time in millipedes. All trees obtained agree on the monophyletic status of the Sphaerotheriida and several of its tribes, each restricted to a modern land mass. The species from Madagascar displaying island gigantism do not form a monophyletic group. The classic division of Giant Pill-Millipedes into two families, Sphaerotheriidae and Zephronidae, was not reflected in the analysis. The genus Procyliosoma is the sister-group to all other Sphaerotheriida, rendering the family Sphaerotheriidae paraphyletic. A new family-level classification of Giant Pill-Millipedes, based on the current phylogeny, is introduced. The new family Procyliosomatidae contains only the genus Procyliosoma, distributed in Australia and New Zealand. The family Zephronidae remains unchanged, while the family Sphaerotheriidae now incorporates only the African Giant Pill-Millipede genera. All genera from southern India and Madagascar form a monophyletic group and are placed in the new family Arthrosphaeridae. The Malagasy genus Sphaeromimus is more closely related to the Indian Arthrosphaera species than to other genera from Madagascar. A biogeographical analysis identifies the group as a Gondwana taxon (with a notable absence from South America). The current phylogeny of Giant Pill-Millipede families mirrors perfectly the suggested break-up of Gondwana fragments 160-90 Ma. No evidence for a dispersal event could be found, highlighting the importance of Giant Pill-Millipedes as a potential model taxon.


Remission of rheumatoid arthritis in clinical practice: application of the American College of Rheumatology/European League Against Rheumatism 2011 remission criteria.
To describe use of the American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) rheumatoid arthritis (RA) remission criteria in clinical practice. Remission was examined using data on 1,341 patients with RA (91% men) from the US Department of Veterans Affairs RA (VARA) registry (total of 9,700 visits) and 1,153 patients with RA (25.8% men) in a community rheumatology practice (Arthritis and Rheumatology Clinics of Kansas [ARCK]) (total of 6,362 visits). Cross-sectional and cumulative probabilities were studied, and agreement between the various remission criteria was assessed. Aspects of reliability of the criteria were determined using Boolean-based definitions, as well as the Clinical Disease Activity Index (CDAI) and Simplified Disease Activity Index (SDAI) scoring methods proposed by the ACR/EULAR joint committee. When the 3-variable ACR/EULAR definition of remission recommended for use in community practice (swollen and tender joint counts ≤1, and visual analog scale score for patient's global assessment of disease activity ≤1) was applied, cross-sectional remission was 7.5% (95% confidence interval [95% CI] 6.4, 8.7%) for ARCK and 8.9% (95% CI 7.9, 9.9%) for VARA, and cumulative remission (remission at any observation) was 18.0% (for ARCK) and 24.4% (for VARA), over a mean followup of ∼2.2 years. Addition of the erythrocyte sedimentation rate or C-reactive protein level to the criteria set reduced remission to 5.0-6.2%, and use of the CDAI/SDAI increased the proportions to 6.9-10.1%. Moreover, 1.8-4.6% of the patients met remission criteria at ≥2 visits. Agreement between criteria definitions was good, as assessed by kappa statistics and Jaccard coefficients. Among patients in remission, the probability of a remission lasting 2 years was 6.0-14.1%. Among all patients, the probability of a remission lasting 2 years was <3%. Remission status and examination results for each patient varied substantially among physicians, as determined by multilevel analyses. Cross-sectional remission occurred in 5.0-10.1% of the patients in these cohorts, with cumulative remission being 2-3 times greater; however, long-term remission was rare. Problems with reliability and agreement limit the usefulness of these criteria in the individual patient. However, the criteria can be an effective method for measuring clinical status and treatment effect in groups of patients in the community.


Cardiotrophin-1 is inversely associated with obesity in non-diabetic individuals.
Cardiotrophin-1 is known to be a key regulator of energy homeostasis, as well as glucose and lipid metabolism in vivo. However, there are inconsistent results of the association between cardiotrophin-1 and obesity in humans, possibly confounded by hyperglycemia. Therefore, the aim of this study was to investigate the relationships among cardiotrophin-1 levels, overweight and obese individuals without diabetes in a Chinese population. The median (inter-quarter range) serum cardiotrophin-1 levels were 447.9 (230.9, 913.9), 350.6 (201.1, 666.5), and 288.1 (162.3, 572.4) pg/ml in non-diabetic subjects who were of normal weight (n = 522), overweight (n = 203), and obese (n = 93), respectively (trend test p < 0.001). Subjects who were overweight and obese had significantly lower cardiotrophin-1 levels than those with normal weight. The multivariate linear regression analyses showed that overweight (beta = -338.718, 95% CI = -552.786 ~ -124.651, p < 0.01), obese (beta = -530.275, 95% CI = -832.967 ~ -227.583, p < 0.01), and smoking (beta = -377.375, 95% CI = -654.353 ~ -100.397, p < 0.01) were negatively related to cardiotrophin-1 after adjusting for age, gender, HOMA-IR, hypertension, total cholesterol, HDL, triglyceride, eGFR, ALT, and alcohol drinking. The results of this study provided epidemiological evidence that non-diabetic subjects who were overweight or obesity had significantly lower cardiotrophin-1 concentrations than those with normal weight, and both obesity and being overweight were inversely associated with cardiotrophin-1 levels.


High burden of private mutations due to explosive human population growth and purifying selection.
Recent studies have shown that human populations have experienced a complex demographic history, including a recent epoch of rapid population growth that led to an excess in the proportion of rare genetic variants in humans today. This excess can impact the burden of private mutations for each individual, defined here as the proportion of heterozygous variants in each newly sequenced individual that are novel compared to another large sample of sequenced individuals. We calculated the burden of private mutations predicted by different demographic models, and compared with empirical estimates based on data from the NHLBI Exome Sequencing Project and data from the Neutral Regions (NR) dataset. We observed a significant excess in the proportion of private mutations in the empirical data compared with models of demographic history without a recent epoch of population growth. Incorporating recent growth into the model provides a much improved fit to empirical observations. This phenomenon becomes more marked for larger sample sizes, e.g. extrapolating to a scenario in which 10,000 individuals from the same population have been sequenced with perfect accuracy, still about 1 in 400 heterozygous sites (or about 6,000 variants) at the 10,001 st individual are predicted to be novel, 18-times as predicted in the absence of recent population growth. The proportion of private mutations is additionally increased by purifying selection, which differentially affect mutations of different functional annotations. The burden of private mutations for each individual, which are singletons (i.e. appearing in a single copy) in a larger sample that includes this individual, is predicted to be greatly increased by recent population growth, as well as by purifying selection. Comparison with empirical data supports that European populations have experienced recent rapid population growth, consistent with previous studies. These results have important implications to the design and analysis of sequencing-based association studies of complex human disease as they pertain to private and very rare variants. They also imply that personalized genomics will indeed have to be very personal in accounting for the large number of private mutations.


Convergence of Ets- and notch-related structural motifs in a heteromeric DNA binding complex.
Analysis of the heteromeric DNA binding protein GABP has revealed the interaction of two distinct peptide sequence motifs normally associated with proteins located in different cellular compartments. The alpha subunit of GABP contains an 85-amino acid segment related to the Ets family of DNA binding proteins. The ETS domain of GABP alpha facilitates weak binding to DNA and, together with an adjacent segment of 37 amino acids, mediates stable interaction with GABP beta. The beta subunit of GABP contains four imperfect repeats of a sequence present in several transmembrane proteins including the product of the Notch gene of Drosophila melanogaster. These amino-terminal repeats of GABP beta mediate stable interaction with GABP alpha and, when complexed with GABP alpha, directly contact DNA. These observations provide evidence for a distinct biochemical role for the 33-amino acid repeats, and suggest that they may serve as a module for the generation of specific dimerization interfaces.


Antigenic role of the endosymbionts of filarial nematodes: IgG response against the Wolbachia surface protein in cats infected with Dirofilaria immitis.
Filarial nematodes harbour intracellular endosymbiotic bacteria, which have been assigned to the genus Wolbachia. These bacteria appear to play an important role in the pathogenesis of filarial diseases through their lipopolysaccharides. In view of the presence of Wolbachia endosymbionts in the body of filarial nematodes, one might also expect that proteins from these bacteria play an antigenic role in humans and animals affected by filariases. To test this hypothesis, we produced in recombinant form the surface protein WSP and a portion of the cell-cycle protein FTSZ from the Wolbachia of Dirofilaria immitis. Western immunoblot assays were then performed using cat sera to test the immunogenicity of these proteins. Sera were collected from owners' cats, which were either sero-negative or sero-positive for D. immitis and from cats before and after experimental infection with D. immitis. FTSZ was recognized in Western blots by sera from both positive and negative cats and from both uninfected and experimentally infected cats. WSP was recognized only by sera from positive cats and from cats experimentally infected with D. immitis; this protein was not recognized by sera from negative cats and from cats before experimental infection with D. immitis. The results of Western blot assays on WSP thus support the hypothesis that infection with filarial nematodes induces the production of antibodies against Wolbachia proteins.


The Diffraction Pattern Calculator (DPC) toolkit: a user-friendly approach to unit-cell lattice parameter identification of two-dimensional grazing-incidence wide-angle X-ray scattering data.
The DPC toolkit is a simple-to-use computational tool that helps users identify the unit-cell lattice parameters of a crystal structure that are consistent with a set of two-dimensional grazing-incidence wide-angle X-ray scattering data. The input data requirements are minimal and easy to assemble from data sets collected with any position-sensitive detector, and the user is required to make as few initial assumptions about the crystal structure as possible. By selecting manual or automatic modes of operation, the user can either visually match the positions of the experimental and calculated reflections by individually tuning the unit-cell parameters or have the program perform this process for them. Examples that demonstrate the utility of this program include determining the lattice parameters of a polymorph of a fluorinated contorted hexabenzocoronene in a blind test and refining the lattice parameters of the thin-film phase of 5,11-bis(triethylsilylethynyl)anthradithiophene with the unit-cell dimensions of its bulk crystal structure being the initial inputs.


New Signal Readout Principle for Solid-Contact Ion-Selective Electrodes.
A novel approach to signal transduction concerning solid-contact ion-selective electrodes (SC-ISE) with a conducting polymer (CP) as the solid contact is investigated. The method presented here is based on constant potential coulometry, where the potential of the SC-ISE vs the reference electrode is kept constant using a potentiostat. The change in the potential at the interface between the ion-selective membrane (ISM) and the sample solution, due to the change in the activity of the primary ion, is compensated with a corresponding but opposite change in the potential of the CP solid contact. This enforced change in the potential of the solid contact results in a transient reducing/oxidizing current flow through the SC-ISE. By measuring and integrating the current needed to transfer the CP to a new state of equilibrium, the total cumulated charge that is linearly proportional to the change of the logarithm of the primary ion activity is obtained. In this work, different thicknesses of poly(3,4-ethylenedioxythiophene) (PEDOT) doped with poly(styrenesulfonate) (PSS) were used as solid contact. Also, coated wire electrodes (CWEs) were included in the study to show the general validity of the new approach. The ISM employed was selective for K(+) ions, and the selectivity of the membrane under implementation of the presented transduction mechanism was confirmed by measurements performed with a constant background concentration of Na(+) ions. A unique feature of this signal readout principle is that it allows amplification of the analytical signal by increasing the capacitance (film thickness) of the solid contact of the SC-ISE.


A comparison of different QSAR approaches to modeling CYP450 1A2 inhibition.
Prediction of CYP450 inhibition activity of small molecules poses an important task due to high risk of drug-drug interactions. CYP1A2 is an important member of CYP450 superfamily and accounts for 15% of total CYP450 presence in human liver. This article compares 80 in-silico QSAR models that were created by following the same procedure with different combinations of descriptors and machine learning methods. The training and test sets consist of 3745 and 3741 inhibitors and noninhibitors from PubChem BioAssay database. A heterogeneous external test set of 160 inhibitors was collected from literature. The studied descriptor sets involve E-state, Dragon and ISIDA SMF descriptors. Machine learning methods involve Associative Neural Networks (ASNN), K Nearest Neighbors (kNN), Random Tree (RT), C4.5 Tree (J48), and Support Vector Machines (SVM). The influence of descriptor selection on model accuracy was studied. The benefits of "bagging" modeling approach were shown. Applicability domain approach was successfully applied in this study and ways of increasing model accuracy through use of applicability domain measures were demonstrated as well as fragment-based model interpretation was performed. The most accurate models in this study achieved values of 83% and 68% correctly classified instances on the internal and external test sets, respectively. The applicability domain approach allowed increasing the prediction accuracy to 90% for 78% of the internal and 17% of the external test sets, respectively. The most accurate models are available online at http://ochem.eu/models/Q5747 .


Photoreactions of 2-(furan-2-yl)-3-hydroxy-4H-chromen-4-one and 3-hydroxy-2-(thiophene-2-yl)-4H-chromen-4-one using cyclohexane and acetonitrile as solvents.
Photolysis of the titled chromenones was carried out at their longest absorption band (∼360 nm) using cyclohexane (CH) and acetonitrile (ACN) as solvents, in both aerated and de-aerated solutions. Different dimeric photoproducts were formed with both chromenones in aerated solutions. On photolysing 2-(furan-2-yl)-3-hydroxy-4H-chromen-4-one (FHC) in aerated cyclohexane, 2-(furan-2-yl)-2-{[2-(furan-2yl)-4-oxo-4H-chromen-3-yl]oxy}-2H-chromene-3,4-dione (a dehydrodimer) was formed, and on photolysing 3-hydroxy-2-(thiophene-2-yl)-4H-chromen-4-one (THC) in aerated ACN, a different dimeric product was isolated and identified. The corresponding 3-aryl-3-hydroxy-1,2-indandiones were also detected with FHC in ACN and with THC in CH, in addition to the dimeric products in both cases. On the other hand, in the de-aerated solutions, only the corresponding 1,2-indandiones were detected. 3-(Furan-2-yl)isobenzofuran-1(3H)-one as a secondary product was also detected with FHC in both solvents. An attempt was made to isolate the spectra of the photoproducts in situ. Excited State Intramolecular Proton Transfer (ESIPT) and Excited State Intramolecular Charge Transfer (ESICT) processes complicate the photodynamics of the reaction, making it difficult to predict the mechanisms of the photoreactions. However, tentative mechanisms have been proposed for the formation of the photoproducts.


Inflammatory systemic biomarkers in setting acute coronary syndromes--effects of the diurnal variation.
Clinicians have used additional tools to aid clinical assessment and to enhance their ability to identify the "vulnerable" patient at risk for cardiovascular diseases. Circulating biomarkers are one such tool used for identifying better high-risk individuals and to prognosticate effectively and treat patients with disease. A persistent immune activation is a main feature of atherosclerosis. The inflammatory activity is not only detectable in the vascular wall, but also in peripheral blood. Patients with coronary artery disease show increased numbers of neutrophils and T cells as well as elevated levels of several inflammatory mediators. On the other hand, several cardiovascular disease states show a daily cycle of activity, i.e. a peak incidence of cerebrovascular and cardiovascular events has been documented in the early morning hours. Several studies have shown diurnal variations in inflammatory systemic markers in patients with acute coronary syndrome. Diurnal variations can alter the analysis of blood-derived samples. Prior to the analysis of a blood sample, multiple steps are necessary to generate the desired specimen. The knowledge of diurnal variations is a prerequisite to understand and control their impact. This brief review comments the effect of the diurnal variation on the most important inflammatory systemic biomarkers in the setting acute coronary syndrome: interleukin-6, neopterin, matrix metalloproteinases, vascular cell adhesion molecule-1, intercellular adhesión molecule-1, soluble CD40 ligand, and C-reactive protein.


Prospects for treatment of human retrovirus-associated diseases.
Overwhelming evidence has been obtained over the past 2 years that human T-cell leukemia virus type III (HTLV-III) is the primary cause of acquired immunodeficiency syndrome. The direct cytopathic effect of the virus on T4 helper cells demonstrated in vitro is probably responsible for the depletion of this critical population of cells in affected individuals which results in a series of devastating immunological abnormalities. It follows that any approaches effective in suppressing the virulent virus would be of benefit to the infected patient. The vast amount of knowledge about retroviruses in general, and the rapidly accumulating information about HTLV-III in particular, makes it likely that a targeted approach for development of antiviral agents will bear fruit. General strategies further making use of chemical and biological agents as well as genetic manipulations are discussed.


Whole-grain food consumption in Singaporean children aged 6-12 years.
Public health bodies in many countries are attempting to increase population-wide habitual consumption of whole grains. Limited data on dietary habits exist in Singaporean children. The present study therefore aimed to assess whole grain consumption patterns in Singaporean children and compare these with dietary intake, physical activity and health parameters. Dietary intake (assessed by duplicate, multipass, 24-h food recalls), physical activity (by questionnaire) and anthropometric measurements were collected from a cross-section of 561 Singaporean children aged 6-12 years. Intake of whole grains was evaluated using estimates of portion size and international food composition data. Only 38·3 % of participants reported consuming whole grains during the dietary data collection days. Median intake of whole grains in consumers was 15·3 (interquartile range 5·4-34·8) g/d. The most commonly consumed whole-grain food groups were rice (29·5 %), wholemeal bread (28·9 %) and ready-to-eat breakfast cereals (18·8 %). A significantly lower proportion of Malay children (seven out of fifty-eight; P < 0·0001) consumed whole grains than children of other ethnicities. Only 6 % of all children consumed the amount of whole grains most commonly associated with improved health outcomes (48 g/d). There was no relationship between whole grain consumption patterns and BMI, waist circumference or physical activity but higher whole grain intake was associated with increased fruit, vegetable and dairy product consumption (P < 0·001). These findings demonstrate that consumption of whole grain foods is low at a population level and infrequent in Singaporean children. Future drives to increase whole-grain food consumption in this population are likely to require input from multiple stakeholders.


Ionic mechanisms underlying history-dependence of conduction delay in an unmyelinated axon.
Axonal conduction velocity can change substantially during ongoing activity, thus modifying spike interval structures and, potentially, temporal coding. We used a biophysical model to unmask mechanisms underlying the history-dependence of conduction. The model replicates activity in the unmyelinated axon of the crustacean stomatogastric pyloric dilator neuron. At the timescale of a single burst, conduction delay has a non-monotonic relationship with instantaneous frequency, which depends on the gating rates of the fast voltage-gated Na+ current. At the slower timescale of minutes, the mean value and variability of conduction delay increase. These effects are because of hyperpolarization of the baseline membrane potential by the Na+/K+ pump, balanced by an h-current, both of which affect the gating of the Na+ current. We explore the mechanisms of history-dependence of conduction delay in axons and develop an empirical equation that accurately predicts this history-dependence, both in the model and in experimental measurements.


Discussions about clinical trials among patients with newly diagnosed lung and colorectal cancer.
Clinical trials are essential to establish the effectiveness of new cancer therapies, but less than 5% of adults with cancer enroll in trials. In addition to ineligibility or lack of available trials, barriers to enrollment may include limited patient awareness about the option of participation. We surveyed a multiregional cohort of patients with lung or colorectal cancer (or their surrogates) three to six months after diagnosis. We assessed whether respondents reported learning that clinical trial participation might be an option, and, if so, with whom they discussed trials. We used logistic regression to assess the association of patient characteristics with discussing trial participation and enrolling in trials. All statistical tests were two-sided. Of 7887 respondents, 1114 (14.1%) reported discussing the possibility of clinical trial participation; most learned about trials from their physicians, and 287 patients (3.6% of all patients, 25.8% of trial discussants) enrolled. Among 2173 patients who received chemotherapy for advanced (stage III/IV lung or stage IV colorectal) cancer, 25.7% discussed trials, and 7.6% (29.5% of trial discussants) enrolled. Discussions were less frequent among older patients, African American or Asian vs white patients, and those with lower incomes and more comorbidity. Enrollment was higher among patients reporting shared vs physician-driven decisions (all P < .05). In this population-based cohort, only 14% of patients discussed participation in clinical trials. Discussions were more frequent among advanced cancer patients but were still reported by a minority of patients. Strategies to expand access to trials and facilitate patient-provider communication about participation may accelerate development of better cancer therapeutics.


[A couple suffering acute respiratory illness due to waterproofing spray exposure].
The patients were a 28-year-old man and a his 27-year-old wife. The husband smoked a cigarette immediately after using a waterproofing spray, and developed fever, cough, and dyspnea 15 min later. The wife had nausea 2 hours later. Nine hours later, the husband visited a local clinic, and was referred to our hospital because of hypoxemia. In addition, chest CT showed ill-defined areas of increased density, predominantly in the bilateral upper lung fields, with interlobular septal thickening, and he was hospitalized. Although the wife was asymptomatic at the time of examination, she had chest CT findings similar to those of her husband, and was also hospitalized. After admission, the husband received steroid pulse therapy and oxygen inhalation for his symptoms and hypoxemia, with return of arterial blood gas analysis results to normal on the third day. The wife had no symptoms or hypoxemia during her hospital stay. Their chest CT findings improved on the seventh day after admission, and they were discharged. Thus, it appears that the couple suffered from acute respiratory illness due to waterproofing spray exposure, and probably heat degradation products from cigarette smoking caused the husband to have severe symptoms.


Effect of Foeniculum vulgare Mill. (fennel) on menopausal symptoms in postmenopausal women: a randomized, triple-blind, placebo-controlled trial.
Preliminary data suggest that Foeniculum vulgare (fennel) can be an effective treatment for menopausal symptoms. This trial was designed to assess the efficacy of fennel in the management of menopausal symptoms in postmenopausal women. In this triple-blind, placebo-controlled trial, 90 postmenopausal women aged 45 to 60 years in Tehran were randomly assigned to treatment (n = 45) or placebo (n = 45) groups. The participants received 8 weeks of treatment with soft capsules containing 100 mg fennel or a placebo (2 per day for each group). The participants were followed for 2 weeks postintervention to assess the continuance of the effect of intervention. The Menopause Rating Scale (MRS) questionnaire was used to assess changes in menopausal symptoms at baseline and at 4, 8, and 10 weeks after onset of intervention. The groups recorded similar mean scores on the MRS questionnaire before intervention. After intervention, the treatment group showed a significant decrease in the mean MRS score. The results of the Friedman test showed significant differences between the mean score at baseline and those at 4, 8, and 10 weeks after onset of intervention in the treatment group (P < 0.001), whereas there were no significant differences in the placebo group. When the fennel and the placebo groups were compared, the independent t test showed significant differences in mean scores between groups at 4, 8, and 10 weeks (2 weeks postintervention; P < 0.001). Fennel is an effective and safe treatment to reduce menopausal symptoms in postmenopausal women without serious side effects. More clinical trials with larger populations are required to confirm this result.


Myocardial inotrophy of CO2 in water- and air-breathing vertebrates.
A negative-inotropic effect of CO2 on myocardial contractility presumably occurs because increasing H+ concentration competes with Ca2+ at cellular membranes and proteins. Since air-breathing vertebrates have higher blood and tissue CO2 concentration than water breathers the question was raised whether the cardiac cell has a modified sensitivity to CO2 correlated with the evolutionary transition of vertebrates from water breathers to air breathers. The water-breathing fish, Salmo gairdneri, and the air-breathing turtle, Pseudemys scripta, were selected as experimental animals, since their total CO2 concentration differs markedly (3.0 and 16.0 mmol.kg-1). Electrically paced isometric ventricular strips from both species were subjected to a stepwise increase in PCO2 from 25 to 114 Torr (pH0 7.80 to 7.0; HCO3- 30 mM). Trout were additionally exposed to the same pH0 changes at 5 mM HCO3- by a stepwise increase in PCO2 (4.5-12 Torr). At each increase in PCO2 the turtle heart showed a lesser negative inotropic effect than trout. The present findings offer direct evidence that the negative inotropic effect of CO2 on heart muscle is inversely proportional to the in vivo levels of tissue CO2 concentration. The results obtained are discussed in relation to phylogenetical and ecological aspects of acid-base balance.


A perinucleolar compartment contains several RNA polymerase III transcripts as well as the polypyrimidine tract-binding protein, hnRNP I.
We have investigated the subcellular organization of the four human Y RNAs. These RNAs, which are transcribed by RNA polymerase III, are usually found complexed with the Ro autoantigen, a 60-kD protein. We designed 2'-OMe oligoribonucleotides that were complementary to accessible single-stranded regions of Y RNAs within Ro RNPs and used them in fluorescence in situ hybridization. Although all four Y RNAs were primarily cytoplasmic, oligonucleotides directed against three of the RNAs hybridized to discrete structures near the nucleolar rim. We have termed these structures "perinucleolar compartments" (PNCs). Double labeling experiments with appropriate antisera revealed that PNCs are distinct from coiled bodies and fibrillar centers. Co-hybridization with a genomic DNA clone spanning the human Y1 and Y3 genes showed that PNCs are not stably associated with the transcription site for these Y RNAs. Although 5S rDNA was often located near the nucleolar periphery, PNCs are not associated with 5S gene loci. Two additional pol III transcripts, the RNA components of RNase P and RNase MRP, did colocalize within PNCs. Most interestingly, the polypyrimidine tract-binding protein hnRNP I/PTB was also concentrated in this compartment. Possible roles for this novel nuclear subdomain in macromolecular assembly and/or nucleocytoplasmic shuttling of these five pol III transcripts, along with hnRNP I/PTB, are discussed.


Effects of tyrosine kinase inhibitors on cell death induced by sodium fluoride and pertussis toxin in the pancreatic beta-cell line, RINm5F.
1. Sodium fluoride causes apoptosis of pancreatic beta-cells and this response is enhanced by pre-treatment with pertussis toxin. In the present study, tyrosine kinase inhibitors were used to investigate the mechanisms of action of NaF and pertussis toxin in the beta-cell line, RINm5F. 2. Exposure of RINm5F cells to low concentrations of genistein or tyrphostin A25 resulted in significant inhibition of cell death induced by 5 mM NaF. Higher concentrations (>25 microM) were cytotoxic in the absence of NaF but, paradoxically, the combination of genistein and NaF induced less cell death than when each agent was used alone. 3. The increase in cell death induced by 100 microM genistein was markedly inhibited by ciprofloxacin, a drug which binds to topoisomerase II. Etoposide (which inhibits topoisomerase II but has no effect on tyrosine kinase activity) also caused an increase in RINm5F cell death. Neither etoposide nor ciprofloxacin altered the response to 5 mM NaF. 4. Pertussis toxin markedly enhanced the extent of RINm5F cell death induced by NaF and this effect was completely prevented by 25 microM genistein. The inhibition caused by genistein was not affected by ciprofloxacin but was reproduced by a structurally dissimilar tyrosine kinase inhibitor, herbimycin A. 5. The results demonstrate that RINm5F beta-cells express a pertussis toxin sensitive pathway that is anti-apoptotic. The activity of this pathway is most evident in cells exposed to pro-apoptotic stimuli where the effects of pertussis toxin can be blocked by inhibitors of tyrosine kinase enzymes. A genistein-sensitive tyrosine kinase does not appear to be involved in RINm5F cell survival under basal conditions.


A flexible and efficient template format for circular consensus sequencing and SNP detection.
A novel template design for single-molecule sequencing is introduced, a structure we refer to as a SMRTbell template. This structure consists of a double-stranded portion, containing the insert of interest, and a single-stranded hairpin loop on either end, which provides a site for primer binding. Structurally, this format resembles a linear double-stranded molecule, and yet it is topologically circular. When placed into a single-molecule sequencing reaction, the SMRTbell template format enables a consensus sequence to be obtained from multiple passes on a single molecule. Furthermore, this consensus sequence is obtained from both the sense and antisense strands of the insert region. In this article, we present a universal method for constructing these templates, as well as an application of their use. We demonstrate the generation of high-quality consensus accuracy from single molecules, as well as the use of SMRTbell templates in the identification of rare sequence variants.


Androgens suppress EZH2 expression via retinoblastoma (RB) and p130-dependent pathways: a potential mechanism of androgen-refractory progression of prostate cancer.
Androgens and the androgen receptor are important for both normal prostate development and progression of prostate cancer (PCa). However, the underlying mechanisms are not fully understood. The Polycomb protein enhancer of zeste homolog 2 (EZH2) functions as an epigenetic gene silencer and plays a role in oncogenesis by promoting cell proliferation and invasion. EZH2 has been implicated in human PCa progression, because its expression is often elevated in hormone-refractory PCa. Here, we demonstrated that expression of EZH2 is lower in androgen-sensitive LNCaP PCa cells compared with Rf and C4-2 cells, two androgen-refractory sublines that are derived from LNCaP cells. Androgen ablation by castration increased the level of EZH2 proteins in LNCaP xenografts in mice. In contrast, treatment of LNCaP cells in culture with the synthetic androgen methyltrieolone (R1881) at doses of 1 nm or higher suppressed EZH2 expression. Moreover, our data suggest that androgen repression of EZH2 requires a functional androgen receptor and this effect is mediated through the retinoblastoma protein and its related protein p130. We further showed that androgen treatment not only increases expression of EZH2 target genes DAB2IP and E-cadherin but also affects LNCaP cell migration. Our results reveal that androgens function as an epigenetic regulator in prostatic cells by repression of EZH2 expression through the retinoblastoma protein and p130-dependent pathways. Our findings also suggest that blockade of EZH2 derepression during androgen deprivation therapy may represent an effective tactic for the treatment of androgen-refractory PCa.


Nebulized solvent ablation of aligned PLLA fibers for the study of neurite response to anisotropic-to-isotropic fiber/film transition (AFFT) boundaries in astrocyte-neuron co-cultures.
Developing robust in vitro models of in vivo environments has the potential to reduce costs and bring new therapies from the bench top to the clinic more efficiently. This study aimed to develop a biomaterial platform capable of modeling isotropic-to-anisotropic cellular transitions observed in vivo, specifically focusing on changes in cellular organization following spinal cord injury. In order to accomplish this goal, nebulized solvent patterning of aligned, electrospun poly-l-lactic acid (PLLA) fiber substrates was developed. This method produced a clear topographic transitional boundary between aligned PLLA fibers and an isotropic PLLA film region. Astrocytes were then seeded on these scaffolds, and a shift between oriented and non-oriented astrocytes was created at the anisotropic-to-isotropic fiber/film transition (AFFT) boundary. Orientation of chondroitin sulfate proteoglycans (CSPGs) and fibronectin produced by these astrocytes was analyzed, and it was found that astrocytes growing on the aligned fibers produced aligned arrays of CSPGs and fibronectin, while astrocytes growing on the isotropic film region produced randomly-oriented CSPG and fibronectin arrays. Neurite extension from rat dissociated dorsal root ganglia (DRG) was studied on astrocytes cultured on anisotropic, aligned fibers, isotropic films, or from fibers to films. It was found that neurite extension was oriented and longer on PLLA fibers compared to PLLA films. When dissociated DRG were cultured on the astrocytes near the AFFT boundary, neurites showed directed orientation that was lost upon growth into the isotropic film region. The AFFT boundary also restricted neurite extension, limiting the extension of neurites once they grew from the fibers and into the isotropic film region. This study reveals the importance of anisotropic-to-isotropic transitions restricting neurite outgrowth by itself. Furthermore, we present this scaffold as an alternative culture system to analyze neurite response to cellular boundaries created following spinal cord injury and suggest its usefulness to study cellular responses to any aligned-to-unorganized cellular boundaries seen in vivo.


Anion-templated hexagonal nanotubes.
Hydrogen bonding between bromide anions and a tetrahydroxytriptycene ligand was used to assemble crystalline hexagonal tubes with nanometer diameters in good yield. Use of a hexahydroxytriptycene ligand again gave hexagonal nanotubes, but containing the spontaneously-oxidised quinone-tetrahydroxy ligand. The surprisingly robust nanotubes are stable to heat, vacuum and water, and represent an unprecedented use of O-H···anion coordination to assemble complex three-dimensional structures.


Predicting rehospitalization and outpatient services from administration and clinical databases.
The study tests whether psychiatric services utilization may be predicted from administrative databases without clinical variables equally as well as from databases with clinical variables. Persons with a psychiatric hospitalization at an urban medical center were followed for 1 year postdischarge (N = 1384.) Dependent variables included statewide rehospitalization and the number of hours of outpatient services received. Three linear and logistic regression models were developed and cross-validated: a basic model with limited administrative independent variables, an intermediate model with diagnostic and limited clinical indicators, and a full model containing additional clinical predictors. For rehospitalization, the clinical cross-validated model accounted for twice the variance accounted by the basic model (adjusted R2 = .13 and .06, respectively). For outpatient hours, the basic cross-validated model performed as well as the clinical model (adjusted R2 = .36 and .34, respectively). Clinical indicators such as assessment of functioning and co-occurring substance use disorder should be considered for inclusion in predicting rehospitalization.


Managing the risk of germ cell tumourigenesis in disorders of sex development patients.
The risk of germ cell cancer (GCC) is elevated in many disorders of sex development (DSD) patients, although not to the same extent. A number of risk factors have been identified recently, but their interplay and relative impact is currently not fully clear. This paper offers guidance on how theoretical knowledge on GCC risk can be translated to the clinical setting, taking into account individual patient characteristics. Guidelines for decision making in different patient groups, based on a literature review, epidemiological evidence, pathological and clinical research, and personal experience are offered. Until the advent of reliable screening tools for the detection of pre-invasive cancer lesions, managing germ cell tumour risk focuses on the question of if and when to perform biopsy or gonadectomy in most patients, and how to interpret the histological findings.


Contrast-Enhanced Cardiac Magnetic Resonance Imaging With a Manganese-Based Alternative to Gadolinium for Tissue Characterization of Acute Myocardial Infarction.
Background Late gadolinium enhancement cardiac magnetic resonance imaging is an effective and reproducible method for characterizing myocardial infarction. However, gadolinium-based contrast agents are contraindicated in patients with acute and chronic renal insufficiency. In addition, several recent studies have noted tissue deposition of free gadolinium in patients who have undergone serial contrast-enhanced magnetic resonance imaging. There is a clinical need for alternative forms of magnetic resonance imaging contrast agents that are acceptable in the setting of renal insufficiency. Methods and Results Three days after 80 minutes of ischemia/reperfusion of the left anterior descending coronary artery, cardiac magnetic resonance imaging was performed to assess myocardial lesion burden using both contrast agents. Late gadolinium enhancement cardiac magnetic resonance imaging was examined 10 and 15 minutes after contrast injection. Contrast agents were administered in alternating manner with a 2- to 3-hour washout period between contrast agent injections. Lesion evaluation and image processing were performed using Segment Medviso software. Mean infarct size and transmurality, measured using RVP-001, were not different compared with those measured using late gadolinium enhancement images. Bland-Altman analysis demonstrated a nominal bias of 0.13 mL (<1% of average total lesion volume) for RVP-001 in terms of gross infarct size measurement. Conclusions The experimental manganese-based contrast agent RVP-001 appears to be an effective agent for assessment of myocardial infarction location, size, and transmurality, and it may be useful as an alternative to gadolinium-based agents.


Seed treatment with Trichoderma harzianum alleviates biotic, abiotic, and physiological stresses in germinating seeds and seedlings.
Trichoderma spp. are endophytic plant symbionts that are widely used as seed treatments to control diseases and to enhance plant growth and yield. Although some recent work has been published on their abilities to alleviate abiotic stresses, specific knowledge of mechanisms, abilities to control multiple plant stress factors, their effects on seed and seedlings is lacking. We examined the effects of seed treatment with T. harzianum strain T22 on germination of seed exposed to biotic stress (seed and seedling disease caused by Pythium ultimum) and abiotic stresses (osmotic, salinity, chilling, or heat stress). We also evaluated the ability of the beneficial fungus to overcome physiological stress (poor seed quality induced by seed aging). If seed were not under any of the stresses noted above, T22 generally had little effect upon seedling performance. However, under stress, treated seed germinated consistently faster and more uniformly than untreated seeds whether the stress was osmotic, salt, or suboptimal temperatures. The consistent response to varying stresses suggests a common mechanism through which the plant-fungus association enhances tolerance to a wide range of abiotic stresses as well as biotic stress. A common factor that negatively affects plants under these stress conditions is accumulation of toxic reactive oxygen species (ROS), and we tested the hypothesis that T22 reduced damages resulting from accumulation of ROS in stressed plants. Treatment of seeds reduced accumulation of lipid peroxides in seedlings under osmotic stress or in aged seeds. In addition, we showed that the effect of exogenous application of an antioxidant, glutathione, or application of T22, resulted in a similar positive effect on seed germination under osmotic stress or in aged seed. This evidence supports the model that T. harzianum strain T22 increases seedling vigor and ameliorates stress by inducing physiological protection in plants against oxidative damage.


Supplementation with calcium salts of linoleic and trans-octadecenoic acids improves fertility of lactating dairy cows.
Objectives were to evaluate effects of feeding a calcium salt rich in linoleic and trans-octadecenoic acids (LTFA) on synthesis of prostaglandin F(2alpha) based on its metabolite (PGFM), uterine involution and pregnancy rates in lactating dairy cows. Five hundred and eleven Holstein cows were blocked according to parity, body condition score and milk yield in the previous lactation. Primiparous and multiparous cows were randomly assigned to one of the two treatments consisting of calcium salt (2% diet dry matter) of either palm oil (PO) or LTFA from 25 days prepartum to 80 days of lactation. Cows were time-inseminated at 70 +/- 3 days postpartum. Feeding LTFA tended (p = 0.08) to decrease the incidence of puerperal metritis (15.1% vs 8.8%). Primiparous cows supplemented with LTFA showed larger increase in plasma PGFM concentration at day 1 postpartum (17018 vs 6897 pm). Pregnancy rate after first insemination tended (p = 0.07) to be greater at 27 days after insemination (37.9% vs 28.6%), and was greater (p = 0.05) at 41 days after insemination (35.5% vs 25.8%) for cows fed LTFA compared with PO. These results indicate that unsaturated fatty acids fed in a rumen inert form have the potential to modulate reproductive events and improve pregnancy rates in lactating dairy cows.


Action of C-type natriuretic peptide in isolated canine arteries and veins.
C-type natriuretic peptide (CNP) is a hormone that shares structural homology to atrial natriuretic peptide (ANP); however, distinct receptors have been found for each in cultured aortic endothelial and smooth muscle cells. CNP in vivo reduces arterial pressure, atrial pressures, and cardiac output. These actions are consistent with a decrease in cardiac preload. Therefore, the current studies were designed to test the hypothesis that CNP is a vasodilator distinct from ANP. Rings of canine renal and saphenous arteries and renal, saphenous, and femoral veins with and without endothelium were suspended to measure isometric force in an organ chamber. CNP caused significant concentration-dependent relaxations in veins with and without endothelium contracted with phenylephrine (10(-6) log M). In marked contrast, ANP caused no significant relaxation in veins either with or without endothelium. In arterial rings, responses to the peptides were heterogeneous. ANP caused relaxation in renal but not saphenous arteries. CNP induced modest and comparable relaxation in rings of saphenous but not renal arteries with and without endothelium. These results demonstrate that: 1) CNP and not ANP is a relaxing factor of isolated peripheral canine veins, and 2) the responses of arteries to ANP and CNP are heterogeneous. These findings support a distinct biological role for CNP in the regulation of cardiovascular tone.


Bone morphogenetic protein 8B promotes the progression of non-alcoholic steatohepatitis.
Non-alcoholic steatohepatitis (NASH) is characterized by lipotoxicity, inflammation and fibrosis, ultimately leading to end-stage liver disease. The molecular mechanisms promoting NASH are poorly understood, and treatment options are limited. Here, we demonstrate that hepatic expression of bone morphogenetic protein 8B (BMP8B), a member of the transforming growth factor beta (TGFβ)-BMP superfamily, increases proportionally to disease stage in people and animal models with NASH. BMP8B signals via both SMAD2/3 and SMAD1/5/9 branches of the TGFβ-BMP pathway in hepatic stellate cells (HSCs), promoting their proinflammatory phenotype. In vivo, the absence of BMP8B prevents HSC activation, reduces inflammation and affects the wound-healing responses, thereby limiting NASH progression. Evidence is featured in primary human 3D microtissues modelling NASH, when challenged with recombinant BMP8. Our data show that BMP8B is a major contributor to NASH progression. Owing to the near absence of BMP8B in healthy livers, inhibition of BMP8B may represent a promising new therapeutic avenue for NASH treatment.


Characterization and validation studies of powerPlex 2.1, a nine-locus short tandem repeat (STR) multiplex system and penta D monoplex.
In order to increase the power of discrimination for human identification purposes, a nine-locus short tandem repeat (STR) multiplex, the GenePrint PowerPlex 2.1 system (PowerPlex 2.1) developed by Promega Corporation and a separate pentanucleotide-repeat locus, Penta D, were tested. This megaplex system includes the highly polymorphic loci FGA, TPOX, D8S1179, vWA, Penta E, D18S51, D21S11, TH01, and D3S1358 and may be used in combination with the eight-locus STR multiplex, the GenePrint PowerPlex 1.1 system (PowerPlex 1.1) that has been previously developed. Three of the loci, TPOX, TH01 and vWA, have been included in both systems for quality control purposes. As with PowerPlex 1.1, PowerPlex 2.1 is also based on a two-color detection of fluorescent-labeled DNA products amplified by polymerase chain reaction (PCR) and provides a valuable tool for accurate and rapid allele determination. The primer sequences used in the PowerPlex 2.1/Penta D system are also presented in this report. To meet the "Quality Assurance Standards for Forensic DNA Testing Laboratories" (FBI), we tested the efficiency and reproducibility of the PowerPlex 2.1/PentaD system by several validation studies that were conducted as a joint project among seven laboratories. Validation tests included concordance studies, sensitivity, and species specificity determination, as well as performance in forensic and environmentally impacted samples. The results produced from these tests demonstrated the consistency and reliability of the PowerPlex 2.1/Penta D system.


Long-term efficacy of metformin in overweight-obese PCOS: longitudinal follow-up of retrospective cohort.
Long-term efficacy of metformin in polycystic ovarian syndrome (PCOS) apart from in those with impaired glucose tolerance or diabetes remains unproven. We aimed to evaluate the impact of metformin in overweight-obese patients with PCOS and normal baseline glycemic homeostasis. A 10-year longitudinal follow-up of a retrospective cohort comprising 159 patients with PCOS defined by Rotterdam criteria, BMI ≥25 kg/m2 and normal initial glucose homeostasis (age 28.4 ± 6.4 years, BMI 34.9 ± 6.6 kg/m2) that had been receiving metformin 1000 mg BID. Collection data contained 6085 time-points including anthropometric, hormonal and metabolic parameters. After the first year body mass (BM) decreased for 3.9 ± 6.8 kg (P < 0.001) and remained stable during the following 3 years. Menstrual frequency (MF) increased to 3.0 ± 3.9 bleeds/year (P < 0.001) after first year to over 11 bleeds/year in the following years. The total testosterone and androstenedione decreased to 15.4 ± 47.9% and 11.3 ± 46.4% within first year, with further decrease in total testosterone and androstenedione to 37.8 ± 61.8 and 24.8 ± 40.5% at the fifth year of the follow-up. The total conversion rate to prediabetes and diabetes was extremely low throughout observation period. Less than 25% of patients continued with metformin for more than 5 years with further dropout to only 6% on metformin therapy at the tenth year of follow-up. Long-term metformin treatment of overweight-obese women with PCOS and normal baseline glycemic homeostasis resulted in reduction and stabilization of BM, improvements of MF and androgen profile and low conversion rate to diabetes.


Incidence and Risk Factors Associated with Readmission After Surgical Treatment for Adrenocortical Carcinoma.
Adrenocortical carcinoma (ACC) is a rare disease with a poor prognosis. Given the lack of data on readmission after resection of ACC, the objective of the current study was to define the incidence of readmission, as well as identify risk factors associated with readmission among patients with ACC who underwent surgical resection. Two hundred nine patients who underwent resection of ACC between January 1993 and December 2014 at 1 of 13 major centers in the USA were identified. Demographic and clinicopathological data were collected and analyzed relative to readmission. Median patient age was 52 years, and 62 % of the patients were female. Median tumor size was 12 cm, and the majority of patients had an American Society of Anesthesiologists (ASA) class of 3-4 (n = 85, 56 %). The overall incidence of readmission within 90 days from surgery was 18 % (n = 38). Factors associated with readmission included high ASA class (odds ratio (OR), 4.88 (95 % confidence interval (CI), 1.75-13.61); P = 0.002), metastatic disease on presentation (OR, 2.98 (95 % CI, 1.37-6.46); P = 0.006), EBL (>700 mL: OR, 2.75 (95 % CI, 1.16-6.51); P = 0.02), complication (OR, 1.91 (95 % CI, 1.20-3.05); P = 0.007), and prolonged length of stay (LOS; ≥9 days: OR, 4.12 (95 % CI, 1.88-9.01); P < 0.001). On multivariate logistic regression, a high ASA class (OR, 4.01 (95 % CI, 1.44-11.17); P = 0.008) and metastatic disease on presentation (OR, 3.44 (95 % CI, 1.34-8.84); P = 0.01) remained independently associated with higher odds of readmission. Readmission following surgery for ACC was common as one in five patients experienced a readmission. Patients with a high ASA class and metastatic disease on presentation were over four and three times more likely to be readmitted after surgical treatment for ACC, respectively.


The prognostic value of harvested lymph nodes and the metastatic lymph node ratio for gastric cancer patients: results of a study of 1,101 patients.
To investigate whether the recommendation to remove 15 lymph nodes that is used in the staging system is necessary to assess gastric cancer progression and to evaluate whether our metastatic lymph node ratio dividing method, adapted from the AJCC's (American Joint Committee on Cancer) 7(th) TNM staging system, is helpful for the patients with fewer than 15 harvested lymph nodes. We performed a retrospective study of 1101 patients with histologically diagnosed gastric cancer who underwent a D2 gastrectomy at the Sun Yat-sen University Cancer Center between January 2001 and December 2010. The Kappa and Chi-squared tests were employed to compare the clinicopathological variables. The Kaplan-Meier method and Cox regression were employed for the univariate and multivariate survival analyses. In the trial, 346, 601 and 154 patients had 0-14, 15-30 and more than 30 lymph nodes harvested, respectively. The median survival times of patients with different lymph nodes harvested in N0, N1, N2 and N3a groups were 45.43, 54.28 and 66.95 months (p=0.068); 49.22, 44.25 and 56.72 months (p<0.001), 43.94, 47.97 and 35.19 months (p=0.042); 32.88, 42.76 and 23.50 months (p=0.016). Dividing the patients who had fewer than 15 lymph nodes harvested by the metastatic lymph node ratio at 0, 0.13 and 0.40, the median survival times of these 4 groups were 70.6, 50.5, 53.5 and 30.7 months (p<0.001). After re-categorising these 4 groups into the N0, N1, N2, N3a groups, the histological grade, T staging, premier N staging, and restaged N staging were the independent prognostic factors. Large numbers of lymph nodes harvested in radical gastrectomy do not cause stage migration. For those patients with a small number of harvested lymph nodes, their stage should be divided by the metastatic lymph node ratio, referred to in the TNM staging system, to assign them an accurate stage.


Benign neonatal sleep myoclonus in newborn infants of opioid dependent mothers.
The aim of our study was to evaluate the incidence, duration and risk factors for benign neonatal sleep myoclonus (BNSM) in infants with neonatal abstinence syndrome (NAS) treated with opioids or sedatives, compared with control infants. This is a single centre observational case control study. Seventy-eight near term and term infants with neonatal opiate abstinence syndrome confirmed by meconium analysis were included. Exclusion criteria were cerebral malformation, intracranial haemorrhage and perinatal asphyxia. The babies were assessed eight hourly with a modified Finnegan score that included sleep myoclonus. Seventy-eight infants not exposed to opiates during pregnancy, hospitalized for at least 14 days and matched for gestational age were used as controls. The median gestational age was 38 (1)/(7) (95% CI: 35 (3)/(7)-41 (2)/(7)) weeks, birth weight 2730 (95% CI: 1890-3600) g, umbilical artery pH 7.25 (CI 7.10-7.37) and Apgar score at 5 minutes 9 (95% CI: 7-10). The control infants did not differ in these characteristics. Sleep myoclonus was diagnosed in 52 (67%) of the infants with NAS and 2 (2.6%) of the controls (OR 26 [95% CI: 7-223], p < 0.001). Myoclonus appeared as early as day 2 and as late as day 56 of life (median day 6) and lasted for 1 to 93 days (median 13 days). All infants had serum glucose > 2.5 mmol/L at first occurrence. The neurological examinations as well as cerebral ultrasound scans were normal. An electroencephalogram (EEG) carried out in 18 infants showed no signs of epileptic activity. BNSM has a high incidence in infants with NAS. The diagnosis can be made clinically. In the absence of other neurological symptoms further investigations such as EEG are not necessary and anticonvulsive treatment is not indicated.


Effect of natural organic matter on iron uptake by the freshwater cyanobacterium Microcystis aeruginosa.
The mode of Fe uptake by the cyanobacterium Microcystis aeruginosa cultured in Fraquil* (pH 8) containing Suwannee River fulvic acid (SRFA) was examined using short-term radiolabeled (55)Fe uptake assays and a kinetic model that describes extracellular Fe transformations. Both Fe(II) and Fe(III) uptake rates decreased substantially with increasing SRFA concentration as the availability of unchelated Fe decreased due to complexation by SRFA. Fe uptake rates under illuminated conditions were comparable to or slightly higher than those observed in the dark at the same Fe:SRFA concentration ratio, in contrast to results for systems containing ethylenediaminetetraacetic acid where Fe uptake rates were much greater under illumination than in the dark. The limited effect of light principally resulted from the relatively high rates of thermal dissociation and dark reduction of Fe(III) bound to SRFA and complexation of photogenerated Fe(II) by SRFA. Our findings imply that Fe uptake by M. aeruginosa at a fixed total Fe concentration of 200 nM is close to saturation when fulvic acid is present at concentrations near those typically found in natural waters (< ∼5 mg·L(-1)), with cellular growth likely to be limited by Fe availability only when natural organic matter is present at very high concentrations (>25 mg·L(-1)).


Women's patterns of participation in mammography screening in Denmark.
The objective of the study is to analyse individual women's participation patterns in mammography screening in Denmark. The study is set in the capital of Copenhagen and the county of Fyn representing around 95,000 women aged 50-69. The Central Population Register (CPR) was used to define the total target group, and supply information on migrations and deaths. Invitation and participation data came from the mammography screening programmes in Copenhagen (1991-1999) and Fyn (1993-2001), containing personal identification number, data on invitation date, participation and examination date for each screening round. In Copenhagen the coverage went from 70.5% in the first round to 63.1% in the fourth round, and the equivalent data for Fyn is 84.6% in the first round and 82.8% in the fourth round. Of the women eligible for at least three invitation rounds, 52.6% in Copenhagen and 76.4% in Fyn were faithful users, i.e. had participated in all screenings they were invited to. The conclusion is that the programme participation rates tend to overestimate the protection of the individual women covered by the programme. Behind the urban-rural gradient in programme participation is an even greater gradient in programme protection.


In vitro characterization of nebulizer delivery of liposomal amphotericin B aerosols.
Pharmaceutical aerosols have the potential to prevent pulmonary infectious diseases. Liposomal amphotericin B (LAMB, Ambisome, Astellas Pharma US, Deerfield, IL, USA) is approved as an intravenous infusion for empiric treatment of presumed fungal infections in neutropenic, febrile patients, as well as patients infected with Aspergillus, Cryptococcus, and other fungal pathogens. In this study, four different nebulizers were tested for their ability to deliver LAMB in aerodynamic droplet-size ranges relevant to lung deposition by an inertial sampling technique Mass median aerodynamic diameter (MMAD) and fine particle fraction percent <3.3 μm (FPF(3.3)) and <5.8 μm (FPF(5.8)) were determined by cascade impaction during a 2 min sampling period for each of three trials of all nebulizers. The MMADs for all nebulizers ranged from 1.72 ± 0.11 μm to 2.89 ± 0.12 μm; FPF(3.3) and FPF(5.8) were approximately 80% and 90%, respectively. Although all nebulizers appear acceptable for delivery of LAMB, the Pari LC Star and the Aeroeclipse II were considered the best in terms of delivery of aerosol efficiently and the proportion suitable for lung deposition. Additional research on pulmonary delivery and clinical tolerability is warranted.


Promotion of prescription drugs to consumers and providers, 2001-2010.
Pharmaceutical firms heavily promote their products and may have changed marketing strategies in response to reductions in new product approvals, restrictions on some forms of promotion, and the expanding role of biologic therapies. We used descriptive analyses of annual cross-sectional data from 2001 through 2010 to examine direct-to-consumer advertising (DTCA) (Kantar Media) and provider-targeted promotion (IMS Health and SDI), including: (1) inflation-adjusted total promotion spending ($ and percent of sales); (2) distribution by channel (consumer v. provider); and (3) provider specialty both for the industry as a whole and for top-selling biologic and small molecule therapies. Total promotion peaked in 2004 at US$36.1 billion (13.4% of sales). By 2010 it had declined to $27.7B (9.0% of sales). Between 2006 and 2010, similar declines were seen for promotion to providers and DTCA (both by 25%). DTCA's share of total promotion increased from 12% in 2002 to 18% in 2006, but then declined to 16% and remains highly concentrated. Number of products promoted to providers peaked in 2004 at over 3000, and then declined 20% by 2010. In contrast to top-selling small molecule therapies having an average of $370 million (8.8% of sales) spent on promotion, top biologics were promoted less, with only $33 million (1.4% of sales) spent per product. Little change occurred in the composition of promotion between primary care physicians and specialists from 2001-2010. These findings suggest that pharmaceutical companies have reduced promotion following changes in the pharmaceutical pipeline and patent expiry for several blockbuster drugs. Promotional strategies for biologic drugs differ substantially from small molecule therapies.


Utilizing novel diversity estimators to quantify multiple dimensions of microbial biodiversity across domains.
Microbial ecologists often employ methods from classical community ecology to analyze microbial community diversity. However, these methods have limitations because microbial communities differ from macro-organismal communities in key ways. This study sought to quantify microbial diversity using methods that are better suited for data spanning multiple domains of life and dimensions of diversity. Diversity profiles are one novel, promising way to analyze microbial datasets. Diversity profiles encompass many other indices, provide effective numbers of diversity (mathematical generalizations of previous indices that better convey the magnitude of differences in diversity), and can incorporate taxa similarity information. To explore whether these profiles change interpretations of microbial datasets, diversity profiles were calculated for four microbial datasets from different environments spanning all domains of life as well as viruses. Both similarity-based profiles that incorporated phylogenetic relatedness and naïve (not similarity-based) profiles were calculated. Simulated datasets were used to examine the robustness of diversity profiles to varying phylogenetic topology and community composition. Diversity profiles provided insights into microbial datasets that were not detectable with classical univariate diversity metrics. For all datasets analyzed, there were key distinctions between calculations that incorporated phylogenetic diversity as a measure of taxa similarity and naïve calculations. The profiles also provided information about the effects of rare species on diversity calculations. Additionally, diversity profiles were used to examine thousands of simulated microbial communities, showing that similarity-based and naïve diversity profiles only agreed approximately 50% of the time in their classification of which sample was most diverse. This is a strong argument for incorporating similarity information and calculating diversity with a range of emphases on rare and abundant species when quantifying microbial community diversity. For many datasets, diversity profiles provided a different view of microbial community diversity compared to analyses that did not take into account taxa similarity information, effective diversity, or multiple diversity metrics. These findings are a valuable contribution to data analysis methodology in microbial ecology.


Influence of the hormonal changes during the normal menstrual cycle in healthy young women on soluble adhesion molecules, plasma homocysteine, free radical markers and lipoprotein fractions.
Female sex hormones are known to exert a protective role on the vascular endothelial function, but the exact mechanisms of such protection is not known. We aimed to study the possible regulatory role of the female sex hormones changes during the normal menstrual cycle on soluble adhesion molecules E-selectin and ICAM-1, plasma homocyteine, free radical markers and lipoproteins in healthy young women. a cross sectional study of healthy female volunteers studied during a single normal menstrual cycle at 3 specific time points. North Staffordshire Hospitals NHS Trust. 20 healthy young menstruating women, aged (mean +/- SEM) 34 +/- 1 years, with normal menstruation, defined as a menstrual cycle of 21-35 days were studied at 3 time points of the same menstrual cycle. First in the early follicular phase (M-phase), at mid-follicular phase (F-phase), and during the luteal phase (L-phase). none. serum levels of soluble E-selectin, ICAM-1, plasma homocysteine, vitamin E and malondialdehyde (MDA), as well as lipoprotein fractions were measured at each time points. The mean percentage change for E-selectin between the M-phase and L-phase, F-phase and L-phase were 6% and 4%, respectively, p<0.005, p<0.066. Levels of ICAM-1, vitamin E and malondialdehyde did not vary through the cycle. Homocysteine was not different between M-phase and F-phase (10.39 +/- 0.68 micromol/l vs 10.33 +/- 0.65), nor between M-phase and L-phase (10.39+/-0.68 vs 9.77 +/- 0.75 micromol/l). Although the mean percentage decrease in homocysteine between F- and L-phases was significant (5.36 +/- 0.53%, p=0.029), the absolute decrease in concentrations was not (p=0.07). There were no cyclical changes in total, LDL, HDL cholesterol, triglycerides, apo A-I, apo B or Lp(a). Using a linear regression model, after correction for age, smoking, body mass index (BMI) and waist/hip ratio (WHR), oestrogen levels were the only predictor of E-selectin during the L-phase p<0.005. There were no significant correlations between oestrogen with lipids, apolipoproteins or homocysteine. There was an interesting significant univariate correlation between homocysteine with low-density-lipoprotein (LDL) cholesterol and apo B throughout all phases of the cycle, which persisted after correction for the effects of age, BMI, WHR and smoking history. Multiple regression analysis with all these factors showed homocysteine to be a significant predictor of apo B concentration during M (p=0.030) and L-phases (p=0.023) of the cycle and of LDL cholesterol in the M-phase (p=0.033). Female sex hormones may have small, though significant modulating role on E-selectin and homocysteine metabolism in healthy premenopausal women. Furthermore, the correlation between homocysteine, LDL and apo B levels suggests that induction of cholesterol synthesis by homocysteine, shown previously in vitro, may be of relevance in vivo.


Unidirectional ring-laser operation using sum-frequency mixing.
A technique enforcing unidirectional operation of ring lasers is proposed and demonstrated. The approach relies on sum-frequency mixing between a single-pass laser and one of the two counterpropagating intracavity fields of the ring laser. Sum-frequency mixing introduces a parametric loss for the intracavity field copropagating with the single-pass field, effectively generating a loss difference between the copropagating and counterpropagating intracavity fields. This loss mechanism ensures stable unidirectional lasing. The approach is generic and can be implemented at any desired lasing wavelength where lossless second-order nonlinear materials are available. Numerical modeling and experimental demonstration of parametric-induced unidirectional operation of a diode-pumped solid-state 1342nm cw ring laser are presented.


Cooperative Role of MAPK Pathways in the Interaction of Candida albicans with the Host Epithelium.
Candida albicans is an important human fungal pathogen responsible for tens of millions of infections as well as hundreds of thousands of severe life-threatening infections each year. MAP kinase (MAPK) signal transduction pathways facilitate the sensing and adaptation to external stimuli and control the expression of key virulence factors such as the yeast-to-hypha transition, the biogenesis of the cell wall, and the interaction with the host. In the present study, we have combined molecular approaches and infection biology to analyse the role of C. albicans MAPK pathways during an epithelial invasion. Hog1 was found to be important for adhesion to abiotic surfaces but was dispensable for damage to epithelial cells. The Mkc1 cell wall integrity (CWI) and Cek1 pathways, on the other hand, were both required for oral epithelial damage. Analysis of the ability to penetrate nutrient-rich semi-solid media revealed a cooperative role for Cek1 and Mkc1 in this process. Finally, cek2Δ (as well as cek1Δ) but not mkc1Δ or hog1Δ mutants, exhibited elevated β-glucan unmasking as revealed by immunofluorescence studies. Therefore, the four MAPK pathways play distinct roles in adhesion, epithelial damage, invasion and cell wall remodelling that may contribute to the pathogenicity of C. albicans.


The effects of P-glycoprotein inhibitor zosuquidar on the sex and time-dependent pharmacokinetics of parenterally administered talinolol in mice.
P-glycoprotein (P-gp) is an efflux protein that forms a tissue barrier and plays a role in the pharmacokinetics of drugs, limiting the influx of them and other xenobiotics into the cells, as expressed in various tissues such as liver, brain, intestinal mucosa and kidneys. Circadian clock controls many biological functions in mammals including xenobiotic metabolism and detoxification. Circadian rhythms of biological functions may affect the pharmacokinetics, and thus efficacy and/or toxicity of drugs. Aim of this study is to determine how the intraperitoneally administered pharmacokinetics of talinolol, as the probe substrate of P-gp, will change depending on the circadian time and sex in the presence of P-gp inhibitor zosuquidar. 20 mg/kg talinolol with or without 30 mg/kg zosuquidar was administred intraperitoneally to male and female mice at day period (ZT3) and night period (ZT15). Plasma and tissue concentrations of talinolol were determined by using validated HPLC/UV method. The protein levels of P-gp in the liver and small intestine in male and female mice were determined by PCR and Western blot techniques. P-gp protein levels in liver and ileum tissues were not different in female mice but higher in ZT15 as compared to ZT3 in male mice (p<0.05). There was no statistically significant difference in talinolol concentration depending on time and sex in the plasma and liver. There was significant time-dependent difference between ZT3 and ZT15 groups in ileum AUC0-5 h of talinolol (p<0.01). Talinolol plasma and liver AUC0-5 h were increased by zosuquidar administration regardless of dosing-time and sex (p<0.05). Our study findings are considerable in terms of revealing changes in pharmacokinetic profiles of P-gp substrates due to the time of administration in combination with P-gp inhibitors/modulators in managing polypharmacy.


Variations in Healthy Swallowing Mechanics During Various Bolus Conditions Using Computational Analysis of Swallowing Mechanics (CASM).
Bolus properties such as volume, consistency, and density have been shown to influence swallowing through the analysis of kinematics and timing in both normal and disordered swallowing. However, inherent intra- and inter-person variability of swallowing cloud interpretation of group data. Computational analysis of swallow mechanics (CASM) is an established methodology that uses coordinate tracking to map structural movements during swallowing and yields statistically powerful analyses at both the group and individual levels. In this study, the CASM method was used to determine how different bolus properties (volume, consistency, and density) altered swallow mechanics in healthy young adults at the group and individual levels. Videofluoroscopic swallow studies of 10 (4 females) healthy young adults were analyzed using CASM. Five bolus types were administered in each study (3 × 5 ml 40% w/v nectar, 3 × 5 ml 22% w/v thin, 3 × 5 ml 40% w/v thin, 3 × 10 ml 22% w/v thin, and 3 × 20 ml 22% w/v thin). Canonical variate analyses demonstrated that bolus condition did not affect swallowing mechanics at the group level, but bolus condition did affect pharyngeal swallow mechanics at the individual level. Functional swallow adaptations (e.g., hyoid movement) to bolus conditions were not uniform across participants, consistent with the nonsignificant group finding. These results suggest that individual swallowing systems of healthy young individuals vary in how they respond to bolus different conditions, highlighting the intrinsic variability of the swallow mechanism and the importance of individually tailored evaluation and treatment of swallowing. Findings warrant further investigation with different bolus conditions and aging and disordered populations.


Duodenal trauma: experience of a trauma center.
In the past decade 93 patients with duodenal injury were treated at a trauma center. By chart review, the age, sex, mechanism of injury, time to initial exploration (and the reason for delay), laboratory results, associated injury, extent of duodenal injury, operative repair, use of drains and tube decompression, morbidity, and cause of death were tabulated in order to improve management of these injuries. Of 87 patients surviving until the time of operative repair 73% required no repair (four) or primary closure (59). The remainder had either resection with primary anastomosis (ten), diverticulization (12), or pancreaticoduodenectomy (two). All patients with penetrating trauma were immediately explored. Patients with blunt trauma were explored on the basis of the judgment of house staff and faculty. Overall mortality was 18%. Significant morbidity occurred in 49% of survivors. This urban experience was heavily weighted toward penetrating injury. In this group early death usually resulted from associated vascular injuries. Blunt duodenal injury was less frequently associated with immediate exsanguination. Mortality associated with blunt duodenal injury was usually the result of delayed diagnosis. In blunt duodenal trauma peritoneal lavage is not diagnostic and may often be misleading; in this series 50% of lavages were false negatives. Blunt duodenal trauma, particularly when combined with pancreatic injury or delayed repair, was a lethal combination. A high index of suspicion and aggressive diagnostic evaluation (CT contrast study/amylase) in the emergency department is required in equivocal cases to avoid morbidity and mortality.


Self-rated health as a predictor of outcomes of type 2 diabetes patient education programmes in Denmark.
To explore if self-rated health (SRH) can predict differences in outcomes of patient education programmes among patients with type 2 diabetes over time. This is an observational cohort study conducted among 83 patients with type 2 diabetes participating in patient education programmes in the Capital Region of Denmark. Questionnaire data were collected by telephone interview at baseline and 2 weeks (77 participants, 93%) and 12 months (66, 80%) after the patient education ended. The seven-scale Health Education Impact Questionnaire (HeiQ) was the primary outcome. The independent variable was SRH, which was dichotomized into optimal or poor SRH. Changes over time were assessed using mean values and standard deviation (SD) at each time point and Cohen effect sizes. Odds ratios and 95% confidence intervals were calculated for the likelihood of having poor SRH for each baseline sociodemographic and health-related variable. Twelve months after patient education programmes, 60 (72%) patients with optimal SRH at baseline demonstrated increased self-management skills, overall acceptance of chronic illness, positive social interaction with others, and improved emotional well-being. Participants with poor SRH (23, 28%) reported no improvements over time. Not being married (odds ratio [OR] 7.79, P < 0.001), living alone (OR 4.93, P = 0.003), having hypertension (OR 8.00, P = 0.031), and being severely obese (OR 4.07, P = 0.009) were significantly associated with having poor SRH. After adjusting for sex, age and vocational training, marital status (OR 9.35, P < 0.001), cohabitation status (OR = 4.96, P = 0.005) and hypertension (OR 10.9, P = 0.03) remained associated with poor SRH. We found a strong association between SRH and outcomes of patient education, as measured by the HeiQ, at 12 months. Only participants with optimal SRH appeared to benefit from patient education. Other patient characteristics may be responsible to explain the observed difference between patients with optimal and poor SRH.


Levisunguis subaequalis n. g., n. sp., a tongue worm (Pentastomida: Porocephalida: Sebekidae) infecting softshell turtles, Apalone spp. (Testudines: Trionychidae) in the southeastern United States.
A new tongue worm (Pentastomida) belonging to the Sebekidae Sambon, 1922 (Porocephaloidea Sambon, 1922) is described based on exemplars collected from softshell terrapins Apalone spinifera aspera (Agassiz) and Apalone ferox (Schneider) in the southeastern United States; a new genus is erected to accommodate the new species. The new species belongs in the Sebekidae because adults possess four simple hooks arranged in a trapezoid pattern on the ventral surface of the cephalothorax, a mouth opening between the anterior and posterior pairs of hooks, a terminal anus, an elongated uterus with preanal uterine pore, and a Y-shaped seminal vesicle. Nymphs possess geminate hooks, and the new species has an aquatic life-cycle in which nymphs become encapsulated in the body cavity of a freshwater fish and mature in the lungs of a terrapin. The new genus is distinct from other genera in the Sebekidae primarily by differences in hook morphology and the fact that representatives use a terrapin as a definitive host. Nymphs infecting fish and presumed to be the new species matured as postlarval juveniles conspecific with the new species when they were fed to the eastern mud turtle, Kinosternon subrubrum (Lacépède). Nymphs of the new species are anatomically similar to but larger than nymphs of Sebekia mississippiensis Overstreet, Self & Vliet, 1985 found in the mesentery of fishes captured in Florida, U.S.A. Adults of the new species differ from those of S. mississippiensis based on hook features, chloride cell pore pattern on annuli, body size, and use of a turtle rather than crocodilian definitive host. The new species is the third North American member of the Sebekidae.


Mechanism of low-level microwave radiation effect on nervous system.
The aim of this study is to explain the mechanism of the effect of low-level modulated microwave radiation on brain bioelectrical oscillations. The proposed model of excitation by low-level microwave radiation bases on the influence of water polarization on hydrogen bonding forces between water molecules, caused by this the enhancement of diffusion and consequences on neurotransmitters transit time and neuron resting potential. Modulated microwave radiation causes periodic alteration of the neurophysiologic parameters and parametric excitation of brain bioelectric oscillations. The experiments to detect logical outcome of the mechanism on physiological level were carried out on 15 human volunteers. The 450-MHz microwave radiation modulated at 7, 40 and 1000 Hz frequencies was applied at the field power density of 0.16 mW/cm2. A relative change in the EEG power with and without radiation during 10 cycles was used as a quantitative measure. Experimental data demonstrated that modulated at 40 Hz microwave radiation enhanced EEG power in EEG alpha and beta frequency bands. No significant alterations were detected at 7 and 1000 Hz modulation frequencies. These results are in good agreement with the theory of parametric excitation of the brain bioelectric oscillations caused by the periodic alteration of neurophysiologic parameters and support the proposed mechanism. The proposed theoretical framework has been shown to predict the results of experimental study. The suggested mechanism, free of the restrictions related to field strength or time constant, is the first one providing explanation of low-level microwave radiation effects.