18F-FDG PET/CT and Urothelial Carcinoma: Impact on Management and Prognosis-A Multicenter Retrospective Study.
Objectives: To evaluate the ability of 18F-labeled fluoro-2-deoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) to predict survivorship of patients with bladder cancer (BC) and/or upper urinary tract carcinoma (UUTC). Materials: Data from patients who underwent FDG PET/CT for suspicion of recurrent urothelial carcinoma (UC) between 2007 and 2015 were retrospectively collected in a multicenter study. Disease management after the introduction of FDG PET/CT in the diagnostic algorithm was assessed in all patients. Kaplan-Meier and log-rank analysis were computed for survival assessment. A Cox regression analysis was used to identify predictors of recurrence and death, for BC, UUTC, and concomitant BC and UUTC. Results: Data from 286 patients were collected. Of these, 212 had a history of BC, 38 of UUTC and 36 of concomitant BC and UUTC. Patient management was changed in 114/286 (40%) UC patients with the inclusion of FDG PET/CT, particularly in those with BC, reaching 74% (n = 90/122). After a mean follow-up period of 21 months (Interquartile range: 4-28 mo.), 136 patients (47.4%) had recurrence/progression of disease. Moreover, 131 subjects (45.6%) died. At Kaplan-Meier analyses, patients with BC and positive PET/CT had a worse overall survival than those with a negative scan (log-rank < 0.001). Furthermore, a negative PET/CT scan was associated with a lower recurrence rate than a positive examination, independently from the primary tumor site. At multivariate analysis, in patients with BC and UUTC, a positive FDG PET/CT resulted an independent predictor of disease-free and overall survival (p < 0,01). Conclusions: FDG PET/CT has the potential to change patient management, particularly for patients with BC. Furthermore, it can be considered a valid survival prediction tool after primary treatment in patients with recurrent UC. However, a firm recommendation cannot be made yet. Further prospective studies are necessary to confirm our findings.


Isolation and characterization of epinectin, a novel adhesion protein for epithelial cells.
A 70,000-mol-wt protein was isolated from A431 carcinoma cell extracellular matrix that promotes cell substratum adhesion of these epidermoid tumor cells. Extracellular matrix was isolated by a modification of a procedure described by Hedman et al. (Hedman, K., M. Kurkinen, K. Alitalo, A. Vaheri, S. Johansson, and M. Höök, 1979 J. Cell Biol., 81:83-91) and Yamada and Weston (Yamada, K., and J. A. Weston, 1974, Proc. Natl. Acad. Sci. USA, 71:3492-3496). Cells were solubilized with 0.5% deoxycholate, 10 mM Tris, 0.9% NaCl, and 1 mM phenylmethylsulfonyl fluoride, pH 8.0. The residual matrix was then removed from the plates with 6 M urea and 1 mM phenylmethylsulfonyl fluoride and phosphate-buffered saline. SDS PAGE gels of the 6 M urea extract showed one major band at 70,000-mol-wt by Coomassie Blue staining. A 70,000-mol-wt isotopically-labeled band could also be extracted from the matrix of cells incubated with [35S]methionine. Because of the presence of this protein on squamous-derived epithelial cells we have called the 70,000-mol-wt molecule epinectin. Indirect immunofluorescence with polyclonal rabbit antibodies against epinectin stained A431 cells pericellularly in dense punctate accumulations and along the plasma membrane. Enzyme-linked immunoassays and gel-transfer immunolocalization studies showed that the extract did not cross-react with antibodies to fibronectin, laminin, serum-spreading factor, epibolin, or keratin. Additionally, antibodies to epinectin did not cross-react with these proteins. Further studies showed that epinectin does not bind to gelatin. Cell-adhesion assay, using radiolabeled A431 carcinoma cells on various adhesion-promoting substrates, showed that epinectin has similar adhesion-promoting capacity as serum-spreading factor, was somewhat less active than fibronectin, but more effective than laminin or epibolin. Epinectin appears to be a unique protein isolated from epidermoid tumor cells that is distinct from other known adhesion proteins.


[Myelotoxicity of paclitaxel (mitotax)].
Paclitaxel (single intravenous injection in a maximum tolerated dose of 4.6 mg/kg) to white outbred rats causes bone marrow hypoplasia, increased granulocyte and erythroid cell mitosis (metaphase-anaphase transition), and moderate pancytopenia developments in peripheral blood (hypoplastic anemia, deep, short-term neutropenia, lymphopenia and thrombocytopenia) in the first hours after injection. A considerable increase of polyploidy (4n) cells and a moderate increase in the structural changes (chromatid deletions) of chromosomes was observed on bone marrow metaphase plates in 24 h. The drug introduction causes earlier increase in the rate of thymus cells mitosis, a growth in the number of thymocytes with apoptosis signs, and a moderate decrease in the thymus and spleen weight. All changes are reversible. Long-term (90 days after injection) observation revealed decreased lymphocyte count in the peripheral blood and bone marrow and earlier thymus involution.


Mortality from pandemic A/H1N1 2009 influenza in England: public health surveillance study.
To establish mortality from pandemic A/H1N1 2009 influenza up to 8 November 2009. Investigation of all reported deaths related to pandemic A/H1N1 in England. Mandatory reporting systems established in acute hospitals and primary care. Physicians responsible for the patient. Numbers of deaths from influenza combined with mid-range estimates of numbers of cases of influenza to calculate age specific case fatality rates. Underlying conditions, time course of illness, and antiviral treatment. With the official mid-range estimate for incidence of pandemic A/H1N1, the overall estimated case fatality rate was 26 (range 11-66) per 100 000. It was lowest for children aged 5-14 (11 (range 3-36) per 100 000) and highest for those aged >or=65 (980 (range 300-3200) per 100 000). In the 138 people in whom the confirmed cause of death was pandemic A/H1N1, the median age was 39 (interquartile range 17-57). Two thirds of patients who died (92, 67%) would now be eligible for the first phase of vaccination in England. Fifty (36%) had no, or only mild, pre-existing illness. Most patients (108, 78%) had been prescribed antiviral drugs, but of these, 82 (76%) did not receive them within the first 48 hours of illness. Viewed statistically, mortality in this pandemic compares favourably with 20th century influenza pandemics. A lower population impact than previous pandemics, however, is not a justification for public health inaction. Our data support the priority vaccination of high risk groups. We observed delayed antiviral use in most fatal cases, which suggests an opportunity to reduce deaths by making timely antiviral treatment available, although the lack of a control group limits the ability to extrapolate from this observation. Given that a substantial minority of deaths occur in previously healthy people, there is a case for extending the vaccination programme and for continuing to make early antiviral treatment widely available.


Modifications of glass microelectrodes: a self-filling and a semifloating glass microelectrode.
A simple and inexpensive method for constructing a double capillary was developed for preparing a self-filling glass microelectrode. The modified method uses a thin capillary instead of a bundle of glass fibers and fuses it to the outer capillary wall with flame before pulling the latter with a puller. The validity of the new electrode was proven by measuring the proximal cell PD of the rat kidney slices (mean of the stable cell PDs lasting longer than 30 min,--69 mV). A semifloating electrode was modified from the original floating electrode to be applicable to the in vivo kidney preparation whose pulsatile and respiratory movements had hindered yielding a stable cell PD. This electrode is constructed simply by replacing the shaft of the Ling-Gerard glass microelectrode with a length of flexible silcone-rubber tubing. The cell PD recorded by this electrode is found to be more stable and can withstand minor tissue distortion in mammalian kidney cells in vivo and also in such a contractile tissue as the secretory coil of the eccrine sweat gland.


Improved metabolic status and insulin sensitivity in obese fatty (fa/fa) Zucker rats and Zucker Diabetic Fatty (ZDF) rats treated with the thiazolidinedione, MCC-555.
1. We examined the effect of chronic (21 days) oral treatment with the thiazolidinedione, MCC-555 ((+)-5-[[6-(2-fluorbenzyl)-oxy-2-naphy]methyl]-2,4-thiazo lid inedione) on metabolic status and insulin sensitivity in obese (fa/fa) Zucker rats and Zucker Diabetic Fatty (ZDF) rats which display an impaired glucose tolerance (IGT) or overt diabetic symptoms, respectively. 2. MCC-555 treatment to obese Zucker rats (10 and 30 mg kg(-1)) and diabetic ZDF rats (10 mg kg(-1)) reduced non-esterified fatty acid concentrations in both rat strains and reduced plasma glucose and triglyceride concentrations in the obese Zucker rats. Liver glycogen concentrations were significantly increased by chronic MCC-555 treatment in both obese Zucker rats (30 mg kg(-1) day(-1)) and diabetic ZDF rats (10 mg kg(-1) day(-1)), as compared with vehicle-treated lean and obese rats and there was a significant increase in hepatic glycogen synthase activity in MCC-555-treated diabetic ZDF rats as compared to vehicle-treated controls. 3. During a euglycaemic hyperinsulinaemic clamp, MCC-555-treated obese Zucker rats and diabetic ZDF rats required significantly higher glucose infusion rates to maintain stable glucose concentrations (2.01+/-0.19 mg min(-1) and 6.42+/-1.03 mg min(-1), respectively) than vehicle-treated obese controls (0.71+/-0.17 mg min(-1) and 2.09+/-0.71 mg min(-1); P<0.05), demonstrating improved insulin sensitivity in both Zucker and ZDF rats. MCC-555 treatment also enhanced insulin-induced suppression of hepatic glucose production in ZDF rats as measured using infusions of [6-3H]-glucose under clamp conditions. 4. In conclusion, we have demonstrated that MCC-555 improves metabolic status and insulin sensitivity in obese Zucker and diabetic ZDF rats. MCC-555 may prove a useful compound for alleviating the metabolic disturbances and IGT associated with insulin resistance in man.


Reconstitution of Formylglycine-generating Enzyme with Copper(II) for Aldehyde Tag Conversion.
To further our aim of synthesizing aldehyde-tagged proteins for research and biotechnology applications, we developed methods for recombinant production of aerobic formylglycine-generating enzyme (FGE) in good yield. We then optimized the FGE biocatalytic reaction conditions for conversion of cysteine to formylglycine in aldehyde tags on intact monoclonal antibodies. During the development of these conditions, we discovered that pretreating FGE with copper(II) is required for high turnover rates and yields. After further investigation, we confirmed that both aerobic prokaryotic (Streptomyces coelicolor) and eukaryotic (Homo sapiens) FGEs contain a copper cofactor. The complete kinetic parameters for both forms of FGE are described, along with a proposed mechanism for FGE catalysis that accounts for the copper-dependent activity.


Effect of a single bout of resistance exercise on arterial stiffness following a high-fat meal.
Consumption of a high-fat meal (HFM) causes postprandial lipemia and vascular dysfunction. Acute resistance exercise (RE) alone may also have a negative effect on vascular function. The purpose of this study was to measure arterial stiffness and postprandial lipemia after a HFM with or without acute RE. 9 recreationally active men (age 24±5 years, BMI 25±3 kg/m2) completed both: (1) HFM alone and (2) HFM+RE in a randomized order. Pulse wave velocity (PWV) from carotid to femoral artery and carotid to radial artery were used as measures of central/aortic and peripheral arterial stiffness, respectively. Circulating triglycerides (TRG) were obtained from finger stick samples as a marker of lipemia. There was a significant condition-by-time interaction for TRG (p<0.05). TRG levels increased significantly following both conditions with a significantly attenuated increase following HFM+RE (p<0.05). There was a significant condition-by-time interaction for peripheral PWV as this parameter increased following HFM, but decreased following HFM+RE (p=0.021). Central PWV did not change with HFM or HFM+RE (p>0.05). Following a HFM, acute RE attenuates postprandial lipemia and improves peripheral arterial stiffness without having a negative effect on central arterial stiffness.


Treatment characteristics and illness burden among European Americans, African Americans, and Latinos in the first 2,000 patients of the systematic treatment enhancement program for bipolar disorder.
Across ethnicity/race, prevalence rates of bipolar disorder are similar. However, African Americans and Latinos may receive less specialty mental health treatment and different medications, and may be less adherent to treatment regimens than European American patients. This study compared illness characteristics, treatment history, and overall functioning in a sample of European American, African American, and Latino patients with bipolar disorder. The samples were drawn from the first 2,000 patients enrolled in the Systematic Treatment Enhancement Program for Bipolar Disorder. There were 1,686 European Americans, 65 African Americans, and 77 Latinos. The data were collected upon study entry, with structured interviews, clinicianrated forms, and self report. African Americans had a greater likelihood of psychosis and fewer psychiatric medication prescriptions than did European Americans. Latinos had greater alcohol comorbidity, fewer psychiatric medication prescriptions and specialty treatment visits, and more frequent religious service attendance than did European Americans. Depression and manic episode severity and functional outcomes were similar across groups. Patients with bipolar disorder who are members of ethnic/racial minority groups continue to receive less intensive specialized mental health treatment than do European American patients. These findings may be related to provider, patient, or provider-patient relationship variables. Despite treatment differences and greater comorbidity and sympomatology, there were no differences among the three groups in overall functioning, suggesting additional outcome measurement is warranted.


Exposure to Bisphenol a Substitutes and Gestational Diabetes Mellitus: A Prospective Cohort Study in China.
Background: The association of bisphenol A (BPA) and gestational diabetes mellitus (GDM) has been investigated in only a small number of studies, and research on the associations between BPA substitutes and GDM is scarce. Objective: We aimed to investigate the associations of four bisphenols [bisphenol A (BPA), bisphenol S (BPS), bisphenol F (BPF), and bisphenol AF (BPAF)] levels in urine sample with the risk of gestational diabetes mellitus (GDM) and plasma glucose levels. Methods: A total of 1,841 pregnant women from a cohort study were recruited at their first prenatal examination between 2013 and 2015 in Wuhan, China. Concentrations of four bisphenols (BPA, BPS, BPF, BPAF) were measured in first-trimester urine samples using Ultra-high performance liquid chromatography system coupled to a Triple Quadrupole mass spectrometer (UHPLC-TQMS). An oral glucose tolerance test (OGTT) was performed at 24-28 gestational weeks and GDM was diagnosed post hoc using International Association of Diabetes and Pregnancy Study Groups criteria. We used multivariable logistic regression models to examine the associations of urinary bisphenols with the risk of GDM, and multiple linear regression models to determine the associations between bisphenols exposure and plasma glucose levels. Results: Urinary BPAF was associated with increased odds of GDM among women with normal pre-pregnancy BMI [adjusted odds ratio (aOR) = 1.70 (95% CI: 1.08, 2.67) for the highest group compared to the lowest group], and the association remained significant after additional adjustment for other bisphenols [aOR = 1.68 (95% CI: 1.03, 2.72)]. No significant associations were observed for other bisphenols and GDM. Consistent with the result of GDM, women in the highest BPAF category had a mean of 0.05 mmol/L (95% CI: 0.01, 0.09) higher fasting plasma glucose (FPG) levels than women in the lowest category. For BPA and plasma glucose, non-linear associations were observed between urinary BPA and FPG and the sum of the PG z-score among women who were overweight (p for non-linear association < 0.05). We also found that the per-unit increase in natural log transformed specific gravity adjusted BPS [ln (SG-adj BPS)] was associated with a 0.03 mmol/L (95% CI: 0.01, 0.04) increase in FPG levels and the associations might be modified by fetal sex (p for interaction < 0.05). Among women with female fetus, a per-unit increase in ln (SG-adj BPS) was associated with a 0.04 mmol/L (95% CI: 0.02, 0.06) increase in FPG, a 0.11 mmol/L (95% CI: 0.04, 0.17) increase in 1 h-PG and a 0.19 mmol/L (95% CI: 0.08, 0.30) increase in the sum of PG z-score. Conclusions: Our results provide evidence that BPAF and BPS might be potential risk factors of GDM, which require to be studied further.


Cross-β amyloid nanotubes for hydrolase-peroxidase cascade reactions.
Herein, we report the catalytic potential of short peptide based cross-β amyloid nanotubes with surface exposed histidine capable of binding hemin and showing facile cascade reactions, playing the dual roles of hydrolases and peroxidases, two of the most important classes of enzymes in extant biology. The activity of these simple systems exceeded those of modern and larger proteins like cytochrome C and hemoglobin. Further, evidence suggested that these self-assembled nanotubes foreshadow the process of intermediate channeling, a feature seen in the case of advanced enzymes.


Biotic and abiotic controls of Argentine ant invasion success at local and landscape scales.
Although the ecological success of introduced species hinges on biotic interactions and physical conditions, few experimental studies--especially on animals--have simultaneously investigated the relative importance of both types of factors. The lack of such research may stem from the common assumption that native and introduced species exhibit similar environmental tolerances. Here we combine experimental and spatial modeling approaches (1) to determine the relative importance of biotic and abiotic controls of Argentine ant (Linepithema humile) invasion success, (2) to examine how the importance of these factors changes with spatial scale in southern California (USA), and (3) to assess how Argentine ants differ from native ants in their environmental tolerances. A factorial field experiment that combined native ant removal with irrigation revealed that Argentine ants failed to invade any dry plots (even those lacking native ants) but readily invaded all moist plots. Native ants slowed the spread of Argentine ants into irrigated plots but did not prevent invasion. In areas without Argentine ants, native ant species showed variable responses to irrigation. At the landscape scale, Argentine ant occurrence was positively correlated with minimum winter temperature (but not precipitation), whereas native ant diversity increased with precipitation and was negatively correlated with minimum winter temperature. These results are of interest for several reasons. First, they demonstrate that fine-scale differences in the physical environment can eclipse biotic resistance from native competitors in determining community susceptibility to invasion. Second, our results illustrate surprising complexities with respect to how the abiotic factors limiting invasion can change with spatial scale, and third, how native and invasive species can differ in their responses to the physical environment. Idiosyncratic and scale-dependent processes complicate attempts to forecast where introduced species will occur and how their range limits may shift as a result of climate change.


BIBF 1120: triple angiokinase inhibitor with sustained receptor blockade and good antitumor efficacy.
Inhibition of tumor angiogenesis through blockade of the vascular endothelial growth factor (VEGF) signaling pathway is a novel treatment modality in oncology. Preclinical findings suggest that long-term clinical outcomes may improve with blockade of additional proangiogenic receptor tyrosine kinases: platelet-derived growth factor receptors (PDGFR) and fibroblast growth factor receptors (FGFR). BIBF 1120 is an indolinone derivative potently blocking VEGF receptor (VEGFR), PDGFR and FGFR kinase activity in enzymatic assays (IC(50), 20-100 nmol/L). BIBF 1120 inhibits mitogen-activated protein kinase and Akt signaling pathways in three cell types contributing to angiogenesis, endothelial cells, pericytes, and smooth muscle cells, resulting in inhibition of cell proliferation (EC(50), 10-80 nmol/L) and apoptosis. In all tumor models tested thus far, including human tumor xenografts growing in nude mice and a syngeneic rat tumor model, BIBF 1120 is highly active at well-tolerated doses (25-100 mg/kg daily p.o.), as measured by magnetic resonance imaging of tumor perfusion after 3 days, reducing vessel density and vessel integrity after 5 days, and inducing profound growth inhibition. A distinct pharmacodynamic feature of BIBF 1120 in cell culture is sustained pathway inhibition (up to 32 hours after 1-hour treatment), suggesting slow receptor off-kinetics. Although BIBF 1120 is rapidly metabolized in vivo by methylester cleavage, resulting in a short mean residence time, once daily oral dosing is fully efficacious in xenograft models. These distinctive pharmacokinetic and pharmacodynamic properties may help explain clinical observations with BIBF 1120, currently entering phase III clinical development.


Use of intravenous immunoglobulin for the treatment of autoimmune encephalitis: audit of the NHS experience.
The treatments of limbic and other autoimmune encephalitis include immunosuppression, symptomatic treatment, and in the case of paraneoplastic syndromes, appropriate therapy for underlying neoplasms. When immunotherapy is considered, intravenous immunoglobulin is one option for treatment, either alone or in combination with corticosteroids. To date, however, evidence for the use of intravenous immunoglobulin in this context comes from case series/expert reviews as no controlled trials have been performed. We aimed to analyse the NHS England Database of intravenous immunoglobulin usage, which was designed to log use and guide procurement, to explore usage and therapeutic effect of intravenous immunoglobulin in autoimmune encephalitis in England. We conducted a retrospective audit and review of the NHS England Database on intravenous immunoglobulin use. NHS England Database of intravenous immunoglobulin use which covers secondary and tertiary care prescribing and use of intravenous immunoglobulin for all patients in hospitals in England. Hospital in-patients with confirmed or suspected autoimmune/limbic encephalitis between September 2010 and January 2017. A total of 625 patients who were 18 years of age or older were treated with intravenous immunoglobulin for autoimmune encephalitis, of whom 398 were determined as having 'highly likely' or 'definite' autoimmune/limbic encephalitis. Ninety-six percent were treated with a single course of intravenous immunoglobulin. The availability and accuracy of reporting of outcomes was very poor, with complete data only available in 27% of all cases. This is the first review of data from this unique national database. Whilst there was evidence for clinical improvement in many cases of patients treated with intravenous immunoglobulin, the quality of outcome data was generally inadequate. Methods to improve quality, accuracy and completeness of reporting are crucial to maximise the potential value of this resource as an auditing tool.


Anaesthesia in connective tissue disorders.
Patients with the more common connective tissue disorders require surgical operations more frequently than has been realized. They may present the anaesthetist with many potential problems. A few minutes of careful questioning and examination pre-operatively may prevent a tragic situation. A history of drug therapy is essential pre-operative information, particularly since many of these patients will need augmentation or coverage with steroid drugs. The anaesthetist must be aware of the patient's general state of health and must search for evidence of pulmonary, cardiac or haematological abnormalities.


Investigating the Potential of Amnion-Based Scaffolds as a Barrier Membrane for Guided Bone Regeneration.
Guided bone regeneration is a new concept of large bone defect therapy, which employs a barrier membrane to afford a protected room for osteogenesis and prevent the invasion of fibroblasts. In this study, we developed a novel barrier membrane made from lyophilized multilayered acellular human amnion membranes (AHAM). After decellularization, the AHAM preserved the structural and biomechanical integrity of the amnion extracellular matrix (ECM). The AHAM also showed minimal toxic effects when cocultured with mesenchymal stem cells (MSCs), as evidenced by high cell density, good cell viability, and efficient osteogenic differentiation after 21-day culturing. The effectiveness of the multilayered AHAM in guiding bone regeneration was evaluated using an in vivo rat tibia defect model. After 6 weeks of surgery, the multilayered AHAM showed great efficiency in acting as a shield to avoid the invasion of the fibrous tissues, stabilizing the bone grafts and inducing the massive bone growth. We hence concluded that the advantages of the lyophilized multilayered AHAM barrier membrane are as follows: preservation of the structural and mechanical properties of the amnion ECM, easiness for preparation and handling, flexibility in adjusting the thickness and mechanical properties to suit the application, and efficiency in inducing bone growth and avoiding fibrous tissues invasion.


All-fibre supercontinuum laser for in vivo multispectral photoacoustic microscopy of lipids in the extended near-infrared region.
Among the numerous endogenous biological molecules, information on lipids is highly coveted for understanding both aspects of developmental biology and research in fatal chronic diseases. Due to the pronounced absorption features of lipids in the extended near-infrared region (1650-1850 nm), visualisation and identification of lipids become possible using multi-spectral photoacoustic (optoacoustic) microscopy. However, the spectroscopic studies in this spectral region require lasers that can produce high pulse energies over a broad spectral bandwidth to efficiently excite strong photoacoustic signals. The most well-known laser sources capable of satisfying the multi-spectral photoacoustic microscopy requirements (tunability and pulse energy) are tunable nanosecond optical parametric oscillators. However, these lasers have an inherently large footprint, thus preventing their use in compact microscopy systems. Besides, they exhibit low-repetition rates. Here, we demonstrate a compact all-fibre, high pulse energy supercontinuum laser that covers a spectral range from 1440 to 1870 nm with a 7 ns pulse duration and total energy of 18.3 μJ at a repetition rate of 100 kHz. Using the developed high-pulse energy source, we perform multi-spectral photoacoustic microscopy imaging of lipids, both ex vivo on adipose tissue and in vivo to study the development of Xenopus laevis tadpoles, using six different excitation bands over the first overtone transition of C-H vibration bonds (1650-1850 nm).


The structure of the pharmaceutical market in Iran using concentration indices.
The efficiency and function of the pharmaceutical sector, as a vital portion of the health system, have a significant effect on intermediate and final indices of health. In this research, the structure of the pharmaceutical market in Iran was examined through the calculation of concentration indices in 2011. In this cross-sectional study, the needed data was gathered from the Food and Drug Administration in the year 2011. Data were analyzed using SPSS software version 20 and Microsoft Office Excel software. Finally, two common measures of market concentration, the Concentration Ratio and the Herfindahl-Hirschman Index, were calculated. The largest and the smallest shares of the industry were 5.57% and 0.01%, respectively. The average industry share was 1.09%. The share range was calculated to be 5.56%. The Herfindahl-Hirschman Index was 248.5, which indicates a very low concentration of the pharmaceutical market in Iran. Also, based on the Concentration Ratio of 4 companies (18.39%), the concentration of the pharmaceutical market has been too low. The pharmaceutical market in Iran has a very low concentration and it does not have an exclusive mode in terms of market structure. Therefore, it can be attributed to the competitive model. The policy makers in this area can use this characteristic as a leverage to improve efficiency, fairness, revenue and health indices.


Coupled laboratory and field investigations resolve microbial interactions that underpin persistence in hydraulically fractured shales.
Hydraulic fracturing is one of the industrial processes behind the surging natural gas output in the United States. This technology inadvertently creates an engineered microbial ecosystem thousands of meters below Earth's surface. Here, we used laboratory reactors to perform manipulations of persisting shale microbial communities that are currently not feasible in field scenarios. Metaproteomic and metabolite findings from the laboratory were then corroborated using regression-based modeling performed on metagenomic and metabolite data from more than 40 produced fluids from five hydraulically fractured shale wells. Collectively, our findings show that Halanaerobium, Geotoga, and Methanohalophilus strain abundances predict a significant fraction of nitrogen and carbon metabolites in the field. Our laboratory findings also exposed cryptic predatory, cooperative, and competitive interactions that impact microorganisms across fractured shales. Scaling these results from the laboratory to the field identified mechanisms underpinning biogeochemical reactions, yielding knowledge that can be harnessed to potentially increase energy yields and inform management practices in hydraulically fractured shales.


SHIVA - a web application for drug resistance and tropism testing in HIV.
Drug resistance testing is mandatory in antiretroviral therapy in human immunodeficiency virus (HIV) infected patients for successful treatment. The emergence of resistances against antiretroviral agents remains the major obstacle in inhibition of viral replication and thus to control infection. Due to the high mutation rate the virus is able to adapt rapidly under drug pressure leading to the evolution of resistant variants and finally to therapy failure. We developed a web service for drug resistance prediction of commonly used drugs in antiretroviral therapy, i.e., protease inhibitors (PIs), reverse transcriptase inhibitors (NRTIs and NNRTIs), and integrase inhibitors (INIs), but also for the novel drug class of maturation inhibitors. Furthermore, co-receptor tropism (CCR5 or CXCR4) can be predicted as well, which is essential for treatment with entry inhibitors, such as Maraviroc. Currently, SHIVA provides 24 prediction models for several drug classes. SHIVA can be used with single RNA/DNA or amino acid sequences, but also with large amounts of next-generation sequencing data and allows prediction of a user specified selection of drugs simultaneously. Prediction results are provided as clinical reports which are sent via email to the user. SHIVA represents a novel high performing alternative for hitherto developed drug resistance testing approaches able to process data derived from next-generation sequencing technologies. SHIVA is publicly available via a user-friendly web interface.


Life expectancy after Mohs micrographic surgery in patients aged 90 years and older.
The population of people aged 90 years and older is expected to more than triple by 2050. The incidence of skin cancers is increasing. We sought to determine whether treatment of patients aged 90 years and older with skin cancer by Mohs micrographic surgery (MMS) changed their survival. A group of 214 patients aged 90 years and older who underwent MMS from July 1997 to May 2006 was identified. Patient gender, age, tumor type, size, site, defect size, number of MMS stages, and surgical repair were recorded. Comorbid medical conditions were assessed using the Charlson index. Actual survival was compared with expected length of survival using life tables. Data were analyzed by the Kaplan-Meier method with log rank significance tests. Average patient age was 92.3 years. All patients tolerated the procedures well with no deaths within 1 month after surgery. Median survival after surgery was 36.9 months. Tumor characteristics, defect size, number of surgical stages, and closure type did not affect survival. There was no significant difference in survival based on comorbidities according to Charlson scores. Instantaneous mortality hazard was highest 2 to 3 years after surgery. Specific causes of death were not accessible. This growing section of the population may safely undergo MMS.


Milk oral immunotherapy is effective in school-aged children.
To study the efficacy of oral immunotherapy (OIT) in schoolchildren with cow's milk (CM) allergy (CMA). Twenty-eight children aged 6-14 years with CMA documented by oral challenge were enrolled into a randomized, double-blind, placebo-controlled OIT study. In the active treatment, CM protein amount was increased during 23 weeks from 0.06 mg to a maximum of 6400 mg (200 mL of milk). Twenty-four (86%) patients completed the protocol: 16/18 in active and 8/10 in placebo groups. All children in the active and 2/3 in the placebo group suffered from symptoms considered by the parents as induced by milk. The children were contacted by phone 12 months later, and 13 (81%) used daily CM or milk products corresponding 6400 mg of CM protein. After double-blind OIT, all 10 children in the placebo group completed successfully an open-label OIT by an identical protocol, and all used daily CM or milk products 6 months later. Three to 3.5 years later, one child more had discontinued daily milk use. Thus, the long-term success rate was 22/28 (79%). This placebo-controlled, double-blind study confirmed that OIT was effective in desensitizing schoolchildren with CMA. With occasional exceptions, the reached desensitization sustained for more than 3 years.


Auditory brainstem correlates of perceptual timing deficits.
Children with language-based learning problems often exhibit pronounced speech perception difficulties. Specifically, these children have increased difficulty separating brief sounds occurring in rapid succession (temporal resolution). The purpose of this study was to better understand the consequences of auditory temporal resolution deficits from the perspective of the neural encoding of speech. The findings provide evidence that sensory processes relevant to cognition take place at much earlier levels than traditionally believed. Thresholds from a psychophysical backward masking task were used to divide children into groups with good and poor temporal resolution. Speech-evoked brainstem responses were analyzed across groups to measure the neural integrity of stimulus-time mechanisms. Results suggest that children with poor temporal resolution do not have an overall neural processing deficit, but rather a deficit specific to the encoding of certain acoustic cues in speech. Speech understanding relies on the ability to attach meaning to rapidly fluctuating changes of both the temporal and spectral information found in consonants and vowels. For this to happen properly, the auditory system must first accurately encode these time-varying acoustic cues. Speech perception difficulties that often co-occur in children with poor temporal resolution may originate as a neural encoding deficit in structures as early as the auditory brainstem. Thus, speech-evoked brainstem responses are a biological marker for auditory temporal processing ability.


BAY-8400: A Novel Potent and Selective DNA-PK Inhibitor which Shows Synergistic Efficacy in Combination with Targeted Alpha Therapies.
Eukaryotes have evolved two major pathways to repair potentially lethal DNA double-strand breaks. Homologous recombination represents a precise, DNA-template-based mechanism available during the S and G2 cell cycle phase, whereas non-homologous end joining, which requires DNA-dependent protein kinase (DNA-PK), allows for fast, cell cycle-independent but less accurate DNA repair. Here, we report the discovery of BAY-8400, a novel selective inhibitor of DNA-PK. Starting from a triazoloquinoxaline, which had been identified as a hit from a screen for ataxia telangiectasia and Rad3-related protein (ATR) inhibitors with inhibitory activity against ATR, ATM, and DNA-PK, lead optimization efforts focusing on potency and selectivity led to the discovery of BAY-8400. In in vitro studies, BAY-8400 showed synergistic activity of DNA-PK inhibition with DNA damage-inducing targeted alpha therapy. Combination of PSMA-targeted thorium-227 conjugate BAY 2315497 treatment of human prostate tumor-bearing mice with BAY-8400 oral treatment increased antitumor efficacy, as compared to PSMA-targeted thorium-227 conjugate monotherapy.


Minimizing Stimulus Current in a Wearable Pudendal Nerve Stimulator Using Computational Models.
After spinal cord injury, functions of the lower urinary tract may be disrupted. A wearable device with surface electrodes which can effectively control the bladder functions would be highly beneficial to the patients. A trans-rectal pudendal nerve stimulator may provide such a solution. However, the major limiting factor in such a stimulator is the high level of current it requires to recruit the nerve fibers. Also, the variability of the trajectory of the nerve in different individuals should be considered. Using computational models and an approximate trajectory of the nerve derived from an MRI study, it is demonstrated in this paper that it may be possible to considerably reduce the required current levels for trans-rectal stimulation of the pudendal nerve compared to the values previously reported in the literature. This was corroborated by considering an ensemble of possible and probable variations of the trajectory. The outcome of this study suggests that trans-rectal stimulation of the pudendal nerve is a plausible long term solution for treating lower urinary tract dysfunctions after spinal cord injury.


Inverse relationship between physical activity and arterial stiffness in adults with hypertension.
Physical activity has beneficial effects on arterial stiffness among healthy adults. There is a lack of data on this relationship in adults with hypertension. The majority of studies which have examined physical activity and arterial stiffness have used subjective measures of activity. The aim of this study was to investigate the relationship between objectively measured habitual physical activity and arterial stiffness in individuals with newly diagnosed essential hypertension. Adults attending an outpatient hypertension clinic were recruited into this cross sectional study. Physical activity was measured using a triaxial accelerometer. Pulse wave velocity (PWV) and augmentation index (AIx) were measured using applanation tonometry. Participant's full lipid profile and glucose were determined through the collection of a fasting blood sample. Fifty-three adults [51(14) years, 26 male] participated, 16 of whom had the metabolic syndrome. Inactivity was positively correlated with PWV (r = .53, P < .001) and AIx (r = .48, P < .001). There were significant inverse associations between habitual physical activity of all intensities and both AIx and PWV. In stepwise regression, after adjusting for potential confounders, physical activity was a significant predictor of AIx and PWV. Habitual physical activity of all intensities is associated with reduced arterial stiffness among adults with hypertension.


Effect of culture conditions on microorganism identification by matrix-assisted laser desorption ionization mass spectrometry.
Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) has been used to identify bacteria based upon protein signatures. This research shows that while some different proteins are produced by vegetative bacteria when they are cultured in different growth media, positive identification with MALDI-TOF MS is still possible with the protocol established at the Pacific Northwest National Laboratory (K. H. Jarman, S. T. Cebula, A. J. Saenz, C. E. Petersen, N. B. Valentine, M. T. Kingsley, and K. L. Wahl, Anal. Chem. 72:1217-1223, 2000). A core set of small proteins remain constant under at least four different culture media conditions and blood agar plates, including minimal medium M9, rich media, tryptic soy broth (TSB) or Luria-Bertani (LB) broth, and blood agar plates, such that analysis of the intact cells by matrix-assisted laser desorption/ionization mass spectrometry allows for consistent identification.


Adverse effects on health and wellbeing of working as a doctor: views of the UK medical graduates of 1974 and 1977 surveyed in 2014.
Objective To report on any adverse effects on health and wellbeing of working as a doctor, as described by senior doctors. Design Questionnaires sent in 2014 to all medical graduates of 1974 and 1977. Participants 3695 UK medical graduates. Setting United Kingdom. Main outcome measures Statements about adverse effects upon health, wellbeing and career. Results The aggregated response rate from contactable doctors was 84.6% (3695/4369). In response to the question 'Do you feel that working as a doctor has had any adverse effects on your own health or wellbeing?', 44% of doctors answered 'yes'. More GPs (47%) than hospital doctors (42%) specified that this was the case. Three-quarters of doctors who answered 'yes' cited 'stress/work-life balance/workload' as an adverse effect, and 45% mentioned illness. In response to the statement 'The NHS of today is a good employer when doctors become ill themselves', 28% of doctors agreed, 29% neither agreed nor disagreed and 43% disagreed. More women doctors (49%) than men doctors (40%) disagreed with this statement. More general practitioners (49%) disagreed than hospital doctors (37%). Conclusions Chronic stress and illness, which these doctors attributed to their work, were widely reported. Although recent changes may have alleviated some of these issues, there are lessons for the present and future if the NHS is to ensure that its medical workforce receives the support which enables current doctors to enjoy a full and satisfying career and to contribute fully to health service provision in the UK. Older doctors, in particular, need support to be able to continue successfully in their careers.


A Repeat Validated Population Questionnaire of a Defined Swedish Population Verifies Reduction in Leg Ulcer Prevalence Over Time.
This study was performed to measure if the introduced interventions in leg ulcer care in a selected Swedish county yield a detectable reduction in leg ulcer prevalence in the population. A validated postal questionnaire sent to 10,000 (9,868) randomly selected 30-89 year olds in the Skaraborg county (255,042 inhabitants). All positive responders were telephone interviewed and verified ulcer patients were clinically examined including assessment of arterial/venous circulation with hand-held Doppler and, where indicated, duplex ultrasound scanning. All results were compared with numbers from 1990 (initial study). The response rate was 82% (8,070/9,868), 200 active ulcers and 290 previous ulcers. The calculated prevalence was 0.75% for 30-89 years and 1.05% for 50-89 years (2.1% in 1990). The leg ulcer prevalence was reduced by 32% (0.52% compared to 0.77% in 1990), and the relative risk was reduced by 50% (95%, CI 0.36-0.69). The study shows a true reduction in leg ulcer prevalence detectable in the population supporting a successful care of leg ulcer patients.


Affirmative action programs and network benefits in the number of board positions.
Whereas governments are increasingly considering affirmative action programs to increase corporate board diversity, the effect of such programs can be superficial as they do not address the underlying problem, which is women's access to and inclusion in relevant corporate networks. To address this issue, we study the relationship among affirmative action programs (binding gender quotas and non-binding gender targets), director networks, and the number of board positions individual directors hold given their gender. We use personal, professional, and network characteristics of 25,127 unique directors from 2,435 public firms in 32 European countries over the period of 2000 through 2017. We find that in the absence of affirmative action programs, women directors benefit less from their networks than men directors suggesting the existence of a gender gap in network benefits. After the passage of binding gender quotas, this gender gap in network benefits narrows between women and men directors. Overall, this research suggests that binding gender quotas make director networks a more salient tool for hiring women and may help in leveling the playing field in the way these networks are used for achieving top management positions.


Absolute configuration of the diastereomers of adenosine 5'-O-(1-thiotriphosphate): consequences for the stereochemistry of polymerization by DNA-dependent RNA polymerase from Escherichia coli.
The diastereomers of uridyl-(3'-5')adenyl-O,O-phosphorothioate [Up(S)A] have been separated by high-performance liquid chromatography. Their identification as RP and SP follows from the RNase A digestion of these products. It was then shown, by the same method, that the R isomer is hydrolyzed by snake venom phosphodiesterase (PDEase) approximately 500 times faster than the S isomer. Similarly, the stereoisomer of adenosine 5'-O-(1-thiotriphosphate) (ATPalphaS), until now arbitrarily designated as isomer B, is hydrolyzed ca 400 times faster by PDEase than is isomer A. From these results it is concluded that the R isomers of Up(S)A and ATPalphaS, isomers B, have the same absolute configuration. It then follows that isomer A of ATPalphaS, the preferred of the two isomers as substrate for DNA-dependent RNA polymerase, has the S configuration. The implications for the stereochemistry of action of the latter enzyme are discussed.


Antarctic-wide array of high-resolution ice core records reveals pervasive lead pollution began in 1889 and persists today.
Interior Antarctica is among the most remote places on Earth and was thought to be beyond the reach of human impacts when Amundsen and Scott raced to the South Pole in 1911. Here we show detailed measurements from an extensive array of 16 ice cores quantifying substantial toxic heavy metal lead pollution at South Pole and throughout Antarctica by 1889 - beating polar explorers by more than 22 years. Unlike the Arctic where lead pollution peaked in the 1970s, lead pollution in Antarctica was as high in the early 20(th) century as at any time since industrialization. The similar timing and magnitude of changes in lead deposition across Antarctica, as well as the characteristic isotopic signature of Broken Hill lead found throughout the continent, suggest that this single emission source in southern Australia was responsible for the introduction of lead pollution into Antarctica at the end of the 19(th) century and remains a significant source today. An estimated 660 t of industrial lead have been deposited over Antarctica during the past 130 years as a result of mid-latitude industrial emissions, with regional-to-global scale circulation likely modulating aerosol concentrations. Despite abatement efforts, significant lead pollution in Antarctica persists into the 21(st) century.


Predictive models to determine imagery strategies employed by children to judge hand laterality.
A commonly used paradigm to study motor imagery is the hand laterality judgment task. The present study aimed to determine which strategies young children employ to successfully perform this task. Children of 5 to 8 years old (N = 92) judged laterality of back and palm view hand pictures in different rotation angles. Response accuracy and response duration were registered. Response durations of the trials with a correct judgment were fitted to a-priori defined predictive sinusoid models, representing different strategies to successfully perform the hand laterality judgment task. The first model predicted systematic changes in response duration as a function of rotation angle of the displayed hand. The second model predicted that response durations are affected by biomechanical constraints of hand rotation. If observed data could be best described by the first model, this would argue for a mental imagery strategy that does not involve motor processes to solve the task. The second model reflects a motor imagery strategy to solve the task. In line with previous research, we showed an age-related increase in response accuracy and decrease in response duration in children. Observed data for both back and palm view showed that motor imagery strategies were used to perform hand laterality judgments, but that not all the children use these strategies (appropriately) at all times. A direct comparison of response duration patterns across age sheds new light on age-related differences in the strategies employed to solve the task. Importantly, the employment of the motor imagery strategy for successful task performance did not change with age.


Systematic reviews and meta-analyses on treatment of asthma: critical evaluation.
To evaluate the clinical, methodological, and reporting aspects of systematic reviews and meta-analyses on the treatment of asthma and to compare those published by the Cochrane Collaboration with those published in paper based journals. Analysis of studies identified from Medline, CINAHL, HealthSTAR, EMBASE, Cochrane Library, personal collections, and reference lists. Articles describing a systematic review or a meta-analysis of the treatment of asthma that were published as a full report, in any language or format, in a peer reviewed journal or the Cochrane Library. General characteristics of studies reviewed and methodological characteristics (sources of articles; language restrictions; format, design, and publication status of studies included; type of data synthesis; and methodological quality). 50 systematic reviews and meta-analyses were included. More than half were published in the past two years. Twelve reviews were published in the Cochrane Library and 38 were published in 22 peer reviewed journals. Forced expiratory volume in one second was the most frequently used outcome, but few reviews evaluated the effect of treatment on costs or patient preferences. Forty reviews were judged to have serious or extensive flaws. All six reviews associated with industry were in this group. Seven of the 10 most rigorous reviews were published in the Cochrane Library. Most reviews published in peer reviewed journals or funded by industry have serious methodological flaws that limit their value to guide decisions. Cochrane reviews are more rigorous and better reported than those published in peer reviewed journals.


Terminal diversification of the myocyte lineage generates Purkinje fibers of the cardiac conduction system.
The rhythmic contraction of the vertebrate heart is dependent on organized propagation of electrical excitation through the cardiac conduction system. Because both muscle- and neuron-specific genes are co-expressed in cells forming myocardial conduction tissues, two origins, myogenic and neural, have been suggested for this specialized tissue. Using replication-defective retroviruses, encoding recombinant beta-galactosidase (beta-gal), we have analyzed cell lineage for Purkinje fibers (i.e., the peripheral elements of the conduction system) in the chick heart. Functioning myocyte progenitors were virally tagged at embryonic day 3 of incubation (E3). Clonal beta-gal+ populations of cells, derived from myocytes infected at E3 were examined at 14 (E14) and 18 (E18) days of embryonic incubation. Here, we report that a subset of clonally related myocytes differentiates into conductile Purkinje fibers, invariably in close spatial association with forming coronary arterial blood vessels. These beta-gal+ myogenic clones, containing both working myocytes and Purkinje fibers, did not incorporate cells contributing to tissues of the central conduction system (e.g. atrioventricular ring and bundles). In quantitative analyses, we found that whereas the number of beta-gal+ myocyte nuclei per clone more than doubled between E14 and E18, the number of beta-gal+ Purkinje fiber nuclei remained constant.(ABSTRACT TRUNCATED AT 250 WORDS)


Bcl-2-associated athanogene 3 protects the heart from ischemia/reperfusion injury.
Bcl-2-associated athanogene 3 (BAG3) is an evolutionarily conserved protein expressed at high levels in the heart and the vasculature and in many cancers. While altered BAG3 expression has been associated with cardiac dysfunction, its role in ischemia/reperfusion (I/R) is unknown. To test the hypothesis that BAG3 protects the heart from reperfusion injury, in vivo cardiac function was measured in hearts infected with either recombinant adeno-associated virus serotype 9-expressing (rAAV9-expressing) BAG3 or GFP and subjected to I/R. To elucidate molecular mechanisms by which BAG3 protects against I/R injury, neonatal mouse ventricular cardiomyocytes (NMVCs) in which BAG3 levels were modified by adenovirus expressing (Ad-expressing) BAG3 or siBAG3 were exposed to hypoxia/reoxygenation (H/R). H/R significantly reduced NMVC BAG3 levels, which were associated with enhanced expression of apoptosis markers, decreased expression of autophagy markers, and reduced autophagy flux. The deleterious effects of H/R on apoptosis and autophagy were recapitulated by knockdown of BAG3 with Ad-siBAG3 and were rescued by Ad-BAG3. In vivo, treatment of mice with rAAV9-BAG3 prior to I/R significantly decreased infarct size and improved left ventricular function when compared with mice receiving rAAV9-GFP and improved markers of autophagy and apoptosis. These findings suggest that BAG3 may provide a therapeutic target in patients undergoing reperfusion after myocardial infarction.


Overcoming drug resistance with alginate oligosaccharides able to potentiate the action of selected antibiotics.
The uncontrolled, often inappropriate use of antibiotics has resulted in the increasing prevalence of antibiotic-resistant pathogens, with major cost implications for both United States and European health care systems. We describe the utilization of a low-molecular-weight oligosaccharide nanomedicine (OligoG), based on the biopolymer alginate, which is able to perturb multidrug-resistant (MDR) bacteria by modulating biofilm formation and persistence and reducing resistance to antibiotic treatment, as evident using conventional and robotic MIC screening and microscopic analyses of biofilm structure. OligoG increased (up to 512-fold) the efficacy of conventional antibiotics against important MDR pathogens, including Pseudomonas, Acinetobacter, and Burkholderia spp., appearing to be effective with several classes of antibiotic (i.e., macrolides, β-lactams, and tetracyclines). Using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM), increasing concentrations (2%, 6%, and 10%) of alginate oligomer were shown to have a direct effect on the quality of the biofilms produced and on the health of the cells within that biofilm. Biofilm growth was visibly weakened in the presence of 10% OligoG, as seen by decreased biomass and increased intercellular spaces, with the bacterial cells themselves becoming distorted and uneven due to apparently damaged cell membranes. This report demonstrates the feasibility of reducing the tolerance of wound biofilms to antibiotics with the use of specific alginate preparations.


Casuarina root exudates alter the physiology, surface properties, and plant infectivity of Frankia sp. strain CcI3.
The actinomycete genus Frankia forms nitrogen-fixing symbioses with 8 different families of actinorhizal plants, representing more than 200 different species. Very little is known about the initial molecular interactions between Frankia and host plants in the rhizosphere. Root exudates are important in Rhizobium-legume symbiosis, especially for initiating Nod factor synthesis. We measured differences in Frankia physiology after exposure to host aqueous root exudates to assess their effects on actinorhizal symbioses. Casuarina cunninghamiana root exudates were collected from plants under nitrogen-sufficient and -deficient conditions and tested on Frankia sp. strain CcI3. Root exudates increased the growth yield of Frankia in the presence of a carbon source, but Frankia was unable to use the root exudates as a sole carbon or energy source. Exposure to root exudates caused hyphal "curling" in Frankia cells, suggesting a chemotrophic response or surface property change. Exposure to root exudates altered Congo red dye binding, which indicated changes in the bacterial surface properties at the fatty acid level. Fourier transform infrared spectroscopy (FTIR) confirmed fatty acid changes and revealed further carbohydrate changes. Frankia cells preexposed to C. cunninghamiana root exudates for 6 days formed nodules on the host plant significantly earlier than control cells. These data support the hypothesis of early chemical signaling between actinorhizal host plants and Frankia in the rhizosphere.


Evaluation of an Emergency Bulk Chlorination Project Targeting Drinking Water Vendors in Cholera-Affected Wards of Dar es Salaam and Morogoro, Tanzania.
In August 2015, an outbreak of cholera was reported in Tanzania. In cholera-affected areas of urban Dar es Salaam and Morogoro, many households obtained drinking water from vendors, who sold water from tanks ranging in volume from 1,000 to 20,000 L. Water supplied by vendors was not adequately chlorinated. The Tanzanian Ministry of Health, Community Development, Gender, Elderly and Children and the U.N. Children's Fund, Tanzania, collaborated to enroll and train vendors to treat their water with 8.68-g sodium dichloroisocyanurate tablets (Medentech, Ireland). The Centers for Disease Control and Prevention (CDC) provided monitoring and evaluation support. Vendors were provided a 3-month supply of chlorine tablets. A baseline assessment and routine monitoring were conducted by ward environmental health officers. Approximately 3 months after chlorine tablet distribution, an evaluation of the program was conducted. The evaluation included a full enumeration of all vendors, an in-depth survey with half of the vendors enumerated, and focus group discussions. In total, 797 (88.9%) vendors were included in the full enumeration and 392 in the in-depth survey. Free residual chlorine (FRC) was detected in 12.0% of tanks at baseline and 69.6% of tanks during the evaluation; however, only 17.4% of these tanks had FRC ≥ 0.5 mg/L. The results suggest high acceptability and use of the chlorine tablets by water vendors. However, given variation in the water source used and longer storage times, dosing could be increased in future programming. Bulk chlorination using chlorine tablets offers an efficient community-level approach to treating water closer to the point of use.


Odour-based context reinstatement effects with indirect measures of memory: the curious case of rosemary.
Previous studies examining environmental context-dependent memory (ECDM) effects using indirect measures of memory have produced inconsistent findings. We report three experiments that examined ECDM in an indirect memory paradigm (word-fragment completion) using ambient odours as environmental contexts. Expt 1 manipulated the odour present at learning and testing (rosemary or lemon) to produce reinstated-context or switched-context conditions. Reinstating rosemary led to a striking ECDM effect, indicating that indirect memory testing can be sensitive to ECDM manipulations. Odour ratings also indicated that rosemary induced a more unpleasant mood in participants than lemon. Expt 2 assessed the influence on indirect retrieval of odour-based mood induction as well as odour distinctiveness, and indicated that rosemary's capacity to promote ECDM effects appears to arise from an additive combination of its unpleasantness-inducing properties and its distinctiveness. Expt 3 partially supported these proposals. Overall, our findings indicate that some odours are capable of producing ECDM effects using indirect testing procedures. Moreover, it appears that it is the inherent proprieties of odours on dimensions such as unpleasantness and distinctiveness that mediate the emergence of ECDM effects, thereby explaining the particular potency of rosemary's mnemonic influence when it is reinstated.


Integrating HIV and mental health interventions to address a global syndemic among men who have sex with men.
In this Series paper, we review evidence on the co-occurring and synergistic epidemics (syndemic) of HIV and mental health problems worldwide among men who have sex with men (MSM). The multilevel determinants of this global syndemic include structural factors that enable stigma, systematic bias, and violence towards MSM across geographical and cultural contexts. Cumulative exposure to these factors over time results in population-level inequities in the burden of HIV infections and mental health problems among MSM. Evidence for this syndemic among MSM is strongest in the USA, Canada, western Europe, and parts of Asia and Latin America, with emerging evidence from sub-Saharan Africa. Integrated interventions are needed to address syndemics of HIV and mental health problems that challenge the wellbeing of MSM populations worldwide, and such interventions should consider various mental health conditions (eg, depression, anxiety, trauma, and suicidality) and their unique expressions and relationships with HIV outcomes depending on cultural contexts. In addition, interventions should identify and intervene with locally relevant structural factors that result in HIV and mental health vulnerabilities among MSM.


Mutually exclusive expression of the Strongylocentrotus purpuratus Spec1 gene and its Lytechinus pictus homologue in cells of hybrid embryos.
The expression of the Spec1 gene of Strongylocentrotus purpuratus and its Lytechinus pictus homologue LpS1 was analyzed in reciprocal hybrid embryos of these two species of sea urchin. While the time course of accumulation of Spec1 mRNA was nearly normal in hybrid embryo populations, the accumulation of LpS1 mRNA was not. This was particularly evident in plutei, where the level of LpS1 mRNA was less than 5% that in normal L. pictus plutei. In situ hybridization analysis of serial sections indicated that LpS1 mRNA was detectable in only about 2% of hybrid plutei in either cross, whereas Spec1 mRNA was present in nearly all hybrid plutei; expression of either homologue was appropriately restricted to the aboral ectoderm. In crosses of L. pictus eggs with S. purpuratus sperm (LpSp), about 1% of hybrid plutei expressed LpS1 RNA in most or all aboral ectoderm cells at normal levels, and did not express Spec1 RNA; in another 1% of the LpSp hybrid plutei the Spec1 and LpS1 transcripts were present at normal levels in complementary, non-overlapping patches of contiguous aboral ectoderm cells. In the reciprocal SpLp cross, each hybrid pluteus expressed either only the LpS1 gene (about 2%) or only the Spec1 gene throughout the aboral ectoderm. In SpLp hybrid gastrulae the level of LpS1 mRNA was less restricted; about 2% of the embryos contained only LpS1 RNA, and about half expressed only Spec1 transcripts, but in the remaining embryos Spec1 and LpS1 transcripts were coexpressed in the same aboral ectoderm cells.(ABSTRACT TRUNCATED AT 250 WORDS)


A pharmacogenetic versus a clinical algorithm for warfarin dosing.
The clinical utility of genotype-guided (pharmacogenetically based) dosing of warfarin has been tested only in small clinical trials or observational studies, with equivocal results. We randomly assigned 1015 patients to receive doses of warfarin during the first 5 days of therapy that were determined according to a dosing algorithm that included both clinical variables and genotype data or to one that included clinical variables only. All patients and clinicians were unaware of the dose of warfarin during the first 4 weeks of therapy. The primary outcome was the percentage of time that the international normalized ratio (INR) was in the therapeutic range from day 4 or 5 through day 28 of therapy. At 4 weeks, the mean percentage of time in the therapeutic range was 45.2% in the genotype-guided group and 45.4% in the clinically guided group (adjusted mean difference, [genotype-guided group minus clinically guided group], -0.2; 95% confidence interval, -3.4 to 3.1; P=0.91). There also was no significant between-group difference among patients with a predicted dose difference between the two algorithms of 1 mg per day or more. There was, however, a significant interaction between dosing strategy and race (P=0.003). Among black patients, the mean percentage of time in the therapeutic range was less in the genotype-guided group than in the clinically guided group. The rates of the combined outcome of any INR of 4 or more, major bleeding, or thromboembolism did not differ significantly according to dosing strategy. Genotype-guided dosing of warfarin did not improve anticoagulation control during the first 4 weeks of therapy. (Funded by the National Heart, Lung, and Blood Institute and others; COAG ClinicalTrials.gov number, NCT00839657.).


Differential pharmacological effects on brain reactivity and plasticity in Alzheimer's disease.
Acetylcholinesterase inhibitors (AChEIs) are the most commonly prescribed monotherapeutic medications for Alzheimer's disease (AD). However, their underlying neurophysiological effects remain largely unknown. We investigated the effects of monotherapy (AChEI) and combination therapy (AChEI and memantine) on brain reactivity and plasticity. Patients treated with monotherapy (AChEI) (N = 7) were compared to patients receiving combination therapy (COM) (N = 9) and a group of age-matched, healthy controls (HCs) (N = 13). Cortical reactivity and plasticity of the motor cortex were examined using transcranial magnetic stimulation. Cognitive functions were assessed with the cognitive subscale of the Alzheimer Disease Assessment Scale-Cognitive Subscale (ADAS-Cog), activities of daily living (ADLs) with the ADCS-ADL. In addition we assessed the degree of brain atrophy by measuring brain-scalp distances in seven different brain areas. Patient groups differed in resting motor threshold and brain atrophy, with COM showing a lower motor threshold but less atrophy than AChEI. COM showed similar plasticity effects as the HC group, while plasticity was reduced in AChEI. Long-interval intracortical inhibition (LICI) was impaired in both patient groups when compared to HC. ADAS-Cog scores were positively correlated with LICI measures and with brain atrophy, specifically in the left inferior parietal cortex. AD patients treated with mono- or combination-therapy show distinct neurophysiological patterns. Further studies should investigate whether these measures might serve as biomarkers of treatment response and whether they could guide other therapeutic interventions.


Reaction-diffusion basis of retroviral infectivity.
Retrovirus particle (virion) infectivity requires diffusion and clustering of multiple transmembrane envelope proteins (Env3) on the virion exterior, yet is triggered by protease-dependent degradation of a partially occluding, membrane-bound Gag polyprotein lattice on the virion interior. The physical mechanism underlying such coupling is unclear and only indirectly accessible via experiment. Modelling stands to provide insight but the required spatio-temporal range far exceeds current accessibility by all-atom or even coarse-grained molecular dynamics simulations. Nor do such approaches account for chemical reactions, while conversely, reaction kinetics approaches handle neither diffusion nor clustering. Here, a recently developed multiscale approach is considered that applies an ultra-coarse-graining scheme to treat entire proteins at near-single particle resolution, but which also couples chemical reactions with diffusion and interactions. A model is developed of Env3 molecules embedded in a truncated Gag lattice composed of membrane-bound matrix proteins linked to capsid subunits, with freely diffusing protease molecules. Simulations suggest that in the presence of Gag but in the absence of lateral lattice-forming interactions, Env3 diffuses comparably to Gag-absent Env3 Initial immobility of Env3 is conferred through lateral caging by matrix trimers vertically coupled to the underlying hexameric capsid layer. Gag cleavage by protease vertically decouples the matrix and capsid layers, induces both matrix and Env3 diffusion, and permits Env3 clustering. Spreading across the entire membrane surface reduces crowding, in turn, enhancing the effect and promoting infectivity.This article is part of the themed issue 'Multiscale modelling at the physics-chemistry-biology interface'.


Extending the shine-through effect to classical masking paradigms.
A vernier, presented for a short time, shines through a following grating if the grating contains nine and more elements but remains largely invisible for smaller gratings. Therefore, extended grating masks yield, surprisingly, less masking than smaller ones. Here, we show that this mask size effect is not unique to grating masks. Masking diminishes if the size of classical pattern-, noise-, light-, and metacontrast masks increases and if these masks are regular, i.e. highly ordered.


The latency-associated nuclear antigen of Kaposi sarcoma-associated herpesvirus induces B cell hyperplasia and lymphoma.
Kaposi sarcoma-associated herpesvirus (KSHV) is a human lymphotropic herpesvirus. It is implicated in B cell neoplasias such as primary effusion lymphoma and multicentric Castleman disease in AIDS patients. The KSHV latency-associated nuclear antigen (LANA) is consistently expressed in all KSHV-associated tumor cells and was shown to bind the tumor suppressor proteins p53 and pRb. To test LANA's contribution to lymphomagenesis in vivo we generated transgenic mice expressing LANA under the control of its own promoter, which is B cell specific. All of the transgenic mice developed splenic follicular hyperplasia due to an expansion of IgM+ IgD+ B cells and showed increased germinal center formation. We also observed lymphomas, implying that LANA can activate B cells and provide the first step toward lymphomagenesis.


Adaptation to Low Salinity Promotes Genomic Divergence in Atlantic Cod (Gadus morhua L.).
How genomic selection enables species to adapt to divergent environments is a fundamental question in ecology and evolution. We investigated the genomic signatures of local adaptation in Atlantic cod (Gadus morhua L.) along a natural salinity gradient, ranging from 35‰ in the North Sea to 7‰ within the Baltic Sea. By utilizing a 12 K SNPchip, we simultaneously assessed neutral and adaptive genetic divergence across the Atlantic cod genome. Combining outlier analyses with a landscape genomic approach, we identified a set of directionally selected loci that are strongly correlated with habitat differences in salinity, oxygen, and temperature. Our results show that discrete regions within the Atlantic cod genome are subject to directional selection and associated with adaptation to the local environmental conditions in the Baltic- and the North Sea, indicating divergence hitchhiking and the presence of genomic islands of divergence. We report a suite of outlier single nucleotide polymorphisms within or closely located to genes associated with osmoregulation, as well as genes known to play important roles in the hydration and development of oocytes. These genes are likely to have key functions within a general osmoregulatory framework and are important for the survival of eggs and larvae, contributing to the buildup of reproductive isolation between the low-salinity adapted Baltic cod and the adjacent cod populations. Hence, our data suggest that adaptive responses to the environmental conditions in the Baltic Sea may contribute to a strong and effective reproductive barrier, and that Baltic cod can be viewed as an example of ongoing speciation.


Integrating mechanisms of response and resistance against the tubulin binding agent Eribulin in preclinical models of osteosarcoma.
Osteosarcoma is the most frequently occurring bone cancer in children and adolescents. Unfortunately, treatment failures are common. Eribulin is a synthetic microtubule inhibitor that has demonstrated activity in preclinical osteosarcoma models. The effects of eribulin were evaluated in two human osteosarcoma cell lines as well as in eribulin-sensitive and -resistant osteosarcoma xenograft tumors of the Pediatric Preclinical Testing Program (PPTP) by characterizing cell viability, microtubule destabilization, mitotic arrest and mechanism of cell death. Eribulin demonstrated cytotoxic activity in vitro, through promotion of microtubule dynamic instability, arrest of cells in the G2/M phase, mitotic catastrophe and cell death. The microtubule-destabilizing protein stathmin-1 (STMN1) was coimmunoprecipitated with the cyclin-dependent kinase inhibitor p27 indicating that these cytoplasmic complexes can protect cells from the microtubule destabilizing effect of eribulin. Increased tumoral expression of P-glycoprotein (P-gp) and TUBB3 were also associated with lower drug sensitivity. In summary, eribulin successfully blocked cells in G2/M phase but interfered with mitochondria activity to inhibit proteins involved in apoptosis. Understanding the complex and inter-related mechanisms involved in the overall drug response to eribulin may help in the design of therapeutic strategies that enhance drug activity and improve benefits of eribulin in pediatric patients with osteosarcoma.


Suspended particles in wastewater: their optical, sedimentation and acoustical characterization and modeling.
Wastewater regulation and treatment is still a major concern in planetary pollution management. Some pollutants, referred to as particulate matter, consist of very small particles just suspended in the water. Various techniques are used for the suspended particles survey. Few of them are able to provide real-time data. The development of new, real time instruments needs the confrontation with real wastewater. Due its instability, the modeling of wastewater in terms of suspended solids was explored. Knowing the description of real wastewater, we tried to produce a synthetic mixture made of basic organic ingredients. A good agreement in terms of turbidity and settling velocity was observed between the artificial wastewater matrix and the real one. The investigation of the individual contribution of the different compounds to the acoustical signal showed a more complex dependence. Thus the modeling of wastewater with reference to turbidity and settling velocity is not sufficient to describe it acoustically. Further studies should lead to a good comparison of the acoustical and turbidity behavior of wastewater.


[Accuracy of measurement of radiochromic film density for high-energy X-rays].
Radiochromic film (RC-film) is of great interest as a film-type dosimeter for radiation oncology applications. We present a two-dimensional image-based evaluation of the measurement accuracy of a commercial RC-film product (Gafchromic MD-55-2 film, ISP TECHNOLOGIES, Inc.) by using a commercial Laser Densitometer (Model 1710, Computerized Medical Systems, Inc.) as an optical density imaging system. The coefficient of variation of the density (pixel-value) in one sample was approximately 3% to 11% at 3 Gy or less, and 3% or less at 4 to 60 Gy. Although the coefficient of variation between three samples at the same dose was about 14% at 1 Gy, it decreased as the dose increased, reaching several percent. In 1 to 6 Gy samples, geometric imaging artifacts [interference (moire) patterns] were observed, and it was found that scan-sampling pitch influenced the accuracy of measurement of the density of the sample. To improve the accuracy of density measurement, sufficient knowledge about characteristic features of the density measuring system is essential.


High-performance liquid chromatographic determination of diazepam in plasma of children with severe malaria.
A sensitive, selective and reproducible reversed-phase HPLC method with ultraviolet detection was developed for the quantification of diazepam in small plasma samples from children with severe malaria. The method involves plasma deproteinization with acetonitrile, followed by liquid-liquid extraction with ethyl acetate-n-hexane. Diazepam was eluted at ambient temperatures from a reversed-phase C18 column with an acidic (pH 3.5) aqueous mobile phase (10 mM KH2PO4-acetonitrile, 69:31, v/v). Calibration curves in spiked plasma were linear from 10 to 200 ng (r2 > or = 0.99). The limit of detection was 5.0 ng/ml, and relative recoveries at 25 and 180 ng were >87%. Intra- and inter-assay relative standard deviations were <15%. There was no interference from drugs commonly administered to children with severe malaria (phenobarbitone, phenytoin, chloroquine, quinine, sulfadoxine, pyrimethamine, halofantrine, cycloguanil, chlorcycloguanil, acetaminophen and salicylate). This method has been used for monitoring plasma diazepam concentrations in children with seizures associated with severe malaria.


Levels of heavy metals and essential minerals in hair samples of children with autism in Oman: a case-control study.
Toxic levels of heavy metals and low levels of essential minerals have been suggested to play a critical role in the pathogenesis of autism spectrum disorders (ASD). This study documents the levels of heavy metals and essential minerals in hair samples of children with ASD in Muscat, the urbanized capital of Oman, Muscat. The study included 27 children with ASD and 27 matched non-ASD controls. Parental interviews were held and dietary intake questionnaires completed in conjunction with the collection of hair samples. Analysis of heavy metals and essential minerals was carried out by inductively coupled plasma mass spectrometry. Chi-square analysis and non-parametric Fisher's exact tests were used to assess statistical significance. Children with ASD had significantly higher levels of all 11 analyzed heavy metals in their hair samples (P < 0.05), ranging from 150 to 365 % of control levels. ASD children also had significantly higher levels of essential minerals sulfur, sodium, magnesium, potassium, zinc, and iron, but lower levels of calcium and copper in their hair samples. This study corroborates data from previous studies in different parts of the world indicating the presence of elevated levels of heavy metals and selective depletion of essential minerals in the hair of children with ASD.


Subsuns and rainbows during solar eclipses.
A photographic observation sequence was obtained of a subsun before, during, and after the total phase of the 2016 solar eclipse. The time-resolved images were obtained from a high-altitude jet aircraft. The image sequence was searched for the possible presence of a solar corona-generated subsun during totality. Although the subsun-creating conditions apparently persisted during totality, the drop in signal intensity compared to the local background prevented its detection. Separately, we document a visual observation from the 1977 total solar eclipse of a rainbow that faded, in the last a few seconds before totality, from being normally multicolored to monochromatic red from water drops then predominantly illuminated by light from the solar chromosphere. A similar transition in the final seconds before, and after, totality is expected to occur for parhelia. The posited short-living monochromatic red parhelion resulting from the momentary illumination of ice crystals by the solar chromosphere is still waiting to be observed.


Impact of the acquired subgenome on the transcriptional landscape in Brettanomyces bruxellensis allopolyploids.
Gene expression variation can provide an overview of the changes in regulatory networks that underlie phenotypic diversity. Certain evolutionary trajectories such as polyploidization events can have an impact on the transcriptional landscape. Interestingly, the evolution of the yeast species Brettanomyces bruxellensis has been punctuated by diverse allopolyploidization events leading to the coexistence of a primary diploid genome associated with various haploid acquired genomes. To assess the impact of these events on gene expression, we generated and compared the transcriptomes of a set of 87 B. bruxellensis isolates, selected as being representative of the genomic diversity of this species. Our analysis revealed that acquired subgenomes strongly impact the transcriptional patterns and allow discrimination of allopolyploid populations. In addition, clear transcriptional signatures related to specific populations have been revealed. The transcriptional variations observed are related to some specific biological processes such as transmembrane transport and amino acids metabolism. Moreover, we also found that the acquired subgenome causes the overexpression of some genes involved in the production of flavor-impacting secondary metabolites, especially in isolates of the beer population.


Genotyping and Axenic Growth of Coxiella burnetii Isolates Found in the United States Environment.
Coxiella burnetii is a gram-negative bacterium that is the etiologic agent of the zoonotic disease Q fever. Common reservoirs of C. burnetii include sheep, goats, and cattle. These animals shed C. burnetii into the environment, and humans are infected by inhalation of aerosols. A survey of 1622 environmental samples taken across the United States in 2006-2008 found that 23.8% of the samples contained C. burnetii DNA. To identify the strains circulating in the U.S. environment, DNA from these environmental samples was genotyped using an SNP-based approach to derive sequence types (ST) that are also compatible with multispacer sequence typing methods. Three different sequence types were observed in 31 samples taken from 19 locations. ST8 was associated with goats and ST20 with dairy cattle. ST16/26 was detected in locations with exposure to various animals and also in locations with no direct animal contact. Viable isolates were obtained for all three sequence types, but only the ST20 and ST16/26 isolates grew in acidified citrate cysteine medium (ACCM)-2 axenic media. Examination of a variety of isolates with different sequence types showed that ST8 and closely related isolates did not grow in ACCM-2. These results suggest that a limited number of C. burnetii sequence types are circulating in the U.S. environment and these strains have close associations with specific reservoir species. Growth in ACCM-2 may not be suitable for isolation of many C. burnetii strains.


The influence of deer browsing on the reproductive biology of Canada yew (Taxus canadensis marsh.) : I. Direct effect on pollen, ovule, and seed production.
Canada yew (Taxus canadensis) populations currently browsed by white-tailed deer (Odocoileus virginianus) or browsed by deer in the past had significantly lower production of male strobili, female strobili, and seeds than unbrowsed yew populations. Exclosure studies showed that protected yews produced significantly more male and female strobili than unprotected yews, but only after several years of protection. Seed production did not respond as readily to protection from deer perhaps because of reduced pollination levels in browsed yew populations. Previously unbrowsed yews were clipped at different levels of removal of available browse (control (no removal), 25%, 50%, 75%, and 100% removal) to simulate deer browsing. Reduction in male strobilus production was linearly related to clipping intensity in three years of observation. Female strobilus production was significantly reduced only at the 100% level of removal. Intermediate levels of clipping may have even stimulated production of female strobili. Analysis of covariance, with previous year's branch production as the covariate, showed no significant effect of clipping on male strobilus production except in the 100% removal group. Female strobilus production showed no such covariance with branch production. Effects of clipping on seed production could not be reliably assessed in 1984 and 1985 due to low seed production. Seed production in 1986 was significantly reduced only in the 100% removal group. Field observations of deer browsing of Canada yew indicate that 100% levels of removal are typical of natural levels of browsing.


Experimental study of the water-to-air stopping power ratio of monoenergetic carbon ion beams for particle therapy.
Reference dosimetry with ionization chambers requires a number of chamber-specific and beam-specific calibration factors. For carbon ion beams, IAEA report TRS-398 yields a total uncertainty of 3% in the determination of the absorbed dose to water, for which the biggest contribution arises from the water-to-air stopping power ratio (s(w, air)), with an uncertainty of 2%. The variation of (s(w, air)) along the treatment field has been studied in several Monte Carlo works presented over the last few years. Their results were, in all cases, strongly dependent on the choice of mean ionization potentials (I-values) for air and water. A smaller dependence of (s(w, air)) with penetration depth was observed. Since a consensus on I(w, air) and I(air) has not yet been reached, the validity of such studies for clinical use cannot be assessed independently. Our approach is based on a direct experimental measurement of water-equivalent thicknesses of different air gaps at different beam energies. A theoretical expression describing the variation of the stopping power ratio with kinetic energy, s(w,air)(E), was derived from the Bethe-Bloch formula and fit to the measured data, yielding a coherent pair of I(w) and I(air) values with I(air)/I(w) = 1.157 ± 0.023. Additionally, the data from five different beam energies were combined in an average value of s(w,air) = 1.132 ± 0.003 (statistical) ± 0.003 (variation over energy range), valid for monoenergetic carbon ion beams at the plateau area of the depth dose distribution. A detailed uncertainty analysis was performed on the data, in order to assess the limitations of the method, yielding an overall standard uncertainty below 1% in s(w,air)(E). Therefore, when properly combined with the appropriate models for the fragment spectra, our experimental work can contribute to narrow the uncertainty margins currently in use in absorbed dose to water determination for dosimetry of carbon ion beam radiotherapy.


Comparison of two methods of pediatric resuscitation and critical care management.
We compare time to drug delivery and the incidence of dosage error between 2 different systems of medication administration: The Broselow Pediatric Emergency Tape and a standardized volume/weight-based dose reformulation of resuscitation and critical care medications (reformulated to 0.1 mL/kg). This was a randomized crossover trial, in which volunteers (n=16) from emergency department (ED) pediatric resuscitation teams from the ED of a large, urban, teaching hospital in Australia were assigned to manage simulated (Advanced Pediatric Life Support scenario) patients. The volunteers were each presented with 3 case scenarios (brady-asystolic arrest, status epilepticus, and rapid sequence intubation requiring administration of 4, 5, and 4 medications, respectively). The order of presentation was randomized for the 2 methods. The volunteers were then asked to manage 3 case scenarios using one and then the other method (resulting in a total of 6 cases managed per participant). The dosage of each medication ordered, as well as the time to the simulated administration of that medication, was recorded for all scenarios. The expected dosages were compared with the actual dosages delivered to determine which system provided greater accuracy in medication administration. Statistical analysis was undertaken using the Wilcoxon signed rank test and McNemars test for paired proportions. Compared with the Broselow tape, the standardized volume/weight-based dose reformulation significantly reduced median time to medication delivery for all clinical scenarios (147 versus 72 seconds; 197 versus 87 seconds; 146 versus 64 seconds; P<.001). The proportion of dosing errors with Broselow tape across the 3 scenarios was greater than with volume/weight-based dosing (0.08 versus 0, 0 versus 0, and 0.08 versus 0.02, respectively). Use of a standardized volume/weight-based dose reformulation method is a simple, fast, and accurate method of medication delivery for the pediatric patient that eliminates the need for memorization and/or calculation. The standardized volume/weight-based dose reformulation method performs better than the Broselow tape in speed of delivery of medications used for pediatric resuscitation and critical care without any reduction in dosing accuracy.


Hemodynamic impairment, venous segmental disease, and clinical severity scoring in limbs with Klippel-Trenaunay syndrome.
Klippel-Trenaunay syndrome (KTS) is a complex congenital anomaly featuring two or more of the following: (1) capillary malformations (port-wine stains), (2) soft tissue or bony hypertrophy (or both), and (3) varicose veins or venous malformations. With the purpose of determining the actual significance of venous impairment in patients with KTS, we quantified the venous valvular competency and calf muscle pump function and examined their effect on clinical severity. Included were patients with near-normal function of affected limb(s) and minimal/small foot hypertrophy. Excluded were those with deep venous hypoplasia, aplasia or thrombosis, lymphedema, limb length discrepancy (>2.5 cm), peripheral arterial (ankle-brachial index <1.0), or cardiac disease and walking impairment. Venous duplex scanning, ascending venography, magnetic resonance imaging, strain gauge plethysmography, and a bone scanogram were performed. We studied eight men and seven women aged 15 to 51 years (median, 24 years). The KTS involved 17 limbs (unilateral in 13 patients and bilateral in 2). Contralateral limbs in patients with unilateral KTS acted as controls (n = 13). Venous clinical severity was graded according to the CEAP and venous clinical severity score (VCSS), and reflux complexity was classified according to the venous segmental disease score. Outflow obstruction (outflow fraction at 1 and 4 seconds; OF(1) and OF(4), respectively), reflux (venous filling index), calf muscle pump function (ejection fraction), and hypertension (residual volume fraction) were determined in both limbs with strain gauge plethysmography. Data, reported as median and interquartile range, were analyzed with the Mann-Whitney test. Varicose veins or venous malformations occurred in the medial, posterior, or anterolateral limb segments of the ankle (7/17, 7/17, and 9/17), calf (10/17, 8/17, and 12/17), knee (9/17, 8/17, and 8/17), and thigh (10/17, 6/17, and 8/17, respectively). Venous malformations occupied the subcutaneous space (17/17) and extended into the subfascial space in 6 (35.3%) of 17 limbs. Abnormal reflux (>0.5 seconds) was distributed in the great (64.7%; 11/17) and small (5.9%; 1/17) saphenous veins and the common femoral (23.5%; 4/17), femoral (41.1%; 7/17), popliteal (29.4%; 5/17), perforator (70.6%; 12/17), and axial calf (35.3%; 6/17) veins. There was no difference in the OF(1) and OF(4) between the affected limbs and the controls. Limbs with KTS had a fivefold greater venous filling index (0.133-0.46 mL . 100 mL(-1) . s(-1); 0.258 mL . 100 mL(-1) . s(-1)) than the controls (0.034-0.055 mL . 100 mL(-1) . s(-1); 0.046 mL . 100 mL(-1) . s(-1); P < .0001), and this was linked to a higher venous segmental disease score (3 [2-4] vs 0 [0-1]; P < .0001). Limbs with KTS had half the ejection fraction (20.8%; 12.3%-24%) of the controls (39.3%; 30.9%-64.6%) and twice as high a residual venous fraction (77% [69.6%-84.5%] vs 40.9% [20.6%-60%]; both P < .004). Patients complained of swelling (100%; 15/15), aching (100%; 15/15), pain (93.3%; 14/15) and heaviness (100%; 15/15), tiredness (66.7%; 10/15), and tightness (33.3%; 5/15) of the limb(s) with KTS. Limbs with KTS had a worse (1) venous clinical severity by 11 VCSS points (11 [8-12] vs 0 [0-1]) and (2) clinical status by 3 CEAP classes (C3 [C3-C4] vs C0 [C0-C2]) than the control limbs (both P < .0001). Venous disease in limbs with KTS is a major source of morbidity in affected patients. Limbs with KTS are characterized by complex reflux patterns, severe valvular incompetence, calf muscle pump impairment, and venous hypertension, thus explaining the advanced clinical severity (VCSS) and CEAP grade.


A holistic phylogeny of the coronin gene family reveals an ancient origin of the tandem-coronin, defines a new subfamily, and predicts protein function.
Coronins belong to the superfamily of the eukaryotic-specific WD40-repeat proteins and play a role in several actin-dependent processes like cytokinesis, cell motility, phagocytosis, and vesicular trafficking. Two major types of coronins are known: First, the short coronins consisting of an N-terminal coronin domain, a unique region and a short coiled-coil region, and secondly the tandem coronins comprising two coronin domains. 723 coronin proteins from 358 species have been identified by analyzing the whole-genome assemblies of all available sequenced eukaryotes (March 2011). The organisms analyzed represent most eukaryotic kingdoms but also cover every taxon several times to provide a better statistical sampling. The phylogenetic tree of the coronin domains based on the Bayesian method is in accordance with the most recent grouping of the major kingdoms of the eukaryotes and also with the grouping of more recently separated branches. Based on this "holistic" approach the coronins group into four classes: class-1 (Type I) and class-2 (Type II) are metazoan/choanoflagellate specific classes, class-3 contains the tandem-coronins (Type III), and the new class-4 represents the coronins fused to villin (Type IV). Short coronins from non-metazoans are equally related to class-1 and class-2 coronins and thus remain unclassified. The coronin class distribution suggests that the last common eukaryotic ancestor possessed a single and a tandem-coronin, and most probably a class-4 coronin of which homologs have been identified in Excavata and Opisthokonts although most of these species subsequently lost the class-4 homolog. The most ancient short coronin already contained the trimerization motif in the coiled-coil domain.


Pre-existing cross-reactive antibodies to avian influenza H5N1 and 2009 pandemic H1N1 in US military personnel.
We studied cross-reactive antibodies against avian influenza H5N1 and 2009 pandemic (p) H1N1 in 200 serum samples from US military personnel collected before the H1N1 pandemic. Assays used to measure antibodies against viral proteins involved in protection included a hemagglutination inhibition (HI) assay and a neuraminidase inhibition (NI) assay. Viral neutralization by antibodies against avian influenza H5N1 and 2009 pH1N1 was assessed by influenza (H5) pseudotyped lentiviral particle-based and H1N1 microneutralization assays. Some US military personnel had cross-neutralizing antibodies against H5N1 (14%) and 2009 pH1N1 (16.5%). The odds of having cross-neutralizing antibodies against 2009 pH1N1 were 4.4 times higher in subjects receiving more than five inactivated whole influenza virus vaccinations than those subjects with no record of vaccination. Although unclear if the result of prior vaccination or disease exposure, these pre-existing antibodies may prevent or reduce disease severity.


A role for locus coeruleus in Parkinson tremor.
We analyzed rest tremor, one of the etiologically most elusive hallmarks of Parkinson disease (PD), in 12 consecutive PD patients during a specific task activating the locus coeruleus (LC) to investigate a putative role of noradrenaline (NA) in tremor generation and suppression. Clinical diagnosis was confirmed in all subjects by reduced dopamine reuptake transporter (DAT) binding values investigated by single photon computed tomography imaging (SPECT) with [(123)I] N-ω-fluoropropyl-2β-carbomethoxy-3β-(4-iodophenyl) tropane (FP-CIT). The intensity of tremor (i.e., the power of Electromyography [EMG] signals), but not its frequency, significantly increased during the task. In six subjects, tremor appeared selectively during the task. In a second part of the study, we retrospectively reviewed SPECT with FP-CIT data and confirmed the lack of correlation between dopaminergic loss and tremor by comparing DAT binding values of 82 PD subjects with bilateral tremor (n = 27), unilateral tremor (n = 22), and no tremor (n = 33). This study suggests a role of the LC in Parkinson tremor.