The associations of leptin, adiponectin and resistin with incident atrial fibrillation in women.
Higher body mass index (BMI) is an important risk factor for atrial fibrillation (AF). The adipokines leptin, adiponectin and resistin are correlates of BMI, but their association with incident AF is not well known. We explored this relationship in a large cohort of postmenopausal women. We studied an ethnically diverse cohort of community-dwelling postmenopausal women aged 50-79 who were nationally recruited at 40 clinical centres as part of the Women's Health Initiative investigation. Participants underwent measurements of baseline serum leptin, adiponectin and resistin levels and were followed for incident AF. Adipokine levels were log transformed and normalised using inverse probability weighting. Cox proportional hazard regression models were used to estimate associations with adjustment for known AF risk factors. Of the 4937 participants included, 892 developed AF over a follow-up of 11.1 years. Those with AF had higher mean leptin (14.9 pg/mL vs 13.9 pg/mL), adiponectin (26.3 ug/mL vs 24.5 ug/mL) and resistin (12.9 ng/mL vs 12.1 ng/mL) levels. After multivariable adjustment, neither log leptin nor log adiponectin levels were significantly associated with incident AF. However, log resistin levels remained significantly associated with incident AF (HR=1.57 per 1 log (ng/mL) increase, p=0.006). Additional adjustment for inflammatory cytokines only partially attenuated the association between resistin and incident AF (HR=1.43, p=0.06 adjusting for C-reactive protein (CRP); HR=1.39, p=0.08 adjusting for IL-6). Adjusting for resistin partially attenuated the association between BMI and incident AF (HR=1.14 per 5 kg/m(2), p=0.006 without resistin; HR=1.12, p=0.02 with resistin). In women, elevated levels of serum resistin are significantly associated with higher rates of incident AF and partially mediate the association between BMI and AF. In the same population, leptin and adiponectin levels are not significantly associated with AF.


Future innovations in anti-platelet therapies.
Platelets have long been recognized to be of central importance in haemostasis, but their participation in pathological conditions such as thrombosis, atherosclerosis and inflammation is now also well established. The platelet has therefore become a key target in therapies to combat cardiovascular disease. Anti-platelet therapies are used widely, but current approaches lack efficacy in a proportion of patients, and are associated with side effects including problem bleeding. In the last decade, substantial progress has been made in understanding the regulation of platelet function, including the characterization of new ligands, platelet-specific receptors and cell signalling pathways. It is anticipated this progress will impact positively on the future innovations towards more effective and safer anti-platelet agents. In this review, the mechanisms of platelet regulation and current anti-platelet therapies are introduced, and strong, and some more speculative, potential candidate target molecules for future anti-platelet drug development are discussed.


The maternal hemodynamic effect of indomethacin in normal pregnancy.
The vasodilation of pregnancy is thought by many to be due to increased endothelial production of prostacyclin, a vasodilatory prostanoid. Indomethacin, a potent inhibitor of prostaglandin synthesis, is known to increase the maternal blood pressure response to angiotensin II infusion. We sought to measure directly the hemodynamic effects of a short course of indomethacin. Twenty-three healthy pregnant women with uncomplicated pregnancies between 26-32 weeks' gestation completed the study. Using Doppler technology, we determined cardiac output, stroke volume, and total peripheral resistance before and after three 25-mg doses of indomethacin. Although blood pressure did not change, peripheral resistance rose and stroke volume fell following indomethacin administration. Our findings support the hypothesis that indomethacin interferes with tonic prostaglandin-induced vasodilation in pregnancy. However, the increase in vascular resistance was very slight, suggesting that other vasodilators are also at work in pregnancy. We recommend that indomethacin be used judiciously in hypertensive pregnant patients until more information concerning possible adverse hemodynamic effects becomes available.


Gain-of-function amino acid substitutions drive positive selection of FGFR2 mutations in human spermatogonia.
Despite the importance of mutation in genetics, there are virtually no experimental data on the occurrence of specific nucleotide substitutions in human gametes. C>G transversions at position 755 of FGF receptor 2 (FGFR2) cause Apert syndrome; this mutation, encoding the gain-of-function substitution Ser252Trp, occurs with a birth rate elevated 200- to 800-fold above background and originates exclusively from the unaffected father. We previously demonstrated high levels of both 755C>G and 755C>T FGFR2 mutations in human sperm and proposed that these particular mutations are enriched because the encoded proteins confer a selective advantage to spermatogonial cells. Here, we examine three corollaries of this hypothesis. First, we show that mutation levels at the adjacent FGFR2 nucleotides 752-754 are low, excluding any general increase in local mutation rate. Second, we present three instances of double-nucleotide changes involving 755C, expected to be extremely rare as chance events. Two of these double-nucleotide substitutions are shown, either by assessment of the pedigree or by direct analysis of sperm, to have arisen in sequential steps; the third (encoding Ser252Tyr) was predicted from structural considerations. Finally, we demonstrate that both major alternative spliceforms of FGFR2 (Fgfr2b and Fgfr2c) are expressed in rat spermatogonial stem cell lines. Taken together, these observations show that specific FGFR2 mutations attain high levels in sperm because they encode proteins with gain-of-function properties, favoring clonal expansion of mutant spermatogonial cells. Among FGFR2 mutations, those causing Apert syndrome may be especially prevalent because they enhance signaling by FGF ligands specific for each of the major expressed isoforms.


Final report of the safety assessment of cosmetic ingredients derived from Zea mays (corn).
Many cosmetic ingredients are derived from Zea mays (corn). While safety test data were not available for most ingredients, similarities in preparation and the resulting similar composition allowed extrapolation of safety data to all listed ingredients. Animal studies included acute toxicity, ocular and dermal irritation studies, and dermal sensitization studies. Clinical studies included dermal irritation and sensitization. Case reports were available for the starch as used as a donning agent in medical gloves. Studies of many other endpoints, including reproductive and developmental toxicity, use corn oil as a vehicle control with no reported adverse effects at levels used in cosmetics. While industry should continue limiting ingredient impurities such as pesticide residues before blending into a cosmetic formulation, the CIR Expert Panel determined that corn-derived ingredients are safe for use in cosmetics in the practices of use and concentration described in the assessment.


High GUD incidence in the early 20 century created a particularly permissive time window for the origin and initial spread of epidemic HIV strains.
The processes that permitted a few SIV strains to emerge epidemically as HIV groups remain elusive. Paradigmatic theories propose factors that may have facilitated adaptation to the human host (e.g., unsafe injections), none of which provide a coherent explanation for the timing, geographical origin, and scarcity of epidemic HIV strains. Our updated molecular clock analyses established relatively narrow time intervals (roughly 1880-1940) for major SIV transfers to humans. Factors that could favor HIV emergence in this time frame may have been genital ulcer disease (GUD), resulting in high HIV-1 transmissibility (4-43%), largely exceeding parenteral transmissibility; lack of male circumcision increasing male HIV infection risk; and gender-skewed city growth increasing sexual promiscuity. We surveyed colonial medical literature reporting incidences of GUD for the relevant regions, concentrating on cities, suffering less reporting biases than rural areas. Coinciding in time with the origin of the major HIV groups, colonial cities showed intense GUD outbreaks with incidences 1.5-2.5 orders of magnitude higher than in mid 20(th) century. We surveyed ethnographic literature, and concluded that male circumcision frequencies were lower in early 20(th) century than nowadays, with low rates correlating spatially with the emergence of HIV groups. We developed computer simulations to model the early spread of HIV-1 group M in Kinshasa before, during and after the estimated origin of the virus, using parameters derived from the colonial literature. These confirmed that the early 20(th) century was particularly permissive for the emergence of HIV by heterosexual transmission. The strongest potential facilitating factor was high GUD levels. Remarkably, the direct effects of city population size and circumcision frequency seemed relatively small. Our results suggest that intense GUD in promiscuous urban communities was the main factor driving HIV emergence. Low circumcision rates may have played a role, probably by their indirect effects on GUD.


Model Evaluation in Generalized Structured Component Analysis Using Confirmatory Tetrad Analysis.
Generalized structured component analysis (GSCA) is a component-based approach to structural equation modeling (SEM). GSCA regards weighted composites or components of indicators as proxies for latent variables and estimates model parameter via least squares without resorting to a distributional assumption such as multivariate normality of indicators. As with other SEM approaches, model evaluation is a crucial procedure in GSCA that is used to examine whether a hypothesized model is consistent with the data in hand. However, the few descriptive measures of model evaluation available for GSCA are limited to evaluating models in a more confirmatory manner. This study integrates confirmatory tetrad analysis (CTA) into GSCA for model evaluation or comparison. Although CTA has been used in factor-based SEM as an inferential statistic, CTA is actually more compatible with GSCA because it is completely free of the multivariate normality assumption. Utilizing empirical data collected for 18,174 students' social skills in an early childhood longitudinal study of 2010-11 kindergarten cohort, we demonstrate the capability and applicability of CTA in GSCA and compare its performance with existing measures for GSCA.


Cytotoxicity assessments of Portulaca oleracea and Petroselinum sativum seed extracts on human hepatocellular carcinoma cells (HepG2).
The Pharmacological potential, such as antioxidant, anti-inflammatory, and antibacterial activities of Portulaca oleracea (PO) and Petroselinum sativum (PS) extracts are well known. However, the preventive properties against hepatocellular carcinoma cells have not been explored so far. Therefore, the present investigation was designed to study the anticancer activity of seed extracts of PO and PS on the human hepatocellular carcinoma cells (HepG2). The HepG2 cells were exposed with 5-500 μg/ml of PO and PS for 24 h. After the exposure, cell viability by 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assay, neutral red uptake (NRU) assay, and cellular morphology by phase contrast inverted microscope were studied. The results showed that PO and PS extracts significantly reduced the cell viability of HepG2 in a concentration dependent manner. The cell viability was recorded to be 67%, 31%, 21%, and 17% at 50, 100, 250, and 500 μg/ml of PO, respectively by MTT assay and 91%, 62%, 27%, and 18% at 50, 100, 250, and 500 μg/ml of PO, respectively by NRU assay. PS exposed HepG2 cells with 100 μg/ml and higher concentrations were also found to be cytotoxic. The decrease in the cell viability at 100, 250, and 500 μg/ml of PS was recorded as 70%, 33%, and 15% by MTT assay and 63%, 29%, and 17%, respectively by NRU assay. Results also showed that PO and PS exposed cells reduced the normal morphology and adhesion capacity of HepG2 cells. HepG2 cells exposed with 50 μg/ml and higher concentrations of PO and PS lost their typical morphology, become smaller in size, and appeared in rounded bodies. Our results demonstrated preliminary screening of anticancer activity of Portulaca oleracea and Petroselinum sativum extracts against HepG2 cells, which can be further used for the development of a potential therapeutic anticancer agent.


The AcrAB-TolC pump is involved in macrolide resistance but not in telithromycin efflux in Enterobacter aerogenes and Escherichia coli.
The role of the AcrAB-TolC pump in macrolide and ketolide susceptibility in Escherichia coli and Enterobacter aerogenes was studied. Efflux pump inhibitor restored erythromycin, clarithromycin, and telithromycin susceptibilities to multidrug-resistant isolates. No modification of telithromycin accumulation was detected in E. aerogenes acrAB or tolC derivatives compared to that in the parental strain. Two independent efflux pumps, inhibited by phenylalanine arginine beta-naphthylamide, expel macrolides and telithromycin in E. aerogenes.


When is it MODY? Challenges in the Interpretation of Sequence Variants in MODY Genes.
The genomics revolution has raised more questions than it has provided answers. Big data from large population-scale resequencing studies are increasingly deconstructing classic notions of Mendelian disease genetics, which support a simplistic correlation between mutational severity and phenotypic outcome. The boundaries are being blurred as the body of evidence showing monogenic disease-causing alleles in healthy genomes, and in the genomes of individu-als with increased common complex disease risk, continues to grow. In this review, we focus on the newly emerging challenges which pertain to the interpretation of sequence variants in genes implicated in the pathogenesis of maturity-onset diabetes of the young (MODY), a presumed mono-genic form of diabetes characterized by Mendelian inheritance. These challenges highlight the complexities surrounding the assignments of pathogenicity, in particular to rare protein-alerting variants, and bring to the forefront some profound clinical diagnostic implications. As MODY is both genetically and clinically heterogeneous, an accurate molecular diagnosis and cautious extrapolation of sequence data are critical to effective disease management and treatment. The biological and translational value of sequence information can only be attained by adopting a multitude of confirmatory analyses, which interrogate variant implication in disease from every possible angle. Indeed, studies which have effectively detected rare damaging variants in known MODY genes in normoglycemic individuals question the existence of a sin-gle gene mutation scenario: does monogenic diabetes exist when the genetic culprits of MODY have been systematical-ly identified in individuals without MODY?


Association analysis of the ACTN3 R577X polymorphism and complex quantitative body composition and performance phenotypes in adolescent Greeks.
The functional allele (577R) of ACTN3, which encodes human alpha-actinin-3, has been reported to be associated with elite athletic status and with response to resistance training, while the nonfunctional allele (577X) has been proposed as a candidate metabolically thrifty allele. In a study of 992 adolescent Greeks, we show that there is a significant association (P=0.003) between the ACTN3 R577X polymorphism and 40 m sprint time in males that accounts for 2.3% of phenotypic variance, with the 577R allele contributing to faster times in an additive manner. The R577X polymorphism is not associated with other power phenotypes related to 40 m sprint, nor with an endurance phenotype. Furthermore, the polymorphism is not associated with obesity-related phenotypes in our population, suggesting that the 577X allele is not a thrifty allele, and thus the persistence of this null allele must be explained in other terms.


Pyrophosphate ion-triggered competitive displacement of ssDNA from a metal-organic framework and its application in fluorescent sensing of alkaline phosphatase.
Nanomaterial/aptamer assembly has been extensively explored in the detection of various targets, but some limitations still exist in its practical applications, especially time consumption and low-efficient detachment of the aptamer from the nanomaterial surface. In this work, we demonstrated the ligand role of pyrophosphate ion (PPi) in the competitive displacement of ssDNA from the nanoscaffold surface. For this purpose, a fluorescein-labeled ssDNA (F-DNA) and a mixed valence state cerium (Ce3+/Ce4+)-based MOF (MVCM) were employed as the signal response unit and nanoscaffold, respectively. Benefiting from the existence of Ce4+, the MVCM exhibits an ultrahigh quenching efficiency (more than 90%) to F-DNA fluorescence, which is 3-fold higher than that of the MOF with Ce3+ only. However, it was found that PPi can effectively suppress the quenching effect of the MVCM by competitive coordination with the MVCM to displace F-DNA. Different from the conventional target-induced conformational change of aptamers, the PPi-triggered displacement assay is independent of the ssDNA sequence and can be rapidly completed in just 2 min. The displacement assay is also highly sensitive, even at a PPi concentration as low as 55 nM. In contrast to PPi, however, the phosphate ions and other anions cannot displace F-DNA from the MVCM surface to switch on the F-DNA fluorescence. Inspired by this fact, the PPi-triggered displacement assay was further applied in the detection of alkaline phosphatase (ALP). The detection limit toward ALP was obtained at 0.18 mU mL-1. Moreover, the accurate determination of ALP concentration in serum samples indicates the applicability of this sensing system in detecting real samples.


Instantaneous and quantitative functionalization of gold nanoparticles with thiolated DNA using a pH-assisted and surfactant-free route.
The attachment of thiolated DNA to gold nanoparticles (AuNPs) has enabled many landmark works in nanobiotechnology. This conjugate chemistry is typically performed using a salt-aging protocol where, in the presence of an excess amount of DNA, NaCl is gradually added to increase DNA loading over 1-2 days. To functionalize large AuNPs, surfactants need to be used, which may generate difficulties for downstream biological applications. We report herein a novel method using a pH 3.0 citrate buffer to complete the attachment process in a few minutes. More importantly, it allows for quantitative DNA adsorption, eliminating the need to quantify the number of adsorbed DNA and allowing the adsorption of multiple DNAs with different sequences at predetermined ratios. The method has been tested for various DNAs over a wide range of AuNP sizes. Our work suggests a synergistic effect between pH and salt in DNA attachment and reveals the fundamental kinetics of AuNP aggregation versus DNA adsorption, providing a novel means to modulate the interactions between DNA and AuNPs.


Adaptive mutations alter antibody structure and dynamics during affinity maturation.
While adaptive mutations can bestow new functions on proteins via the introduction or optimization of reactive centers, or other structural changes, a role for the optimization of protein dynamics also seems likely but has been more difficult to evaluate. Antibody (Ab) affinity maturation is an example of adaptive evolution wherein the adaptive mutations may be identified and Abs may be raised to specific targets that facilitate the characterization of protein dynamics. Here, we report the characterization of three affinity matured Abs that evolved from a common germline precursor to bind the chromophoric antigen (Ag), 8-methoxypyrene-1,3,6-trisulfonate (MPTS). In addition to characterizing the sequence, molecular recognition, and structure of each Ab, we characterized the dynamics of each complex by determining their mechanical response to an applied force via three-pulse photon echo peak shift (3PEPS) spectroscopy and deconvoluting the response into elastic, anelastic, and plastic components. We find that for one Ab, affinity maturation was accomplished via the introduction of a single functional group that mediates a direct contact with MPTS and results in a complex with little anelasticity or plasticity. In the other two cases, more mutations were introduced but none directly contact MPTS, and while their effects on structure are subtle, their effects on anelasticity and plasticity are significant, with the level of plasticity correlated with specificity, suggesting that the optimization of protein dynamics may have contributed to affinity maturation. A similar optimization of structure and dynamics may contribute to the evolution of other proteins.


Antihypertensive agents for preventing diabetic kidney disease.
Various blood pressure-lowering agents, and particularly inhibitors of the renin-angiotensin system (RAS), are widely used for people with diabetes to prevent the onset of diabetic kidney disease (DKD) and adverse cardiovascular outcomes. This is an update of a Cochrane review first published in 2003 and updated in 2005. This systematic review aimed to assess the benefits and harms of blood pressure lowering agents in people with diabetes mellitus and a normal amount of albumin in the urine (normoalbuminuria). In January 2011 we searched the Cochrane Renal Group's Specialised Register through contact with the Trials Search Co-ordinator. Randomised controlled trials (RCTs) comparing any antihypertensive agent with placebo or another agent in hypertensive or normotensive patients with diabetes and no kidney disease (albumin excretion rate < 30 mg/d) were included. Two investigators independently extracted data on kidney and other patient-relevant outcomes (all-cause mortality and serious cardiovascular events), and assessed study quality. Analysis was by a random effects model was applied to analyse results which were expressed as risk ratio (RR) and 95% confidence intervals (CI). We identified 26 studies that enrolling 61,264 participants. Angiotensin-converting enzyme inhibitors (ACEi) reduced the risk of new onset of microalbuminuria, macroalbuminuria or both when compared to placebo (8 studies, 11,906 patients: RR 0.71, 95% CI 0.56 to 0.89), with similar benefits in people with and without hypertension (P = 0.74), and when compared to calcium channel blockers (5 studies, 1253 participants: RR 0.60, 95% CI 0.42 to 0.85). ACEi reduced the risk of death when compared to placebo (6 studies, 11,350 participants: RR 0.84, 95% CI 0.73 to 0.97). No effect was observed for angiotensin receptor blockers (ARB) when compared to placebo for new microalbuminuria, macroalbuminuria or both (5 studies, 7653 participants: RR 0.90, 95% CI 0.68 to 1.19) or death (5 studies, 7653 participants: RR 1.12, 95% CI 0.88 to 1.41); however, meta-regression suggested possible benefits from ARB for preventing kidney disease in high risk patients. There was a trend towards benefit from use of combined ACEi and ARB for prevention of DKD compared with ACEi alone (2 studies, 4171 participants: RR 0.88, 95% CI 0.78 to 1.00).The risk of cough was significantly increased with ACEi when compared to placebo (6 studies, 11,791 patients: RR 1.84, 95% CI 1.24 to 2.72), however there was no significant difference in the risk of headache or hyperkalaemia. There was no significant difference in the risk of cough, headache or hyperkalaemia when ARB was to placebo. On average risk of bias was judged to be either low (27% to 69%) or unclear (i.e. no information available) (8% to 73%). Blinding of participants, incomplete outcome data and selective reporting were judged to be high in 23%, 31% and 31% of studies, respectively. ACEi were found to prevent new onset DKD and death in normoalbuminuric people with diabetes, and could therefore be used in this population. More data are needed to clarify the role of ARB and other drug classes in preventing DKD.


Uncertainty in the response of terrestrial carbon sink to environmental drivers undermines carbon-climate feedback predictions.
Terrestrial ecosystems play a vital role in regulating the accumulation of carbon (C) in the atmosphere. Understanding the factors controlling land C uptake is critical for reducing uncertainties in projections of future climate. The relative importance of changing climate, rising atmospheric CO2, and other factors, however, remains unclear despite decades of research. Here, we use an ensemble of land models to show that models disagree on the primary driver of cumulative C uptake for 85% of vegetated land area. Disagreement is largest in model sensitivity to rising atmospheric CO2 which shows almost twice the variability in cumulative land uptake since 1901 (1 s.d. of 212.8 PgC vs. 138.5 PgC, respectively). We find that variability in CO2 and temperature sensitivity is attributable, in part, to their compensatory effects on C uptake, whereby comparable estimates of C uptake can arise by invoking different sensitivities to key environmental conditions. Conversely, divergent estimates of C uptake can occur despite being based on the same environmental sensitivities. Together, these findings imply an important limitation to the predictability of C cycling and climate under unprecedented environmental conditions. We suggest that the carbon modeling community prioritize a probabilistic multi-model approach to generate more robust C cycle projections.


Sexual risk reduction interventions for patients attending sexually transmitted disease clinics in the United States: a meta-analytic review, 1986 to early 2009.
Sexually transmitted disease (STD) patients are more likely to experience a future STD including human immunodeficiency virus (HIV). The aim of this study was to examine the efficacy of behavioral interventions to reduce sexual risk behavior and incident STDs among patients attending STD clinics in the United States. A meta-analysis of 32 studies with 48 separate interventions targeting STD patients (N = 67,538) was conducted. Independent raters coded study, sample, and intervention characteristics. Effect sizes, using both fixed- and random-effects models, were calculated. Potential moderators of intervention efficacy were assessed. Relative to controls, intervention participants increased their condom use and had fewer incident STDs, including HIV, across assessment intervals (d (+)s ranging from 0.05 to 0.64). Several sample (e.g., age and ethnicity) and intervention features (e.g., targeting intervention to a specific group) moderated the efficacy of the intervention. Behavioral interventions targeted to STD clinic patients reduce sexual risk behavior and prevent HIV/STDs. Widespread use of behavioral interventions in STD clinics should be a public health priority.


Antimelanogenic, Antioxidant and Antiproliferative Effects of Antrodia camphorata Fruiting Bodies on B16-F0 Melanoma Cells.
Antrodia camphorata is a fungus that is endemic to Taiwan, and its fruiting body has been used as a folk medicine for the prevention or treatment of diverse diseases. The present study is aimed at investigating the antimelanogenesis and antioxidation effect of the ethanolic extract of Antrodia camphorata fruiting body (EE-AC), as well as its antiproliferation effects in B16-F0 melanoma cells. Regarding antimelanogenic effects, EE-AC had effective cupric ions reducing capacity and expressed more potent inhibitory effect than kojic acid on mushroom tyrosinase activity. Moreover, EE-AC significantly inhibited cellular tyrosinase activity and the melanin content in B16-F0 cells at 12.5 μg/mL concentration without cell toxicities. Regarding antioxidant effects, EE-AC exhibited potent DPPH radical- and SOD-like-scavenging activities. Regarding antiproliferative effects, EE-AC exhibited a selective cytotoxic effect and markedly inhibited the migration ability of B16-F0 cells. EE-AC increased the population of B16-F0 cells at sub-G1 phase of the cell cycle. EE-AC also caused the increase of early apoptotic cells and chromatin condensation, which indicated the apoptotic effects in B16-F0 cells. We demonstrated that EE-AC possessed antimelanogenic, antioxidant and anti-skin cancer actions. The results would contribute to the development and application of cosmetics, healthy food and pharmaceuticals.


Relationship between sodium clearance and proximal tubular fluid output in conscious unoperated rats adapted to different dietary sodium contents.
The relationship between the proximal tubular fluid output (Vprox) and the sodium clearance (CNa) was examined in conscious unoperated diabetes insipidus rats adapted to different dietary sodium contents ranging from 5 to 300 mmol/kg. The lithium clearance (CLi) and the urine flow (V) were used as measures of Vprox, CLi being equal to and V being about one-third of Vprox. Variations in the dietary sodium content had almost no influence on CLi and V. However, repeated 2-hour clearance measurements on different days in the same rat showed considerable intraindividual variations of CLi and V, and at each dietary sodium level CNa was linearly correlated to CLi (r = 0.83-0.90) and to V (r = 0.57-0.89). Errors in bladder emptying could not explain these correlations. The slopes of the regression lines increased with increasing dietary sodium content, and the regression lines all intercepted the abscissa at approximately the same value of CLi, i.e. on the average about 40 microliter/min./100 g body weight. The results suggest that Vprox is almost independent of the dietary sodium content. Spontaneous intraindividual variation of Vprox is considerable and is associated with linearly related variation of CNa. The change of CNa for a given change of Vprox increases with the dietary sodium content.


Bacopa Monnieri Protects the Directly Affected Organ as Well as Distant Organs Against I/R Injury by Modulating Anti-Inflammatory and Anti-Nitrosative Pathways in A Rat Model for Infra-Renal Aortic Occlusion.
To investigate the protective effect and underlying mechanisms of B. monnieri, a medicinal plant, on kidney and skeletal muscle injury induced by infra-renal abdominal aorta clamping for 2-hours (ischemia) and following removal of the clamp (reperfusion, 2-hours). Rats were divided into four groups (n = 6): (I) animals given only saline (sham-control); (II) animals given B. monnieri extract for 10-days (300 mg/kg/day) (Bacopa-treated sham); (III) animals subjected to ischemia/reperfusion (I/R); (IV) animals given B. monnieri extract and then subjected to I/R. Kidneys and lower extremity muscles were examined for GPx, CAT, iNOS, 3-NT, IL-1β and TNF-α. Apoptosis and injury were evaluated by TUNEL and H&E staining, respectively. I/R resulted in TUNEL positive cells, periarterial edema and glomerular capillary dilatation, decreased GPx activity, unchanged CAT, iNOS, 3-NT, IL-1β and TNF-α in kidney. B. monnieri minimized renal remote reperfusion injury, and Group IV showed a lower degree of renal histopathology score when compared to the others. B. monnieri mitigated muscle I/R injury, decreased muscle hypertrophy, myofibril abnormalities and apoptosis. Muscle 3-NT and cytokine levels were increased by I/R, and B. monnieri inhibited iNOS and 3-NT both in sham-control and I/R groups. Muscle GPx unaffected by I/R or B. monnieri, but CAT was inhibited only in B. monnieri-treated I/R group. Muscle iNOS, 3-NT, IL-1β, TNF-α levels and CAT activity of B. monnieri-treated I/R rats were lower than those in sham-control or Bacopa-treated sham. B. monnieri can protect the directly affected organ as well as distant organs against I/R injury by modulating anti-inflammatory and anti-nitrosative pathways.


Automated cosmic spike filter optimized for process Raman spectroscopy.
Despite the existence of various methods to remove cosmic spikes from Raman data, only a few of them are suitable for process Raman spectroscopy. The disadvantages of these algorithms include increased analysis time, low accuracy of spike detection, or reliance on variable parameters that must be chosen by trial and error in each case. We demonstrate a novel approach to detecting cosmic spikes in process Raman data and validate it using a wide range of experimental data. This new method features a multistage spike recognition algorithm that is based on tracking sharp changes of intensity in the time domain. The algorithm effectively distinguishes cosmic spikes from random spectral noise and abrupt variations of Raman peaks, allowing accurate detection of both high and low intensity cosmic spikes. The procedure is free from variable user-defined parameters and operates reliably in a fully automated manner with a wide range of time-series process Raman data sets containing more than 40 to 50 spectra.


Going beyond the textbook: The need to integrate open access primary literature into the Chemistry curriculum.
Unrestricted, open access to scholarly scientific literature provides an opportunity for chemistry educators to go beyond the textbook, introducing students to the real work of scientists. Despite the best efforts of textbook authors to provide information about recent research results, textbooks are not a substitute for learning to use the primary literature. Chemical educators can use open access articles to develop research-related skills, to foster curiosity, and to cultivate the next generation of scientists. It is becoming increasingly important for chemical educators to teach undergraduates how online journals are changing the nature of chemical research. Some institutions can not afford online subscription costs, and open access journals can be an important resource to provide practical experience. Open access publications eliminate the barriers to the central work of scientists providing chemistry educators (whether at well-endowed or economically limited colleges) with the key resources for enhancing student learning through current, relevant research.


Tuning the electronic structure of thiolate-protected 25-atom clusters by co-substitution with metals having different preferential sites.
Trimetallic Au24-xAgxPd and tetrametallic Au24-x-yAgxCuyPd clusters were synthesized by the subsequential metal exchange reactions of dodecanethiolate-protected Au24Pd clusters. EXAFS measurements revealed that Pd, Ag, and Cu dopants preferentially occupy the center and edge sites of the core, and staple sites, respectively. Spectroscopic and theoretical studies demonstrated that the synergistic effects of multiple substitutions on the electronic structures are additive in nature.


Evidence-based therapy of diabetic retinopathy.
Evidence-based medicine is often misunderstood as 'cookbook medicine with standard recipes' that does not take clinical experience into account. It is, however, supposed to be a basis for decision making in caring for individual patients under consideration of patients' preferences. This seems to be very important, since diabetic retinopathy continues to be the most frequent cause of vision loss in working age adults with negative consequences for patients' quality of life and for health economics. The most important evidence-based therapy for diabetic retinopathy and maculopathy is laser coagulation. Vitrectomy for proliferative stages has also been proven effective by clinical studies. For more recent treatment options like triamcinolone injection and vitrectomy for diabetic macular edema there is a lower level of evidence so far. The Diabetic Retinopathy Study was the first to show the effectiveness of panfundus laser coagulation for a larger group of patients. The Early Treatment Diabetic Retinopathy Study in turn serves as a basis for laser coagulation of retinopathy and maculopathy. The Diabetic Retinopathy Vitrectomy Study could show the advantages of timely vitrectomy. Both the Diabetes Control and Complications Trial and the United Kingdom Prospective Diabetes Study could show the value of intensive blood glucose control. Evidence-based medicine on the basis of the studies mentioned above is practiced quite self-evidently in ophthalmo-diabetology. It should be regarded as a helpful tool for special therapeutic situations which still leaves room for one's personal clinical experience to be included. It is somewhat problematic that the term evidence-based medicine seems to be restricted to the results of large randomized studies, because even special problems and very individual, difficult therapeutic questions can be placed on an evidence-based foundation, although at a lower level of evidence, using today's modern means of literature research.


A novel mechanism for regulating activity of a transcription factor that controls the unfolded protein response.
Cells respond to an accumulation of unfolded proteins in the endoplasmic reticulum (ER) by increasing transcription of genes encoding ER-resident proteins. The information is transmitted from the ER lumen to the nucleus by an intracellular signaling pathway, the unfolded protein response (UPR). We have identified a basic-leucine zipper transcription factor, Hac1p, that is required for the UPR and binds to the UPR element in the promoter of UPR-regulated genes. Surprisingly, Hac1p is found in UPR-activated cells only, and its level is controlled by regulated splicing of its mRNA. Splicing replaces the C-terminal tail of Hac1p with a different peptide that renders Hac1p more resistant to an otherwise extremely rapid ubiquitin-dependent degradation. We propose that the complex regulation of Hac1p expression serves to provide multiple levels at which the UPR can be controlled.


Parvovirus B19 associated acute cholestatic hepatitis.
There are few reports in the literature of hepatitis as a manifestation of Parvovirus B19 infection. We describe a case of Parvovirus B19 associated acute cholestatic hepatitis diagnosed based on a positive serologic test (IgM) and molecular detection of parvovirus B19 DNA in peripheral blood. Parvovirus B19 infection should be considered in the differential diagnosis of patient presenting with acute hepatitis of unknown etiology.


The sources of variability in saccadic eye movements.
Our movements are variable, but the origin of this variability is poorly understood. We examined the sources of variability in human saccadic eye movements. In two experiments, we measured the spatiotemporal variability in saccade trajectories as a function of movement direction and amplitude. One of our new observations is that the variability in movement direction is smaller for purely horizontal and vertical saccades than for saccades in oblique directions. We also found that saccade amplitude, duration, and peak velocity are all correlated with one another. To determine the origin of the observed variability, we estimated the noise in motor commands from the observed spatiotemporal variability, while taking into account the variability resulting from uncertainty in localization of the target. This analysis revealed that uncertainty in target localization is the major source of variability in saccade endpoints, whereas noise in the magnitude of the motor commands explains a slightly smaller fraction. In addition, there is temporal variability such that saccades with a longer than average duration have a smaller than average peak velocity. This noise model has a large generality because it correctly predicts the variability in other data sets, which contain saccades starting from very different initial locations. Because the temporal noise most likely originates in movement planning, and the motor command noise in movement execution, we conclude that uncertainty in sensory signals and noise in movement planning and execution all contribute to the variability in saccade trajectories. These results are important for understanding how the brain controls movement.


Lysozyme and bilirubin bind to ACE and regulate its conformation and shedding.
Angiotensin I-converting enzyme (ACE) hydrolyzes numerous peptides and is a critical participant in blood pressure regulation and vascular remodeling. Elevated tissue ACE levels are associated with increased risk for cardiovascular and respiratory disorders. Blood ACE concentrations are determined by proteolytic cleavage of ACE from the endothelial cell surface, a process that remains incompletely understood. In this study, we identified a novel ACE gene mutation (Arg532Trp substitution in the N domain of somatic ACE) that increases blood ACE activity 7-fold and interrogated the mechanism by which this mutation significantly increases blood ACE levels. We hypothesized that this ACE mutation disrupts the binding site for blood components which may stabilize ACE conformation and diminish ACE shedding. We identified the ACE-binding protein in the blood as lysozyme and also a Low Molecular Weight (LMW) ACE effector, bilirubin, which act in concert to regulate ACE conformation and thereby influence ACE shedding. These results provide mechanistic insight into the elevated blood level of ACE observed in patients on ACE inhibitor therapy and elevated blood lysozyme and ACE levels in sarcoidosis patients.


Assessing the fall risks of community-dwelling stroke survivors using the Short-form Physiological Profile Assessment (S-PPA).
Fall is common after stroke. The Short-form Physiological Profile Assessment (S-PPA) was developed to assess the fall risks and underlying physiological factors, and it has been used in healthy older adults and older adults with stroke. This study aimed to establish the psychometric properties of the S-PPA among cognitively intact and ambulant community-dwelling older adults with stroke. The S-PPA, Chinese version of the Activities-specific Balance Confidence (ABC-C) scale and 3 balance measures, namely the Berg Balance Scale (BBS), Timed "Up & Go" (TUG) and Functional Reach Test (FRT), were administered. Inter- and intra-rater reliability were assessed at baseline and after a 1-week interval, respectively. The validity of the S-PPA was assessed through correlations with balance measures and the ABC-C and by comparing the S-PPA scores between the stroke and healthy groups and between fallers and non-fallers in the stroke group. A receiver operating characteristic (ROC) curve analysis was used to assess the ability of balance measures to distinguish fall status in the stroke group. The S-PPA yielded good inter-rater (intraclass correlation coefficient (ICC) = 0.83) and moderate intra-rater reliability (ICC = 0.74), correlated significantly with the 3 balance measures (rho = 0.49-0.70) and ABC-C (rho = 0.35) and revealed significant differences between stroke survivors and controls and between stroke survivors with and without a fall history. However, the ROC analysis revealed that the S-PPA had a poor ability to distinguish the fall statuses of community-dwelling stroke survivors. The S-PPA is reliable and valid for evaluating fall risks but cannot adequately distinguish the fall statuses of stroke survivors.


A prescription for sustaining community engagement in malaria elimination on Aneityum Island, Vanuatu: an application of Health Empowerment Theory.
Community engagement has contributed to disease control and elimination in many countries. Community engagement in malaria elimination (ME) on Aneityum Island has been sustained since its introduction in the early 1990s. Capacity developed within this population has led to a health empowered community response. Health Empowerment Theory (HET) can account for the innovative community actions and capacity development efforts taken to realize and sustain meaningful changes in well-being. This study used the HET framework to investigate participant perceptions of ME efforts on the island focusing on two HET elements, personal and social-contextual resources. The purpose of this study was to explore the role of empowerment as a critical element of community engagement. Six focus group discussions, ten key informant interviews and 17 in-depth interviews were conducted in July 2012 on Aneityum. Both deductive and inductive approaches to qualitative content analysis were used to identify themes, which were condensed, coded and classified based on the HET elements above. Awareness and use of personal and social-contextual resources played an important role in ME efforts. Most participants shared their knowledge to prevent malaria reintroduction. Many participants reported their skills needed for behavioral maintenance, problem-solving or leadership. Participants who perceived a threat took preventive actions even in the dry season. Community leaders focused on second generation capacity development. A local health coalition provided ME services. Members of networks were sources of information and assistance. Face-to-face was the preferred method of communication. Barriers to engagement (e.g., financial difficulties, health literacy issues and underdeveloped infrastructure) were minimized through active collaboration and mutual assistance. In the community engagement continuum, health empowerment develops incrementally overtime as people gain their knowledge and skills, form coalitions and develop collaborative networks (social capital) to make decisions and take action for change. Community engagement, which facilitates local personal and social-contextual resource development, has potential for ME and multilevel empowerment through community-based capacity development processes. These self-empowered communities have written and will continue to write a 'prescription' for sustaining high levels of engagement.


Pazopanib in Locally Advanced or Metastatic Renal Cell Carcinoma: Results of a Randomized Phase III Trial.
Pazopanib is an oral angiogenesis inhibitor targeting vascular endothelial growth factor receptor, platelet-derived growth factor receptor, and c-Kit. This randomized, double-blind, placebo-controlled phase III study evaluated efficacy and safety of pazopanib monotherapy in treatment-naive and cytokine-pretreated patients with advanced renal cell carcinoma (RCC). Adult patients with measurable, locally advanced, and/or metastatic RCC were randomly assigned 2:1 to receive oral pazopanib or placebo. The primary end point was progression-free survival (PFS). Secondary end points included overall survival, tumor response rate (Response Evaluation Criteria in Solid Tumors), and safety. Radiographic assessments of tumors were independently reviewed. Of 435 patients enrolled, 233 were treatment naive (54%) and 202 were cytokine pretreated (46%). PFS was significantly prolonged with pazopanib compared with placebo in the overall study population (median, PFS 9.2 v 4.2 months; hazard ratio [HR], 0.46; 95% CI, 0.34 to 0.62; P < .0001), the treatment-naive subpopulation (median PFS 11.1 v 2.8 months; HR, 0.40; 95% CI, 0.27 to 0.60; P < .0001), and the cytokine-pretreated subpopulation (median PFS, 7.4 v 4.2 months; HR, 0.54; 95% CI, 0.35 to 0.84; P < .001). The objective response rate was 30% with pazopanib compared with 3% with placebo (P < .001). The median duration of response was longer than 1 year. The most common adverse events were diarrhea, hypertension, hair color changes, nausea, anorexia, and vomiting. There was no evidence of clinically important differences in quality of life for pazopanib versus placebo. Pazopanib demonstrated significant improvement in PFS and tumor response compared with placebo in treatment-naive and cytokine-pretreated patients with advanced and/or metastatic RCC.


[Catatonia and neuroleptic malignant syndrome: two disorders on a same spectrum? Three case reports].
The authors present the cases of three bipolar patients who developed Neuroleptic Malignant Syndrome (NMS) after antipsychotic treatment, both typical and atypical, focusing on relationship between NMS and catatonia. In all three cases, administration of antipsychotics has been stopped at once, when fever and autonomic disturbances occurred. A supportive therapy (including rehydration, electrolyte restoration and blood pressure aids, together with antipyretics, antibiotics and anticoagulants) was prescribed in order to stabilize general conditions. Every patient started then Electroconvulsive Therapy (ECT) in combination with benzodiazepines. High risk of complications and lethal outcome, associated with NMS, were successfully reduced by the tempestive adoption of a supportive care, while combination between ECT and BDZ was effective in resolution of clinical picture. DISCUSSIONS; These cases may provide further evidences about hypothesis of catatonia and NMS as disorders on the same spectrum. In one patient, NMS occurred overlapping with a previous catatonic state, while two others exhibited catatonic features after resolution of NMS. However, catatonic symptoms arose or worsened with administration of antipsychotics, supporting hypothesis of neuroleptic-induced catatonia as a step of progressive development of NMS. Our experience also confirms efficacy and safety of ECT in combination with BDZ as treatment of NMS and residual catatonia.


Invited Commentary: Cassel's "The Contribution of the Social Environment to Host Resistance"-A Modern Classic.
John Cassel's 1976 paper "The Contribution of the Social Environment to Host Resistance" (Am J Epidemiol. 1976;104(2):107-123) is widely regarded as a classic in epidemiology. He makes the compelling argument that the quality of a person's social relationships, that is, the degree to which her relationships are more stressful than supportive (or vice versa) influences her susceptibility to disease independent of genetic endowment, diet, physical activity, etc. Cassel's provocative thesis was anchored in a cogent synthesis of findings from animal experiments and observational studies on diverse human populations. Beginning in the late 1970s, the paper stimulated an explosion of epidemiologic research on social support and human health. Beyond advancing epidemiologic theory, Cassel showed how findings from various epidemiologic study designs could be marshalled to build a persuasive causal argument that impaired social bonds increase the risk of premature disease and death. The paper also foreshadowed core ideas of later theoretical constructs, such as weathering and allostatic load, regarding the power of chronic environmental stressors to accelerate biological aging across multiple organ systems. Cassel's assessment of the research and practice implications of his conclusions has remarkable contemporary resonance for the field of epidemiology.


Patient perspectives on provision of drug information services in Japan.
Today pharmacists provide indispensable drug information to patients, however, few studies have examined the kind of information that patients want, or their satisfaction with pharmacists' advice. Therefore, we have examined a variety of factors, including individual patient characteristics, in order to identify ways to provide useful and appropriate drug information on an individual basis. In short, the aim of this study was to develop a drug information service that addresses patient needs. A prospective survey of patient satisfaction with the level of care provided by pharmacists, before and after a telephone counseling session, was performed over a 9 month period (2000-2001). Along with satisfaction ratings, the content of the interviews was assessed, along with patient characteristics. Correspondence analysis was used to classify the content of the consultations, and cluster analysis was performed to classify caller characteristics. In total, 2022 people were counseled. It should be noted that, on occasion, family members were interviewed instead of the patients themselves. The average counseling session was 11.5 minutes (n = 1876). Patients expressing the least dissatisfaction prior to counseling tended to have the highest satisfaction levels after counseling. Almost all patients were satisfied with the counseling they received. An association was found between levels of pre-counseling dissatisfaction and time spent counseling patients over the telephone. No dimensions of high inertia were identified by correspondence analysis of consultation content and patient characteristics (n = 1667), however, questions regarding "efficacy and indications", "dosages and administration", "anxiety of adverse events", "realization of adverse events", and "interactions" were deemed similar by correspondence analysis. This study shows that patient perceptions regarding drug information services differ from those of pharmacists. Furthermore, several subtypes of patients were identified, based on their responsiveness to counseling. Pharmacists should take this into consideration when standing face-to-face with patients in the setting of their pharmacy practice.


Individual-based modelling of the development and transport of a Karenia mikimotoi bloom on the North-west European continental shelf.
In 2006, a large and prolonged bloom of the dinoflagellate Karenia mikimotoi occurred in Scottish coastal waters, causing extensive mortalities of benthic organisms including annelids and molluscs and some species of fish (Davidson et al., 2009). A coupled hydrodynamic-algal transport model was developed to track the progression of the bloom around the Scottish coast during June-September 2006 and hence investigate the processes controlling the bloom dynamics. Within this individual-based model, cells were capable of growth, mortality and phototaxis and were transported by physical processes of advection and turbulent diffusion, using current velocities extracted from operational simulations of the MRCS ocean circulation model of the North-west European continental shelf. Vertical and horizontal turbulent diffusion of cells are treated using a random walk approach. Comparison of model output with remotely sensed chlorophyll concentrations and cell counts from coastal monitoring stations indicated that it was necessary to include multiple spatially distinct seed populations of K. mikimotoi at separate locations on the shelf edge to capture the qualitative pattern of bloom transport and development. We interpret this as indicating that the source population was being transported northwards by the Hebridean slope current from where colonies of K. mikimotoi were injected onto the continental shelf by eddies or other transient exchange processes. The model was used to investigate the effects on simulated K. mikimotoi transport and dispersal of: (1) the distribution of the initial seed population; (2) algal growth and mortality; (3) water temperature; (4) the vertical movement of particles by diurnal migration and eddy diffusion; (5) the relative role of the shelf edge and coastal currents; (6) the role of wind forcing. The numerical experiments emphasized the requirement for a physiologically based biological model and indicated that improved modelling of future blooms will potentially benefit from better parameterisation of temperature dependence of both growth and mortality and finer spatial and temporal hydrodynamic resolution.


Hepatoprotective activity and phenolic profile of Zanthoxylum alatum Roxb. fruit extract.
Zanthoxylum alatum Roxb. (Rutaceae) is a medicinal plant, which abundantly grows in the hilly areas of Pakistan. In the present study, the hepatoprotective activity of the Z. alatum whole fruit was evaluated. The hepatoprotective effect of the aqueous extracts was studied on mice liver damage using a single dose of paracetamol (640 mg/kg) orally by monitoring biochemical parameters. Pre-treatment of mice with Z. alatum (100 mg/kg) aqueous extract orally for four days prevented the paracetamol induced rise in serum transaminases (ALT and AST), restored the altered levels of antioxidant enzyme (catalase), thiobarbituric acid reactive substances (TBARS), non-protein thiol and ascorbic acid to near normal levels. The major phenolic acids, some flavonoid aglycone and glycosides were identified in fruit by high performance liquid chromatography. Ellagic acid (24.57±0.01mg/g), chlorogenic acid (10.65±0.01mg/g), gallic acid (9.15±0.02mg/g), chrysin (16.81±0.03), quercetin (16.81±0.03mg/g) and epicatechin (10.93±0.01) were prédominant in infusion of Z. alatum. The result substianted the hepatoprotective activity of Z. alatum which may be associated with its potential use in liver disease.


Acute fulminant carditis presenting with sustained ventricular tachycardia, and recovery after extracorporeal cardiopulmonary resuscitation.
A 12-year-old boy consulted a local physician with complaints of cough, abdominal pain, shortness of breath and general malaise. Medications for symptomatic relief and bed rest were suggested. The flu-like symptoms were relieved on the 2nd day, and the general malaise with repeated vomiting, chest pain and chest tightness attenuated on the 3rd day. A chest x ray showed multiple pneumonic patches with borderline cardiomegaly. Poor left ventricular function was noted, and the left ventricular ejection fraction was reduced to 21%. Although multiple episodes of sustained ventricular tachycardia were converted by six repeated cardiac defibrillations and a xylocaine (intravenous) bolus infusion, his general condition went downhill to shock and proceeded to several episodes of heart standstill that necessitated cardiopulmonary resuscitation. Extracorporeal cardiopulmonary resuscitation was installed via femoral cannulation. Cardiac function progressively recovered to normal, and extracorporeal membrane oxygenation was removed on the 7th day. The patient completely recovered and was discharged on the 15th day with no neurological sequelae.


Is asymptomatic hemorrhagic transformation really innocuous?
Asymptomatic hemorrhagic transformation (HT) is not associated with immediate deterioration of patients with acute ischemic stroke. However, it is unclear whether it is clinically innocuous with respect to long-term outcome. The aim of this study was to determine the impact of asymptomatic HT on 3-month outcome. A consecutive series of 1,618 patients, hospitalized between January 2004 and August 2007 for ischemic stroke within 7 days from symptom onset were identified in a prospective stroke registry database. Those who had no evidence of acute cerebral ischemia on diffusion-weighted MRI, who did not undergo T2-weighted gradient echo MRI, whose modified Rankin Scale (mRS) score at 3 months after stroke onset was not available, or who had symptomatic HT were excluded. The odds ratio (OR) of asymptomatic HT was calculated for the full distribution of mRS score and adjusted for variables with p < 0.25 with respect to their associations with asymptomatic HT or functional outcome. Of 1,412 patients eligible for the study, 100 (7.1%) had asymptomatic HT. Patients who experienced asymptomatic HT were more likely to have cardioembolic stroke, to receive thrombolytic therapy, to receive anticoagulation with heparin, and to have a higher initial NIH Stroke Scale score. The crude and adjusted ORs of asymptomatic HT for an increment of mRS score at 3 months were 2.94 (95% confidence interval 2.05-4.24) and 1.90 (1.27-2.82), respectively. Our study shows that the odds of a worse outcome are increased by a factor of 2 in patients with asymptomatic HT compared with those without HT after acute ischemic stroke.


NMR conformational study of the cytoplasmic domain of the canine Sec61 gamma protein from the protein translocation pore of the endoplasmic reticulum membrane.
Conformational studies of the synthesized N-terminal cytoplasmic domain of the canine Sec61 gamma protein, an essential protein from the translocation pore of secretory proteins across the endoplasmic reticulum membrane, were performed using two-dimensional proton NMR spectroscopy. This canine domain is one of the smallest domains within the homologous protein family and may thus constitute the minimal functional structure. The peptide was solubilized in pure aqueous solution or in the presence of dodecylphosphocholine micelles mimicking a membrane-solution interface. In pure aqueous solution, the peptide is remarkably unfolded. Forming a stable complex with dodecylphosphocholine micelles, it acquires a well-defined alpha-helix-loop-alpha-helix secondary structure, with the helix, highly amphipathic, lying at the micelle surface. The loop comprising four residues is delimited by two flanking helix-capping structures, highly conserved in the whole homologous protein family. No tertiary structure, which could have been revealed by interhelix NOE contacts, was observed. From these experimental results and using general arguments based on sequence information and knowledge of peptide-membrane interactions, a structure of the entire Sec61 gamma protein in membrane bilayers is proposed.


Covalent binding of a reactive metabolite derived from propranolol and its active metabolite 4-hydroxypropranolol to hepatic microsomal proteins of the rat.
Repeated administration of propranolol (PL) to rats causes the inhibition of cytochrome P450-2D (P450-2D) enzyme. We recently found that 4-hydroxypropranolol (4-OH-PL) was biotransformed to 1,4-naphthoquinone (1,4-NQ) by superoxide (SO) anions in medium containing rat liver microsomes and NADPH and proposed that the binding of the quinone to P450-2D apoproteins might be one of mechanisms for the enzyme inhibition [Narimatsu et al. (1995) Chem. Res. Toxicol. 8, 721-728]. In this study, we have searched for possible sources of SO for the conversion of 4-OH-PL to 1,4-NQ in rat liver microsomes and determined the radioactivity covalently bound to microsomal proteins after incubation of radioactive PL and 4-OH-PL with rat liver microsomes. Elimination of 4-OH-PL from a mixture containing microsomes and NADPH was suppressed by carbon monoxide. Antibodies raised to P450-2B1 and -3A2 partially, and antibody against NADPH-cytochrome P450 reductase (fp2) markedly suppressed the reaction. 1,4-NQ was formed concomitantly with 4-OH-PL elimination by a reconstituted preparation of fp2. Binding studies using naphthalene ring (NR)- and side chain (SC)-radiolabeled PL and 4-OH-PL showed that radioactivity covalently bound to microsomal proteins was much higher from 4-OH-PL than from PL for the NR-labeled compounds, but higher from PL than from 4-OH-PL for the SC-labeled compounds. These results suggest that the 4-OH-PL formed from PL by P450-2D enzyme is converted to 1,4-NQ with loss of the side chain, and the 1,4-NQ accounts for most of the radioactivity covalently bound to microsomal proteins, including the P450-2D enzymes. The SO for conversion of 4-OH-PL to 1,4-NQ is supplied mainly by fp2 with some contribution by P450 enzymes.


Assessment of adolescent body perception: development and characterization of a novel tool for morphing images of adolescent bodies.
We developed a computer-based method of distorting adolescent body images, which incorporates the covariation between body parts found during growth and sexual maturation. An adolescent body-shape database (AdoBSD) and adolescent body morphing tool (AdoBMT) are described; the AdoBSD comprises real (n=320) and morphed (n approximately 41,000) images (front and side view) of 160 adolescents (9-17 years). We used a point distribution model, based upon principal components analysis, to characterize the covariation between predefined body tag-points manually positioned on the body images and to morph the body images in a realistic manner. Eight principal components (PCs) were found to characterize 96.3% of the covariation between body tag-point. Application of the PCs to the body images resulted in the manipulation of body parts including shoulder width, waist, hip, belly, thigh and calf sizes. The AdoBMT and AdoBSD may be used to investigate changes in body perception during adolescence, and the role of body perception in adolescent obesity and eating disorders. The AdoBSD is available to the research community (www.brainbody.nottingham.ac.uk).


Epistasis: obstacle or advantage for mapping complex traits?
Identification of genetic loci in complex traits has focused largely on one-dimensional genome scans to search for associations between single markers and the phenotype. There is mounting evidence that locus interactions, or epistasis, are a crucial component of the genetic architecture of biologically relevant traits. However, epistasis is often viewed as a nuisance factor that reduces power for locus detection. Counter to expectations, recent work shows that fitting full models, instead of testing marker main effect and interaction components separately, in exhaustive multi-locus genome scans can have higher power to detect loci when epistasis is present than single-locus scans, and improvement that comes despite a much larger multiple testing alpha-adjustment in such searches. We demonstrate, both theoretically and via simulation, that the expected power to detect loci when fitting full models is often larger when these loci act epistatically than when they act additively. Additionally, we show that the power for single locus detection may be improved in cases of epistasis compared to the additive model. Our exploration of a two step model selection procedure shows that identifying the true model is difficult. However, this difficulty is certainly not exacerbated by the presence of epistasis, on the contrary, in some cases the presence of epistasis can aid in model selection. The impact of allele frequencies on both power and model selection is dramatic.


Connecting Our Gut Feeling and How Our Gut Feels: The Role of Well-being Attributes in Irritable Bowel Syndrome.
There is a close relationship between the mind and gut in the pathogenesis of functional bowel disorders. Common psychological disturbances such as depression and anxiety are not uncommon in those with irritable bowel syndrome (IBS). There is little research investigating the role of positive psychology and gastrointestinal (GI) conditions. In this pilot study we investigated the well-being attributes in those with and without IBS. We used an anonymous online survey and recruited 416 study subjects using social media as the main method of recruitment. We gathered demographic information, GI symptoms, history of mental health issues such as anxiety and depression, assessed several well-being attributes, and finally assessed subjective well-being. We hypothesized that those with GI symptoms and IBS have lower scores in their well-being attributes compared to healthy controls. We observed that a history of anxiety and depression is significantly associated with GI symptoms and IBS. In addition, sense of subjective well-being and several well-being attributes are negatively associated with GI symptoms and/or IBS. Of interest, the household income showed a negative correlation with the prevalence of GI symptoms and IBS. Subjective well-being, and several well-being attributes that contribute to the sense of overall contentment, are negatively associated with GI symptoms and IBS. The link between subjective well-being, and GI symptoms and IBS are independent of anxiety and depression. Well-being attributes and sense of subjective well-being may be a contributory factor in clinical expression of GI symptoms or IBS consistent with the biopsychosocial model of the disease.


An optimised patient information sheet did not significantly increase recruitment or retention in a falls prevention study: an embedded randomised recruitment trial.
Randomised controlled trials are generally regarded as the 'gold standard' experimental design to determine the effectiveness of an intervention. Unfortunately, many trials either fail to recruit sufficient numbers of participants, or recruitment takes longer than anticipated. The current embedded trial evaluates the effectiveness of optimised patient information sheets on recruitment of participants in a falls prevention trial. A three-arm, embedded randomised methodology trial was conducted within the National Institute for Health Research-funded REducing Falls with ORthoses and a Multifaceted podiatry intervention (REFORM) cohort randomised controlled trial. Routine National Health Service podiatry patients over the age of 65 were randomised to receive either the control patient information sheet (PIS) for the host trial or one of two optimised versions, a bespoke user-tested PIS or a template-developed PIS. The primary outcome was the proportion of patients in each group who went on to be randomised to the host trial. Six thousand and nine hundred patients were randomised 1:1:1 into the embedded trial. A total of 193 (2.8%) went on to be randomised into the main REFORM trial (control n = 62, template-developed n = 68; bespoke user-tested n = 63). Information sheet allocation did not improve recruitment to the trial (odds ratios for the three pairwise comparisons: template vs control 1.10 (95% CI 0.77-1.56, p = 0.60); user-tested vs control 1.01 (95% CI 0.71-1.45, p = 0.94); and user-tested vs template 0.92 (95% CI 0.65-1.31, p = 0.65)). This embedded methodology trial has demonstrated limited evidence as to the benefit of using optimised information materials on recruitment and retention rates in the REFORM study. International Standard Randomised Controlled Trials Number registry, ISRCTN68240461 . Registered on 01 July 2011.


Analysis of turnover dynamics of the submembranous actin cortex.
The cell cortex is a thin network of actin, myosin motors, and associated proteins that underlies the plasma membrane in most eukaryotic cells. It enables cells to resist extracellular stresses, perform mechanical work, and change shape. Cortical structural and mechanical properties depend strongly on the relative turnover rates of its constituents, but quantitative data on these rates remain elusive. Using photobleaching experiments, we analyzed the dynamics of three classes of proteins within the cortex of living cells: a scaffold protein (actin), a cross-linker (α-actinin), and a motor (myosin). We found that two filament subpopulations with very different turnover rates composed the actin cortex: one with fast turnover dynamics and polymerization resulting from addition of monomers to free barbed ends, and one with slow turnover dynamics with polymerization resulting from formin-mediated filament growth. Our data suggest that filaments in the second subpopulation are on average longer than those in the first and that cofilin-mediated severing of formin-capped filaments contributes to replenishing the filament subpopulation with free barbed ends. Furthermore, α-actinin and myosin minifilaments turned over significantly faster than F-actin. Surprisingly, only one-fourth of α-actinin dimers were bound to two actin filaments. Taken together, our results provide a quantitative characterization of essential mechanisms under-lying actin cortex homeostasis.


Pseudomonas aeruginosa: an uncommon cause of diabetic foot infection.
Pseudomonas aeruginosa has traditionally been considered a common pathogen in diabetic foot infection (DFI), yet the 2012 Infectious Diseases Society of America guideline for DFI states that "empiric therapy directed at P aeruginosa is usually unnecessary." The objective of this study was to evaluate the frequency of P aeruginosa isolated from bone or tissue cultures from patients with DFI. This study is a cross-sectional survey of diabetic patients presenting with a foot infection to an urban county hospital between July 1, 2012, and December 31, 2013. All of the patients had at least one debridement procedure during which tissue or bone cultures from operative or bedside debridements were obtained. The χ(2) test and the t test of means were used to determine relationships between variables and the frequency of P aeruginosa in culture. The median number of bacteria isolated from DFI was two. Streptococcus spp and Staphylococcus aureus were the most commonly isolated organisms; P aeruginosa was isolated in only five of 112 patients (4.5%). The presence of P aeruginosa was not associated with the patient's age, glycosylated hemoglobin level, tobacco abuse, the presence of osteomyelitis, a prescription for antibiotic drugs in the preceding 3 months, or the type of operative procedure. Pseudomonas aeruginosa was an infrequent isolate from DFI in this urban, underserved diabetic population. The presence of P aeruginosa was not associated with any measured risk factors. By introducing a clinical practice guideline, we hope to discourage frontline providers from using routine antipseudomonal antibiotic drugs for DFI.


The role of chemical antifouling defence in the invasion success of Sargassum muticum: A comparison of native and invasive brown algae.
Competition and fouling defence are important traits that may facilitate invasions by non-indigenous species. The 'novel weapons hypothesis' (NWH) predicts that the invasive success of exotic species is closely linked to the possession of chemical defence compounds that the recipient community in the new range is not adapted to. In order to assess whether chemical defence traits contribute to invasion success, anti-bacterial, anti-quorum sensing, anti-diatom, anti-larval and anti-algal properties were investigated for the following algae: a) the invasive brown alga Sargassum muticum from both, its native (Japan) and invasive (Germany) range, b) the two non- or weak invasive species Sargassum fusiforme and Sargassum horneri from Japan, and c) Fucus vesiculosus, a native brown alga from Germany. Crude and surface extracts and lipid fractions of active extracts were tested against common fouling organisms and zygotes of a dominant competing brown alga. Extracts of the native brown alga F. vesiculosus inhibited more bacterial strains (75%) than any of the Sargassum spp. (17 to 29%). However, Sargassum spp. from Japan exhibited the strongest settlement inhibition against the diatom Cylindrotheca closterium, larvae of the bryozoan Bugula neritina and zygotes of the brown alga F. vesiculosus. Overall, extracts of S. muticum from the invasive range were less active compared to those of the native range suggesting an adaptation to lower fouling pressure and competition in the new range resulting in a shift of resource allocation from costly chemical defence to reproduction and growth. Non-invasive Sargassum spp. from Japan was equally defended against fouling and competitors like S. muticum from Japan indicating a necessity to include these species in European monitoring programs. The variable antifouling activity of surface and crude extracts highlights the importance to use both for an initial screening for antifouling activity.


Immunization and protection against malaria during murine pregnancy.
Normal and immune mice were evaluated for their ability to resist infection to the rodent malaria parasite, Plasmodium yoelii, during pregnancy. Parasitemia levels were slightly higher and time-to-death shorter in the nonimmunized pregnant group infected with virulent parasites relative to virgin controls. Subinoculation experiments revealed that numerous virulent organisms were present in the placentas of unprotected gravida but were absent from the fetal livers of their conceptuses. It was also found that mice preimmunized with irradiated P. yoelii survived a usually lethal challenge infection during mid-gestation and delivered healthy newborns. Associated with this protection against transplacental spread of parasites was the additional key finding that placental macrophages were as effective as peritoneal exudate cells in phagocytosing parasite derived material in vitro. This murine malaria-pregnancy model should provide new insights on the various factors (virulence, immunogenicity) of microbial infections affecting the fetal-maternal relationship, as well as on the expression of immune effector mechanisms and immunoregulation, during the reproductive process.


Model dependence in quantification of spike interdependence by joint peri-stimulus time histogram.
Multineuronal recordings have enabled us to examine context-dependent changes in the relationship between the activities of multiple cells. The joint peri-stimulus time histogram (JPSTH) is a much-used method for investigating the dynamics of the interdependence of spike events between pairs of cells. Its results are often taken as an estimate of interaction strength between cells, independent of modulations in the cells' firing rates. We evaluate the adequacy of this estimate by examining the mathematical structure of how the JPSTH quantifies an interaction strength after excluding the contribution of firing rates. We introduce a simple probabilistic model of interacting point processes to generate simulated spike data and show that the normalized JPSTH incorrectly infers the temporal structure of variations in the interaction parameter strength. This occurs because, in our model, the correct normalization of firing-rate contributions is different than that used in Aertsen, Gerstein, Habib, and Palm's (1989) effective connectivity model. This demonstrates that firing-rate modulations cannot be corrected for in a model-independent manner, and therefore the effective connectivity does not represent a universal characteristic that is independent of modulation of the firing rates. Aertsen et al.'s (1989) effective connectivity may still be used in the analysis of experimental data, provided we are aware that this is simply one of many ways of describing the structure of interdependence. We also discuss some measure-independent characteristics of the structure of interdependence.


Electronic transitions in perovskite: possible nonconvecting layers in the lower mantle.
We measured the spin state of iron in magnesium silicate perovskite (Mg(0.9),Fe(0.1))SiO(3) at high pressure and found two electronic transitions occurring at 70 gigapascals and at 120 gigapascals, corresponding to partial and full electron pairing in iron, respectively. The proportion of iron in the low spin state thus grows with depth, increasing the transparency of the mantle in the infrared region, with a maximum at pressures consistent with the D" layer above the core-mantle boundary. The resulting increase in radiative thermal conductivity suggests the existence of nonconvecting layers in the lowermost mantle.


Block copolymer micelles as switchable templates for nanofabrication.
Block copolymer inverse micelles from polystyrene-block-poly-2-vinylpyridine (PS-b-P2VP) deposited as monolayer films onto surfaces show responsive behavior and are reversibly switchable between two states of different topography and surface chemistry. The as-coated films are in the form of arrays of nanoscale bumps, which can be transformed into arrays of nanoscale holes by switching through exposure to methanol. The use of these micellar films to act as switchable etch masks for the structuring of the underlying material to form either pillars or holes depending on the switching state is demonstrated.


17p13.3 genomic rearrangement in a Chinese family with split-hand/foot malformation with long bone deficiency: report of a complicated duplication with marked variation in phenotype.
Split hand/foot malformation (SHFM) is a genetically heterogeneous limb malformation with variable expressivity. SHFM with tibia or femur aplasia is called SHFM with long bone deficiency (SHFLD). 17p13.3 duplications containing BHLHA9 are associated with SHFLD. Cases with variable SHFLD phenotype and different 17p13.3 duplicated regions are reported. The severity of long bone defect could not be simply explained by BHLHA9 overdosage or 17p13.3 duplication. A four-generation Chinese SHFM family was recruited. Three family members have long bone defects, one male was severely affected with hypoplasia or aplasia in three of four limbs. Linkage analysis and direct sequencing of candidate genes were used to exclude six responsible genes/loci for isolated SHFM. Array comparative genomic hybridization (CGH) was performed to detect copy number variations on a genome-wide scale, and quantitative real-time polymerase chain reaction (qPCR) assays were designed to validate the identified copy number variation in the index and other family members. No mutations were found in genes or loci linked to isolated SHFM. A ~ 966 kb duplication was identified in 17p13.3 by array CGH, in which BHLHA9 surrounding region presented as triplication. The qPCR assays confirmed the indicated 17p13.3 duplication as well as BHLHA9 triplication in all available affected family members and other two asymptomatic carriers. Given the incomplete penetrance in SHFLD, those two carriers were regarded as non-penetrant, which suggested that the genomic rearrangement was co-segregated with malformation in this family. The present study reports an additional SHFLD family case with 17p13.3 genomic rearrangement. To our knowledge, the 966 kb genomic rearrangement is larger in size than any previously reported SHFLD-associated 17p13.3 duplication, and the present family shows marked phenotypic variability with two asymptomatic carriers and one patient with an extremely severe phenotype. This rare case provides the opportunity to identify underlying genotype-phenotype correlations between SHFLD and 17p13.3 genomic rearrangement.


Esophageal Pharmacobezoar in a Patient with Achalasia: A Case Report.
A few cases with esophageal bezoar have been reported in achalasia. We describe here a rare case of esophageal pharmacobezoar after ingestion of ferrous sulfate capsules in a patient with achalasia. A 29-year-old woman presented with severe dysphagia since five days earlier. She had history of achalasia since 3 years ago but had refused any treatment option. After about 3 weeks of ferrous sulfate capsules ingestion, she developed severe dysphagia and was referred to a gastroenterologist. Physical examination was unremarkable. A barium swallow revealed dilated esophagus and bird's beak appearance. Esophagogastroduodenoscopy (EGD) showed dilated esophagus and soft black color bezoar in distal part of esophagus. The bezoar was retrieved with basket. In the next endoscopic session, achalasia balloon dilation was successfully applied. Ferrous sulfate capsules can cause pharmacobezoar in patients with achalasia. Esophageal bezoar should be considered in differential diagnosis of untreated achalasia and acute exacerbation of dysphagia.


Contribution of symptom clusters to multiple sclerosis consequences.
There were two objectives in this study: (1) to identify, among women and men with MS, the extent to which different MS-related symptoms, including fatigue, pain, sleep disturbance, depression, anxiety, irritability, cognitive impairment, spasticity, and poor balance, cluster and (2) to compare the contribution of generated symptom clusters to MS consequences including functional walking capacity, perceived health, illness intrusiveness, and quality of life (QOL). This was a cross-sectional study. A center-stratified random sample comprising 139 women and 49 men was recruited from three major MS clinics in Montreal. Subjects completed several self-report and performance-based measures that assessed symptoms and downstream MS consequences. Hierarchical and K-means cluster analyses were used to create clusters. Three symptom clusters were identified. Cluster 1, labeled the "emotional/cognitive symptom cluster," comprised of depression, anxiety, cognitive impairments, and irritability. The second cluster, labeled the "physical symptom cluster," included pain, fatigue, and sleep disorders. Cluster 3, labeled the "motor symptom cluster," included spasticity and poor balance. Furthermore, the motor symptom cluster had a strong effect on functional walking capacity, while it did not affect significantly illness intrusiveness and QOL. On the other hand, the physical symptom clusters and emotional/cognitive symptom clusters showed a significant contribution to prediction of illness intrusiveness and QOL. All symptom clusters showed a significant effect in predicting the overall variability of perceived health status. The findings of this study provide useful information to help healthcare professionals, clinicians, and researchers to target symptoms that are often in the same cluster when one or two of them are present. Identification of the strength of the contributions of each symptom clusters to the targeted MS consequences would further help to prioritize treatment approaches for the MS population.


Caregiver-Focused, Web-Based Interventions: Systematic Review and Meta-Analysis (Part 2).
Approaches to support the health and well-being of family caregivers of adults with chronic conditions are increasingly important given the key roles caregivers play in helping family members to live in the community. Web-based interventions to support caregivers have the potential to lessen the negative health impacts associated with caregiving and result in improved health outcomes. The primary objective of this systematic review and meta-analysis was to examine the effect of caregiver-focused, Web-based interventions, compared with no or minimal Web-based interventions, on caregiver outcomes. The secondary objective was to assess the effect of different types of Web-based interventions (eg, education, peer and professional psychosocial support, and electronic monitoring of the care recipient), compared with no or minimal Web-based interventions, on caregiver outcomes. MEDLINE, EMBASE, CIHAHL, PsychInfo, Cochrane, and AgeLine were searched from January 1995 to April 2017 for relevant randomized controlled trials (RCTs) or controlled clinical trials (CCTs) that compared caregiver-focused, Web-based intervention programs with no or minimal Web-based interventions for caregivers of adults with at least one chronic condition. Studies were included if they involved: adult family or friend caregivers (aged ≥18 years) of adults living in the community with a chronic condition; a caregiver-focused, Web-based intervention of education or psychosocial support or electronic monitoring of the care recipient; and general caregiver outcomes (ie, burden, life satisfaction, self-efficacy or mastery, reaction to problem behavior, self-esteem, strain, and social support). Title and abstract as well as full-text screening were completed in duplicate. Data were extracted by a single reviewer and verified by a second reviewer, and risk of bias assessments were completed accordingly. Where possible, data for these caregiver outcomes were meta-analyzed. The search yielded 7927 unique citations, of which 294 studies were screened at full text. Of those, 14 studies met the inclusion criteria; 12 were RCTs and 1 study was a CCT. One study used an RCT design in 1 country and a CCT design in 2 other countries. The beneficial effects of any Web-based intervention program, compared with no or minimal Web-based intervention, resulted in a mean increase of 0.85 points (95% CI 0.12 to 1.57) for caregiver self-esteem, a mean increase of 0.36 points (95% CI 0.11 to 0.62) for caregiver self-efficacy or mastery, and a mean decrease of 0.32 points (95% CI -0.54 to -0.09) for caregiver strain. However, the results are based on poor-quality studies. The review found evidence for the positive effects of Web-based intervention programs on self-efficacy, self-esteem, and strain of caregivers of adults living with a chronic condition. Further high-quality research is needed to inform the effectiveness of specific types of Web-based interventions on caregiver outcomes. PROSPERO CRD42018091715; https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=91715 (Archived by WebCite at http://www.webcitation.org/738zAa5F5).


Marching to a Different Drummer: A Cross-Cultural Comparison of Young Adolescents Who Challenge Gender Norms.
Little is known about how gender norms regulate adolescents' lives across different cultural settings. This study aims to illustrate what is considered as violating gender norms for boys and girls in four urban poor sites as well as the consequences that follow the challenging of gender norms. Data were collected as part of the Global Early Adolescent Study, a 15-country collaboration to explore gender norms and health in early adolescence. The current study analyzed narrative and in-depth interviews conducted in urban poor sites in two middle-income (Shanghai, China; and New Delhi, India) and two high-income countries (Baltimore, U.S.; and Ghent, Belgium). A total of 238 participants, 59 boys and 70 girls aged 11-13 years old and 109 of their parents/guardians (28 male adults and 81 female adults), were interviewed. A thematic analysis was conducted across sites using Atlas.Ti 7.5 software. Findings revealed that although most perceptions and expressions about gender were regulated by stereotypical norms, there was a growing acceptability for girls to wear boyish clothes and engage in stereotypical masculine activities such as playing soccer/football. However, there was no comparable acceptance of boys engaging in traditional feminine behaviors. Across all sites, challenging gender norms was often found to lead to verbal, physical, and/or psychological retribution. While it is sometimes acceptable for young adolescents to cross gender boundaries, once it becomes clear that a behavior is socially defined as typical for the other sex, and the adolescent will face more resistance. Researchers, programmers, and clinicians working in the field of adolescent health need not only attend to those who are facing the consequences of challenging prevailing gender norms, but also to address the environment that fosters exclusion and underscores differences.


Interleukin-3 and bryostatin 1 mediate rapid nuclear envelope protein phosphorylation in growth factor-dependent FDC-P1 hematopoietic cells. A possible role for nuclear protein kinase C.
Interleukin-3 (IL-3) is a lymphokine which stimulates the proliferation of normal and transformed multilineage hematopoietic cells. Recently we reported that bryostatin 1, a macrocyclic lactone and potent activator of protein kinase C, could stimulate normal multipotential hematopoietic progenitor cells in vitro in the absence of added polypeptide growth factors. We have now used the murine IL-3-dependent cell line FDC-P1, derived from normal murine marrow cells, to examine the early biochemical events associated with stimulation of hematopoietic cells. We find that both IL-3 and bryostatin 1 are mitogenic and stimulate the growth of FDC-P1 cells. Cells grown for extended periods in the presence of bryostatin 1 (1 nM) alone retain IL-3 responsiveness, indicating that bryostatin 1 does not induce an IL-3-independent state. Protein phosphorylation studies in cells treated with either IL-3 or bryostatin 1 indicate that both stimulators can mediate the rapid (within 5 min) serine-specific phosphorylation of several nuclear envelope polypeptides, including lamin B. Both IL-3- and bryostatin 1-mediated nuclear envelope phosphorylation is dose-dependent, occurring at concentrations which are mitogenic to FDC-P1 cells. The extent of nuclear envelope phosphorylation mediated by IL-3 and bryostatin 1 correlates with the mitogenic response. Furthermore, both mitogens mediate the rapid immunologic translocation of protein kinase C to the nuclear envelope where phosphorylation occurs. These data indicate that the early mitogenic signal(s) generated by IL-3 and bryostatin 1 may converge at the level of the nuclear envelope, perhaps through a protein kinase C-like activity which mediates phosphorylation of specific nuclear envelope polypeptides such as lamin B.


Toxicity of cadmium and its health risks from leafy vegetable consumption.
Cadmium (Cd) is a highly toxic heavy metal and has spread widely in the environment in recent decades. This review summarizes current knowledge about Cd contamination of leafy vegetables, its toxicity, exposure, health risks, and approaches to reducing its toxicity in humans. Leafy vegetable consumption has been identified as a dominant exposure pathway of Cd in the human body. An overview of Cd pollution in leafy vegetables as well as the main sources of Cd is given. Notable estimated daily intakes and health risks of Cd exposure through vegetable consumption for humans are revealed in occupational exposure areas and even in some reference areas. Vegetable consumption is one of the most significant sources of exposure to Cd, particularly in occupational exposure regions. Therefore, numerous approaches have been developed to minimize the accumulation of Cd in leafy vegetables, among which the breeding of Cd pollution-safe cultivars is one of the most effective tools. Furthermore, dietary supplements from leafy vegetables perform positive roles in alleviating Cd toxicity in humans with regard to the effects of essential mineral elements, vitamins and phytochemicals taken into the human body via leafy vegetable consumption.


Comparative ultrasonographic examination and measurements of the urethra and penis of castrated and intact male lambs.
Early castration of male small ruminants is regarded as a risk factor for urolithiasis, although the underlying correlations are still unclear. One possible reason is a deferred development of the penis and the urethra after castration. Therefore, we examined the penis and urethra of castrated and intact lambs by ultrasonography to determine the correlation between urethral area and pe- nile cross-sectional area. Ultrasonography was performed in 6-month-old Lacaune crossbred lambs (early castrated, late castrated, and intact; each group, n = 11). Sectional images at 5 loca- tions (glans penis, penile urethra, distal and proximal sigmoid flexure, and ischial arch) were ob- tained to determine the urethral and penile diameters. Urethral and penile cross-sectional areas were calculated. Grey-scale analysis of ultrasound images was performed to evaluate possible differences in the penile texture between the groups. Correlation analyses between both cross-sectional areas showed a significant general correlation for location 2 in all lambs (R = 0.52; P = 0.003), for location 3 in late-castrated lambs, and for location 5 in early-castrated lambs. Statistically significant correlations between the penile and the urethral area of castrated and intact lambs were not evident. Therefore, measurement of the penile cross-sectional area alone does not allow for accurate estimation of urethral size. Statistically significant differences con- cerning the grey-scale analysis between the groups were also not detectable. Thus, simplification of the formerly presented ultrasonographic examination of the urethra is not recommended. In animals at a risk of obstructive urolithiasis, complete urethral examina- tion is essential.


MindSurf: a pilot study to assess the usability and acceptability of a smartphone app designed to promote contentment, wellbeing, and goal achievement.
The Method of Levels (MOL) is a transdiagnostic cognitive therapy that promotes contentment, wellbeing, and goal achievement through the resolution of internal conflicts underlying psychological distress. MOL, based on Perceptual Control Theory (PCT), was developed in routine clinical practice and has been used effectively across different health services by different practitioners. Access to MOL-style questions through a smartphone app could, potentially, help both the general public maintain robust mental health, and also be a useful adjunct to therapy for clinical populations. The app is called MindSurf because of its focus on helping people explore their thinking. Prior to developing the app and using it with different populations it was necessary to determine whether such an idea would be usable for and acceptable to potential app users. Therefore, a pilot study was conducted with a non-clinical sample to assess the usability and acceptability of the app including monitoring whether the questions delivered in this way were associated with any adverse events. A pilot study using quantitative as well as qualitative methods and incorporating a repeated measures, A-B design was conducted. The 23 participants were healthy adult volunteers who were all either undergraduate students, postgraduate students, or staff of the University of Manchester. They received MOL-style questions on their mobile phones over a 1-week period. Qualitative results were encouraging and indicated that the format and style of questioning were acceptable to participants and did not lead to increased worry or concern. A one-way, repeated measures ANOVA indicated that there was a nonsignificant decrease in scores on the 21-item Depression, Anxiety, and Stress Scale (DASS21) over a 2 week period. The results of the pilot study justified development of MindSurf and further testing once it is available for use. A power analysis indicated that the pilot study was underpowered to detect significant effects but provided important information regarding the appropriate sample size for future research. The pilot study also indicated that future research should investigate the effects of receiving more than three questions per day. Results of the pilot study indicate that MindSurf will be a usable and acceptable app. Its benefits should be further explored through longer studies with larger sample of both the general population as well as clinical populations.


RNA velocity of single cells.
RNA abundance is a powerful indicator of the state of individual cells. Single-cell RNA sequencing can reveal RNA abundance with high quantitative accuracy, sensitivity and throughput1. However, this approach captures only a static snapshot at a point in time, posing a challenge for the analysis of time-resolved phenomena such as embryogenesis or tissue regeneration. Here we show that RNA velocity-the time derivative of the gene expression state-can be directly estimated by distinguishing between unspliced and spliced mRNAs in common single-cell RNA sequencing protocols. RNA velocity is a high-dimensional vector that predicts the future state of individual cells on a timescale of hours. We validate its accuracy in the neural crest lineage, demonstrate its use on multiple published datasets and technical platforms, reveal the branching lineage tree of the developing mouse hippocampus, and examine the kinetics of transcription in human embryonic brain. We expect RNA velocity to greatly aid the analysis of developmental lineages and cellular dynamics, particularly in humans.