The classification of antiseptic products to be administered to wounds--another borderline case between medicinal products and medical devices?
The importance of a correct demarcation between a Medicinal Product (MP) and a Medical Device (MD) is undisputedly one of the major topics related to the development and launch of a new healthcare product. However, for some products the correct demarcation between MPs and MDs can turn out to be somewhat complicated. This article aims to provide an overview on the existing legislation and its adequate application based on a suitable example at hand. Article 2 (2) of the European Directive 2001/83/EC as amended by Directive 2004/27/EC on the Community code relating to medicinal products for human use stipulates that the respective Medicinal Products Legislation must be applied whenever a product can be covered by both the definitions for MPs and for products regulated by other legal provisions enacted by the European Community, e.g. Cosmetic Products (CPs) or MDs. This basic principle implies that the decision to base the risk-benefit assessment of the product in question on the Medical Device Directive (MDD) would contradict the aforementioned constitutional principle, pursuant to which the stricter of the regulatory procedures theoretically possible is to apply in cases of doubt. In contrast to the approval procedure established for MPs, the MDD requires a Conformity Assessment Procedure to be performed by the manufacturer himself and a "Notified Body". Thus, in the majority of cases the responsibility for the risk assessment of MDs lies solely with the manufacturer and is prior to launch not subject to further scrutiny by regulators. Only in specific cases, i.e. for the Conformity Assessment Procedure of Class III MDs which contain an Active Pharmaceutical Ingredient one of the Member States competent authorities designated in accordance with Directive 65/ 65/EEC has to be involved before taking a decision. It is therefore important that the classification of the product is carried out carefully in full compliance with existing legal provisions, also taking into account the related guidance documents issued by the European Commission. The adequate application of these rules is explained using the example of the antiseptic compound polihexanide, which is used both in approved medicinal products (wound antiseptics) and wound irrigation solutions labelled as medical devices.


A cell cycle phosphoproteome of the yeast centrosome.
Centrosomes organize the bipolar mitotic spindle, and centrosomal defects cause chromosome instability. Protein phosphorylation modulates centrosome function, and we provide a comprehensive map of phosphorylation on intact yeast centrosomes (18 proteins). Mass spectrometry was used to identify 297 phosphorylation sites on centrosomes from different cell cycle stages. We observed different modes of phosphoregulation via specific protein kinases, phosphorylation site clustering, and conserved phosphorylated residues. Mutating all eight cyclin-dependent kinase (Cdk)-directed sites within the core component, Spc42, resulted in lethality and reduced centrosomal assembly. Alternatively, mutation of one conserved Cdk site within γ-tubulin (Tub4-S360D) caused mitotic delay and aberrant anaphase spindle elongation. Our work establishes the extent and complexity of this prominent posttranslational modification in centrosome biology and provides specific examples of phosphorylation control in centrosome function.


Check-up examination: recommendations in adults.
Check-up examinations, or periodic health examinations (PHEs), have gained in importance during the last decades and are nowadays among the most common reasons for consultations in primary care settings. The aim of PHEs is to identify risk factors and early signs of disease, but also to prevent future illness by early intervention. Therefore, each PHE should include counselling, immunisation and physical examination according to the patient's age and gender. However, deciding whether to screen a patient and choosing the most appropriate screening method can be challenging for general practitioners. The U.S. Preventive Service Task Force (USPSTF) provides updated recommendations on different existing preventive care measures based on relevant literature review. The aim of this review is to provide an updated statement of recommendations regarding preventive care measures based mostly on the guidelines derived from the USPSTF and the Swiss Medical Board. Among the major updates, there is no recommendation anymore to routinely screen for breast cancer and prostate cancer in asymptomatic adults. Since 2013, however, the USPSTF recommends annual screening for lung cancer with low-dose CT in patients aged 55 to 80 years with a smoking history of ≥30 pack years. During PHEs, the physician should be alert to the patients' hidden agendas, which are the reason for one third of all consultations in primary care.


Polyclonal Antibody Production for Membrane Proteins via Genetic Immunization.
Antibodies are essential for structural determinations and functional studies of membrane proteins, but antibody generation is limited by the availability of properly-folded and purified antigen. We describe the first application of genetic immunization to a structurally diverse set of membrane proteins to show that immunization of mice with DNA alone produced antibodies against 71% (n = 17) of the bacterial and viral targets. Antibody production correlated with prior reports of target immunogenicity in host organisms, underscoring the efficiency of this DNA-gold micronanoplex approach. To generate each antigen for antibody characterization, we also developed a simple in vitro membrane protein expression and capture method. Antibody specificity was demonstrated upon identifying, for the first time, membrane-directed heterologous expression of the native sequences of the FopA and FTT1525 virulence determinants from the select agent Francisella tularensis SCHU S4. These approaches will accelerate future structural and functional investigations of therapeutically-relevant membrane proteins.


Strong Light-Matter Coupling between Plasmons in Individual Gold Bi-pyramids and Excitons in Mono- and Multilayer WSe2.
Monolayer transition-metal dichalcogenides (TMDCs) have attracted a lot of research attention recently, motivated by their remarkable optical properties and potential for strong light-matter interactions. Realization of strong plasmon-exciton coupling is especially desirable in this context because it holds promise for the enabling of room-temperature quantum and nonlinear optical applications. These efforts naturally require investigations at a single-nanoantenna level, which, in turn, should possess a compact optical mode interacting with a small amount of excitonic material. However, standard plasmonic nanoantenna designs such as nanoparticle dimers or particle-on-film suffer from misalignment of the local electric field in the gap with the in-plane transition dipole moment of monolayer TMDCs. Here, we circumvent this problem by utilizing gold bi-pyramids (BPs) as very efficient plasmonic nanoantennas. We demonstrate strong coupling between individual BPs and tungsten diselenide (WSe2) monolayers at room temperature. We further study the coupling between multilayers of WSe2 and BPs to elucidate the effect of the number of layers on the coupling strength. Importantly, BPs adopt a reduced-symmetry configuration when deposited on WSe2, such that only one sharp antenna tip efficiently interacts with excitons. Despite the small interaction area, we manage to achieve strong coupling, with Rabi splitting exceeding ∼100 meV. Our results suggest a feasible way toward realizing plasmon-exciton polaritons involving nanoscopic areas of TMDCs, thus pointing toward quantum and nonlinear optics applications at ambient conditions.


Dynamic exchange of myosin VI on endocytic structures.
The actin-based molecular motor myosin VI functions in the endocytic uptake pathway, both during the early stages of clathrin-mediated uptake and in later transport to/from early endosomes. This study uses fluorescence recovery after photobleaching (FRAP) to examine the turnover rate of myosin VI during endocytosis. The results demonstrate that myosin VI turns over dynamically on endocytic structures with a characteristic half-life common to both the large insert isoform of myosin VI on clathrin-coated structures and the no-insert isoform on early endosomes. This half-life is shared by the myosin VI-binding partner Dab2 and is identical for full-length myosin VI and the cargo-binding tail region. The 4-fold slower half-life of an artificially dimerized construct of myosin VI on clathrin-coated structures suggests that wild type myosin VI does not function as a stable dimer, but either as a monomer or in a monomer/dimer equilibrium. Taken together, these FRAP results offer insight into both the basic turnover dynamics and the monomer/dimer nature of myosin VI.


Multisite phosphorylation of voltage-gated sodium channel alpha subunits from rat brain.
Reversible phosphorylation of ion channels underlies cellular plasticity in mammalian neurons. Voltage-gated sodium or Nav channels underlie action potential initiation and propagation, dendritic excitability, and many other aspects of neuronal excitability. Various protein kinases have been suggested to phosphorylate the primary or alpha subunit of Nav channels, affecting diverse aspects of channel function. Previous studies of Nav alpha subunit phosphorylation have led to the identification of a small set of phosphorylation sites important in mediating diverse aspects of Nav channel function. Here we use nanoflow liquid chromatography tandem mass spectrometry (nano-LC MS/MS) on Nav alpha subunits affinity-purified from rat brain with two distinct monoclonal antibodies to identify 15 phosphorylation sites on Nav1.2, 12 of which have not been previously reported. We also found 3 novel phosphorylation sites on Nav1.1. In general, commonly used phosphorylation site prediction algorithms did not accurately predict these novel in vivo phosphorylation sites. Our results demonstrate that specific Nav alpha subunits isolated from rat brain are highly phosphorylated, and suggest extensive modulation of Nav channel activity in mammalian brain. Identification of phosphorylation sites using monoclonal antibody-based immunopurification and mass spectrometry is an effective approach to define the phosphorylation status of Nav channels and other important membrane proteins in mammalian brain.


RBM5 is a male germ cell splicing factor and is required for spermatid differentiation and male fertility.
Alternative splicing of precursor messenger RNA (pre-mRNA) is common in mammalian cells and enables the production of multiple gene products from a single gene, thus increasing transcriptome and proteome diversity. Disturbance of splicing regulation is associated with many human diseases; however, key splicing factors that control tissue-specific alternative splicing remain largely undefined. In an unbiased genetic screen for essential male fertility genes in the mouse, we identified the RNA binding protein RBM5 (RNA binding motif 5) as an essential regulator of haploid male germ cell pre-mRNA splicing and fertility. Mice carrying a missense mutation (R263P) in the second RNA recognition motif (RRM) of RBM5 exhibited spermatid differentiation arrest, germ cell sloughing and apoptosis, which ultimately led to azoospermia (no sperm in the ejaculate) and male sterility. Molecular modelling suggested that the R263P mutation resulted in compromised mRNA binding. Within the adult mouse testis, RBM5 localises to somatic and germ cells including spermatogonia, spermatocytes and round spermatids. Through the use of RNA pull down coupled with microarrays, we identified 11 round spermatid-expressed mRNAs as putative RBM5 targets. Importantly, the R263P mutation affected pre-mRNA splicing and resulted in a shift in the isoform ratios, or the production of novel spliced transcripts, of most targets. Microarray analysis of isolated round spermatids suggests that altered splicing of RBM5 target pre-mRNAs affected expression of genes in several pathways, including those implicated in germ cell adhesion, spermatid head shaping, and acrosome and tail formation. In summary, our findings reveal a critical role for RBM5 as a pre-mRNA splicing regulator in round spermatids and male fertility. Our findings also suggest that the second RRM of RBM5 is pivotal for appropriate pre-mRNA splicing.


The role of dietary patterns and exceptional parental longevity in healthy aging.
Individuals with exceptional longevity and their offspring manifest a lower prevalence of age-related diseases than families without longevity. However, the contribution of dietary habits to protection from disease has not been systematically assessed in families with exceptional longevity. The aim of this study is to compare dietary patterns between individuals with parental longevity and individals without parental longevity. Dietary intake was evaluated using the Block Brief Food Frequency Questionnaire in 234 community dwelling Ashkenazi Jewish adults aged 65 years and older who were participants of the LonGenity study, which enrolls the offspring of parents with exceptional longevity (OPEL) and offspring of parents with usual survival (OPUS). OPEL constituted 38% of the subjects. The two groups had similar daily intake of total calories (1119 vs. 1218 kcal, p = 0.83), grams of cholesterol (141 g vs. 143 g, p = 0.19), and grams of sodium (1324 g vs.1475 g, p = 0.45), in OPEL vs. OPUS respectively. There were also no significant differences in the intake of other macronutrients, micronutrients, nutritional supplements and consumption of various food groups between OPEL and OPUS after adjustment for age and sex. A healthy diet is associated with a lower risk of several chronic diseases. Our study revealed that dietary intake did not differ between OPEL and OPUS; thus, pointing to the role of longevity genes in protecting from disease among individuals with familial longevity. The offspring of long-lived parents do not differ in their dietary patterns compared to individuals without parental longevity.


Role of inflammasome regulation on immune modulators.
Inflammatory responses are essential in eliminating harmful substrates from damaged tissue and inducing recovery. Several cytokines participate in and facilitate this response. Certain cytokines such as interleukin (IL)-1β and IL-18 are initially produced in precursor form in response to toll-like receptor (TLR) ligands and undergo maturation by inflammasomes, which are cytosolic multi-protein complexes containing nucleotide-binding oligomerization domain (NOD)-containing protein 2-like receptors (NLRs). Immune modulators targeting inflammasomes have been investigated to control inflammatory diseases such as metabolic syndrome. However, most immune modulators possessing anti-inflammasome properties attenuate production of other cytokines, which are essential for host defense. In this review, we analyzed the effect of anti-inflammasome agents on the production of cytokines which are not regulated by inflammasome and involving in initial immune responses. As a result, the inflammasome inhibitors are put into three categories: non-effector, stimulator, or inhibitor of cytokine production. Even the stimulator of cytokine production ameliorated symptoms resulting from inflammasome activation in mouse models. Thus, we suggest ideal immune modulators targeting inflammasomes in order to enhance cytokine production while inhibiting cytokine maturation.


Effect of beat frequency on the measured phase of laser-diode heterodyne interferometry.
An optical heterodyne interferometer with a frequency-ramped laser diode has been constructed. The effect of the beat frequency on the measured phase has been theoretically investigated in the frequency domain and experimentally verified. Phase errors caused by the difference between the ramp frequency and the beat frequency alter sinusoidally in accordance with the π periodicity of the interferogram. The error can be eliminated by the electronic calibration technique of the beat frequency.


Evaluation of effectiveness of a computerized notification system for reporting critical values.
Failure to adequately communicate a critical laboratory value is a potential cause of adverse events. Accreditation requirements specify that clinical laboratories must undertake assessments and appropriate measures to improve the timeliness of critical value reporting and prompt receipt by the responsible caregiver. Documentation and communication processes must be regularly monitored and implemented under ongoing systems for quality monitoring. Critical value reporting is an important phase of the clinical laboratory testing process, and notifications of results outside the target time can indicate ineffectiveness of the process. In the present study, we report data obtained in a 12-month period of critical values analysis and describe a computerized communication system conducive to improving the quality of critical value reporting at a university hospital. Automated communication improves the timeliness of notification and avoids the potential errors for which accreditation programs require read-back of the result. The communication also improves the likelihood of reaching the physician on call and may provide important decision support.


Eicosapentaenoic acid regulates brown adipose tissue metabolism in high-fat-fed mice and in clonal brown adipocytes.
Brown adipose tissue (BAT) plays a key role in energy expenditure through its specialized thermogenic function. Therefore, BAT activation may help prevent and/or treat obesity. Interestingly, subcutaneous white adipose tissue (WAT) also has the ability to differentiate into brown-like adipocytes and may potentially contribute to increased thermogenesis. We have previously reported that eicosapentaenoic acid (EPA) reduces high-fat (HF)-diet-induced obesity and insulin resistance in mice. Whether BAT mediates some of these beneficial effects of EPA has not been determined. We hypothesized that EPA activates BAT thermogenic program, contributing to its antiobesity effects. BAT and WAT were harvested from B6 male mice fed HF diets supplemented with or without EPA. HIB 1B clonal brown adipocytes treated with or without EPA were also used. Gene and protein expressions were measured in adipose tissues and H1B 1B cells by quantitative polymerase chain reaction and immunoblotting, respectively. Our results show that BAT from EPA-supplemented mice expressed significantly higher levels of thermogenic genes such as PRDM16 and PGC1α and higher levels of uncoupling protein 1 compared to HF-fed mice. By contrast, both WATs (subcutaneous and visceral) had undetectable levels of these markers with no up regulation by EPA. HIB 1B cells treated with EPA showed significantly higher mRNA expression of PGC1α and SIRT2. EPA treatment significantly increased maximum oxidative and peak glycolytic metabolism in H1B 1B cells. Our results demonstrate a novel and promising role for EPA in preventing obesity via activation of BAT, adding to its known beneficial anti-inflammatory effects.


Membrane ion channels and diabetes.
Type-2, or non-insulin-dependent diabetes mellitus is a serious disease that is now widespread throughout Western society. Glucose intolerance, or failure of glucose to stimulate insulin secretion, is a primary factor in the manifestation of this disease and is likely to be due to the failure of glucose metabolism to stimulate pancreatic beta-cell electrical activity, calcium influx, and insulin secretion. In this review we describe how ion channels regulate the electrical behaviour of the beta-cell and how the membrane potential depolarises in response to a rise in glucose metabolism. Central to these electrical events is the inhibition of ATP-sensitive potassium channel by ATP, and we summarise recent advances in our understanding of the properties of this ion channel in coupling beta-cell metabolism to electrical activity. We discuss the mechanism, specificity, and clinical implications of the pharmacological inhibition of KATP channels by sulphonyureas and other antidiabetic drugs. The roles of other ion channels in regulating electrical activity are considered, and also their potential use as targets for drug action in treating beta-cell disorders.


Assessment of atrial electromechanical interval and P wave dispersion in patients with polycystic ovary syndrome.
Polycystic ovary syndrome (PCOS) is associated with increased cardiovascular risk, including ischemic stroke. Prolonged atrial electromechanical interval (EMI) is related to increased atrial fibrillation (AF) risk. The aim of the study is to evaluate atrial EMI and electrocardiographic P-wave indices related to increased AF risk in patients with PCOS. Forty PCOS patients diagnosed on the basis of the Rotterdam criteria and 20 age-matched controls were prospectively included. patients with atrioventricular or intraventricular conduction abnormalities, dysrhythmia or taking antiarrhythmic drugs, atherosclerotic heart disease, cardiomyopathies, valvular lesions, pericardial disease, a history of pulmonary emboli or pulmonary hypertension, and abnormal thyroid function were excluded. Intra and interatrial EMI were measured by tissue Doppler imaging and P-wave dispersion (Pd) was calculated on 12-lead electrocardiography (ECG). The Isovolumetric relaxation time was the interval between the aortic valve closure artifact at the end of the LV outflow envelope and the mitral valve opening artifact at the beginning of the mitral E wave. Patients with PCOS had significantly higher interatrial [38 (24-65) ms vs. 16 (9-19) ms p<0.001], left-sided intra-atrial (14.8±6.1 vs. 7±1.7 ms, p<0.001), and right-sided intra-atrial (22.3±8.1 vs. 8.6±3.6 ms, p<0.001) EMI compared with the control group. Pd was significantly greater in the PCOS group compared with control group [45 (27-60) ms vs. 30 (26-38) ms, p<0.001]. Echocardiographic parameters of atrial EMI were significantly correlated with body mass index, Pd, and isovolumetric relaxation time in patients with PCOS. PCOS is associated with prolonged inter- and intra-atrial conduction times, which are related to increased AF risk.


Off-hours admission and acute stroke care quality: a nationwide study of performance measures and case-fatality.
Studies have reported higher risks of death and other adverse outcomes in acute stroke patients admitted off-hours; however, little is known about the underlying mechanisms. According to time of admission, our aim was to examine compliance with performance measures for acute stroke care processes, including the effect of a systematic quality improvement program, and to examine 30 days case-fatality. A population-based historical cohort study, including patients admitted to Danish hospitals with a first ever acute stroke (January 1, 2003, to December 31, 2011; N=64 975). Off-hours were weekends and evening and nighttime shifts on weekdays. Compliance with performance measures was compared using general linear modeling, and odds ratios for 30 days case-fatality were obtained using multivariable logistic regression. Patients admitted off-hours had a lower chance of compliance with 8 out of 10 performance measures; however, these differences diminished over time. Unadjusted odds ratio for 30 days case-fatality, for patients admitted off-hours compared with patients admitted on-hours, was 1.15 (95% confidence interval, 1.09-1.21). Adjusting for patient characteristics (in particular, stroke severity) decreased the odds ratio to 1.03 (95% confidence interval, 0.97-1.10). Additional adjustment for hospital characteristics and compliance with performance measures had no effect on the odds ratio. Patients admitted off-hours received a poorer quality of care. However, the admission time-related differences in care were substantially reduced over time, and the differences in 30 days case-fatality appeared primarily to be explained by differences in stroke severity.


Half-time strategies to enhance second-half performance in team-sports players: a review and recommendations.
A number of intermittent team sports require that two consecutive periods of play (lasting for ~30-45 min) are separated by a 10-20 min half-time break. The half-time practices employed by team-sports players generally include returning to the changing rooms, temporarily relaxing from the cognitive and physical demands of the first half, rehydration and re-fuelling strategies, addressing injury or equipment concerns, and receiving tactical instruction and coach feedback. However, the typically passive nature of these actions has been associated with physiological changes that impair performance during the second half. Both physical and cognitive performances have been found to decline in the initial stages of subsequent exercise that follows half-time. An increased risk of injury has also been observed during this period. Therefore, half-time provides sports scientists and strength and conditioning coaches with an opportunity to optimise second-half performance. An overview of strategies thought to benefit team-sports athletes is presented; specifically, the efficacy of heat maintenance strategies (including passive and active methods), post-activation potentiation, hormonal priming, and modified hydro-nutritional practices are discussed. A theoretical model of applying these strategies in a manner that compliments current practice is also offered.


A Systematic Review of Bright Light Therapy for Eating Disorders.
Bright light therapy is a noninvasive biological intervention for disorders with nonnormative circadian features. Eating disorders, particularly those with binge-eating and night-eating features, have documented nonnormative circadian eating and mood patterns, suggesting that bright light therapy may be an efficacious stand-alone or adjunctive intervention. The purpose of this systematic literature review, using PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines, was (1) to evaluate the state of the empirical treatment outcome literature on bright light therapy for eating disorders and (2) to explore the timing of eating behavior, mood, and sleep-related symptom change so as to understand potential mechanisms of bright light therapy action in the context of eating disorder treatment. A comprehensive literature search using PsycInfo and PubMed/MEDLINE was conducted in April 2016 with no date restrictions to identify studies published using bright light therapy as a treatment for eating disorders. Keywords included combinations of terms describing disordered eating (eating disorder, anorexia nervosa, bulimia nervosa, binge eating, binge, eating behavior, eating, and night eating) and the use of bright light therapy (bright light therapy, light therapy, phototherapy). After excluding duplicates, 34 articles were reviewed for inclusion. 14 published studies of bright light therapy for eating disorders met inclusion criteria (included participants with an eating disorder/disordered-eating behaviors; presented as a case study, case series, open-label clinical trial, or randomized/nonrandomized controlled trial; written in English; and published and available by the time of manuscript review). Results suggest that bright light therapy is potentially effective at improving both disordered-eating behavior and mood acutely, although the timing of symptom response and the duration of treatment effects remain unknown. Future research should systematically control for placebo response, assess symptom change frequently and across a broad range of systems, and evaluate the longer-term efficacy of bright light therapy for eating disorders.


Physical activity and lipidomics in a population at high risk of type 2 diabetes mellitus.
The aim was to investigate how measurements of the lipidome differ according to the level and intensity of physical activity in a population at high risk of type 2 diabetes (T2DM). A targeted metabolomics platform provided quantitative molecular data on lipid species. Linear regression examined the associations between plasma lipid concentrations, particle size and time spent in objectively measured physical activity intensity domains, in increments of 500 counts per minute (cpm) (up to >4500 cpm (~>5.6METs)). Results are presented as % difference in the concentration (lower/higher) or particle size (smaller/larger) per 10 min of activity within each intensity. Five hundred and nine participants were included. Time spent in the lowest physical activity intensity domain (<500 cpm) was unfavourably associated with VLDL (2%), HDL (-2%) and Apolipoprotein A-1 particle concentrations (-2%) and HDL diameter (-2%). Conversely, time spent in intensities ≥1000 cpm were favourably associated with HDL subclass concentrations; with stronger associations seen at moderate intensities (2000-3999 cpm (~4.5METs)). For Apolipoprotein-B concentration and VLDL particle concentration and size, a negative association was consistently observed at the highest physical activity intensity only. If these associations are causal, HDL subclasses appear sensitive to light-intensities whereas only the high category of physical activity intensity was consistently associated with VLDL subclasses.


Mechanisms of smokeless tobacco-induced oral mucosa inflammation: role of bradykinin.
The purpose of this study was to determine whether an aqueous extract of smokeless tobacco (moist snuff) increases clearance of macromolecules from postcapillary venules in the in situ oral mucosa and, if so, whether bradykinin mediated this response. Using intravital microscopy, we found that 20-min suffusion of the extract elicited significant concentration-dependent leaky site formation and increase in clearance of FITC-dextran (molecular mass, 70 kDa) from the hamster cheek pouch (p < 0.05). These responses were associated with a significant increase in bradykinin-like immunoreactivity in the suffusate. Smokeless tobacco extract-induced leaky site formation and increase in clearance of FITC-dextran were significantly attenuated by NPC 17647 and Hoe 140 (p < 0.05), two bradykinin B2 receptor antagonists, but not by desArg9,[Leu8]bradykinin, a bradykinin B1 receptor antagonist. Both bradykinin B2 receptor antagonists had no significant effects on adenosine-induced responses. Indomethacin had no significant effects on smokeless tobacco extract-induced responses. Exposure to smokeless tobacco extract was associated with a significant decrease in angiotensin I-converting enzyme activity and a small, but significant, increase in neutral endopeptidase 24.11 activity in the cheek pouch, two peptidases widely distributed in the microcirculation that cleave and inactivate bradykinin (p < 0.05). Overall, these data suggest that smokeless tobacco elicits plasma exudation in the oral mucosa in vivo in a specific fashion, and that this response is mediated by bradykinin.


Human endogenous retrovirus type W envelope protein inhibits oligodendroglial precursor cell differentiation.
Differentiation of oligodendroglial precursor cells is crucial for central nervous system remyelination and is influenced by both extrinsic and intrinsic factors. Recent studies showed that human endogenous retrovirus type W (HERV-W) contributes significantly to brain damage. In particular, its envelope protein ENV can mediate injury to specific cell types of the brain and immune system. Here, we investigated whether ENV protein affects oligodendroglial differentiation. Immunostaining and gene expression analyses were performed to establish the expression and regulation of the known ENV receptor, Toll-like receptor 4 (TLR4), on oligodendroglial precursor cells in human brain tissue and in culture. Cultured primary oligodendroglial precursor cells were stimulated with ENV protein to determine the effects of this ligand/receptor interaction. We demonstrated that the ENV protein is present in close proximity to TLR4-expressing oligodendroglial precursor cells adjacent to multiple sclerosis lesions. Human and rat oligodendroglial precursor cells expressed TLR4, and the ENV-mediated activation of TLR4 led to the induction of proinflammatory cytokines and inducible nitric oxide synthase as well as the formation of nitrotyrosine groups and a subsequent reduction in myelin protein expression. Our findings suggest that ENV-mediated induction of nitrosative stress via activation of TLR4 results in an overall reduction of the oligodendroglial differentiation capacity, thereby contributing to remyelination failure. Therefore, pharmacological or antibody-mediated inhibition of ENV may prevent the blockade of myelin repair in the diseased or injured central nervous system.


Washing with contaminated bar soap is unlikely to transfer bacteria.
Recent reports of the isolation of microorganisms from used soap bars have raised the concern that bacteria may be transferred from contaminated soap bars during handwashing. Since only one study addressing this question has been published, we developed an additional procedure to test this concern. In our new method prewashed and softened commercial deodorant soap bars (0.8% triclocarban) not active against Gram-negative bacteria were inoculated with Escherichia coli and Pseudomonas aeruginosa to give mean total survival levels of 4.4 X 10(5) c.f.u. per bar which was 70-fold higher than those reported on used soap bars. Sixteen panelists were instructed to wash with the inoculated bars using their normal handwashing procedure. After washing, none of the 16 panelists had detectable levels of either test bacterium on their hands. Thus, the results obtained using our new method were in complete agreement with those obtained with the previously published method even though the two methods differ in a number of procedural aspects. These findings, along with other published reports, show that little hazard exists in routine handwashing with previously used soap bars and support the frequent use of soap and water for handwashing to prevent the spread of disease.


Coffee and its waste repel gravid Aedes albopictus females and inhibit the development of their embryos.
Dengue is a prevalent arboviral disease and the development of insecticide resistance among its vectors impedes endeavors to control it. Coffee is drunk by millions of people daily worldwide, which is associated with the discarding of large amounts of waste. Coffee and its waste contain large amounts of chemicals many of which are highly toxic and none of which have a history of resistance in mosquitoes. Once in solution, coffee is brownish in colour, resembling leaf infusion, which is highly attractive to gravid mosquitoes. To anticipate the environmental issues related to the increasing popularity of coffee as a drink, and also to combat insecticide resistance, we explored the deterrence potentials of coffee leachates against the ovipositing and embryonic stages of the dengue vector, Aedes albopictus. In a series of choice, no-choice, and embryo toxicity bioassays, we examined changes in the ovipositional behaviours and larval eclosion of Ae. albopictus in response to coffee extracts at different concentrations. Oviposition responses were extremely low when ovicups holding highly concentrated extract (HCE) of coffee were the only oviposition sites. Gravid females retained increased numbers of mature eggs until 5 days post-blood feeding. When provided an opportunity to oviposit in cups containing coffee extracts and with water, egg deposition occurred at lower rates in those containing coffee, and HCE cups were far less attractive to females than those containing water only. Females that successfully developed in a coffee environment preferentially oviposited in such cups when in competition with preferred oviposition sites (water cups), but this trait did not continue into the fourth generation. Larval eclosion occurred at lower rates among eggs that matured in a coffee environment, especially among those that were maintained on HCE-moistened substrates. The observations of the present study indicate a pronounced vulnerability of Ae. albopictus to the presence of coffee in its habitats during the early phases of its life cycle. The observations that coffee repels gravid females and inhibits larval eclosion provide novel possibilities in the search for novel oviposition deterrents and anti-larval eclosion agents against dengue vectors.


Trends in smoking among adults with mental illness and association between mental health treatment and smoking cessation.
Significant progress has been made in reducing the prevalence of tobacco use in the United States. However, tobacco cessation efforts have focused on the general population rather than individuals with mental illness, who demonstrate greater rates of tobacco use and nicotine dependence. To assess whether declines in tobacco use have been realized among individuals with mental illness and examine the association between mental health treatment and smoking cessation. Use of nationally representative surveys of noninstitutionalized US residents to compare trends in smoking rates between adults with and without mental illness and across multiple disorders (2004-2011 Medical Expenditure Panel Survey [MEPS]) and to compare rates of smoking cessation among adults with mental illness who did and did not receive mental health treatment (2009-2011 National Survey of Drug Use and Health [NSDUH]).The MEPS sample included 32,156 respondents with mental illness (operationalized as reporting severe psychological distress, probable depression, or receiving treatment for mental illness) and 133,113 without mental illness. The NSDUH sample included 14,057 lifetime smokers with mental illness. Current smoking status (primary analysis; MEPS sample) and smoking cessation, operationalized as a lifetime smoker who did not smoke in the last 30 days (secondary analysis; NSDUH sample). Adjusted smoking rates declined significantly among individuals without mental illness (19.2% [95% CI, 18.7-19.7%] to 16.5% [95% CI, 16.0%-17.0%]; P < .001) but changed only slightly among those with mental illness (25.3% [95% CI, 24.2%-26.3%] to 24.9% [95% CI, 23.8%- 26.0%]; P = .50), a significant difference in difference of 2.3% (95% CI, 0.7%-3.9%) (P = .005). Individuals with mental illness who received mental health treatment within the previous year were more likely to have quit smoking (37.2% [95% CI, 35.1%-39.4%]) than those not receiving treatment (33.1% [95% CI, 31.5%-34.7%]) (P = .005). Between 2004 and 2011, the decline in smoking among individuals with mental illness was significantly less than among those without mental illness, although quit rates were greater among those receiving mental health treatment. This suggests that tobacco control policies and cessation interventions targeting the general population have not worked as effectively for persons with mental illness.


D-tryptophan from probiotic bacteria influences the gut microbiome and allergic airway disease.
Chronic immune diseases, such as asthma, are highly prevalent. Currently available pharmaceuticals improve symptoms but cannot cure the disease. This prompted demands for alternatives to pharmaceuticals, such as probiotics, for the prevention of allergic disease. However, clinical trials have produced inconsistent results. This is at least partly explained by the highly complex crosstalk among probiotic bacteria, the host's microbiota, and immune cells. The identification of a bioactive substance from probiotic bacteria could circumvent this difficulty. We sought to identify and characterize a bioactive probiotic metabolite for potential prevention of allergic airway disease. Probiotic supernatants were screened for their ability to concordantly decrease the constitutive CCL17 secretion of a human Hodgkin lymphoma cell line and prevent upregulation of costimulatory molecules of LPS-stimulated human dendritic cells. Supernatants from 13 of 37 tested probiotic strains showed immunoactivity. Bioassay-guided chromatographic fractionation of 2 supernatants according to polarity, followed by total ion chromatography and mass spectrometry, yielded C11H12N2O2 as the molecular formula of a bioactive substance. Proton nuclear magnetic resonance and enantiomeric separation identified D-tryptophan. In contrast, L-tryptophan and 11 other D-amino acids were inactive. Feeding D-tryptophan to mice before experimental asthma induction increased numbers of lung and gut regulatory T cells, decreased lung TH2 responses, and ameliorated allergic airway inflammation and hyperresponsiveness. Allergic airway inflammation reduced gut microbial diversity, which was increased by D-tryptophan. D-tryptophan is a newly identified product from probiotic bacteria. Our findings support the concept that defined bacterial products can be exploited in novel preventative strategies for chronic immune diseases.


A tracer methodology for identifying ambient flows in boreholes.
Identifying flows into, out of, and across boreholes is important for characterizing aquifers, determining the depth at which water enters boreholes, and determining the locations and rates of outflow. This study demonstrates how Single Borehole Dilution Tests (SBDTs) carried out under natural head conditions provide a simple and cheap method of identifying vertical flow within boreholes and determining the location of in-flowing, out-flowing, and cross-flowing fractures. Computer simulations were used to investigate the patterns in tracer profiles that arise from different combinations of flows. Field tracer tests were carried out using emplacements of a saline tracer throughout the saturated length of boreholes and also point emplacements at specific horizons. Results demonstrated that SBDTs can be used to identify flowing fractures at the top and bottom of sections of vertical flow, where there is a change in vertical flow rate within a borehole, and also where there are consistent decreases in tracer concentration at a particular depth. The technique enables identification of fractures that might be undetected by temperature and electrical conductance logging, and is a simple field test that can be carried out without pumping the borehole.


Chronic Diarrhea in Dogs - Retrospective Study in 136 Cases.
Chronic diarrhea (CD) is common in dogs, and information on frequency and distribution of primary and secondary causes is lacking. To evaluate underlying causes and predictors of outcome in dogs with CD. One hundred and thirty-six client-owned dogs with CD (≥3 weeks duration). Retrospective review of medical records (Small Animal Clinic, Freie Universität Berlin, Germany, 09/2009-07/2011). Quantification of final diagnoses and comparison of clinical aspects including disease severity and clinicopathological abnormalities among dogs with clinical remission (either complete [gastrointestinal signs absent] or partial [clinical improvement of gastrointestinal signs and reduced episodes with shortened duration]), and those without recovery. Ninety percent of dogs were diagnosed with a primary enteropathy: inflammatory (71%; of those 66% dietary responsive, 23% idiopathic, 11% antibiotic responsive), infectious (13%), neoplastic (4%), and in one dog each mechanical disease or systemic vasculitis. Secondary causes were diagnosed in 10% of dogs: exocrine pancreatic (6%), endocrine (2%), and in one dog each hepatic, renal, and cardiac disease. In total, 87% of dogs had clinical remission, whereas 13% died or did not respond to treatment: Lack of recovery was frequently recorded for dogs with primary inflammatory (idiopathic) or neoplastic disease and was significantly associated with increased disease severity scores (P = .005), anemia (hematocrit < 40%, P < .001), severe hypoalbuminemia (serum albumin <2.0 g/dL, P = .008), and severe hypocobalaminemia (serum cobalamin concentration <200 pg/mL, P = .006). Inflammatory enteropathies and particularly those of dietary origin were the most common causes of CD in dogs. Findings support the usefulness of hematocrit, and serum albumin and cobalamin concentration as prognostic markers in dogs with CD.


Properties and photochemistry of valence-induced-Ti(3+) enriched (Nb,N)-codoped anatase TiO2 semiconductors.
Nb and N codoped TiO2s are outstandingly versatile semiconductor oxides. Their high conductivity makes them valid alternatives to commercially available, but very expensive, conductive oxides. They show increased photonic efficiencies compared to the cases of solely Nb or N doped TiO2, when used as visible light sensitised photocatalysts. Furthermore, they are excellent materials for O2 sensors at very low temperature. Despite these remarkable properties, a clear picture of the electronic and optical mechanisms induced by the simultaneous presence of the dopants has just begun to be understood. Using a combination of electron paramagnetic resonance (EPR) spectroscopy, electrochemical impedance spectroscopy (EIS) and optical spectroscopy, we present here novel fundamental insights into the mechanisms responsible for the enhanced conductivity and visible light photochemistry.


Methyl (E)-2-[(3-chloro-4-cyano-phenyl)-imino]-4-(4-chloro-phen-yl)-6-methyl-1,2,3,4-tetra-hydro-pyrimidine-5-carboxyl-ate.
In the title compound, C(20)H(16)Cl(2)N(4)O(2), the dihedral angles between the planes of the chloro-phenyl, chloro-cyano-phenyl-imine and ester groups and the plane of the six-membered tetra-hydro-pyrimidine ring are 86.9 (2), 72.6 (2) and 7.9 (2)°, respectively. The Cl atom substituent on the cyano-phenyl ring is disordered over two rotationally related sites [occupancy factors 0.887 (2):0.113 (2)], while the mol-ecular conformation is stabilized by the presence of an intra-molecular aromatic C-H⋯π inter-action. Both N-H groups participate in separate inter-molecular hydrogen-bonding associations with centrosymmetric cyclic motifs [graph sets R(2) (2)(8) and R(2) (2)(12)], resulting in ribbons parallel to [010]. Further weak C-H⋯O hydrogen bonds link these ribbons into a two-dimensional mol-ecular assembly.


Pubertal Suppression for Transgender Youth and Risk of Suicidal Ideation.
Gonadotropin-releasing hormone analogues are commonly prescribed to suppress endogenous puberty for transgender adolescents. There are limited data regarding the mental health benefits of this treatment. Our objective for this study was to examine associations between access to pubertal suppression during adolescence and adult mental health outcomes. Using a cross-sectional survey of 20 619 transgender adults aged 18 to 36 years, we examined self-reported history of pubertal suppression during adolescence. Using multivariable logistic regression, we examined associations between access to pubertal suppression and adult mental health outcomes, including multiple measures of suicidality. Of the sample, 16.9% reported that they ever wanted pubertal suppression as part of their gender-related care. Their mean age was 23.4 years, and 45.2% were assigned male sex at birth. Of them, 2.5% received pubertal suppression. After adjustment for demographic variables and level of family support for gender identity, those who received treatment with pubertal suppression, when compared with those who wanted pubertal suppression but did not receive it, had lower odds of lifetime suicidal ideation (adjusted odds ratio = 0.3; 95% confidence interval = 0.2-0.6). This is the first study in which associations between access to pubertal suppression and suicidality are examined. There is a significant inverse association between treatment with pubertal suppression during adolescence and lifetime suicidal ideation among transgender adults who ever wanted this treatment. These results align with past literature, suggesting that pubertal suppression for transgender adolescents who want this treatment is associated with favorable mental health outcomes.


Self-cytoplasmic DNA upregulates the mutator enzyme APOBEC3A leading to chromosomal DNA damage.
Foreign and self-cytoplasmic DNA are recognized by numerous DNA sensor molecules leading to the production of type I interferons. Such DNA agonists should be degraded otherwise cells would be chronically stressed. Most human APOBEC3 cytidine deaminases can initiate catabolism of cytoplasmic mitochondrial DNA. Using the human myeloid cell line THP-1 with an interferon inducible APOBEC3A gene, we show that cytoplasmic DNA triggers interferon α and β production through the RNA polymerase III transcription/RIG-I pathway leading to massive upregulation of APOBEC3A. By catalyzing C→U editing in single stranded DNA fragments, the enzyme prevents them from re-annealing so attenuating the danger signal. The price to pay is chromosomal DNA damage in the form of CG→TA mutations and double stranded DNA breaks which, in the context of chronic inflammation, could drive cells down the path toward cancer.


Lumbar epidural steroid injections: indications, contraindications, risks, and benefits.
The clinical use of lumbar epidural steroid injections has increased dramatically. Although there are certainly beneficial effects to using epidural steroid injections in a treatment regimen for lumbar radicular pain, there is a lack of well designed, placebo-controlled studies to define conclusively specific indications and techniques for different spinal diagnoses. This article reviews the pathophysiology of lumbar radiculopathy and the use of epidural steroid injections as one treatment option, as well as describes their risks and benefits. Based on current literature, we offer an evidence-based perspective regarding rational use of lumbar epidural steroid injections for certain indications and treatment goals.


A randomised controlled trial of the Neuro Emotional Technique (NET) for childhood Attention Deficit Hyperactivity Disorder (ADHD): a protocol.
An abundance of literature is dedicated to research for the treatment of Attention Deficit Hyperactivity Disorder (ADHD). Most, is in the area of pharmacological therapies with less emphasis in psychotherapy and psychosocial interventions and even less in the area of complementary and alternative medicine (CAM).The use of CAM has increased over the years, especially for developmental and behavioral disorders, such as ADHD. 60-65% of parents with children with ADHD have used CAM. Medical evidence supports a multidisciplinary approach (i.e. pharmacological and psychosocial) for the best clinical outcomes. The Neuro Emotional Technique (NET), a branch of Chiropractic, was designed to address the biopsychosocial aspects of acute and chronic conditions including non-musculoskeletal conditions. Anecdotally, it has been suggested that ADHD may be managed effectively by NET. A placebo controlled, double blind randomised clinical trial was designed to assess the effectiveness of NET on a cohort of children with medically diagnosed ADHD. Children aged 5-12 years who met the inclusion criteria were randomised to one of three groups. The control group continued on their existing medical regimen and the intervention and placebo groups had the addition of the NET and sham NET protocols added to their regimen respectively. These two groups attended a clinical facility twice a week for the first month and then once a month for six months. The Conners' Parent and Teacher Rating Scales (CRS) were used at the start of the study to establish baseline data and then in one month and in seven months time, at the conclusion of the study. The primary outcome measures chosen were the Conners' ADHD Index and Conners' Global Index. The secondary outcome measures chosen were the DSM-IV: Inattentive, the DSM-IV:Hyperactive-Impulsive, and the DSM-IV:Total subscales from the Conners' Rating Scales, monitoring changes in inattention, hyperactivity and impulsivity. Calculations for the sample size were set with a significance level of 0.05 and the power of 80%, yielding a sample size of 93. The present study should provide information as to whether the addition of NET to an existing medical regimen can improve outcomes for children with ADHD. Australian New Zealand Clinical Trial Registration Number: ANZCTRN 012606000332527.


A realistic pelvic phantom for electrical impedance measurement.
To design and fabricate an anatomically and conductively accurate phantom for electrical impedance studies of non-invasive bladder volume monitoring. A modular pelvic phantom was designed and fabricated, consisting of a mechanically and conductively stable boundary wall, a background medium, and bladder phantoms. The wall and bladders are made of conductive polyurethane. The background material is an ultrasound gel-based mixture, with conductivity matched to a weighted average of the pelvic cavity organs, bone, muscle and fat. The phantom boundary is developed using a computer tomography model of a male human pelvis. The bladder phantoms were designed to correlate with human bladder dimensions. Electrical impedance measurements of the phantom were recorded, and images produced using six different bladder phantoms and a realistic finite element model. Five different bladder volumes were successfully imaged using an empty bladder as a reference. The average conductivity index from the reconstructed images showed a strong positive correlation with the bladder phantom volumes. A conductively and anatomically accurate pelvic phantom was developed for non-invasive bladder volume monitoring using electrical impedance measurements. Several bladders were designed to correlate with actual human bladder volumes, allowing for accurate volume estimation. The conductivity of the phantom is accurate over 50-250 kHz. This phantom can allow changeable electrode location, contact and size; multi-layer electrodes configurations; increased complexity by addition of other organ or bone phantoms; and electrode movement and deformation. Overall, the pelvic phantom enables greater scope for experimentation and system refinement as a precursor to in-man clinical studies.


Tooth discoloration induced by endodontic materials: a laboratory study.
To investigate the discoloration potential of endodontic materials using a bovine tooth model. Two hundred and 10 dentine-enamel cuboid blocks (10 × 10 × 3.5 mm) were prepared out of the middle thirds of bovine tooth crowns. Standardized cavities were prepared in the walls of the pulp chamber leaving 2 mm of enamel and dentine on the labial wall of the crown. The specimens were randomly assigned to 14 groups (n = 15). Endodontic materials were placed into the cavities as follows: group A: empty, group B: blood, group C: calcium hydroxide, group D: ApexCal, group E: Ultracal XS, group F: Ledermix, group G: triple antibiotic paste (3Mix), group H: grey MTA(GMTA), group I: GMTA + blood, group J: white MTA (WMTA), group K: WMTA + blood, group L: Portland cement (PC), group M: PC + blood and group N: AH Plus. The cavities were sealed with composite and stored in water. Standardized colour measurement (VITA Easyshade compact) was performed at the following intervals: prior to (T0) and after placement of the filling (T1), 1 week (T2), 1 month (T3), 3 months (T4), 6 months (T5) and after 1 year (T6). Colour change (ΔE) values were calculated. A two-way analysis of variance was used to assess significant differences between the endodontic materials. The mean values of all groups were compared using the Tukey multiple comparison test (α = 0.05). Significant differences were detected amongst the experimental groups after 12 months (P < 0.0001). The lowest colour change values were observed in the groups N (AH Plus, 3.2 ± 1.5), A (empty, 3.8 ± 1.4), L (PC, 4.1 ± 1.7), C (calcium hydroxide, 4.7 ± 1.5), E (Ultracal XS, 5.1 ± 1.9) and J (WMTA, 7.9 ± 6.7). The most discoloration was measured in groups G (3Mix, 66.2 ± 9.9) and F (Ledermix, 46.2 ± 11.6). PC showed the best colour stability amongst the Portland cement-based materials; however, when contaminated with blood (group M), a significantly higher ΔE value (13.6 ± 4.2) was detected (P = 0.032). Materials used in endodontics may stain teeth. Therefore, the choice of material should not rely solely on biological and functional criteria, but also take aesthetic considerations into account.


Antidepressant mechanism of ketamine: perspective from preclinical studies.
A debilitating mental disorder, major depressive disorder is a leading cause of global disease burden. Existing antidepressant drugs are not adequate for the majority of depressed patients, and large clinical studies have demonstrated their limited efficacy and slow response onset. Growing evidence of low-dose ketamine's rapid and potent antidepressant effects offers strong potential for future antidepressant agents. However, ketamine has considerable drawbacks such as its abuse potential, psychomimetic effects, and increased oxidative stress in the brain, thus limiting its widespread clinical use. To develop superior antidepressant drugs, it is crucial to better understand ketamine's antidepressant mechanism of action. Recent preclinical studies indicate that ketamine's antidepressant mechanism involves mammalian target of rapamycin pathway activation and subsequent synaptogenesis in the prefrontal cortex, as well as glycogen synthase kinase-3 beta (GSK-3β) inactivation. Adjunct GSK-3β inhibitors, such as lithium, can enhance ketamine's efficacy by augmenting and prolonging its antidepressant effects. Given the potential for depressive relapses, lithium in addition to ketamine is a promising solution for this clinical issue.


Conversion to resectability using transcatheter arterial chemoembolization alternating with mFOLFOX6 in patients with colorectal liver metastases.
Colorectal cancer is one of the most common malignancies in the world, and about 25% of colorectal cancer patients present with colorectal cancer liver metastases (CRCLM) even at new diagnosis. The study was to evaluate the safety and efficacy of transcatheter arterial chemoembolization (TACE) alternating with mFOLFOX6 in Chinese patients with unresectable CRCLM. In this study, by combining the systemic and regional treatment, the resectability rate, overall survival, and progression-free survival were measured with addition of TACE. Included patients had Eastern Cooperative Oncology Group performance status 0-2. Sixty-two patients received mFOLFOX6 plus one TACE after 2 weeks of chemotherapy; after 2 weeks, the next periodical treatment repeated. Patients received operation when the liver metastases were converted to resectability or severe tumor-associated complications occurred. We found that 28 patients (45.2%) patients received operation after the treatment of TACE combined with systemic chemotherapy. The median time from initial treatment to the operation was 6 months. The median follow-up period was 41 months in all the patients. The 3-year survival rate of resected patients and unresected patients was 54% and 17%, respectively. Post-TACE syndrome was the major adverse reaction (81%). Other adverse reactions were neutropenia, nausea, and neurotoxicity. No patient died of the adverse reactions. The resection rate was related to hepatic segments and vasculature involvement. Taken together, TACE alternating with mFOLFOX6 has been proved to be safe and effective for CRCLM treatment to improve resection rate and prolong the survival time.


Virulence of non-type 1-fimbriated and nonfimbriated nonflagellated Salmonella typhimurium mutants in murine typhoid fever.
The virulence of Salmonella typhimurium mutants that were unable to synthesize type 1 fimbriae was tested in a murine typhoid fever model. Nonfimbriated mutants (fim) exhibited a lower 50% lethal dose than a wild-type (fim+) strain and produced significantly higher mortality (fim, 55%; fim+, 37% [P less than 0.002]) in mice that were challenged orally. There was no difference in virulence when the wild-type and mutant strains were injected intraperitoneally into mice. The progress of a short-term lethal infection was monitored after oral inoculation of mice with a mixture containing equivalent numbers of fim+ wild-type and fim mutant bacteria. The results indicated that while both strains colonized the intestinal tract equally well and invaded internal organs, the S. typhimurium fim mutant proliferated in the blood of the mice faster than the fim+ strain. The results of the mixed oral challenge suggested that bacteremia caused by fim+ S. typhimurium was reduced or delayed by the sequestration of the fimbriated bacteria in the spleen, liver, and kidneys. Thus, type 1 fimbriae were not virulence factors for S. typhimurium in this model, and the fimbriae may be an impediment to the pathogen in this setting. An S. typhimurium double mutant lacking type 1 fimbriae and flagella (fla) also was tested in mice. The virulence of the fim fla mutant was greatly reduced compared with that of the wild-type strain (mortality from fim fla challenge, 11% [P less than 0.0005]). The significance of this latter result is discussed in relation to host adaptation by pathogenic salmonellae.


The right posterior paravermis and the control of language interference.
Auditory and written language in humans' comprehension necessitates attention to the message of interest and suppression of interference from distracting sources. Investigating the brain areas associated with the control of interference is challenging because it is inevitable that activation of the brain regions that control interference co-occurs with activation related to interference per se. To isolate the mechanisms that control verbal interference, we used a combination of structural and functional imaging techniques in Italian and German participants who spoke English as a second language. First, we searched structural MRI images of Italian participants for brain regions in which brain structure correlated with the ability to suppress interference from the unattended dominant language (Italian) while processing heard sentences in their weaker language (English). This revealed an area in the posterior paravermis of the right cerebellum in which gray matter density was higher in individuals who were better at controlling verbal interference. Second, we found functional activation in the same region when our German participants made semantic decisions on written English words in the presence of interference from unrelated words in their dominant language (German). This combination of structural and functional imaging therefore highlights the contribution of the right posterior paravermis to the control of verbal interference. We suggest that the importance of this region for language processing has previously been missed because most fMRI studies limit the field of view to increase sensitivity, with the lower part of the cerebellum being the region most likely to be excluded.


Effectiveness of land-based physiotherapy exercise following hospital discharge following hip arthroplasty for osteoarthritis: an updated systematic review.
Existing review required updating. To evaluate the effectiveness of physiotherapy exercise after discharge from hospital on function, walking, range of motion, quality of life and muscle strength, for patients following elective primary total hip arthroplasty for osteoarthritis. Systematic review from January 2007 to November 2013. AMED, CINAHL, EMBASE, MEDLINE, Kingsfund Database, and PEDro. Cochrane CENTRAL, BioMed Central (BMC), The Department of Health National Research Register and Clinical Trials.gov register. Searches were overseen by a librarian. Authors were contacted for missing information. No language restrictions were applied. Trials comparing physiotherapy exercise vs usual/standard care, or comparing two types of relevant exercise physiotherapy, following discharge from hospital after elective primary total hip replacement for osteoarthritis were reviewed. Functional activities of daily living, walking, quality of life, muscle strength and joint range of motion. Quality and risk of bias for studies were evaluated. Data were extracted and meta-analyses considered. 11 trials are included in the review. Trial quality was mixed. Newly included studies were assessed as having lower risk of bias than previous studies. Narrative review indicates that physiotherapy exercise after discharge following total hip replacement may potentially benefit patients in terms of function, walking and muscle strengthening. The overall quality and quantity of trials, and their diversity, prevented meta-analyses. Disappointingly, insufficient evidence still prevents the effectiveness of physiotherapy exercise following discharge to be determined for this patient group. High quality, adequately powered, trials with long term follow up are required.


No effect of insecticide treated curtain deployment on aedes infestation in a cluster randomized trial in a setting of low dengue transmission in Guantanamo, Cuba.
The current study evaluated the effectiveness and cost-effectiveness of Insecticide Treated Curtain (ITC) deployment for reducing dengue vector infestation levels in the Cuban context with intensive routine control activities. A cluster randomized controlled trial took place in Guantanamo city, east Cuba. Twelve neighborhoods (about 500 households each) were selected among the ones with the highest Aedes infestation levels in the previous two years, and were randomly allocated to the intervention and control arms. Long lasting ITC (PermaNet) were distributed in the intervention clusters in March 2009. Routine control activities were continued in the whole study area. In both study arms, we monitored monthly pre- and post-intervention House Index (HI, number of houses with at least 1 container with Aedes immature stages/100 houses inspected), during 12 and 18 months respectively. We evaluated the effect of ITC deployment on HI by fitting a generalized linear regression model with a negative binomial link function to these data. At distribution, the ITC coverage (% of households using ≥1 ITC) reached 98.4%, with a median of 3 ITC distributed/household. After 18 months, the coverage remained 97.4%. The local Aedes species was susceptible to deltamethrin (mosquito mortality rate of 99.7%) and the residual deltamethrin activity in the ITC was within acceptable levels (mosquito mortality rate of 73.1%) after one year of curtain use. Over the 18 month observation period after ITC distribution, the adjusted HI rate ratio, intervention versus control clusters, was 1.15 (95% CI 0.57 to 2.34). The annualized cost per household of ITC implementation was 3.8 USD, against 16.8 USD for all routine ACP activities. Deployment of ITC in a setting with already intensive routine Aedes control actions does not lead to reductions in Aedes infestation levels.


Genetic variation in gonadal impairment in female survivors of childhood cancer: a PanCareLIFE study protocol.
Improved risk stratification, more effective therapy and better supportive care have resulted in survival rates after childhood cancer of around 80% in developed countries. Treatment however can be harsh, and three in every four childhood cancer survivors (CCS) develop at least one late effect, such as gonadal impairment. Gonadal impairment can cause involuntary childlessness, with serious consequences for the well-being of CCS. In addition, early menopause increases the risk of comorbidities such as cardiovascular disease and osteoporosis. Inter-individual variability in susceptibility to therapy related gonadal impairment suggests a role for genetic variation. Currently, only one candidate gene study investigated genetic determinants in relation to gonadal impairment in female CCS; it yielded one single nucleotide polymorphism (SNP) that was previously linked with the predicted age at menopause in the general population of women, now associated with gonadal impairment in CCS. Additionally, one genome wide association study (GWAS) evaluated an association with premature menopause, but no GWAS has been performed using endocrine measurements for gonadal impairment as the primary outcome in CCS. As part of the PanCareLIFE study, the genetic variability of chemotherapy induced gonadal impairment among CCS will be addressed. Gonadal impairment will be determined by anti-Müllerian hormone (AMH) levels or alternatively by fertility and reproductive medical history retrieved by questionnaire. Clinical and genetic data from 837 non-brain or non-bilateral gonadal irradiated long-term CCS will result in the largest clinical European cohort assembled for this late-effect study to date. A candidate gene study will examine SNPs that have already been associated with age at natural menopause and DNA maintenance in the general population. In addition, a GWAS will be performed to identify novel allelic variants. The results will be validated in an independent CCS cohort. This international collaboration aims to enhance knowledge of genetic variation which may be included in risk prediction models for gonadal impairment in CCS.


Forced expression of the motor neuron determinant HB9 in neural stem cells affects neurogenesis.
In contrast to mouse embryonic stem cells and in spite of overlapping gene expression profiles, neural stem cells (NSCs) isolated from the embryonic spinal cord do not respond to physiological morphogenetic stimuli provided by Sonic hedgehog and retinoic acid and do not generate motor neurons upon differentiation. Transcription factors expressed in motor neuron progenitors during embryogenesis include Pax6, Ngn2, Nkx6.1 and Olig2, whose expression precedes that of factors specifying motor neuron fate, including HB9, Islet1 and LIM3. We showed that all these factors were present in neural progenitors derived from mouse ES cells, whereas NSCs derived from the rat embryonic spinal cord expressed neither HB9 nor Islet1 and contained low levels of Nkx6.1 and LIM3. We constructed a lentivirus vector to express HB9 and GFP in NSCs and examined the consequences of HB9 expression on other transcription factors and cell differentiation. Compared to cell expressing GFP alone, NSCs expressing GFP and HB9 cycled less rapidly, downregulated Pax6 and Ngn2 mRNA levels, produced higher proportions of neurons in vitro and lower numbers of neurons after transplantation in the spinal cord of recipient rats. Oligodendrocytic and astrocytic differentiations were not affected. HB9 expressing NSCs did not express Islet1 or upregulate LIM3. They neither responded to Sonic hedgehog and retinoic acid nor produced cholinergic neurons. We concluded that forced HB9 expression affected neurogenesis but was not sufficient to confer motor neuron fate to NSCs.


Zinc ionophore activity of quercetin and epigallocatechin-gallate: from Hepa 1-6 cells to a liposome model.
Labile zinc, a tiny fraction of total intracellular zinc that is loosely bound to proteins and easily interchangeable, modulates the activity of numerous signaling and metabolic pathways. Dietary plant polyphenols such as the flavonoids quercetin (QCT) and epigallocatechin-gallate act as antioxidants and as signaling molecules. Remarkably, the activities of numerous enzymes that are targeted by polyphenols are dependent on zinc. We have previously shown that these polyphenols chelate zinc cations and hypothesized that these flavonoids might be also acting as zinc ionophores, transporting zinc cations through the plasma membrane. To prove this hypothesis, herein, we have demonstrated the capacity of QCT and epigallocatechin-gallate to rapidly increase labile zinc in mouse hepatocarcinoma Hepa 1-6 cells as well as, for the first time, in liposomes. In order to confirm that the polyphenols transport zinc cations across the plasma membrane independently of plasma membrane zinc transporters, QCT, epigallocatechin-gallate, or clioquinol (CQ), alone and combined with zinc, were added to unilamellar dipalmitoylphosphocholine/cholesterol liposomes loaded with membrane-impermeant FluoZin-3. Only the combinations of the chelators with zinc triggered a rapid increase of FluoZin-3 fluorescence within the liposomes, thus demonstrating the ionophore action of QCT, epigallocatechin-gallate, and CQ on lipid membrane systems. The ionophore activity of dietary polyphenols may underlay the raising of labile zinc levels triggered in cells by polyphenols and thus many of their biological actions.


Fox society, contact rate and rabies epizootiology.
The social behaviour of red foxes Vulpes vulpes is flexible and is adapted to their environment. Consequently social organization varies from one habitat to another under the influence of factors such as the availability of food and the pattern of mortality. Variations in social behaviour between fox populations are mirrored in different frequencies of encounters between individual foxes and hence in potentially different 'contact rates' for rabies. We present a computer model whose simulations indicate that such variation in contact rate is of overriding importance in the epizootiology of vulpine rabies, and so emphasize the importance to rabies control of understanding the behavioural ecology of foxes.


Harmonized patient-reported data elements in the electronic health record: supporting meaningful use by primary care action on health behaviors and key psychosocial factors.
Electronic health records (EHR) have the potential to improve patient care through efficient access to complete patient health information. This potential may not be reached because many of the most important determinants of health outcome are rarely included. Successful health promotion and disease prevention requires patient-reported data reflecting health behaviors and psychosocial issues. Furthermore, there is a need to harmonize this information across different EHR systems. To fill this gap a three-phased process was used to conceptualize, identify and recommend patient-reported data elements on health behaviors and psychosocial factors for the EHR. Expert panels (n=13) identified candidate measures (phase 1) that were reviewed and rated by a wide range of health professionals (n=93) using the grid-enabled measures wiki social media platform (phase 2). Recommendations were finalized through a town hall meeting with key stakeholders including patients, providers, researchers, policy makers, and representatives from healthcare settings (phase 3). Nine key elements from three areas emerged as the initial critical patient-reported elements to incorporate systematically into EHR--health behaviors (eg, exercise), psychosocial issues (eg, distress), and patient-centered factors (eg, demographics). Recommendations were also made regarding the frequency of collection ranging from a single assessment (eg, demographic characteristics), to annual assessment (eg, health behaviors), or more frequent (eg, patient goals). There was strong stakeholder support for this initiative reflecting the perceived value of incorporating patient-reported elements into EHR. The next steps will include testing the feasibility of incorporating these elements into the EHR across diverse primary care settings.


Magnesium deficit ? overlooked cause of low vitamin D status?
Like vitamin D deficit, magnesium deficit is considered to be a risk factor for cardiovascular disease. Several steps in the vitamin D metabolism, such as vitamin D binding to its transport protein and the conversion of vitamin D into the hormonal form 1,25-dihydroxyvitamin D by hepatic and renal hydroxylation, depend on magnesium as a cofactor. A new analysis of two National Health and Nutrition Examination Surveys data sets, published in BMC Medicine, investigated potential interactions between magnesium intake, circulating 25-hydroxyvitamin D, which is the generally accepted indicator of vitamin D status, and mortality. Data indicate a reduced risk of insufficient/deficient vitamin D status at high magnesium intake and an inverse association between circulating 25-hydroxyvitamin D and mortality, particularly cardiovascular mortality, among those with magnesium intake above the median. The study provides important findings concerning potential metabolic interactions between magnesium and vitamin D and its clinical relevance. However, results should be considered preliminary since biochemical data on individual magnesium status were lacking, confounding cannot be excluded and questions on the dose?response relationship still remain to be answered. Please see related research article: http://www.biomedcentral.com/1741-7015/11/187.


Genomic and phylogenetic analyses of murine adenovirus 2.
Murine adenoviruses (MAdV) are supposedly the oldest members of the genus Mastadenovirus. Currently, there are three distinct MAdV types known with rather different tropism and pathology. Here we report and annotate the DNA sequence of the full genome of MAdV-2. It was found to consist of 35,203 bp thus being considerably larger than the genomes of the other two MAdV types. The increased size of the MAdV-2 genome is generally due to larger genes and ORFs, although some differences in the number of ORFs were observed for the early regions E1, E3 and E4. The homologue of the 19K gene of E1B from MAdV-2 codes for 330 amino acids (aa) and is almost twice as large as from other mastadenoviruses. Accordingly, only the N-terminal half (155aa) has homology to the 19K protein. A homologue of the gene of the 12.5K protein was identified in the E3 region of MAdV-2, but not in MAdV-1 or MAdV-3. The other gene of yet unknown function in the E3 region of MAdV-2 seems to be unique. The E4 region of MAdV-2 contains three ORFs. One has similarity to the 34K gene of other AdVs. Two unique ORFs in the E4 region of MAdV-2 have no homology to any of the five and six ORFs in the E4 region of MAdV-1 or MAdV-3, respectively. Phylogenetic analyses showed that the three murine AdVs have a close common ancestor. They likely formed the first branching of the lineage of mastadenoviruses, and seem to be the most ancient representatives of this genus.


Transcriptional activation by nuclear receptors.
Transcriptional activation by nuclear receptors (NRs) involves the recruitment of distinct classes of co-activators and other transcription-related factors to target promoters in the chromatin environment of the nucleus. Chromatin has a general repressive effect on transcription, but also provides opportunities for NRs to regulate transcription by directing specific patterns of chromatin remodelling and histone modification. Ultimately, the transcription of hormone-regulated genes by NRs is critically dependent on co-ordinated physical and functional interactions among the receptors, chromatin, co-activators with chromatin-, histone- and factor-modifying activities, and the RNA polymerase II transcriptional machinery. In addition, several mechanisms exist to terminate or attenuate NR-dependent signalling, including modification, recycling, subcellular redistribution and degradation of the receptors or their associated cofactors. The complexity of NR-dependent transcription provides multiple targets for regulatory inputs, thus allowing each hormone-responsive cell to direct its transcriptional output in a physiologically appropriate manner.


Acupuncture for residual insomnia associated with major depressive disorder: a placebo- and sham-controlled, subject- and assessor-blind, randomized trial.
To evaluate the efficacy and safety of acupuncture for residual insomnia and other residual symptoms associated with major depressive disorder (MDD). 150 participants having significant insomnia for more than 3 months and a history of MDD (both based on DSM-IV-TR criteria) were recruited from 4 psychiatric outpatient clinics in Hong Kong from May 2011 to August 2013 to receive 9 sessions of treatment over 3 weeks. They were randomized to receive acupuncture, minimal acupuncture, or placebo acupuncture. Primary outcome was sleep diary-derived sleep efficiency. Secondary outcomes included other sleep diary parameters, actigraphy, anxiety and depressive symptoms, daytime functioning, and adverse events. The mean difference in sleep diary-derived sleep efficiency at 1-week posttreatment was -1.40 (95% CI, -7.08 to 4.28) between the acupuncture and minimal acupuncture groups and was 3.10 (95% CI, -3.64 to 9.84) between the acupuncture and placebo acupuncture groups. A χ(2) test showed that acupuncture produced a significantly higher proportion of participants achieving sleep-onset latency ≤ 30 minutes than did minimal acupuncture at 1-week posttreatment (P = .04). However, there was no significant between-group difference in most of the other outcomes. Treatment blinding was successful, as a majority of participants did not know which treatment they had received. Acupuncture was well tolerated, but the efficacy was only mild and similar to that of minimal acupuncture and placebo acupuncture. A high proportion of patients remained clinically significantly affected by insomnia after treatment. The finding raises certain doubts about the value of acupuncture and underscores the difficulties in the treatment of residual insomnia in MDD. ClinicalTrials.gov identifier: NCT01707706.


Alpha-2 adrenoceptors on nerves and muscles of rat uterus.
To test the hypothesis that functional postsynaptic alpha-2 adrenoceptors exist in rat myometrium, we examined whether specific binding sites for [3H]rauwolscine were present on microsomal membranes from myometrium of nonpregnant, day 16 pregnant and delivering rats and whether an alpha-2 adrenoceptor agonist has functional effects. The myometrium of rats at term undergoes a physiological denervation, confirmed in this study by ultrastructural examination of uterine samples and by [3H]saxitoxin binding studies. Binding sites for rauwolscine of similar Kd (11-15 nM) were present in all groups of myometrium and were localized on plasma membranes. There was no significant change in the density of rauwolscine binding sites in membranes from day 16 animals compared to nonpregnant ones, but a significant fall (38%) from these values at term. Strips of uterine circular or longitudinal muscle from nonpregnant, day-16 or day-22 pregnant rats failed to respond to the selective alpha-2 agonist, BHT-920, in the presence of propranolol; i.e., BHT-920 neither caused contraction nor inhibited contractions induced by oxytocin. BHT-920 did not affect the inhibitory effects of isoproterenol which were antagonized by propranolol. However, it antagonized contractions to norepinephrine in the presence of propranolol with a pKB value of 5.6 to 5.7. These contractions were phentolamine-sensitive. BHT-920 displaced rauwolscine from binding to all groups of myometria (IC50 = 2 to 3 x 10(-6) M) and displaced prazosin (Kd = 0.65 nM) from binding to myometria of nonpregnant rats (IC50 value congruent to 2 x 10(-4) M). Phentolamine also displaced rauwolscine from binding (IC50 = 2 x 10(-8) M). 5-Hydroxytryptamine displaced rauwolscine from binding only at higher concentrations (IC50 greater than 10(-5) M). We conclude that binding sites for alpha-2 adrenoceptor antagonists in myometrial microsomes were located primarily on smooth muscle plasma membrane. A smooth muscle alpha-2 adrenoceptor agonist appeared to occupy a site on muscle with the same affinity as it displayed toward rauwolscine binding site and competitively inhibited effects of an alpha-1 agonist. Our data suggest that alpha-2 adrenoceptor binding sites may exist on smooth muscle without coupling to contractile function, but their occupancy competitively prevents occupancy of alpha-1 agonist receptor activation sites.


Cross talk in implicit assignment of error information during bimanual visuomotor learning.
When a neural movement controller, called an "internal model," is adapted to a novel environment, the movement error needs to be appropriately associated with the controller. However, their association is not necessarily guaranteed for bimanual movements in which two controllers--one for each hand--result in two movement errors. Considering the implicit nature of the adaptation process, the movement error of one hand can be erroneously associated with the controller of the other hand. Here, we investigated this credit-assignment problem in bimanual movement by having participants perform bimanual, symmetric back-and-forth movements while displaying the position of the right hand only with a cursor. In the training session, the cursor position was gradually rotated clockwise, such that the participants were unaware of the rotation. The movement of the right hand gradually rotated counterclockwise as a consequence of adaptation. Although the participants knew that the cursor reflected the movement of the right hand, such gradual adaptation was also observed for the invisible left hand, especially when the cursor was presented on the left side of the display. Thus the movement error of the right hand was implicitly assigned to the left-hand controller. Such cross talk in credit assignment might influence motor adaptation performance, even when two cursors are presented; the adaptation was impaired when the rotations imposed on the cursors were opposite compared with when they were in the same direction. These results indicate the inherent presence of cross talk in the process of associating action with consequence in bimanual movement.


Health care workers' ability and willingness to report to duty during catastrophic disasters.
Catastrophic disasters create surge capacity needs for health care systems. This is especially true in the urban setting because the high population density and reliance on complex urban infrastructures (e.g., mass transit systems and high rise buildings) could adversely affect the ability to meet surge capacity needs. To better understand responsiveness in this setting, we conducted a survey of health care workers (HCWs) (N =6,428) from 47 health care facilities in New York City and the surrounding metropolitan region to determine their ability and willingness to report to work during various catastrophic events. A range of facility types and sizes were represented in the sample. Results indicate that HCWs were most able to report to work for a mass casualty incident (MCI) (83%), environmental disaster (81%), and chemical event (71%) and least able to report during a smallpox epidemic (69%), radiological event (64%), sudden acute respiratory distress syndrome (SARS) outbreak (64%), or severe snow storm (49%). In terms of willingness, HCWs were most willing to report during a snow storm (80%), MCI (86%), and environmental disaster (84%) and least willing during a SARS outbreak (48%), radiological event (57%), smallpox epidemic (61%), and chemical event (68%). Barriers to ability included transportation problems, child care, eldercare, and pet care obligations. Barriers to willingness included fear and concern for family and self and personal health problems. The findings were consistent for all types of facilities. Importantly, many of the barriers identified are amenable to interventions.


Absence of symbiotic leghemoglobins alters bacteroid and plant cell differentiation during development of Lotus japonicus root nodules.
During development of legume root nodules, rhizobia and their host plant cells undergo profound differentiation, which is underpinned by massive changes in gene expression in both symbiotic partners. Oxygen concentrations in infected and surrounding uninfected cells drop precipitously during nodule development. To assess what effects this has on plant and bacterial cell differentiation and gene expression, we used a leghemoglobin-RNA-interference (LbRNAi) line of Lotus japonicus, which is devoid of leghemoglobins and has elevated levels of free-oxygen in its nodules. Bacteroids in LbRNAi nodules showed altered ultrastructure indicating changes in bacterial differentiation. Transcript analysis of 189 plant and 192 bacterial genes uncovered many genes in both the plant and bacteria that were differentially regulated during nodulation of LbRNAi plants compared with the wild type (containing Lb and able to fix nitrogen). These included fix and nif genes of the bacteria, which are involved in microaerobic respiration and nitrogen fixation, respectively, and plant genes involved in primary and secondary metabolism. Metabolite analysis revealed decreased levels of many amino acids in nodules of LbRNAi plants, consistent with the defect in symbiotic nitrogen fixation of this line.


Respiratory syncytial virus epidemiology in Turkey.
Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract infections in infants and young children worldwide. This study was conducted to determine the prevalence of RSV among high-risk children admitted with respiratory symptoms in a developing country. This is a multicenter study conducted among children less than 24 months of age and admitted to the hospital with respiratory symptoms. The inclusion criteria included: lower respiratory tract symptoms on admission, gestational age less than 35 weeks, and admission age less than six months, or children less than 24 months of age with a diagnosis of bronchopulmonary dysplasia requiring medical treatment or intervention during the last six months or with an uncorrected congenital heart disease (other than patent ductus arteriosus). Nasopharyngeal samples were obtained with one of the three standard methods: nasopharyngeal aspirate, nasopharyngeal wash or nasopharyngeal swab. RSV antigen was determined by enzyme immunoassay using Abbott TESTPACK RSV (No. 8100/2027-16). Statistical analysis was performed using Student's t-test and chi-square test. In this study, 332 children (135 females, 40.7%; 197 males, 59.3%) were included, and the nasopharyngeal specimens of 98 (29.5%) children were determined to be RSV-positive. There were no differences in sex, age of gestation, age of admission, family education, number of siblings and smoking at home for RSV-positive and -negative cases. Furthermore, underlying disease and duration of hospital and intensive care unit stay were similar among groups. Only otitis media was more common among RSV-positive cases. No fatality at hospital was recorded. Frozen samples revealed more negative results. Most cases presented during winter and the number of RSV-positive cases was higher in cold and economically poor areas. Premature children and children with underlying medical condition acquire RSV irrespective of other sociodemographic risk factors, and most of them are hospitalized. Thus, an RSV vaccine seems the most effective mode of protection to decrease morbidity and mortality.


Proteomic revelation: SUMO changes partners when the heat is on.
Dynamic changes in the posttranslational modification of proteins govern most cellular signaling pathways. Work over the past decade has connected many of these processes with the covalent attachment of the small ubiquitin-like modifier (SUMO) protein to target proteins, but a global view of the dynamics of SUMOylation was missing. A system-level proteomics approach has now been used to describe quantitative changes in protein modification with the SUMO-2 paralog during the response to heat shock. The SUMOylation status of more than 700 proteins was monitored in HeLa cells during the induction of hyperthermic stress and the recovery period. A massive redistribution of SUMO-2 was observed that affected many biological pathways that are important for the heat shock response, including cell cycle regulation, transcription, translation, protein folding, and DNA repair. Collectively, these data suggest a wide-ranging role for SUMOylation in the cellular response to hyperthermic stress. The strategies that were developed to provide this global view of SUMOylation should guide future approaches to probing quantitative changes in protein modification.


Demethylation in erythrocytes: a reaction involving hemoglobin.
A reaction is described in erythrocytes of rat and of man whereby O-methylated metabolites of the catecholamines are demethylated to the corresponding catechols. The reaction was studied by incubating aliquots of erythrocyte lysates with radiolabeled O-methylated compounds and isolating the catechol product by alumina adsorption chromatography. The demethylating activity was located in the cytosol of the erythrocytes. Evidence was strong that oxyhemoglobin was responsible for the reaction: the demethylase activity was inseparable from oxyhemoglobin in several chromatographic separations. In addition, although commercially available hemoglobins were inactive in the reactions, after their conversion to oxyhemoglobin and purification, they did effect demethylation. Methemoglobin did not demethylate guaiacols and in fact inhibited demethylation by oxyhemoglobin. The reaction was inhibited by the addition of reduced pyridine nucleotides and of the methyl acceptor tetrahydrofolic acid.


Equity in aid allocation and distribution: A qualitative study of key stakeholders in Northern Uganda.
The Sustainable Development Goals have spurred a growing interest in and focus on equitable development. In theory, donors can play an important role in promoting equity within a country by providing services, influencing government policies and incorporating equity into decision-making. However, we know little about whether this actually happens on the ground. We conduct what we believe is the first study to explore the extent to which equity is prioritised in the allocation and distribution of aid, based on in-depth interviews with government officials, bilateral and international donors, and implementing partners operating in Northern Uganda. We find that a broad category of people are perceived to be marginalised/vulnerable, with a substantial segment largely untargeted by major donor programmes. Various stakeholders employ a wide range of strategies to identify the most vulnerable individuals and groups, including the use of available data and statistics, consultation and engagement with relevant stakeholders, and undertaking primary data collection. The strategies used to incorporate equity in aid allocation and distribution include: targeting the regions of Northern Uganda and Karamoja in particular, targeting both refugees and host populations in refugee-hosting districts, prioritising the critically vulnerable in any aid distribution process, and using specific tools and consultants to ensure that major equity issues are addressed in proposals. Challenges undermining the process include poor understanding of the concept of equity among some implementing partners, lack of comprehensively disaggregated data, corruption, and political interference in choice of aid location from government officials and donors.


Mortality among patients with acute myocardial infarction: the influences of patient-centered care and evidence-based medicine.
Recent studies have suggested that there is a positive impact of patient-centered care (PCC) on both the patient-physician relationship and subsequent patient health-related behaviors. One recent prospective study reported a significant relationship between the degree of PCC experienced by patients during their hospitalization for acute myocardial infarction (AMI) and their postdischarge cardiac symptoms. A limitation of this study, however, was a lack of information regarding the technical quality of the AMI care, which might have explained at least part of the differences in outcomes. The present study was undertaken to test the influence of both PCC and technical care quality on outcomes among AMI patients. We analyzed data from a national sample of 1,858 veterans hospitalized for an initial AMI in a Department of Veterans Affairs medical center during fiscal years 2003 and 2004 for whom data had been compiled on evidence-based treatment and who had also completed a Picker questionnaire assessing perceptions of PCC. Cox proportional hazards models were used to estimate the relationship between PCC and survival 1-year postdischarge, controlling for technical quality of care, patient clinical condition and history, admission process characteristics, and patient sociodemographic characteristics. We hypothesized that better PCC would be associated with a lower probability of death 1-year postdischarge, even after controlling for patient characteristics and the technical quality of care. Better PCC was associated with a significantly but modestly lower hazard of death over the 1-year study period (hazard ratio 0.992, 95 percent confidence interval 0.986-0.999). Providing PCC may result in important clinical benefits, in addition to meeting patient needs and expectations.


Reaching for the Holy Grail: insights from infection/cure models on the prospects for vaccines for Trypanosoma cruzi infection.
Prevention of Trypanosoma cruzi infection in mammals likely depends on either prevention of the invading trypomastigotes from infecting host cells or the rapid recognition and killing of the newly infected cells by T. cruzi-specific T cells. We show here that multiple rounds of infection and cure (by drug therapy) fails to protect mice from reinfection, despite the generation of potent T cell responses. This disappointing result is similar to that obtained with many other vaccine protocols used in attempts to protect animals from T. cruzi infection. We have previously shown that immune recognition of T. cruzi infection is significantly delayed both at the systemic level and at the level of the infected host cell. The systemic delay appears to be the result of a stealth infection process that fails to trigger substantial innate recognition mechanisms while the delay at the cellular level is related to the immunodominance of highly variable gene family proteins, in particular those of the trans-sialidase family. Here we discuss how these previous studies and the new findings herein impact our thoughts on the potential of prophylactic vaccination to serve a productive role in the prevention of T. cruzi infection and Chagas disease.