Effect of microgravity on the pharmacokinetics of Ibuprofen in humans.
The pharmacokinetics of ibuprofen were studied under microgravity (µG) conditions and compared with those at normal gravity (1G) in humans. Six healthy human volunteers were given 600 mg oral dose ibuprofen during 1-day simulated µG antiorthostatic bed rest position, then at normal position (1G) in a sequential design with 7 days washout time. Saliva and plasma samples were obtained up to 8 hours after dosing. Ibuprofen was not detected in all saliva samples. Pharmacokinetic parameters in plasma were calculated by either noncompartmental analysis or a 1- compartment model using the Kinetica program. Absorption kinetic parameters were then predicted by ADAM and PE modules using the Simcyp program. Results have showed an increased rate of ibuprofen dissolution and absorption and hence faster onset of action under µG conditions. However, rate of drug elimination and bioavailability was not affected by µG, suggesting no need for dose adjustment.


Different responses of spontaneous and stimulus-related alpha activity to ambient luminance changes.
Alpha oscillations are particularly important in determining our percepts and have been implicated in fundamental brain functions. Oscillatory activity can be spontaneous or stimulus-related. Furthermore, stimulus-related responses can be phase- or non-phase-locked to the stimulus. Non-phase-locked (induced) activity can be identified as the average amplitude changes in response to a stimulation, while phase-locked activity can be measured via reverse-correlation techniques (echo function). However, the mechanisms and the functional roles of these oscillations are far from clear. Here, we investigated the effect of ambient luminance changes, known to dramatically modulate neural oscillations, on spontaneous and stimulus-related alpha. We investigated the effect of ambient luminance on EEG alpha during spontaneous human brain activity at rest (experiment 1) and during visual stimulation (experiment 2). Results show that spontaneous alpha amplitude increased by decreasing ambient luminance, while alpha frequency remained unaffected. In the second experiment, we found that under low-luminance viewing, the stimulus-related alpha amplitude was lower, and its frequency was slightly faster. These effects were evident in the phase-locked part of the alpha response (echo function), but weaker or absent in the induced (non-phase-locked) alpha responses. Finally, we explored the possible behavioural correlates of these modulations in a monocular critical flicker frequency task (experiment 3), finding that dark adaptation in the left eye decreased the temporal threshold of the right eye. Overall, we found that ambient luminance changes impact differently on spontaneous and stimulus-related alpha expression. We suggest that stimulus-related alpha activity is crucial in determining human temporal segmentation abilities.


Estimations of serum arabinitol for diagnosing invasive candidosis.
Arabinitol concentrations were determined in 157 serum samples from 95 patients with suspected invasive candidosis and in 10 serum samples from healthy laboratory workers. Fifty eight of the 95 patients, subsequently diagnosed as not having invasive candidosis had concentrations of arabinitol below 1.2 micrograms/ml (mean 0.59 (SD) 0.26). Sera from the healthy laboratory workers gave similar results (mean 0.55 (0.05]. Concentrations above the normal range were found in 18 of the 19 cases of confirmed or probable invasive candidosis and in seven of eight patients with infected intravenous lines or cannulas and clinical evidence of systemic infection. Raised concentrations were also seen in 10 other patients, including nine with renal failure who did not have invasive infections. Multiple serum samples obtained from 33 patients showed that sequential estimations were of value for diagnosing a developing infection. Despite some difficulties of interpretation the technique is rapid and specific and is suitable for use in the diagnostic laboratory of a larger general hospital.


Water quality of least-impaired lakes in eastern and southern Arkansas.
A three-phased study identified one least-impaired (reference) lake for each of four Arkansas lake classifications: three classifications in the Mississippi Alluvial Plain (MAP) ecoregion and a fourth classification in the South Central Plains (SCP) ecoregion. Water quality at three of the least-impaired lakes generally was comparable and also was comparable to water quality from Kansas and Missouri reference lakes and Texas least-impaired lakes. Water quality of one least-impaired lake in the MAP ecoregion was not as good as water quality in other least-impaired lakes in Arkansas or in the three other states: a probable consequence of all lakes in that classification having a designated use as a source of irrigation water. Chemical and physical conditions for all four lake classifications were at times naturally harsh as limnological characteristics changed temporally. As a consequence of allochthonous organic material, oxbow lakes isolated within watersheds comprised of swamps were susceptible to low dissolved oxygen concentrations to the extent that conditions would be limiting to some aquatic biota. Also, pH in lakes in the SCP ecoregion was <6.0, a level exceeding current Arkansas water-quality standards but typical of black water systems. Water quality of the deepest lakes exceeded that of shallow lakes. N/P ratios and trophic state indices may be less effective for assessing water quality for shallow lakes (<2 m) than for deep lakes because there is an increased exposure of sediment (and associated phosphorus) to disturbance and light in the former.


Impact of normothermic preservation with extracellular type solution containing trehalose on rat kidney grafting from a cardiac death donor.
The aim of this study was to investigate factors that may improve the condition of a marginal kidney preserved with a normothermic solution following cardiac death (CD) in a model of rat kidney transplantation (RTx). Post-euthanasia, Lewis (LEW) donor rats were left for 1 h in a 23°C room. These critical kidney grafts were preserved in University of Wisconsin (UW), lactate Ringer's (LR), or extracellular-trehalose-Kyoto (ETK) solution, followed by intracellular-trehalose-Kyoto (ITK) solution at 4, 23, or 37°C for another 1 h, and finally transplanted into bilaterally nephrectomized LEW recipient rats (n = 4-6). Grafts of rats surviving to day 14 after RTx were evaluated by histopathological examination. The energy activity of these marginal rat kidneys was measured by high-performance liquid chromatography (HPLC; n = 4 per group) and fluorescence intensity assay (n = 6 per group) after preservation with UW or ETK solutions at each temperature. Finally, the transplanted kidney was assessed by an in vivo luciferase imaging system (n = 2). Using the 1-h normothermic preservation of post-CD kidneys, five out of six recipients in the ETK group survived until 14 days, in contrast to zero out of six in the UW group (p<0.01). Preservation with ITK rather than ETK at 23°C tended to have an inferior effect on recipient survival (p = 0.12). Energy activities of the fresh donor kidneys decreased in a temperature-dependent manner, while those of post-CD kidneys remained at the lower level. ETK was superior to UW in protecting against edema of the post-CD kidneys at the higher temperature. Luminescence intensity of successful grafts recovered within 1 h, while the intensity of grafts of deceased recipients did not change at 1 h post-reperfusion. Normothermic storage with extracellular-type solution containing trehalose might prevent reperfusion injury due to temperature-dependent tissue edema.


Acute rejection and cardiac allograft vascular disease is reduced by suppression of subclinical cytomegalovirus infection.
Anticytomegalovirus (CMV) prophylaxis prevents the acute disease but its impact on subclinical infection and allograft outcome is unknown. We sought to determine whether CMV prophylaxis administered for three months after heart transplant would improve patient outcomes. This prospective cohort study of 66 heart transplant recipients compared aggressive CMV prophylaxis (n = 21, CMV hyperimmune globulin [CMVIG] plus four weeks of intravenous ganciclovir followed by two months of valganciclovir); with standard prophylaxis (n = 45, intravenous ganciclovir for four weeks). Prophylaxis was based on pretransplant donor (D) and recipient (R) CMV serology: R-/D+ received aggressive prophylaxis; R+ received standard prophylaxis. Outcome measures were: CMV infection assessed by DNA-polymerase chain reaction on peripheral blood polymorphonuclear leukocytes, acute rejection, and cardiac allograft vascular disease (CAV) assessed by intravascular ultrasound. All patients completed one year of follow-up. RESULTS.: CMV infection was subclinical in all but four patients (two in each group). Aggressively treated patients had a lower incidence of CMV infection (73 +/- 10% vs. 94 +/- 4%; P = 0.038), and an independent reduced relative risk for acute rejection graded > or =3A (relative risk [95% CI] = 0.55 [0.26-0.96]; P = 0.03), as compared with the standard prophylaxis group. Aggressively prophylaxed patients also showed a slower progression of CAV, in terms of coronary artery lumen loss (lumen volume change=-21 +/- 13% vs. -10+/-14%; P = 0.05); and vessel shrinkage (vessel volume change = -15 +/- 11% vs. -3 +/- 18%; P = 0.03). Prolonged (val)ganciclovir plus CMVIG reduces viral levels, acute rejection, and allograft vascular disease, suggesting a role for chronic subclinical infection in the pathophysiology of the most common diseases affecting heart transplant recipients.


Low Serum Lysosomal Acid Lipase Activity Correlates with Advanced Liver Disease.
Fatty liver has become the most common liver disorder and is recognized as a major health burden in the Western world. The causes for disease progression are not fully elucidated but lysosomal impairment is suggested. Here we evaluate a possible role for lysosomal acid lipase (LAL) activity in liver disease. To study LAL levels in patients with microvesicular, idiopathic cirrhosis and nonalcoholic fatty liver disease (NAFLD). Medical records of patients with microvesicular steatosis, cryptogenic cirrhosis and NAFLD, diagnosed on the basis of liver biopsies, were included in the study. Measured serum LAL activity was correlated to clinical, laboratory, imaging and pathological data. No patient exhibited LAL activity compatible with genetic LAL deficiency. However, serum LAL activity inversely predicted liver disease severity. A LAL level of 0.5 was the most sensitive for detecting both histologic and noninvasive markers for disease severity, including lower white blood cell count and calcium, and elevated γ-glutamyltransferase, creatinine, glucose, glycated hemoglobin, uric acid and coagulation function. Serum LAL activity <0.5 indicates severe liver injury in patients with fatty liver and cirrhosis. Further studies should define the direct role of LAL in liver disease severity and consider the possibility of replacement therapy.


Treatment fidelity plan for an activity intervention designed for persons with dementia.
The testing of psychosocial interventions in a clinical trial poses many challenges to maintaining a rigorous experimental protocol and to delivering the interventions uniformly throughout the project. These challenges directly affect the reported effectiveness of psychosocial interventions. In this article, the authors describe the treatment fidelity plan developed by an interdisciplinary research team from recreational therapy and nursing for implementing recreational activities during a clinical trial funded by the National Institute of Nursing Research. The trial tests the efficacy of activities for responding to the behavioral symptoms of dementia. The authors report treatment fidelity strategies to allow comparison of their intervention with that of other studies, to improve effect size in similar studies, and to facilitate replication and translation of this work into clinical practice.


Multidimensional associative factors for improvement in pain, function, and working capacity after rehabilitation of whiplash associated disorder: a prognostic, prospective outcome study.
Whiplash associated disorders (WAD) have dramatic consequences for individual and public health. Risk factors for better and worse outcomes are important to optimize management. This study aimed to determine short- and mid-term associative co-factors of neck pain relief, improved physical functioning, and improved working capacity (dependent variables) in patients suffering from whiplash associated disorder who participated in a standardized, inpatient pain management program. Naturalistic, observational, prospective cohort study. Outcome was measured by standardized assessment instruments. Co-factors covered sociodemographics, comorbidities, social participation, affective health, and coping abilities. Stepwise, multivariate linear regression analysis was performed at discharge and at the 6-month follow-up. All regression models explained high proportions of variance (53.3% - 72.1%). The corresponding baseline level was significantly associated with a change in every dependent variable (explained variances: 11.4%-56.7%). Pain relief significantly depended on improved function and vice-versa (3.4%-14.8%). Improved ability to decrease pain was associated with pain relief at discharge (9.6%). Functional improvement was associated with decreased catastrophizing (19.4%) at discharge and decreased depression (20.5%) at the 6 month follow-up. Pain relief, improved physical function and working capacity were associated with each other. Improved coping (catastrophizing and ability to decrease pain) and reduced depression may act as important predictors for pain relief and improved function. These findings offer toe-holds for optimized therapy of chronic WAD.


The discrete Laplace exponential family and estimation of Y-STR haplotype frequencies.
Estimating haplotype frequencies is important in e.g. forensic genetics, where the frequencies are needed to calculate the likelihood ratio for the evidential weight of a DNA profile found at a crime scene. Estimation is naturally based on a population model, motivating the investigation of the Fisher-Wright model of evolution for haploid lineage DNA markers. An exponential family (a class of probability distributions that is well understood in probability theory such that inference is easily made by using existing software) called the 'discrete Laplace distribution' is described. We illustrate how well the discrete Laplace distribution approximates a more complicated distribution that arises by investigating the well-known population genetic Fisher-Wright model of evolution by a single-step mutation process. It was shown how the discrete Laplace distribution can be used to estimate haplotype frequencies for haploid lineage DNA markers (such as Y-chromosomal short tandem repeats), which in turn can be used to assess the evidential weight of a DNA profile found at a crime scene. This was done by making inference in a mixture of multivariate, marginally independent, discrete Laplace distributions using the EM algorithm to estimate the probabilities of membership of a set of unobserved subpopulations. The discrete Laplace distribution can be used to estimate haplotype frequencies with lower prediction error than other existing estimators. Furthermore, the calculations could be performed on a normal computer. This method was implemented in the freely available open source software R that is supported on Linux, MacOS and MS Windows.


Will SBRT replace conventional radiotherapy in patients with low-intermediate risk prostate cancer? A review.
Stereotactic body radiation therapy (SBRT) is a novel treatment modality in radiation oncology that delivers a very high dose of radiation to the tumor target with high precision using single or a small number of fractions. SBRT is the result of technological advances in patient/tumor immobilization, image guidance, and treatment planning and delivery. This modality is safe and effective in both early stage primary cancer and oligometastases. Compared to the use of stereotactic radiosurgery for other tumor sites, SBRT is slow to be adopted in the management of genitourinary malignancies. Emerging data show the safety and efficacy of this treatment modality in prostate cancer. Preclinical data, clinical experience, and challenges are reviewed and discussed.


Hallucinations in Healthy Older Adults: An Overview of the Literature and Perspectives for Future Research.
KEY POINTS ➢ Studies suggest a substantial minority of healthy older adults have hallucinatory experiences, in line with existing evidence on hallucinations in other age groups, though it is still unclear if hallucination prevalence increases or declines with age in older cohorts.➢ Stigma attached to both hallucinations and ageing leads to considerable under-reporting of these experiences in healthy older adults and may negatively bias how professionals, family members, and the public respond.➢ Why and when hallucinations in healthy older adults remit, persist, or progress to other clinical disorders remains poorly understood.➢ Current evidence points to a range of factors associated with hallucinations in older adults including decline in sensory or cognitive functioning, poor sleep, and psychosocial stressors (e.g., social isolation, loneliness, and bereavement), highlighting the need for accurate assessment and tailored interventions. Hallucinations, though common in youth and younger adults, are not the preserve of these age groups. Accumulating evidence shows that hallucinatory experiences are also present at surprisingly high rates in healthy older adults in the general community. Furthermore, stigma and misunderstanding of hallucinations, together with ageism, may lead to under-reporting of these experiences by older adults, and misdiagnosis or mismanagement by health and mental health practitioners. Consequently, improved public and professional knowledge is needed about the nature and significance of hallucinations with advancing age. The purpose of this review is to provide a comprehensive overview, and critical analysis, of research on the prevalence, psychosocial, and neurobiological factors associated with hallucinations in people aged 60 years and over. To the best of our knowledge, this is the first review of its kind in the literature. The evidence supports a dynamic conceptualization of hallucinations, in which the emergence of hallucinations is viewed as a balance between the sensory, cognitive, or social impairments accompanying advancing age and the degree to which compensatory processes elicited by these impairments are successful. We briefly summarize the implications of the literature for aged care services and interventions, and stress that far more studies are needed in this important field of research.


Cognitive change in the year after a first manic episode: association between clinical outcome and cognitive performance early in the course of bipolar I disorder.
Cognitive impairments are present immediately following recovery from a first episode of mania, although at a lesser severity than those seen in more chronic patients with bipolar I disorder. Little is known about how deficits evolve over the course of illness, however, and whether these changes are associated with disease progression. Patients with bipolar I disorder (DSM-IV-TR) receiving naturalistic clinical follow-up from the Systematic Treatment Optimization Program for Early Mania (STOP-EM) from July 2004 to May 2013 completed a comprehensive cognitive battery following recovery from their first manic episode and again 1 year later. Performance was compared between patients who experienced a recurrence of a mood episode (BDrecur) (n = 26) versus those that maintained remission (BDwell) (n = 27) over follow-up, as well as healthy comparison subjects (HS) (n = 31). While both BDrecur and BDwell had impairments in overall cognitive performance relative to HS at baseline (mean difference = -0.59, P < .001; mean difference = -0.43, P < .05, respectively), at follow-up BDrecur showed deficits compared to both HS (mean difference = -0.62, P = .001) and BDwell (mean difference = -0.41, P = .05), with BDwell cognition similar to that in HS (mean difference = -0.21, P > .4). BDwell showed larger improvements over follow-up relative to both other groups (P < .05). While changes in BDrecur did not differ from HS, in this group more days in a manic or hypomanic episode was associated with performance declines (r = -0.40, P < .05). While cognitive function improves in patients who sustain remission in the year following a first manic episode, those who experience a recurrence remain impaired, with performance declines being most apparent in those who experienced longer manic or hypomanic episodes.


Comparative phylogeography in a genus of coral reef fishes: biogeographic and genetic concordance in the Caribbean.
Geographic barriers that limit the movement of individuals between populations may create or maintain phylogenetically discrete lineages. Such barriers are often inferred from geographic surveys of a single mitochondrial marker to identify phylogenetic splits. Mitochondrial DNA, however, has an effective population size one-fourth that of nuclear DNA, which can facilitate the rapid evolution of monophyletic mtDNA lineages in the absence of geographic barriers. The identification of geographic barriers will thus be more robust if barriers are proposed a priori, and tested with multiple independent genetic markers in multiple species. Here, we tested two proposed marine biogeographic breaks located at the Mona Passage in the Caribbean Sea and at the southern end of Exuma Sound in the Bahamas. We sequenced mitochondrial cytochrome b (400 bp) and nuclear rag1 (573 bp) for nine species and colour forms (183 individuals total) within the teleost genus Elacatinus (Gobiidae) that span the proposed breaks. Our results showed that Mona Passage separated mtcyb and rag1 lineages, with no genetic exchange between populations separated by just 23 km. However, the Central Bahamas barrier was only weakly supported by our data. Importantly, neither barrier coincided with deep genetic splits. This suggests that these two barriers did not initially isolate regional populations, but instead disrupt ongoing gene flow between regions. Our inferred relationships further suggested a division of the Caribbean region into northwestern and southeastern regions, a pattern reflected by some freshwater and terrestrial vertebrates. Our results, coupled with genetic and demographic data from other reef fishes and corals, provide robust support for the Mona Passage as a long-term biogeographic barrier for Caribbean animals.


The global burden of migraine: measuring disability in headache disorders with WHO's Classification of Functioning, Disability and Health (ICF).
This overview of the published epidemiological evidence of migraine helps to identify the size of the public-health problem that migraine represents. It also highlights the need for further epidemiological studies in many parts of the world to gain full understanding of the scale of clinical, economic and humanistic burdens attributable to it. This paper presents some of the work on migraine undertaken by the World Health Organization (WHO) in the Global Burden of Disease study conducted in 2000 and reported in the World Health Report 2001. Migraine was not included in the first Global Burden of Disease 1990. The paper also discussed the measurement of disability attributable to headache disorders using WHO ICF Classification. Using disability-adjusted life years (DALYs) as a summary measure of population health (which adds disability to mortality), WHO have shown that mental and neurological disorders collectively account for 30.8% of all years of healthy life lost to disability (YLDs) whilst migraine, one amongst these, alone accounts for 1.4% and is in the top 20 causes of disability worldwide. This information is combined with the increasingly widely accepted belief that disability and functioning are relevant parameters for monitoring the health of nations and that there is an increasing need to measure them. WHO's Classification of Functioning, Disability and Health (ICF) provides a model of human functioning and disability, as well as a classification system, that allows us to highlight and measure all dimensions of disability. ICF applied to headache disorders allows comparability with other health conditions as well as evaluation of the role of the environment as a cause of disability amongst people with headache. Migraine causes a large proportion of the non-fatal disease-related burden worldwide. Our knowledge of headache related burden is incomplete and it is necessary to add to it epidemiological studies in many parts of the world and to combine this with measurements of disability using both DALYs and WHO's ICF Classification. The work described here has been the base for the Global Campaign against Headache disorders: "Lifting the Burden", launched in 2004 jointly by WHO, IHS (International Headache Society), WHA (World Headache Alliance) and EHF (European Headache Federation).


The Norwegian Cognitive impairment after stroke study (Nor-COAST): study protocol of a multicentre, prospective cohort study.
Early and late onset post-stroke cognitive impairment (PCI) contributes substantially to disability following stroke, and is a high priority within stroke research. The aetiology for PCI is complex and related to the stroke itself, brain resilience, comorbid brain diseases, prestroke vulnerability and complications during the hospital stay. The aim of the Norwegian Cognitive Impairment After Stroke study (Nor-COAST) is to quantify and measure levels of cognitive impairments in a general Norwegian stroke population and to identify biological and clinical markers associated with prognosis for cognitive disorders following incident stroke. The study will be organised within five work packages: 1) Incidence and trajectories 2) Pathological mechanisms 3) Development of a risk score 4) Impact of physical activity and 5) Adherence to secondary prevention. Nor-COAST is an ongoing multicentre (five participating hospitals), prospective, cohort study with consecutive inclusion during the acute phase and with follow-up at three and 18 months, and at three years. Inclusion criteria are stroke defined according to the WHO criteria. During the recruitment period from 18.05.2015 to 31.03.2017, 816 participants have been included. Cognitive impairment will be classified according to the DSM-5 criteria using a consensus group. Cognitive function is assessed by a standardised neuropsychological test battery, the Montreal Cognitive Assessment, Trail making A and B, ten-word immediate and delayed recall test, the Controlled Oral Word Association, Global Deterioration Scale and proxy based information by and the Ascertain Dementia 8 item informant questionnaire. Biomarkers include magnetic resonance imaging, routine blood samples and bio-banking. Clinical assessments include characteristics of the stroke, comorbidity, delirium, frailty and tests for cognitive and physical function, sensor based activity monitoring and adherence to secondary prophylaxis. Nor-COAST is the first Norwegian multicentre study to quantify burden of PCI that will provide reliable estimates in a general stroke population. A multidisciplinary approach aiming to identify biomarkers and clinical markers of overall prognosis will add new knowledge about risk profiles, including pre-stroke vulnerability and modifiable factors such as physical activity and secondary prophylaxis of relevance for clinical practice and later intervention studies. ClinicalTrials.gov: NCT02650531 . Retrospectively registered January 8, 2016. First participant included May 18, 2015.


Binding constants of membrane-anchored receptors and ligands depend strongly on the nanoscale roughness of membranes.
Cell adhesion and the adhesion of vesicles to the membranes of cells or organelles are pivotal for immune responses, tissue formation, and cell signaling. The adhesion processes depend sensitively on the binding constant of the membrane-anchored receptor and ligand proteins that mediate adhesion, but this constant is difficult to measure in experiments. We have investigated the binding of membrane-anchored receptor and ligand proteins with molecular dynamics simulations. We find that the binding constant of the anchored proteins strongly decreases with the membrane roughness caused by thermally excited membrane shape fluctuations on nanoscales. We present a theory that explains the roughness dependence of the binding constant for the anchored proteins from membrane confinement and that relates this constant to the binding constant of soluble proteins without membrane anchors. Because the binding constant of soluble proteins is readily accessible in experiments, our results provide a useful route to compute the binding constant of membrane-anchored receptor and ligand proteins.


Cognitive impairment in neuro-Behcet's disease and multiple sclerosis: a comparative study.
Both multiple sclerosis (MS) and neuro-Behcet's disease (NBD) can cause a cognitive dysfunction mainly involving the executive functions. We conducted this study to clarify the probable differential cognitive/behavioral profiles of MS and NBD. Twenty consecutive cases with parenchymal NBD (13 male, seven female), and 20 cases with MS (five male, 15 female) were evaluated. Both groups had a thorough neurological examination; an evaluation for Expanded Disability Status Scale (EDSS), Multiple Sclerosis Functional Composite (MSFC), and Beck's Depression Scale; and a detailed neuropsychological evaluation masked to the diagnosis. Among the two groups, male/female ratio differed significantly while other demographic and clinical features were not different. In California Verbal Learning Test, both short- and long-term delayed recall and cued recognition were worse in neuro-Behcet's cases. They had impaired semantic clustering and increased false positives. Stroop Test was also more impaired in neuro-Behcet's cases. They needed significantly more trials to complete the first category of the Wisconsin Card Sorting Test and had a poorer total Frontal Behavioral Inventory Score. Our results suggest that neuro-Behcet's patients have a more severe "frontal"-executive dysfunction than MS patients.


Body mass index trajectory classes and incident asthma in childhood: results from 8 European Birth Cohorts--a Global Allergy and Asthma European Network initiative.
The causal link between body mass index (BMI) or obesity and asthma in children is still being debated. Analyses of large longitudinal studies with a sufficient number of incident cases and in which the time-dependent processes of both excess weight and asthma development can be validly analyzed are lacking. We sought to investigate whether the course of BMI predicts incident asthma in childhood. Data from 12,050 subjects of 8 European birth cohorts on asthma and allergies were combined. BMI and doctor-diagnosed asthma were modeled during the first 6 years of life with latent growth mixture modeling and discrete time hazard models. Subpopulations of children were identified with similar standardized BMI trajectories according to age- and sex-specific "World Health Organization (WHO) child growth standards" and "WHO growth standards for school aged children and adolescents" for children up to age 5 years and older than 5 years, respectively (BMI-SDS). These types of growth profiles were analyzed as predictors for incident asthma. Children with a rapid BMI-SDS gain in the first 2 years of life had a higher risk for incident asthma up to age 6 years than children with a less pronounced weight gain slope in early childhood. The hazard ratio was 1.3 (95% CI, 1.1-1.5) after adjustment for birth weight, weight-for-length at birth, gestational age, sex, maternal smoking in pregnancy, breast-feeding, and family history of asthma or allergies. A rapid BMI gain at 2 to 6 years of age in addition to rapid gain in the first 2 years of life did not significantly enhance the risk of asthma. Rapid growth in BMI during the first 2 years of life increases the risk of asthma up to age 6 years.


In-depth immunophenotyping data of IL-6R on the human peripheral regulatory T cell (Treg) compartment.
We provide in this paper a detailed characterization of the human peripheral CD4+ CD127lowCD25+ regulatory T cell (Treg) compartment, with a particular emphasis in defining the population expressing higher levels of the IL-6 receptor (IL-6R). We provide a description of the phenotype of this population by assessing both the surface expression by flow cytometry as well as their transcriptional profile and functional features. In addition, we also present functional data describing the responsiveness of these subsets to IL-6 signalling in vitro and to IL-2 in vivo. The data presented in this paper support the research article "Human IL-6RhiTIGIT- CD4+CD127lowCD25+ T cells display potent in vitro suppressive capacity and a distinct Th17 profile" (Ferreira RC et al., 2017; doi: 10.1016/j.clim.2017.03.002) [1].


Inflammation and endothelial function: direct vascular effects of human C-reactive protein on nitric oxide bioavailability.
Circulating concentrations of the sensitive inflammatory marker C-reactive protein (CRP) predict future cardiovascular events, and CRP is elevated during sepsis and inflammation, when vascular reactivity may be modulated. We therefore investigated the direct effect of CRP on vascular reactivity. The effects of isolated, pure human CRP on vasoreactivity and protein expression were studied in vascular rings and cells in vitro, and effects on blood pressure were studied in rats in vivo. The temporal relationship between changes in CRP concentration and brachial flow-mediated dilation was also studied in humans after vaccination with Salmonella typhi capsular polysaccharide, a model of inflammatory endothelial dysfunction. In contrast to some previous reports, highly purified and well-characterized human CRP specifically induced hyporeactivity to phenylephrine in rings of human internal mammary artery and rat aorta that was mediated through physiological antagonism by nitric oxide (NO). CRP did not alter endothelial NO synthase protein expression but increased protein expression of GTP cyclohydrolase-1, the rate-limiting enzyme in the synthesis of tetrahydrobiopterin, the NO synthase cofactor. In the vaccine model of inflammatory endothelial dysfunction in humans, increased CRP concentration coincided with the resolution rather than the development of endothelial dysfunction, consistent with the vitro findings; however, administration of human CRP to rats had no effect on blood pressure. Pure human CRP has specific, direct effects on vascular function in vitro via increased NO production; however, further clarification of the effect, if any, of CRP on vascular reactivity in humans in vivo will require clinical studies using specific inhibitors of CRP.


Multiple factors related to bottle-feeding performance in preterm infants.
A great deal of attention has focused on understanding preterm infant feeding behaviors and on strategies to support the preterm infant during this period; however, comprehensive descriptions of the feeding behavior of preterm infants that incorporate an examination of multiple subsystem levels are lacking. To examine various physical indicators related to preterm infants' bottle-feeding performance. This was a retrospective, descriptive, exploratory study using a convenience sample. Medical records of 116 preterm infants were reviewed from the initiation of bottle-feeding until discharge from the neonatal intensive care unit. This study examined bottle-feeding performance (volume intake in milliliters per minute) as well as postmenstrual age, weight at each observed feed, oral motor skills, signs of distress, feeding techniques, feeding experience, gender, and Apgar scores at 5 minutes. Oral motor skills were measured by the Neonatal Oral Motor Assessment Scale. Linear mixed-effects models were used to examine the relationship between bottle-feeding performance and the remaining variables. Postmenstrual age, weight at each observed feed, oral motor skills, feeding experience, and feeding techniques were found to be significant predictors of feeding performance at the .05 level. Multiple factors, both intrinsic and extrinsic, play a role in determining an infant's bottle-feeding performance. In addition to age and weight, the presently employed conventional criteria, oral motor skills, feeding practice, and feeding techniques also contribute to infants' feeding performance. Arbitrary age (34 weeks gestational age) and weight criteria (1,500 g) should not be the only indicators for oral feeding.


Coping by metaphors: the versatile function of metaphors in blogs about living with advanced cancer.
Living with a life-limiting cancer illness can entail a turmoil of feelings such as constant fear of loss, suffering and dying. Because patients live longer with life-limiting illness, there is a need for enhanced understanding of how people make sense of and cope with the complicated aspects that this life situation brings on. In this article, we explore how bloggers with advanced cancer use metaphors as ways of making sense of their experiences. Our study is theoretically grounded in Conceptual Metaphor Theory, where metaphors are seen as a powerful phenomenon that both reflects and affects our thinking. The data consist of a corpus of blogs written in Swedish by individuals with advanced cancer, and the findings from our linguistic metaphor analysis are consistently interpreted against the backdrop of literature on coping. Our study thus highlights the intersection of linguistic metaphor analysis and psychological theories of coping by illustrating the many and complex functions metaphors can have as part of sense-making processes. Our hermeneutic approach enables us to show some differences among the three most pervasive metaphor domains in our material, battle, journey and imprisonment: the journey and imprisonment domains are more flexible than the battle domain in terms of the different kinds of coping strategies that are actualised by the bloggers' use of metaphors. One particular finding from our analysis is the way in which the bloggers make use of metaphors to compartmentalise experiences and emotions. Our contention is that careful attention to the metaphors used by patients can improve communication in healthcare and enhance understanding of the complex role language use plays in coping processes more generally. By highlighting the relation between metaphor use and coping, our analysis also provides a way to discuss coping strategies based on the patient's own use of language.


Evaluation of revised IPSS cytogenetic risk stratification and prognostic impact of monosomal karyotype in 783 patients with primary myelodysplastic syndromes.
Cytogenetic classification by the revised international prognostic scoring system (IPSS-R) and the prognostic value of monosomal karyotype (MK) were assessed in 783 patients with primary myelodysplastic syndromes (MDS). At 22 months median follow-up, 562 (72%) deaths were recorded. Percentages of patients with IPSS-R "very good," "good," "intermediate," "poor," and "very poor" cytogenetic categories was 5, 63, 18, 4, and 10%, respectively. The corresponding median survivals were 21, 40, 24, 18, and 6.5 months and the inter-group differences (good vs. very good/intermediate/poor vs. very poor; P < 0.01) or similarities (very good vs. intermediate vs. poor; P = 0.79) were not significantly modified in multivariable analysis. Results were similar when analysis was restricted to 602 patients managed by supportive care. MK adversely affected survival in both poor and very poor karyotype groups (P < 0.01). In conclusion, we were unable to confirm the prognostic superiority of IPSS-R-very good karyotype or prognostically distinguish between very good, intermediate and poor karyotypes. Furthermore, we show additional prognostic information from MK in poor/very poor karyotype.


The role of aquaporin 3 in teleost fish.
The aquaporin isoform, AQP3 has now been identified in a number of different teleost fish species, with additional DNA sequence information on AQP3 genes in further fish species available in genome databases. In zebrafish (Danio rerio), the AQP3 gene is present as two duplicate isoforms resulting from a teleostean fish genome-wide duplication. A further splicoform/isoform has also been identified in rainbow trout (Oncorhynchus mykiss). The identification of these AQP3 isoforms in other fish species is consequently explored. The role of AQP3 in physiological/osmoregulatory processes, in various teleost organs is then described. In teleost gill, AQP3 is expressed in 'chloride' cells, and in some species, in other epithelial cell types, where it may have a number of different functions including the prevention of dehydration. In eel esophagus, immunohistochemistry shows that AQP3 is expressed in surface epithelial cells in the anterior esophagus, but in mucus cells within the epithelium of the posterior esophagus. In eel intestine, AQP3 is found in macrophage-like cells and probably plays no part in osmoregulatory processes. In the rectum, as in the posterior esophagus AQP3 is expressed in mucus cells. In eel kidney, AQP3 is expressed in a subset of renal tubules, and localizes to the apical pole of tubule cells. There is no apparent change in the location or protein abundance of renal AQP3 following the acclimation of eels from freshwater to seawater.


Tissue-specific expressed antibody variable gene repertoires.
Recent developments in genetic technologies allow deep analysis of the sequence diversity of immune repertoires, but little work has been reported on the architecture of immune repertoires in mucosal tissues. Antibodies are the key to prevention of infections at the mucosal surface, but it is currently unclear whether the B cell repertoire at mucosal surfaces reflects the dominant antibodies found in the systemic compartment or whether mucosal tissues harbor unique repertoires. We examined the expressed antibody variable gene repertoires from 10 different human tissues using RNA samples derived from a large number of individuals. The results revealed that mucosal tissues such as stomach, intestine and lung possess unique antibody gene repertoires that differed substantially from those found in lymphoid tissues or peripheral blood. Mutation frequency analysis of mucosal tissue repertoires revealed that they were highly mutated, with little evidence for the presence of naïve B cells, in contrast to blood. Mucosal tissue repertoires possessed longer heavy chain complementarity determining region 3 loops than lymphoid tissue repertoires. We also noted a large increase in frequency of both insertions and deletions in the small intestine antibody repertoire. These data suggest that mucosal immune repertoires are distinct in many ways from the systemic compartment.


RNAi functions in cultured mammalian neurons.
In a wide range of organisms, double-stranded RNA triggers posttranscriptional gene silencing or RNA interference (RNAi). Small interfering RNAs, the 21-nt double-stranded RNA intermediates of this natural pathway, have became a powerful tool to knock down specific gene expression in mammalian cell lines and potentially will be useful for the analysis of loss-of-function phenotypes. In mammalian primary neuronal cultures, where genetic manipulations are especially difficult, RNAi might be developed into a highly efficacious tool to study the roles of specific genes in neuron development and functioning. Neurons, however, have been considered the most resistant to RNAi. We report here an application of RNAi to postmitotic primary neuronal cultures. Synthetic siRNA can be readily introduced into neurons and effectively inhibit the expression of endogenous and transfected genes.


Defining background DNA levels found on the skin of children aged 0-5 years.
There are currently no data available regarding the normal levels of DNA found on the skin of children engaging in routine day to day activities to assist with the forensic interpretation of DNA profiles generated from skin surface swabs. To address this deficit, skin surface swab samples were collected from 12 face/neck sites and 20 body sites on 50 children less than 5 years old. After exclusion of spoilt samples, 60 sets of swabs from 47 children (30 face/neck, 30 body) comprising of 944 individual samples were analysed. The number of alleles observed which could have originated from the child and the number which must have come from another source (non-child) were analysed. The following variables were evaluated: age, kissing, feeding and washing practices, number of contacts and application of cream. Overall, extremely small amounts of non-child DNA were retrieved from skin swabs. Child only (46.3%) or no DNA at all (18.6%) was observed for 64.9% of all swabbed samples. Low levels of non-child DNA (1-5 alleles) were observed on 31.6% of all swabs tested with only 3.4% of swabs showing six or more alleles. A great deal of variation between children and between sites in the levels of both child DNA and non-child DNA was observed. A multilevel model, taking account of clustering within children, showed that there was a strong direct association between the amounts of child and non-child DNA observed. There was no relationship between the amount of DNA recovered and the demographic and biographic variables analysed. These background data have the potential to assist the analysis of DNA from the skin of children during criminal investigation.


CodingQuarry: highly accurate hidden Markov model gene prediction in fungal genomes using RNA-seq transcripts.
The impact of gene annotation quality on functional and comparative genomics makes gene prediction an important process, particularly in non-model species, including many fungi. Sets of homologous protein sequences are rarely complete with respect to the fungal species of interest and are often small or unreliable, especially when closely related species have not been sequenced or annotated in detail. In these cases, protein homology-based evidence fails to correctly annotate many genes, or significantly improve ab initio predictions. Generalised hidden Markov models (GHMM) have proven to be invaluable tools in gene annotation and, recently, RNA-seq has emerged as a cost-effective means to significantly improve the quality of automated gene annotation. As these methods do not require sets of homologous proteins, improving gene prediction from these resources is of benefit to fungal researchers. While many pipelines now incorporate RNA-seq data in training GHMMs, there has been relatively little investigation into additionally combining RNA-seq data at the point of prediction, and room for improvement in this area motivates this study. CodingQuarry is a highly accurate, self-training GHMM fungal gene predictor designed to work with assembled, aligned RNA-seq transcripts. RNA-seq data informs annotations both during gene-model training and in prediction. Our approach capitalises on the high quality of fungal transcript assemblies by incorporating predictions made directly from transcript sequences. Correct predictions are made despite transcript assembly problems, including those caused by overlap between the transcripts of adjacent gene loci. Stringent benchmarking against high-confidence annotation subsets showed CodingQuarry predicted 91.3% of Schizosaccharomyces pombe genes and 90.4% of Saccharomyces cerevisiae genes perfectly. These results are 4-5% better than those of AUGUSTUS, the next best performing RNA-seq driven gene predictor tested. Comparisons against whole genome Sc. pombe and S. cerevisiae annotations further substantiate a 4-5% improvement in the number of correctly predicted genes. We demonstrate the success of a novel method of incorporating RNA-seq data into GHMM fungal gene prediction. This shows that a high quality annotation can be achieved without relying on protein homology or a training set of genes. CodingQuarry is freely available ( https://sourceforge.net/projects/codingquarry/ ), and suitable for incorporation into genome annotation pipelines.


Echinacea purpurea and Allium sativum as immunostimulants in fish culture using Nile tilapia (Oreochromis niloticus).
The study was conducted to evaluate the efficiency of echinacea (E) and garlic (G) supplemented diets as immunostimulant for tilapia (Oreochromis niloticus). Seven treatments were designed including a control (C). Fish were fed on 35% protein diet at a rate of 3% body weight per day. Echinacea (1.0 ppt) and garlic (3%) were incorporated in the feed, which was administered for periods of 1, 2 and 3 months (summer season), followed by basal diet for 4 more months (winter season). Neutrophil adherence and haematocrit values increased in both supplemented groups with prolonging period of application. The neutrophils adherence was significantly increased in all treatments except group administered echinacea for 1 month. The lymphocytic counts were significantly (p < 0.004) elevated that resulted in a significant increase in the total leucocytic count in groups administered echinacea for 1 and 2 months when compared with the control and/or other treatments. The gain in the body weight and specific growth rate was significantly increased in all supplemented groups (p < 0.004) during summer, but remained without any significant increase after winter. The survival rate was significantly high (>85%) in all the supplemented groups. The percentage of protection, after challenge infection using pathogenic Aeromonas hydrophila was the highest in groups supplemented with echinacea and garlic for 3 months after summer and winter seasons. It could be concluded that echinacea and garlic improve the gain in body weight, survival rate and resistance against challenge infection. Both compounds showed extended effects after withdrawal and improved resistance to cold stress during the winter season. However, a full commercial cost benefit analysis is necessary before recommending their application in aquaculture.


Altered Value Coding in the Ventromedial Prefrontal Cortex in Healthy Older Adults.
Previous work suggests that aging is associated with changes in risk taking but less is known about their underlying neural basis, such as the potential age differences in the neural processing of value and risk. The goal of the present study was to investigate adult age differences in functional neural responses in a naturalistic risk-taking task. Twenty-six young adults and 27 healthy older adults completed the Balloon Analogue Risk Task while undergoing functional magnetic resonance imaging. Young and older adults showed similar overt risk-taking behavior. Group comparison of neural activity in response to risky vs. control stimuli revealed similar patterns of activation in the bilateral striatum, anterior insula (AI) and ventromedial prefrontal cortex (vmPFC). Group comparison of parametrically modulated activity in response to continued pumping similarly revealed comparable results for both age groups in the AI and, potentially, the striatum, yet differences emerged for regional activity in the vmPFC. At whole brain level, insular, striatal and vmPFC activation was predictive of behavioral risk taking for young but not older adults. The current results are interpreted and discussed as preserved neural tracking of risk and reward in the AI and striatum, respectively, but altered value coding in the vmPFC in the two age groups. The latter finding points toward older adults exhibiting differential vmPFC-related integration and value coding. Furthermore, neural activation holds differential predictive validity for behavioral risk taking in young and older adults.


Characterization of VCC-1, a Novel Ambler Class A Carbapenemase from Vibrio cholerae Isolated from Imported Retail Shrimp Sold in Canada.
One of the core goals of the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) is to monitor major meat commodities for antimicrobial resistance. Targeted studies with methodologies based on core surveillance protocols are used to examine other foods, e.g., seafood, for antimicrobial resistance to detect resistances of concern to public health. Here we report the discovery of a novel Ambler class A carbapenemase that was identified in a nontoxigenic strain of Vibrio cholerae (N14-02106) isolated from shrimp that was sold for human consumption in Canada. V. cholerae N14-02106 was resistant to penicillins, carbapenems, and monobactam antibiotics; however, PCR did not detect common β-lactamases. Bioinformatic analysis of the whole-genome sequence of V. cholerae N14-02106 revealed on the large chromosome a novel carbapenemase (referred to here as VCC-1, for Vibrio cholerae carbapenemase 1) with sequence similarity to class A enzymes. Two copies of blaVCC-1 separated and flanked by ISVch9 (i.e., 3 copies of ISVch9) were found in an acquired 8.5-kb region inserted into a VrgG family protein gene. Cloned blaVCC-1 conferred a β-lactam resistance profile similar to that in V. cholerae N14-02106 when it was transformed into a susceptible laboratory strain of Escherichia coli. Purified VCC-1 was found to hydrolyze penicillins, 1st-generation cephalosporins, aztreonam, and carbapenems, whereas 2nd- and 3rd-generation cephalosporins were poor substrates. Using nitrocefin as a reporter substrate, VCC-1 was moderately inhibited by clavulanic acid and tazobactam but not EDTA. In this report, we present the discovery of a novel class A carbapenemase from the food supply.


What Are People Tweeting About Zika? An Exploratory Study Concerning Its Symptoms, Treatment, Transmission, and Prevention.
In order to harness what people are tweeting about Zika, there needs to be a computational framework that leverages machine learning techniques to recognize relevant Zika tweets and, further, categorize these into disease-specific categories to address specific societal concerns related to the prevention, transmission, symptoms, and treatment of Zika virus. The purpose of this study was to determine the relevancy of the tweets and what people were tweeting about the 4 disease characteristics of Zika: symptoms, transmission, prevention, and treatment. A combination of natural language processing and machine learning techniques was used to determine what people were tweeting about Zika. Specifically, a two-stage classifier system was built to find relevant tweets about Zika, and then the tweets were categorized into 4 disease categories. Tweets in each disease category were then examined using latent Dirichlet allocation (LDA) to determine the 5 main tweet topics for each disease characteristic. Over 4 months, 1,234,605 tweets were collected. The number of tweets by males and females was similar (28.47% [351,453/1,234,605] and 23.02% [284,207/1,234,605], respectively). The classifier performed well on the training and test data for relevancy (F1 score=0.87 and 0.99, respectively) and disease characteristics (F1 score=0.79 and 0.90, respectively). Five topics for each category were found and discussed, with a focus on the symptoms category. We demonstrate how categories of discussion on Twitter about an epidemic can be discovered so that public health officials can understand specific societal concerns within the disease-specific categories. Our two-stage classifier was able to identify relevant tweets to enable more specific analysis, including the specific aspects of Zika that were being discussed as well as misinformation being expressed. Future studies can capture sentiments and opinions on epidemic outbreaks like Zika virus in real time, which will likely inform efforts to educate the public at large.


Early developmental bisphenol-A exposure sex-independently impairs spatial memory by remodeling hippocampal dendritic architecture and synaptic transmission in rats.
Bisphenol-A (BPA, 4, 4'-isopropylidene-2-diphenol), a synthetic xenoestrogen that widely used in the production of polycarbonate plastics, has been reported to impair hippocampal development and function. Our previous study has shown that BPA exposure impairs Sprague-Dawley (SD) male hippocampal dendritic spine outgrowth. In this study, the sex-effect of chronic BPA exposure on spatial memory in SD male and female rats and the related synaptic mechanism were further investigated. We found that chronic BPA exposure impaired spatial memory in both SD male and female rats, suggesting a dysfunction of hippocampus without gender-specific effect. Further investigation indicated that BPA exposure causes significant impairment of dendrite and spine structure, manifested as decreased dendritic complexity, dendritic spine density and percentage of mushroom shaped spines in hippocampal CA1 and dentate gyrus (DG) neurons. Furthermore, a significant reduction in Arc expression was detected upon BPA exposure. Strikingly, BPA exposure significantly increased the mIPSC amplitude without altering the mEPSC amplitude or frequency, accompanied by increased GABAARβ2/3 on postsynaptic membrane in cultured CA1 neurons. In summary, our study indicated that Arc, together with the increased surface GABAARβ2/3, contributed to BPA induced spatial memory deficits, providing a novel molecular basis for BPA achieved brain impairment.


HGVbaseG2P: a central genetic association database.
The Human Genome Variation database of Genotype to Phenotype information (HGVbaseG2P) is a new central database for summary-level findings produced by human genetic association studies, both large and small. Such a database is needed so that researchers have an easy way to access all the available association study data relevant to their genes, genome regions or diseases of interest. Such a depository will allow true positive signals to be more readily distinguished from false positives (type I error) that fail to consistently replicate. In this paper we describe how HGVbaseG2P has been constructed, and how its data are gathered and organized. We present a range of user-friendly but powerful website tools for searching, browsing and visualizing G2P study findings. HGVbaseG2P is available at http://www.hgvbaseg2p.org.


Analysis of DNA Origami Nanostructures Using Capillary Electrophoresis.
DNA origami nanostructures are engineered nanomaterials (ENMs) that possess significant customizability, biocompatibility, and tunable structural and functional properties, making them potentially useful materials in fields, such as medicine, biocomputing, biomedical engineering, and measurement science. Despite the potential of DNA origami as a functional nanomaterial, a major barrier to its applicability is the difficulty associated with obtaining pure, well-folded structures. Therefore, rapid methods of analysis to ensure purity are needed to support the rapid development of this class of nanomaterials. Here, we present the development of capillary electrophoresis (CE) as an analytical tool for DNA origami. CE was investigated under both capillary zone electrophoresis (CZE) and capillary transient isotachophoresis (ctITP) modes. Optimization of both systems yielded baseline resolved separations of folded DNA origami nanostructures from excess staple strands. The ctITP separation mode demonstrated superior performance in terms of peak resolution (Rs = 2.05 ± 0.3), peak efficiency (N = 12,200 ± 230), and peak symmetry (As = 1.29 ± 0.032). The SYBR family dyes (Gold, Green I, and Green II) were investigated as highly efficient, noncovalent fluorophores for on-column labeling of DNA origami and detection using laser-induced fluorescence. Finally, ctITP analysis conditions were also applied to DNA origami nanostructures with different shapes and for the differentiation of DNA origami aggregates.


Estradiol Protects Proopiomelanocortin Neurons Against Insulin Resistance.
Insulin resistance is at the core of the metabolic syndrome, and men exhibit a higher incidence of metabolic syndrome than women in early adult life, but this sex advantage diminishes sharply when women reach the postmenopausal state. Because 17β-estradiol (E2) augments the excitability of the anorexigenic proopiomelanocortin (POMC) neurons, we investigated the neuroprotective effects of E2 against insulin resistance in POMC neurons from diet-induced obese (DIO) female and male mice. The efficacy of insulin to activate canonical transient receptor potential 5 (TRPC5) channels and depolarize POMC neurons was significantly reduced in DIO male mice but not in DIO female mice. However, the insulin response in POMC neurons was abrogated in ovariectomized DIO females but restored with E2 replacement. E2 increased T-type calcium channel Cav3.1 messenger RNA (mRNA) expression and whole-cell currents but downregulated stromal-interaction molecule 1 mRNA, which rendered POMC neurons more excitable and responsive to insulin-mediated TRPC5 channel activation. Moreover, E2 prevented the increase in suppressor of cytokine signaling-3 mRNA expression with DIO as seen in DIO males. As proof of principle, insulin [intracerebroventricular injection into the third ventricle (ICV)] decreased food intake and increased metabolism in female but not male guinea pigs fed a high-fat diet. The uncoupling of the insulin receptor from its downstream effector system was corroborated by the reduced expression of phosphorylated protein kinase B in the arcuate nucleus of male but not female guinea pigs following insulin. Therefore, E2 protects female POMC neurons from insulin resistance by enhancing POMC neuronal excitability and the coupling of insulin receptor to TRPC5 channel activation.


Differences in chemical composition of soil organic carbon resulting from long-term fertilization strategies.
Chemical composition of soil organic carbon (SOC) is central to soil fertility. We hypothesize that change in SOC content resulting from various long-term fertilization strategies accompanies the shift in SOC chemical structure. This study examined the effect of fertilization strategies along with the time of fertilizer application on the SOC composition by 13C nuclear magnetic resonance (NMR) spectroscopy. The soils (Aquic Inceptisol) subjected to seven fertilizer treatments were collected in 1989, 1999 and 2009, representing 0, 10 and 20 years of fertilization, respectively. The seven fertilizer treatments were (1-3) balanced fertilization with application of nitrogen (N), phosphorus (P) and potassium (K) including organic compost (OM), half organic compost plus half chemical fertilizer (1/2OM), and pure chemical NPK fertilizer (NPK); (4-6) unbalanced chemical fertilization without application of one of the major elements including NP fertilizer (NP), PK fertilizer (PK), and NK fertilizer (NK); and (7) an unamended control (CK). The SOC content in the balanced fertilization treatments were 2.3-52.6% and 9.4-64.6% higher than in the unbalanced fertilization/CK treatments in 1999 and 2009, respectively, indicating significant differences in SOC content with time of fertilizer application between the two treatment groups. There was a significantly greater proportion of O-alkyl C and a lower proportion of aromatic C in the balanced fertilization than in unbalanced fertilization/CK treatments in 1999, but not in 2009, because their proportions in the former treatments approached the latter in 2009. Principal component analysis further showed that the C functional groups from various fertilization strategies tended to become compositionally similar with time. The results suggest that a shift in SOC chemical composition may be firstly dominated by fertilization strategies, followed by fertilization duration.


UniProt: a worldwide hub of protein knowledge.
The UniProt Knowledgebase is a collection of sequences and annotations for over 120 million proteins across all branches of life. Detailed annotations extracted from the literature by expert curators have been collected for over half a million of these proteins. These annotations are supplemented by annotations provided by rule based automated systems, and those imported from other resources. In this article we describe significant updates that we have made over the last 2 years to the resource. We have greatly expanded the number of Reference Proteomes that we provide and in particular we have focussed on improving the number of viral Reference Proteomes. The UniProt website has been augmented with new data visualizations for the subcellular localization of proteins as well as their structure and interactions. UniProt resources are available under a CC-BY (4.0) license via the web at https://www.uniprot.org/.


Switchable Plasmonic Metasurfaces with High Chromaticity Containing Only Abundant Metals.
Plasmonic color generation offers several advantages but is also limited by the cost and availability of noble metals like gold. In this work, we present color-tunable metasurfaces with high chromaticity and reflectivity consisting of an aluminum mirror, a dielectric spacer, and a plasmonic nanohole array in copper. Copper is shown to be an excellent alternative to gold when properly protected from oxidation and makes it possible to generate a wide RGB gamut covering 27% of the standard RGB. By patterning the metasurfaces into microscale pixel triplets, color photos can be well reproduced with high resolution over wafer-sized areas. Further, we demonstrate active modulation of the reflected intensity using an electrochromic conductive polymer deposited on top of the nanostructures by screen printing. This technology opens up for ultrathin and flexible reflective displays in full color, that is, plasmonic electronic paper, compatible with large-scale sustainable production.


Benchmark data set for in silico prediction of Ames mutagenicity.
Up to now, publicly available data sets to build and evaluate Ames mutagenicity prediction tools have been very limited in terms of size and chemical space covered. In this report we describe a new unique public Ames mutagenicity data set comprising about 6500 nonconfidential compounds (available as SMILES strings and SDF) together with their biological activity. Three commercial tools (DEREK, MultiCASE, and an off-the-shelf Bayesian machine learner in Pipeline Pilot) are compared with four noncommercial machine learning implementations (Support Vector Machines, Random Forests, k-Nearest Neighbors, and Gaussian Processes) on the new benchmark data set.


Neighborhood environments and objectively measured physical activity in 11 countries.
Environmental changes are potentially effective population-level physical activity (PA) promotion strategies. However, robust multisite evidence to guide international action for developing activity-supportive environments is lacking. We estimated pooled associations of perceived environmental attributes with objectively measured PA outcomes, between-site differences in such associations, and the extent to which perceived environmental attributes explain between-site differences in PA. This was a cross-sectional study conducted in 16 cities located in Belgium, Brazil, Colombia, Czech Republic, Denmark, China, Mexico, New Zealand, Spain, United Kingdom, and United States of America. Participants were 6968 adults residing in administrative units stratified by socioeconomic status and transport-related walkability. Predictors were 10 perceived neighborhood environmental attributes. Outcome measures were accelerometry-assessed weekly minutes of moderate-to-vigorous PA (MVPA) and meeting the PA guidelines for cancer/weight gain prevention (420 min·wk of MVPA). Most perceived neighborhood attributes were positively associated with the PA outcomes in the pooled, site-adjusted, single-predictor models. Associations were generalizable across geographical locations. Aesthetics and land use mix-access were significant predictors of both PA outcomes in the fully adjusted models. Environmental attributes accounted for within-site variability in MVPA, corresponding to an SD of 3 min·d or 21 min·wk. Large between-site differences in PA outcomes were observed; 15.9%-16.8% of these differences were explained by perceived environmental attributes. All neighborhood attributes were associated with between-site differences in the total effects of the perceived environment on PA outcomes. Residents' perceptions of neighborhood attributes that facilitate walking were positively associated with objectively measured MVPA and meeting the guidelines for cancer/weight gain prevention at the within- and between-site levels. Associations were similar across study sites, lending support for international recommendations for designing PA-friendly built environments.


Inadequate environment, resources and values lead to missed nursing care: A focused ethnographic study on the surgical ward using the Fundamentals of Care framework.
To explore the delivery of care from the perspective of patients with acute abdominal pain focusing on the contextual factors at system level using the Fundamentals of Care framework. The Fundamentals of Care framework describes several contextual and systemic factors that can impact the delivery of care. To deliver high-quality, person-centred care, it is important to understand how these factors affect patients' experiences and care needs. A focused ethnographic approach. A total of 20 observations were performed on two surgical wards at a Swedish university hospital. Data were collected using participant observation and informal interviews and analysed using deductive content analysis. The findings, presented in four categories, reflect the value patients place on the caring relationship and a friendly atmosphere on the ward. Patients had concerns about the environment, particularly the high-tempo culture on the ward and its impact on their integrity, rest and sleep, access to information and planning, and need for support in addressing their existential thoughts. The observers also noted that missed nursing care had serious consequences for patient safety. Patients with acute abdominal pain were cared for in the high-tempo culture of a surgical ward with limited resources, unclear leadership and challenges to patients' safety. The findings highlight the crucial importance of prioritising and valuing the patients' fundamental care needs for recovery. Nursing leaders and nurses need to take the lead to reconceptualise the value of fundamental care in the acute care setting. To improve clinical practice, the value of fundamentals of care must be addressed regardless of patient's clinical condition. Providing a caring relationship is paramount to ensure a positive impact on patient's well-being and recovery.


Initial stage growth of GexSi1-x layers and Ge quantum dot formation on GexSi1-x surface by MBE.
Critical thicknesses of two-dimensional to three-dimensional growth in GexSi1-x layers were measured as a function of composition for different growth temperatures. In addition to the (2 × 1) superstructure for a Ge film grown on Si(100), the GexSi1-x layers are characterized by the formation of (2 × n) reconstruction. We measured n for all layers of Ge/GexSi1-x/Ge heterosystem using our software with respect to the video recording of reflection high-energy electron diffraction (RHEED) pattern during growth. The n reaches a minimum value of about 8 for clear Ge layer, whereas for GexSi1-x films, n is increased from 8 to 14. The presence of a thin strained film of the GexSi1-x caused not only the changes in critical thicknesses of the transitions, but also affected the properties of the germanium nanocluster array for the top Ge layer. Based on the RHEED data, the hut-like island form, which has not been previously observed by us between the hut and dome islands, has been detected. Data on the growth of Ge/GexSi1-x/Ge heterostructures with the uniform array of islands in the second layer of the Ge film have been received.


Epidemic transmission of human immunodeficiency virus in renal dialysis centers in Egypt.
In 1993 an epidemic of human immunodeficiency virus (HIV) infection occurred among 39 patients at 2 renal dialysis centers in Egypt. The centers, private center A (PCA) and university center A (UCA) were visited, HIV-infected patients were interviewed, seroconversion rates at UCA were calculated, and relatedness of HIV strains was determined by sequence analysis; 34 (62%) of 55 patients from UCA and 5 (42%) of 12 patients from PCA were HIV-infected. The HIV seroconversion risk at UCA varied significantly with day and shift of dialysis session. Practices that resulted in sharing of syringes among patients were observed at both centers. The analyzed V3 loop sequences of the HIV strain of 12 outbreak patients were >96% related to each other. V3 loop sequences from each of 8 HIV-infected Egyptians unrelated to the 1993 epidemic were only 76%-89% related to those from outbreak strains. Dialysis patients may be at risk for HIV infection if infection control guidelines are not followed.


Effects of Lifelong Ethanol Consumption on Brain Monoamine Transmitters in Alcohol-Preferring Alko Alcohol (AA) Rats.
The purpose of the present study was to examine the combined effects of aging and lifelong ethanol exposure on the levels of monoamine neurotransmitters in different regions of the brain. This work is part of a project addressing interactions of aging and lifelong ethanol consumption in alcohol-preferring AA (Alko Alcohol) line of rats, selected for high voluntary consumption of ethanol. Intake of ethanol on the level of 4.5-5 g/kg/day for about 20 months induced only limited changes in the neurotransmitter levels; the concentration of noradrenaline was significantly reduced in the frontal cortex. There was also a trend towards lower levels of dopamine and 5-hydroxytryptamine (5-HT) in the frontal cortex, and towards a lower noradrenaline level in the dorsal cortex. Aging was associated with a decreased concentration of dopamine in the dorsal cortex and with a declining trend in the striatum. The levels of 5-HT in the limbic forebrain were higher in the aged than in the young animals, and in the striatum, there was a trend towards higher levels in older animals. The data suggest that a continuous intake of moderate amounts of ethanol does not enhance the age-related alterations in brain monoamine neurotransmission, while the decline in the brain level of dopamine associated with aging may be a factor contributing to age-related neurological disorders.


Association of maternal prenatal selenium concentration and preterm birth: a multicountry meta-analysis.
Selenium (Se), an essential trace mineral, has been implicated in preterm birth (PTB). We aimed to determine the association of maternal Se concentrations during pregnancy with PTB risk and gestational duration in a large number of samples collected from diverse populations. Gestational duration data and maternal plasma or serum samples of 9946 singleton live births were obtained from 17 geographically diverse study cohorts. Maternal Se concentrations were determined by inductively coupled plasma mass spectrometry analysis. The associations between maternal Se with PTB and gestational duration were analysed using logistic and linear regressions. The results were then combined using fixed-effect and random-effect meta-analysis. In all study samples, the Se concentrations followed a normal distribution with a mean of 93.8 ng/mL (SD: 28.5 ng/mL) but varied substantially across different sites. The fixed-effect meta-analysis across the 17 cohorts showed that Se was significantly associated with PTB and gestational duration with effect size estimates of an OR=0.95 (95% CI: 0.9 to 1.00) for PTB and 0.66 days (95% CI: 0.38 to 0.94) longer gestation per 15 ng/mL increase in Se concentration. However, there was a substantial heterogeneity among study cohorts and the random-effect meta-analysis did not achieve statistical significance. The largest effect sizes were observed in UK (Liverpool) cohort, and most significant associations were observed in samples from Malawi. While our study observed statistically significant associations between maternal Se concentration and PTB at some sites, this did not generalise across the entire cohort. Whether population-specific factors explain the heterogeneity of our findings warrants further investigation. Further evidence is needed to understand the biologic pathways, clinical efficacy and safety, before changes to antenatal nutritional recommendations for Se supplementation are considered.


Comparison of automated quantification and semiquantitative visual analysis findings of IQ SPECT MPI with conventional coronary angiography in patients with stable angina.
The aim of this study was to assess the validity of automated quantitative and semiquantitative visual analysis of total perfusion deficit (TPD) using the IQ SPECT gamma camera system compared to conventional coronary angiographically detected significant coronary artery disease (CAD). The study included patients with suspected CAD who underwent myocardial perfusion single photon emission computed tomography and conventional coronary angiography. The summed stress score (SSS), summed rest score (SRS), and summed difference score (SDS) (semiquantitative visual analysis results) were assessed using a 5-point scale in a standard 17-segment model, and TPD (stress, rest, and ischemic TPD) was quantified using automated software. In all, 84 patients (Group 1, those who underwent revascularization) had significant coronary artery lesions, and 81 (Group 2) had non-significant lesions. The median interquartile range values were: stress-TPD (sTPD): 16 (3.5- 33.5) vs 9.2 (2-17.9), rest-TPD: 9.4 (2.2-18.8) vs 4 (1-11), and 6.9 (1.9-14.1) vs 3.4 (1-6.1) for ischemic-TPD (iTPD) in Group 1 and Group 2, respectively. To detect ischemia, the optimal cut-off points were 9.5 (sensitivity: 75%, specificity; 60%) for sTPD, and 4.5 (sensitivity: 56%, specificity: 73%) for iTPD. There were significant correlations between quantitative and semi-quantitative methods in detection of significant coronary artery disease (sTPD-SSS: r=0.954, sTPD-SDS: r=0.746, iTPD-SSS: r=0.654, iTPD-SDS: r=0.759; p<0.05 for all). The quantitative analysis and summed stress scores produced by the IQ SPECT system appear to be a useful and valid method to detect significant CAD.


Yin yoga and mindfulness: a five week randomized controlled study evaluating the effects of the YOMI program on stress and worry.
The YOMI program is a psychoeducational training and physical practice-based program that bridges knowledge from evidence-based psychotherapy with the practice of mindfulness and yin yoga. It consists of 10 content-specific sessions and does not include home assignments. The primary purpose of this randomized controlled trial is to evaluate the effects of the five-week YOMI program on perceived stress, worry and mindfulness in a non-clinical sample. In this randomized controlled trial participants were assigned to two groups. Group 1 participated in the five-week intervention twice a week while Group 2 was assigned to a waiting-list condition and participated in the intervention after Group 1. All measures were administered through self-report questionnaires, conducted via a web-based program. The results of the study indicated significant effects of the YOMI program on decreasing stress and worry, and increasing mindfulness. Notably these changes were still present at five-week follow up. Consistent with the hypotheses, results suggested that the YOMI program established a group setting where individuals learned to use tools and methods to facilitate better self-directed practice. The study shows moderate to large effect sizes.


Characterization of Na+ and Ca2+ currents in bag cells of sexually immature aplysia californica
The neurosecretory bag cells of sexually mature Aplysia californica release egg-laying hormones as part of the reproductive process after a train of action potentials termed afterdischarge. Whole-cell voltage-clamp experiments were performed in cultured cells from sexually immature A. californica to characterize the inward voltage-gated currents for Na+ and Ca2+. The goal of these experiments was to investigate the regulation of excitability during sexual maturation. Na+ currents in bag cells of immature A. californica were similar in several ways to those of mature animals. The Na+ currents activated at voltages less negative than -30 mV and peaked at 10-20 mV in artificial sea water. The time course and pharmacology of bag cell Na+ currents were similar to those of bag cells from mature A. californica, although the Na+ current density was lower in immature A. californica. Na+ currents were inhibited by tetrodotoxin (50 nmol l-1). The Na+ current was relatively insensitive to depolarized holding potentials (Vh), maintaining approximately 50 % of peak current amplitude present at Vh=-70 mV throughout the activation range at Vh=-30 mV. In experiments using a 1 s depolarized Vh prior to a test pulse, the half-inactivation voltage (V1/2) was -27 mV. Recovery of immature Na+ current from steady-state inactivation at Vh=-30 mV had a time constant () of 9.5 ms, significantly slower than in mature animals. Ca2+ currents of immature A. californica activated at approximately -30 mV and peaked at approximately 20 mV with 11 mmol l-1 Ba2+ as the charge carrier. The principle differences from mature Ca2+ currents were the low density of the immature Ca2+ currents and their 'run-down' in whole-cell recordings. The pharmacology and V1/2 of bag cell Ca2+ currents were similar to those of L-type Ca2+ currents in mature cells. The Ca2+ currents were inhibited 61+/-10 % by nifedipine (10 micromol l-1) and were unaffected by -conotoxin GVIA (10 micromol l-1). The Ca2+ currents were relatively insensitive to depolarized Vh, activating maximally at Vh=-90, -70 and -50 mV, and maintaining 50 % of this peak current amplitude throughout the activation range at Vh=-30 mV. The V1/2 was -23 mV in experiments in which cells were subjected to a 1 s depolarized Vh prior to a test pulse. Na+ current amplitudes were maintained or increased during 1 min of 4 Hz test pulses in bag cells at Vh=-70 mV and Vh=-30 mV. In contrast, Ca2+ current run-down occurred during 1 min of 4 Hz test pulses in seven of 10 cells at Vh=-70 mV and in 12 of 12 cells at Vh=-30 mV. The observed scarcity of Na+ and Ca2+ currents in immature bag cells as well as the specific characteristics of immature bag cell Ca2+ currents make repetitive action potential firing and hormone release less likely than in mature bag cells.


Legacy effects of grassland management on soil carbon to depth.
The importance of managing land to optimize carbon sequestration for climate change mitigation is widely recognized, with grasslands being identified as having the potential to sequester additional carbon. However, most soil carbon inventories only consider surface soils, and most large-scale surveys group ecosystems into broad habitats without considering management intensity. Consequently, little is known about the quantity of deep soil carbon and its sensitivity to management. From a nationwide survey of grassland soils to 1 m depth, we show that carbon in grassland soils is vulnerable to management and that these management effects can be detected to considerable depth down the soil profile, albeit at decreasing significance with depth. Carbon concentrations in soil decreased as management intensity increased, but greatest soil carbon stocks (accounting for bulk density differences), were at intermediate levels of management. Our study also highlights the considerable amounts of carbon in subsurface soil below 30 cm, which is missed by standard carbon inventories. We estimate grassland soil carbon in Great Britain to be 2097 Tg C to a depth of 1 m, with ~60% of this carbon being below 30 cm. Total stocks of soil carbon (t ha(-1) ) to 1 m depth were 10.7% greater at intermediate relative to intensive management, which equates to 10.1 t ha(-1) in surface soils (0-30 cm), and 13.7 t ha(-1) in soils from 30 to 100 cm depth. Our findings highlight the existence of substantial carbon stocks at depth in grassland soils that are sensitive to management. This is of high relevance globally, given the extent of land cover and large stocks of carbon held in temperate managed grasslands. Our findings have implications for the future management of grasslands for carbon storage and climate mitigation, and for global carbon models which do not currently account for changes in soil carbon to depth with management.


Dynamic self-representation of interdependent Chinese: The effect of bicultural experience.
The process of globalisation has increased the opportunity for Chinese individuals to utilise influences from other cultures that differ from their own collectivistic culture. The explorations of the impact of acculturation on Chinese individuals' self-construals have become intriguing. Thus, we recruited German-Chinese (N = 192) people who live in Germany and had direct bicultural experiences, as well as Mainland-Chinese (N = 192) people who live in Mainland China and had internalised a second culture through more indirect means of exposure. We investigated their accessibility to both interdependent and independent cultural frameworks using temporal self-construal priming. The results showed that priming effects were observed in both cultural groups. However, the self-construals of the German-Chinese participants were more flexible than those of Mainland-Chinese under self-construal priming. Also, the results suggested that women were easier to be influenced by direct bicultural experiences, due to their private and collective self were different between the two cultures. These findings provide evidence regarding the opinion that individuals could use influences from more than one culture. More importantly, these results suggest that the acculturation induced by direct bicultural experiences facilitates cultural frame switching.


The association between coffee intake and breast cancer risk: a meta-analysis and dose-response analysis using recent evidence.
To assess the association between coffee intake and breast cancer risk using data from studies published during the past 15 years. Articles published between January 2005 and May 2020 were collected from PubMed, Embase, and Web of Science databases, and the Cochrane library. Coffee consumption was set as the exposure factor of breast cancer risk, and relative risk (RR) was set as the assessment indicator. Random-effects or fixed-effects models were used for the meta-analysis, depending on the level of heterogeneity. The linear and non-linear dose-response relationship was assessed using the generalized least-squares method and restricted cubic spline model. Articles were evaluated by sensitivity analysis, and Begg's and Egger's tests and funnel plots were used to evaluate publication bias. The process of meta-analysis referred to Meta-analyses of Observational Studies in Epidemiology (MOOSE). We retrieved 26 relevant studies in the dose-response analysis and meta-analysis. A negative correlation was found between coffee consumption and breast cancer risk (RR: 0.95; 95% CI: 0.92-0.99). A linear and marginal dose-response relationship was found for six case-control studies (RR: 0.98; 95% CI: 0.95-1.00) and 10 cohort studies (RR: 0.98; 95% CI: 0.97-1.00). Subgroup analysis showed that the relationship between coffee consumption and breast cancer risk was moderated by menstrual status and geographic region. Overall, the meta-analysis found a negative correlation between coffee intake and breast cancer risk, especially in postmenopausal and European women.


Essential oil of Pterodon polygalaeflorus Benth attenuates nociception in mice.
Essential oils (EO) are volatile liquids responsible for the aroma of plants. Pterodon polygalaeflorus seeds have received widespread use in folk medicine for the treatment of inflammatory diseases. For this reason and because Pterodon polygalaeflorus seeds have great EO content, which is frequently pharmacologically active, the present study aimed to evaluate the antinociceptive effect of EO from Pterodon polygalaeflorus (EOPPgfl) and its acute toxic effects. The EEOPPgfl sample, which was extracted by steam distillation of the seeds, had a yield of 2.4% of the seeds weight and had, as major constituents, beta-elemene (48.19%), trans-caryophyllene (19.51%), and epi-bicyclosesquiphellandrene (12.24%). The EOPPgfl sample showed mild acute toxicity and its calculated median lethal dose (LD50) was 3.38 g/kg. EOPPgfl (20-60 mg/kg) showed antinociceptive activity as evidenced by several tests and inhibited writhing induced by acetic acid. The maximum effect was obtained with the 30 mg/kg dose and at 60 min after its administration. EOPPgfl also decreased formalin-induced nociception, as verified by the inhibition of the first and second phase of the formalin test. At 30 mg/kg, EOPPgfl also decreased thermally stimulated nociception. Nociception may be related to inflammatory and antiedematogenic activity and at doses ranging 10-100 mg/kg, EOPPgfl blocked dextran- and carrageenan-induced edema. The results demonstrated that EOPPgfl presented, at doses approximately 100 times smaller than LD50, an antinociceptive effect that probably was due to anti-inflammatory activities.


Diffusion of beta-lactam antibiotics through the porin channels of Escherichia coli K-12.
Diffusion rates of various beta-lactam antibiotics through the OmpF and OmpC porin channels of Escherichia coli K-12 were measured by the use of reconstituted proteoliposomes. The results can be interpreted on the basis of the gross physicochemical properties of the antibiotics along the following lines. (i) As noted previously (Nikaido et al., J. Bacteriol., 153:232-240, 1983), there was a monotonous dependence of the penetration rate on the hydrophobicity of the molecule among the classical monoanionic beta-lactams, and a 10-fold increase in the octanol-water partition coefficient of the uncharged molecule decreased the penetration rate by a factor of 5 to 6. (ii) Compounds with exceptionally bulky side chains, such as mezlocillin, piperacillin, and cefoperazone, showed much slower penetration rates than expected from their hydrophobicity. (iii) The substituted oxime side chain on the alpha-carbon of the substituent group at position 7 of the cephem nucleus decreased the penetration rate almost by an order of magnitude; this appears to be largely due to the steric effect. (iv) The presence of a methoxy group at position 7 of the cephalosporins also reduced the penetration rate by 20%, probably also due to the steric hindrance. (v) Zwitterionic compounds penetrated very rapidly, and the correlation between the rate and hydrophobicity appeared to be much weaker than with the monoanionic compounds. Imipenem showed the highest permeability among the compounds tested, presumably due, at least in part, to its compact molecular structure. (vi) Compounds with two negative charges penetrated more slowly than did analogs with only one negatively charged group. Among them, only moxalactam, ceftriaxone, and azthreonam showed penetration rates corresponding to, or higher than, 10% of that of imipenem.


Identifying gnostic predictors of the vaccine response.
Molecular predictors of the response to vaccination could transform vaccine development. They would allow larger numbers of vaccine candidates to be rapidly screened, shortening the development time for new vaccines. Gene-expression based predictors of vaccine response have shown early promise. However, a limitation of gene-expression based predictors is that they often fail to reveal the mechanistic basis of their ability to classify response. Linking predictive signatures to the function of their component genes would advance basic understanding of vaccine immunity and also improve the robustness of vaccine prediction. New analytic tools now allow more biological meaning to be extracted from predictive signatures. Functional genomic approaches to perturb gene expression in mammalian cells permit the function of predictive genes to be surveyed in highly parallel experiments. The challenge for vaccinologists is therefore to use these tools to embed mechanistic insights into predictors of vaccine response.


Characterization and application studies of ProxyPhos, a chemosensor for the detection of proximally phosphorylated peptides and proteins in aqueous solutions.
Proximal phosphorylation on proteins appears to have functional significance and has been associated with several diseases, including Alzheimer's and cancer. While much remains to be learned about the role of proximal phosphorylation in biological systems, no simple and/or affordable technique is available for its detection. To this end, we have previously developed a ProxyPhos chemosensor, which detects proximally phosphorylated peptides and proteins over mono- and non-phosphorylated motifs in aqueous solutions. In this follow-up work, we performed extensive characterization of peptide and protein ProxyPhos assay conditions to achieve enhanced detection, and further explored the selectivity of ProxyPhos, and its potential off-targets. As a result of characterization studies, selective sensing of proximally phosphorylated over mono-phosphorylated peptides and proteins was achieved. Moreover, studies demonstrated that ProxyPhos was compatible with the detection of all commonly phosphorylated residues (i.e. tyrosine, serine and threonine residues). Under optimized conditions, ProxyPhos efficiently discriminated between peptides derived from the activated (proximally phosphorylated, disease-relevant) and inactive (mono-phosphorylated) forms of JAK2, SYK and MAPK1 kinases. In addition, ProxyPhos can be used to probe phosphatase activity on peptides and proteins via detecting changes in proximal phosphorylation, demonstrating immediate utility of this chemosensing system.


PZP and PAI-2: Structurally-diverse, functionally similar pregnancy proteins?
Pregnancy zone protein (PZP) and plasminogen activator inhibitor type 2 (PAI-2) are two multifunctional proteins that are elevated in normal pregnancy and numerous other inflammatory states. Both proteins were originally identified as protease inhibitors, but current evidence supports the notion that they may also function as modulators of T-helper cells and/or extracellular chaperones. Exacerbated inflammation, fibrinolytic disturbances and misfolded proteins are all implicated in the pathology of preeclampsia, a leading cause of maternal and foetal mortality and morbidity. Notably, reduced levels of PZP or PAI-2 are associated with preeclampsia and clarification of their diverse functions in normal pregnancy could provide much needed insight regarding the pathogenesis of this disorder. Given that inflammation and protein misfolding underlie the pathology of a very large number of disorders, the contributions of PZP and PAI-2 to extracellular proteostasis and immunoregulation could be broad-reaching.


The C. elegans developmental timing protein LIN-42 regulates diapause in response to environmental cues.
Environmental conditions can have a major impact on developmental progression in animals. For example, when C. elegans larvae encounter harsh conditions they can reversibly halt the passage of developmental time by forming a long-lived dauer larva at the end of the second larval stage. Here, we show that the period homolog lin-42, known to control developmental time, also acts as a component of a switch that mediates dauer entry. Loss of lin-42 function renders animals hypersensitive to dauer formation under stressful conditions, whereas misexpression of lin-42 in the pre-dauer stage inhibits dauer formation, indicating that lin-42 acts as a negative regulator of this life history decision. These phenotypes place LIN-42 in opposition to the ligand-free form of the nuclear receptor DAF-12, which indirectly senses environmental conditions and helps to integrate external cues into developmental decisions. Mutations that impair DAF-12 ligand binding are exquisitely sensitive to the absence of lin-42, whereas overexpression of LIN-42 can suppress the dauer constitutive phenotype of a ligand-insensitive daf-12 mutant, suggesting that LIN-42 and DAF-12 are intimate partners in controlling the decision to become a dauer larva. The functional outputs of Period family proteins and nuclear receptors also converge in other organisms, suggesting that the relationship between lin-42 and daf-12 represents an ancient genetic framework for responding to environmental stimuli.


Centrosomes have a role in regulating the destruction of cyclin B in early Drosophila embryos.
We reported previously that the disappearance of cyclin B at the end of mitosis in early Drosophila embryos starts at centrosomes and spreads into the spindle [1]. Here, we used a novel mutation, centrosome fall off (cfo), to investigate whether centrosomes are required to initiate the disappearance of cyclin B from the spindle. In embryos laid by homozygous cfo mutant mothers, the centrosomes co-ordinately detached from the mitotic spindle during mitosis, and the centrosomeless spindles arrested at anaphase. Cyclin B levels decreased on the detached centrosomes, but not on the arrested centrosomeless spindles, presumably explaining why the spindles arrest in anaphase in these embryos. We found that the expression of a non-degradable cyclin B in embryos also caused an anaphase arrest, but most centrosomes remained attached to the arrested spindles, and non-degradable cyclin B levels remained high on both the centrosomes and spindles. These findings suggest that the disappearance of cyclin B from centrosomes and spindles is closely linked to its destruction, and that a connection between centrosomes and spindles is required for the proper destruction of the spindle-associated cyclin B in early Drosophila embryos. These results may have important implications for the mechanism of the spindle-assembly checkpoint, as they suggest that unattached kinetochores may arrest cells in mitosis, at least in part, by signalling to centrosomes to block the initiation of cyclin B destruction.