Lipopolysaccharide-reactive immunoglobulin E is associated with lower mortality and organ failure in traumatically injured patients.
Antilipopolysaccharide (anti-LPS) immunoglobulin G (IgG) and IgM have been associated with protection from LPS effects in vivo. We investigated the presence of IgE and anti-LPS in 32 patients that had experienced severe traumatic injury and in 35 healthy volunteers; we also investigated whether IgE anti-LPS was associated with important clinical events. Plasma samples were collected daily from patients in the intensive care unit and on one occasion from volunteers; the samples were assayed for IgE anti-LPS. IgE anti-LPS was assayed by enzyme-linked immunosorbent assay with monoclonal anti-human IgE as the capture antibody. Detection was accomplished with biotin-labeled LPS (Escherichia coli J5 mutant) followed by streptavidin-peroxidase with 2,2'-azino(3-ethylbenzthiazoline)sulfonic acid as the substrate. The assay was demonstrated to be specific for IgE and LPS-biotin by nonreactivity of control sera with high-titer anti-LPS IgG and IgM and by inhibition with unlabeled LPS. IgE anti-LPS was detected in 1 of 35 healthy controls (2.9%) and 25 of 32 traumatically injured patients (78%) (P < 0.001). The presence of IgE anti-LPS was associated with a lower incidence of death (P = 0.026) and of renal failure (P = 0.0012). There was no apparent temporal relationship between detection of IgE anti-LPS and clinical events. IgG anti-LPS was detected more frequently in patients that were positive for IgE anti-LPS (P = 0.06) but was not associated with clinical events. The inability to detect IgE anti-LPS may be related to adverse clinical events through depletion of specific IgE due to LPS exposure after trauma or through saturation of the assay by IgE with other specificities. We have reported increased total IgE concentrations in these patients. (J.T. DiPiro, R.G. Hamilton, T. R. Howdieshell, N. F. Adkinson, and A. R. Mansberger, Ann. Surg. 215:460-466, 1992).


Architectural adaptations of muscle to training and injury: a narrative review outlining the contributions by fascicle length, pennation angle and muscle thickness.
The architectural characteristics of muscle (fascicle length, pennation angle muscle thickness) respond to varying forms of stimuli (eg, training, immobilisation and injury). Architectural changes following injury are thought to occur in response to the restricted range of motion experienced during rehabilitation and the associated neuromuscular inhibition. However, it is unknown if these differences exist prior to injury, and had a role in injury occuring (prospectively), or if they occur in response to the incident itself (retrospectively). Considering that the structure of a muscle will influence how it functions, it is of interest to understand how these architectural variations may alter how a muscle acts with reference to the force-length and force-velocity relationships. Our narrative review provides an overview of muscle architectural adaptations to training and injury. Specifically, we (1) describe the methods used to measure muscle architecture; (2) detail the impact that architectural alterations following training interventions, immobilisation and injury have on force production and (3) present a hypothesis on how neuromuscular inhibition could cause maladaptations to muscle architecture following injury.


The Wnt signaling pathway effector TCF7L2 is upregulated by insulin and represses hepatic gluconeogenesis.
Certain single nucleotide polymorphisms (SNPs) in transcription factor 7-like 2 (TCF7L2) are strongly associated with the risk of type 2 diabetes. TCF7L2 and β-catenin (β-cat) form the bipartite transcription factor cat/TCF in stimulating Wnt target gene expression. cat/TCF may also mediate the effect of other signaling cascades, including that of cAMP and insulin in cell-type specific manners. As carriers of TCF7L2 type 2 diabetes risk SNPs demonstrated increased hepatic glucose production, we aimed to determine whether TCF7L2 expression is regulated by nutrient availability and whether TCF7L2 and Wnt regulate hepatic gluconeogenesis. We examined hepatic Wnt activity in the TOPGAL transgenic mouse, assessed hepatic TCF7L2 expression in mice upon feeding, determined the effect of insulin on TCF7L2 expression and β-cat Ser⁶⁷⁵ phosphorylation, and investigated the effect of Wnt activation and TCF7L2 knockdown on gluconeogenic gene expression and glucose production in hepatocytes. Wnt activity was observed in pericentral hepatocytes in the TOPGAL mouse, whereas TCF7L2 expression was detected in human and mouse hepatocytes. Insulin and feeding stimulated hepatic TCF7L2 expression in vitro and in vivo, respectively. In addition, insulin activated β-cat Ser⁶⁷⁵ phosphorylation. Wnt activation by intraperitoneal lithium injection repressed hepatic gluconeogenic gene expression in vivo, whereas lithium or Wnt-3a reduced gluconeogenic gene expression and glucose production in hepatic cells in vitro. Small interfering RNA-mediated TCF7L2 knockdown increased glucose production and gluconeogenic gene expression in cultured hepatocytes. These observations suggest that Wnt signaling and TCF7L2 are negative regulators of hepatic gluconeogenesis, and TCF7L2 is among the downstream effectors of insulin in hepatocytes.


Closing the mind's eye: deactivation of visual cortex related to auditory task difficulty.
Blood oxygen-level-dependent signal decreases relative to baseline (deactivations) can occur with stimulation of an opposing sensory modality. Here, we show the importance of the difficulty of an auditory task on the deactivation of visual cortical areas. Participants performed an auditory temporal-order judgment task in conjunction with sparse-sampling functional MRI at both moderate and high levels of difficulty (adjusted for each individual's own threshold). With moderate difficulty, small deactivations were observed not only in parietal and cingulate cortex, but occipital cortex as well. When the same task was more difficult, deactivations increased significantly to include a greater extent of functionally defined visual cortex. Together, these results suggest that cross-modal deactivations occur in compensation for task difficulty, perhaps acting as an intrinsic filter for nonrelevant information.


Dissociable fronto-striatal effects of dopamine D2 receptor stimulation on cognitive versus motor flexibility.
Genetic and pharmacological studies suggest an important role of the dopamine D2 receptor (DRD2) in flexible behavioral adaptation, mostly shown in reward-based learning paradigms. Recent evidence from imaging genetics indicates that also intentional cognitive flexibility, associated with lateral frontal cortex, is affected by variations in DRD2 signaling. In the present functional magnetic resonance imaging (MRI) study, we tested the effects of a direct pharmacological manipulation of DRD2 stimulation on intentional flexibility in a task-switching context, requiring switches between cognitive task rules and between response hands. In a double blind, counterbalanced design, participants received either a low dose of the DRD2 agonist bromocriptine or a placebo in two separate sessions. Bromocriptine modulated the blood-oxygen-level-dependent (BOLD) signal during rule switching: rule-switching-related activity in the left posterior lateral frontal cortex and in the striatum was increased compared to placebo, at comparable performance levels. Fronto-striatal connectivity under bromocriptine was slightly increased for rule switches compared to rule repetitions. Hand-switching-related activity, in contrast, was reduced under bromocriptine in sensorimotor regions. Our results provide converging evidence for an involvement of DRD2 signaling in fronto-striatal mechanisms underlying intentional flexibility, and indicate that the neural mechanisms underlying different types of flexibility (cognitive vs motor) are affected differently by increased dopaminergic stimulation.


Change in albuminuria and subsequent risk of end-stage kidney disease: an individual participant-level consortium meta-analysis of observational studies.
Change in albuminuria as a surrogate endpoint for progression of chronic kidney disease is strongly supported by biological plausibility, but empirical evidence to support its validity in epidemiological studies is lacking. We aimed to assess the consistency of the association between change in albuminuria and risk of end-stage kidney disease in a large individual participant-level meta-analysis of observational studies. In this meta-analysis, we collected individual-level data from eligible cohorts in the Chronic Kidney Disease Prognosis Consortium (CKD-PC) with data on serum creatinine and change in albuminuria and more than 50 events on outcomes of interest. Cohort data were eligible if participants were aged 18 years or older, they had a repeated measure of albuminuria during an elapsed period of 8 months to 4 years, subsequent end-stage kidney disease or mortality follow-up data, and the cohort was active during this consortium phase. We extracted participant-level data and quantified percentage change in albuminuria, measured as change in urine albumin-to-creatinine ratio (ACR) or urine protein-to-creatinine ratio (PCR), during baseline periods of 1, 2, and 3 years. Our primary outcome of interest was development of end-stage kidney disease after a baseline period of 2 years. We defined an end-stage kidney disease event as initiation of kidney replacement therapy. We quantified associations of percentage change in albuminuria with subsequent end-stage kidney disease using Cox regression in each cohort, followed by random-effects meta-analysis. We further adjusted for regression dilution to account for imprecision in the estimation of albuminuria at the participant level. We did multiple subgroup analyses, and also repeated our analyses using participant-level data from 14 clinical trials, including nine clinical trials not in CKD-PC. Between July, 2015, and June, 2018, we transferred and analysed data from 28 cohorts in the CKD-PC, which included 693 816 individuals (557 583 [80%] with diabetes). Data for 675 904 individuals and 7461 end-stage kidney disease events were available for our primary outcome analysis. Change in ACR was consistently associated with subsequent risk of end-stage kidney disease. The adjusted hazard ratio (HR) for end-stage kidney disease after a 30% decrease in ACR during a baseline period of 2 years was 0·83 (95% CI 0·74-0·94), decreasing to 0·78 (0·66-0·92) after further adjustment for regression dilution. Adjusted HRs were fairly consistent across cohorts and subgroups (ie, estimated glomerular filtration rate, diabetes, and sex), but the association was somewhat stronger among participants with higher baseline ACR than among those with lower baseline ACR (pinteraction<0·0001). In individuals with baseline ACR of 300 mg/g or higher, a 30% decrease in ACR over 2 years was estimated to confer a more than 1% absolute reduction in 10-year risk of end-stage kidney disease, even at early stages of chronic kidney disease. Results were generally similar when we used change in PCR and when study populations from clinical trials were assessed. Change in albuminuria was consistently associated with subsequent risk of end-stage kidney disease across a range of cohorts, lending support to the use of change in albuminuria as a surrogate endpoint for end-stage kidney disease in clinical trials of progression of chronic kidney disease in the setting of increased albuminuria. US National Kidney Foundation and US National Institute of Diabetes and Digestive and Kidney Diseases.


Accurate de novo design of hyperstable constrained peptides.
Naturally occurring, pharmacologically active peptides constrained with covalent crosslinks generally have shapes that have evolved to fit precisely into binding pockets on their targets. Such peptides can have excellent pharmaceutical properties, combining the stability and tissue penetration of small-molecule drugs with the specificity of much larger protein therapeutics. The ability to design constrained peptides with precisely specified tertiary structures would enable the design of shape-complementary inhibitors of arbitrary targets. Here we describe the development of computational methods for accurate de novo design of conformationally restricted peptides, and the use of these methods to design 18-47 residue, disulfide-crosslinked peptides, a subset of which are heterochiral and/or N-C backbone-cyclized. Both genetically encodable and non-canonical peptides are exceptionally stable to thermal and chemical denaturation, and 12 experimentally determined X-ray and NMR structures are nearly identical to the computational design models. The computational design methods and stable scaffolds presented here provide the basis for development of a new generation of peptide-based drugs.


Environmental Income and Rural Livelihoods: A Global-Comparative Analysis.
This paper presents results from a comparative analysis of environmental income from approximately 8000 households in 24 developing countries collected by research partners in CIFOR's Poverty Environment Network (PEN). Environmental income accounts for 28% of total household income, 77% of which comes from natural forests. Environmental income shares are higher for low-income households, but differences across income quintiles are less pronounced than previously thought. The poor rely more heavily on subsistence products such as wood fuels and wild foods, and on products harvested from natural areas other than forests. In absolute terms environmental income is approximately five times higher in the highest income quintile, compared to the two lowest quintiles.


Instrumenting gait assessment using the Kinect in people living with stroke: reliability and association with balance tests.
The Microsoft Kinect has been used previously to assess spatiotemporal aspects of gait; however the reliability of this system for the assessment of people following stroke has not been established. This study examined the reliability and additional information that the Kinect provides when instrumenting a gait assessment in people living with stroke. The spatiotemporal variables of step length, step length asymmetry, foot swing velocity, foot swing velocity asymmetry, peak and mean gait speed and the percentage difference between the peak and mean gait speed were assessed during gait trials in 30 outpatients more than three months post-stroke and able to stand unsupported. Additional clinical assessments of functional reach (FR), step test (ST), 10 m walk test (10MWT) and the timed up and go (TUG) were performed, along with force platform instrumented assessments of center of pressure path length velocity during double-legged standing balance with eyes closed (DLEC), weight bearing asymmetry (WBA) and dynamic medial-lateral weight-shifting ability (MLWS). These tests were performed on two separate occasions, seven days apart for reliability assessment. Separate adjusted multiple regressions models for predicting scores on the clinical and force platform assessments were created using 1) the easily assessed clinically-derived gait variables 10MWT time and total number of steps; and 2) the Kinect-derived variables which were found to be reliable (ICC > 0.75) and not strongly correlated (Spearman's ρ < 0.80) with each other (i.e. non-redundant). Kinect-derived variables were found to be highly reliable (all ICCs > 0.80), but many were redundant. The final regression model using Kinect-derived variables consisted of the asymmetry scores, mean gait velocity, affected limb foot swing velocity and the difference between peak and mean gait velocity. In comparison with the clinically-derived regression model, the Kinect-derived model accounted for >15% more variance on the MLWS, ST and FR tests and scored similarly on all other measures. In conclusion, instrumenting gait using the Kinect is reliable and provides insight into the dynamic balance capacity of people living with stroke. This system provides a minimally intrusive method of examining potentially important gait characteristics in people living with stroke.


Evidence to support that spontaneous preterm labor is adaptive in nature: neonatal RDS is more common in "indicated" than in "spontaneous" preterm birth.
The onset of preterm labor has been proposed to have survival value and to be adaptive in nature. This hypothesis would predict that induced preterm birth may be associated with higher rates of complications than spontaneous preterm birth. The purpose of this study was to determine if there is a difference in the frequency of neonatal respiratory distress syndrome (RDS), the most common neonatal complication, according to the etiology of preterm birth (e.g., preterm labor [PTL], preterm PROM, or pregnancies which ended because of maternal-fetal indications). The relationship between the occurrence of RDS and the obstetrical circumstances leading to preterm birth was examined in 257 consecutive singleton preterm births (gestational age: 24-32 weeks). Cases with major congenital anomalies were excluded. The study population was divided into two groups according to the cause of preterm birth: 1) preterm birth due to PTL with intact membranes or preterm PROM (spontaneous preterm birth group); and 2) preterm birth due to maternal or fetal indications (indicated preterm birth group). 1) RDS was diagnosed in 47% of cases; 2) RDS was more common in patients with indicated preterm birth than in those with spontaneous preterm birth group (58.1% vs. 38.4%, P=0.002); 3) Patients with indicated preterm birth had a significantly higher mean gestational age at birth, but lower mean birth weight, lower rate of histological chorioamnionitis and higher rates of cesarean delivery, 5 min Apgar score of <7, and umbilical arterial blood pH of <7.15 than those with spontaneous preterm birth (P<0.05 for each); 4) Antenatal corticosteroids were used in 73.4% of cases with indicated preterm birth and in 76.9% of those with spontaneous preterm birth; 5) Multivariate analysis demonstrated that indicated preterm birth was associated with an increased risk of RDS after adjusting for confounding variables (OR=2.29, 95% CI 1.22-4.29). 1) The rate of RDS is greater following "indicated" rather than spontaneous preterm birth; 2) This observation supports the view that spontaneous preterm labor is adaptive in nature.


Overview of Risk Factors for Age-Related Macular Degeneration (AMD).
Age related macular degeneration (AMD) is the leading cause of blindness in individuals older than 65 years of age. It is a multifactorial disorder and identification of risk factors enables individuals to make life style choices that may reduce the risk of disease. This review discusses the role of genetics, sunlight, diet, cardiovascular factors, smoking, and alcohol as possible risk factors for AMD. Genetics plays a more significant role in AMD than previously thought, especially in younger patients, histocompatibility locus antigen (HLA) and complement system genes being the most significant. Whether the risk of AMD is increased by exposure to sunlight, cardiovascular risk factors, and diet is more controversial. Smoking is the risk factor most consistently associated with AMD. Current smokers are exposed to a two to three times higher risk of AMD than non-smokers and the risk increases with intensity of smoking. Moderate alcohol consumption is unlikely to increase the risk of AMD. Optometrists as front-line informers and educators of ocular health play a significant role in increasing public awareness of the risks of AMD. Cessation of smoking, the use of eye protection in high light conditions, dietary changes, and regular use of dietary supplements should all be considered to reduce the lifetime risk of AMD.


Measuring medication: do interviews agree with medical record and pharmacy data?
Medication measurement is crucial in assessing quality for chronic conditions yet agreement of alternate data sources remains uncertain. To evaluate medication agreement between interviews, medical records, and pharmacy data; to assess data source contribution to attributing medication exposure; and to describe the impact of combining data sources on models that predict medication use. Prospective cohort study. Probability sample of HIV-infected participants in the HIV Cost and Services Utilization Study. Medications reported in 2267 interviews, 1936 medical records, and 457 pharmacy records were compared using crude agreement, kappa, and the proportion of average positive and negative agreement. The percent of medications reported in each source alone was used to assess their relative contribution to attributing exposure status. We performed weighted logistic regression in alternate data sources. Kappa varied from 0.38 for nucleoside reverse transcriptase inhibitors to 0.70 for protease inhibitors, when comparing drug classes in interview versus medical record, interview versus pharmacy data, and medical record versus pharmacy data. The percentage of medications reported in medical records was greater than that reported in interviews or pharmacy data. Pharmacy data contributed little to the attribution of medication exposure. Adding medication data to interview data did not appreciably change analytic models predicting medication use. For specific medications, agreement between alternative data sources is fair to substantial, but is lower for key drug classes. Relying on one data source may lead to misclassification of drug exposure status, but combining data sources does not change the results of analytic models predicting appropriate medication use.


Exogenous glucocorticoids amplify the costs of infection by reducing resistance and tolerance, but effects are mitigated by co-infection.
Individual variation in parasite defences, such as resistance and tolerance, can underlie heterogeneity in fitness and could influence disease transmission dynamics. Glucocorticoid hormone concentrations often change in response to fluctuating environmental conditions and mediate changes in immune function, resource allocation and tissue repair. Thus, changes in glucocorticoid hormone concentrations might mediate individual variation in investment in resistance versus tolerance. In this study, we experimentally increased glucocorticoid concentrations in red-winged blackbirds ( Agelaius phoeniceus) that were naturally infected with haemosporidian parasites, and assessed changes in resistance and tolerance of infection. Glucocorticoid treatment increased burdens of Plasmodium, the parasite causing avian malaria, but only in the absence of co-infection with another Haemosporidian, Haemoproteus. Thus, glucocorticoids might reduce resistance to infection, but co-infection can mitigate the negative consequences of increased hormone concentrations. Glucocorticoid treatment also decreased tolerance of infection. We found no evidence that the inflammatory immune response or rate of red blood cell production underlie the effects of glucocorticoids on resistance and tolerance. Our findings suggest that exogenous glucocorticoids can increase the costs of haemosporidian infections by both increasing parasite numbers and reducing an individual's ability to cope with infection. These effects could scale up to impact populations of both host and parasite.


An immunofluorescent study of the distribution of fibronectin and laminin during limb regeneration in the adult newt.
Fibronectin and laminin are two extracellular glycoproteins which are involved in various processes of cellular development and differentiation. The present investigation describes changes in their distribution during regeneration of the newt forelimb, as determined by indirect immunofluorescence. The distribution of fibronectin and laminin was similar in normal limb tissue components. These glycoproteins were localized in the pericellular region of the myofibers corresponding to its basement membrane; the perineurium and endoneurium of the nerves; and the basement membranes of blood vessels, skin epithelium, and dermal glands. The cytoplasm of myofibers, axons, skin epithelium, and bone matrix lacked fluorescence for both glycoproteins. After limb amputation in the regenerating blastema, extensive presence of fibronectin, but not laminin, was seen in and around the undifferentiated blastemal cells. Increased fluorescence for fibronectin was also seen during blastema growth, blastemal cell aggregation, and early stages of redifferentiation. As redifferentiation continued, staining for fibronectin slowly disappeared from the cartilage matrix and the myoblast fusion zone. Laminin was first observed around the regenerated myotubes; this was followed by the appearance of fibronectin suggesting a sequential formation of these two components of the new myotube basement membrane. In the regenerated limb, the distribution of laminin and fibronectin was similar to that seen in normal limb. Based on the distribution pattern of these glycoproteins, it is concluded that fibronectin may play an important role in blastemal cell aggregation, cell alignment, and initiation of redifferentiation. After redifferentiation, both laminin and fibronectin may be important in the determination of the architecture of the regenerated limb.


Changes in cannabis use, exposure, and health perceptions following legalization of adult recreational cannabis use in California: a prospective observational study.
Most U.S. states have legalized cannabis for medical and/or recreational use. In a 6-month prospective observational study, we examined changes in adult cannabis use patterns and health perceptions following broadened legalization of cannabis use from medical to recreational purposes in California. Respondents were part of Stanford University's WELL for Life registry, an online adult cohort concentrated in Northern California. Surveys were administered online in the 10 days prior to state legalization of recreational use (1/1/18) and 1-month (2/1/18-2/15/18) and 6-months (7/1/2018-7/15/18) following the change in state policy. Online surveys assessed self-reported past 30-day cannabis use, exposure to others' cannabis use, and health perceptions of cannabis use. Logistic regression models and generalized estimating equations (GEE) examined associations between participant characteristics and cannabis use pre- to 1-month and 6-months post-legalization. The sample (N = 429, 51% female, 55% non-Hispanic White, age mean = 56 ± 14.6) voted 58% in favor of state legalization of recreational cannabis use, with 26% opposed, and 16% abstained. Cannabis use in the past 30-days significantly increased from pre-legalization (17%) to 1-month post-legalization (21%; odds ratio (OR) = 1.28, p-value (p) = .01) and stayed elevated over pre-legalization levels at 6-months post-legalization (20%; OR = 1.28, p = .01). Exposure to others' cannabis use in the past 30 days did not change significantly over time: 41% pre-legalization, 44% 1-month post-legalization (OR = 1.18, p = .11), and 42% 6-months post-legalization (OR = 1.08, p = .61). Perceptions of health benefits of cannabis use increased from pre-legalization to 6-months post-legalization (OR = 1.19, p = .02). Younger adults, those with fewer years of education, and those reporting histories of depression were more likely to report recent cannabis use pre- and post-legalization. Other mental illness was associated with cannabis use at post-legalization only. In a multivariate GEE adjusted for sociodemographic characteristics and diagnoses, favoring legalization and the interaction of time and positive health perceptions were associated with a greater likelihood of using cannabis. Legalized recreational cannabis use was associated with greater self-reported past 30-day use post-legalization, and with more-positive health perceptions of cannabis use. Future research is needed to examine longer-term perceptions and behavioral patterns following legalization of recreational cannabis use, especially among those with mental illness.


CNS live imaging reveals a new mechanism of myelination: the liquid croissant model.
The overall morphology and with it associated the formation of myelin is generally thought to be resolved. Based on electron microscopic findings more than half a century ago, the current model of myelination describes all myelin membranes to run in parallel with the longitudinal axis of the axon and to form a smooth surface, reminiscent of a rolled up carpet. However, different studies in the past demonstrated a distinct myelin morphology with an uneven myelin surface contour that challenges the established concept. Even though the current model of myelination has since been recognized as insufficient, CNS myelin formation has not yet been investigated in real-time with the requisite technique and resolution. We therefore traced myelin growth in murine organotypic cerebellar slice cultures using high-resolution confocal live imaging, light and electron microscopy and assessed myelin morphology in young and adult mice by confocal microscopy. Our data verify that the myelin surface is indeed not smooth but runs in a bidirectional, regularly spaced coil along the axon in both young and adult mice. Time-lapse imaging revealed that the growth of coiled myelin turns emerges during myelin formation. We therefore propose the "liquid croissant" model as a new concept of myelination that overcomes not only some of the incongruences of previous myelination theories, but potentially also explains the development of certain myelin pathologies observed in remyelination and axonopathies.


Alzheimer's presenilin-1 mutation potentiates inositol 1,4,5-trisphosphate-mediated calcium signaling in Xenopus oocytes.
Perturbations in intracellular Ca2+ signaling may represent one mechanism underlying Alzheimer's disease (AD). The presenilin-1 gene (PS1), associated with the majority of early onset familial AD cases, has been implicated in this signaling pathway. Here we used the Xenopus oocyte expression system to investigate in greater detail the role of PS1 in intracellular Ca2+ signaling pathways. Treatment of cells expressing wild-type PS1 with a cell surface receptor agonist to stimulate the phosphoinositide second messenger pathway evoked Ca2+-activated Cl- currents that were significantly potentiated relative to controls. To determine which elements of the signal transduction pathway are responsible for the potentiation, we used photolysis of caged inositol 1,4,5-trisphosphate (IP3) and fluorescent Ca2+ imaging to demonstrate that PS1 potentiates IP3-mediated release of Ca2+ from internal stores. We show that an AD-linked mutation produces a potentiation in Ca2+ signaling that is significantly greater than that observed for wild-type PS1 and that cannot be attributed to differences in protein expression levels. Our findings support a role for PS1 in modulating IP3-mediated Ca2+ liberation and suggest that one pathophysiological mechanism by which PS1 mutations contribute to AD neurodegeneration may involve perturbations of this function.


APC mutation associated with late onset of familial adenomatous polyposis.
Familial adenomatous polyposis is an autosomal dominantly inherited disorder. Mutation studies in the corresponding gene (APC) may provide information for predictive tests for persons at risk in affected families. We report here a new mutation in exon 6 (codon 233) of the APC gene and clinical data in a large family with late onset of the disease in most affected persons.


A chain reaction approach to modelling gene pathways.
BACKGROUND: Of great interest in cancer prevention is how nutrient components affect gene pathways associated with the physiological events of puberty. Nutrient-gene interactions may cause changes in breast or prostate cells and, therefore, may result in cancer risk later in life. Analysis of gene pathways can lead to insights about nutrient-gene interactions and the development of more effective prevention approaches to reduce cancer risk. To date, researchers have relied heavily upon experimental assays (such as microarray analysis, etc.) to identify genes and their associated pathways that are affected by nutrient and diets. However, the vast number of genes and combinations of gene pathways, coupled with the expense of the experimental analyses, has delayed the progress of gene-pathway research. The development of an analytical approach based on available test data could greatly benefit the evaluation of gene pathways, and thus advance the study of nutrient-gene interactions in cancer prevention. In the present study, we have proposed a chain reaction model to simulate gene pathways, in which the gene expression changes through the pathway are represented by the species undergoing a set of chemical reactions. We have also developed a numerical tool to solve for the species changes due to the chain reactions over time. Through this approach we can examine the impact of nutrient-containing diets on the gene pathway; moreover, transformation of genes over time with a nutrient treatment can be observed numerically, which is very difficult to achieve experimentally. We apply this approach to microarray analysis data from an experiment which involved the effects of three polyphenols (nutrient treatments), epigallo-catechin-3-O-gallate (EGCG), genistein, and resveratrol, in a study of nutrient-gene interaction in the estrogen synthesis pathway during puberty. RESULTS: In this preliminary study, the estrogen synthesis pathway was simulated by a chain reaction model. By applying it to microarray data, the chain reaction model computed a set of reaction rates to examine the effects of three polyphenols (EGCG, genistein, and resveratrol) on gene expression in this pathway during puberty. We first performed statistical analysis to test the time factor on the estrogen synthesis pathway. Global tests were used to evaluate an overall gene expression change during puberty for each experimental group. Then, a chain reaction model was employed to simulate the estrogen synthesis pathway. Specifically, the model computed the reaction rates in a set of ordinary differential equations to describe interactions between genes in the pathway (A reaction rate K of A to B represents gene A will induce gene B per unit at a rate of K; we give details in the "method" section). Since disparate changes of gene expression may cause numerical error problems in solving these differential equations, we used an implicit scheme to address this issue. We first applied the chain reaction model to obtain the reaction rates for the control group. A sensitivity study was conducted to evaluate how well the model fits to the control group data at Day 50. Results showed a small bias and mean square error. These observations indicated the model is robust to low random noises and has a good fit for the control group. Then the chain reaction model derived from the control group data was used to predict gene expression at Day 50 for the three polyphenol groups. If these nutrients affect the estrogen synthesis pathways during puberty, we expect discrepancy between observed and expected expressions. Results indicated some genes had large differences in the EGCG (e.g., Hsd3b and Sts) and the resveratrol (e.g., Hsd3b and Hrmt12) groups. CONCLUSIONS: In the present study, we have presented (I) experimental studies of the effect of nutrient diets on the gene expression changes in a selected estrogen synthesis pathway. This experiment is valuable because it allows us to examine how the nutrient-containing diets regulate gene expression in the estrogen synthesis pathway during puberty; (II) global tests to assess an overall association of this particular pathway with time factor by utilizing generalized linear models to analyze microarray data; and (III) a chain reaction model to simulate the pathway. This is a novel application because we are able to translate the gene pathway into the chemical reactions in which each reaction channel describes gene-gene relationship in the pathway. In the chain reaction model, the implicit scheme is employed to efficiently solve the differential equations. Data analysis results show the proposed model is capable of predicting gene expression changes and demonstrating the effect of nutrient-containing diets on gene expression changes in the pathway. One of the objectives of this study is to explore and develop a numerical approach for simulating the gene expression change so that it can be applied and calibrated when the data of more time slices are available, and thus can be used to interpolate the expression change at a desired time point without conducting expensive experiments for a large amount of time points. Hence, we are not claiming this is either essential or the most efficient way for simulating this problem, rather a mathematical/numerical approach that can model the expression change of a large set of genes of a complex pathway. In addition, we understand the limitation of this experiment and realize that it is still far from being a complete model of predicting nutrient-gene interactions. The reason is that in the present model, the reaction rates were estimated based on available data at two time points; hence, the gene expression change is dependent upon the reaction rates and a linear function of the gene expressions. More data sets containing gene expression at various time slices are needed in order to improve the present model so that a non-linear variation of gene expression changes at different time can be predicted.


Boot cAMP: educational outcomes after 4 successive years of preparatory simulation-based training at onset of internship.
Preparatory training for new trainees beginning residency has been used by a variety of programs across the country. To improve the clinical orientation process for our new postgraduate year (PGY)-1 residents, we developed an intensive preparatory training curriculum inclusive of cognitive and procedural skills, training activities considered essential for early PGY-1 clinical management. We define our surgical PGY-1 Boot Camp as preparatory simulation-based training implemented at the onset of internship for introduction of skills necessary for basic surgical patient problem assessment and management. This orientation process includes exposure to simulated patient care encounters and technical skills training essential to new resident education. We report educational results of 4 successive years of Boot Camp training. Results were analyzed to determine if performance evidenced at onset of training was predictive of later educational outcomes. Learners were PGY-1 residents, in both categorical and preliminary positions, at our medium-sized surgical residency program. Over a 4-year period, from July 2007 to July 2010, all 30 PGY-1 residents starting surgical residency at our institution underwent specific preparatory didactic and skills training over a 9-week period. This consisted of mandatory weekly 1-hour and 3-hour sessions in the Simulation Center, representing a 4-fold increase in time in simulation laboratory training compared with the remainder of the year. Training occurred in 8 procedural skills areas (instrument use, knot-tying, suturing, laparoscopic skills, airway management, cardiopulmonary resuscitation, central venous catheter, and chest tube insertion) and in simulated patient care (shock, surgical emergencies, and respiratory, cardiac, and trauma management) using a variety of high- and low-tech simulation platforms. Faculty and senior residents served as instructors. All educational activities were structured to include preparatory materials, pretraining briefing sessions, and immediate in-training or post-training review and debriefing. Baseline cognitive skills were assessed with written tests on basic patient management. Post-Boot Camp tests similarly evaluated cognitive skills. Technical skills were assessed using a variety of task-specific instruments, and expressed as a mean score for all activities for each resident. All measurements were expressed as percent (%) best possible score. Cognitive and technical performance in Boot Camp was compared with subsequent clinical and core curriculum evaluations including weekly quiz scores, annual American Board of Surgery In-Training Examination (ABSITE) scores, program in-training evaluations (New Innovations, Uniontown, Ohio), and operative assessment instrument scores (OP-Rate, Baystate Medical Center, Springfield, Massachusetts) for the remainder of the PGY-1 year. Performance data were available for 30 PGY-1 residents over 4 years. Baseline cognitive skills were lower for the first year of Boot Camp as compared with subsequent years (71 ± 13, 83 ± 9, 84 ± 11, and 86 ± 6, respectively; p = 0.028, analysis of variance; ANOVA). Performance improved between pretests and final testing (81 ± 11 vs 89 ± 7; p < 0.001 paired t test). There was statistically significant correlation between Boot Camp final cognitive test results and American Board of Surgery In-Training Examination scores (p = 0.01; n = 22), but not quite significant for weekly curriculum quiz scores (p = 0.055; n = 22) and New Innovations cognitive assessments (p = 0.09; n = 25). Statistically significant correlation was also noted between Boot Camp mean overall skills and New Innovations technical skills assessments (p = 0.002; n = 25) and OP-Rate assessments (p = 0.01; n = 12). Individual simulation-based Boot Camp performance scores for cognitive and procedural skills assessments in PGY-1 residents correlate with subjective and objective clinical performance evaluations. This concurrent correlation with multiple traditional evaluation methods used to express competency in our residency program supports the use of Boot Camp performance measures as needs assessment tools as well as adjuncts to cumulative resident evaluation data.


Metazoan parasite communities and diet of the velvet belly lantern shark Etmopterus spinax (Squaliformes: Etmopteridae): a comparison of two deep-sea ecosystems.
By combining an examination of stomach contents yielding a snapshot of the most recent trophic niche and the structure of parasite communities reflecting a long-term feeding niche, this study aimed at gaining more comprehensive information on the role of the small-sized deep-water velvet belly lantern shark Etmopterus spinax in the local food webs of the Galicia Bank and the canyon and valley system of the Avilés Canyon, which have been both proposed for inclusion in the Natura 2000 network of protected areas. As far as is known, this study provides the first comparative parasite infracommunity data for a deep-sea shark species. Component parasite communities in E. spinax were relatively rich, whereas the infracommunities were rather depauperate, with similar low diversity at both localities. The significant differences in the composition and structure of both parasite communities and prey assemblages indicate differential effects of the two deep-sea ecosystems on both long-term and most recent trophic niches of E. spinax. These results underline the importance of the use of multivariate analyses for the assessment of geographical variation in shark populations based on parasites and diet data.


Adolescent contraception: survey of attitudes and practice of health professionals.
In Nigeria, there is a persistent reluctance to provide adolescents with contraceptives and this is based largely on the premise that the culture does not support pre-marital sexual activity. However, several studies have revealed that a large number of adolescents are experiencing early sexual debut, some as early as 13 years. Furthermore, mortality resulting from termination of unwanted pregnancies is on the increase among Nigerian adolescents. The aim of this study is to assess the attitudes and practice of health workers in Ibadan regarding adolescent contraception. A structured questionnaire was administered to 735 health workers in selected hospitals and health facilities in Ibadan, Nigeria. Information relating to their practice and attitudes towards adolescent contraception was sought. The results revealed that while 58.9% of the respondents had counselled adolescents, only 30.7% had ever prescribed contraceptives for them. Among health workers that approved of family planning, 52.6% were favourably disposed to adolescent contraception while 23.9% were not. We observed that 40.2% of health workers that did not approve of family planning are not favourably disposed to adolescent contraception. Health workers who counsel adolescents on the use of contraceptives are more likely than those who do not to approve of its use. In addition, those who approve of family planning are 12 times more than those who do not, to have a favourable disposition to adolescent contraception. Perhaps, if providers are more receptive to adolescent contraception, the services will be more accessible to those who are sexually active. This will, no doubt, stem the incidence of unwanted pregnancies in our community.


Ultrafast dynamics of the ESIPT photoswitch N-(3-pyridinyl)-2-pyridinecarboxamide.
Molecular switches based on proton transfer that are photochromic and can be interconverted by light at different wavelengths back and forth between two thermodynamically stable tautomeric states in solution at room temperature are rare to date. We report on a study of the ultrafast conversion of the bistable proton transfer switch N-(3-pyridinyl)-2-pyridinecarboxamide (NPPCA) to a corresponding iminol after photoexcitation at λpump ≈ 265 nm by means of femtosecond time-resolved broad-band and single-colour transient electronic absorption spectroscopy (TEAS), transient fluorescence spectroscopy (TFLS), and transient vibrational absorption spectroscopy (TVAS) in acetonitrile solution. The interpretation of the data was accompanied by ab initio quantum chemical calculations of the excited electronic states and the vibrational frequencies of the reactant and product in their ground electronic state. The TEAS experiments provided four time constants, τ1 = 0.09 ± 0.01 ps, τ2 = 0.61 ± 0.01 ps, τ3 = 5.10 ± 0.80 ps, and τ4 = 20.0 ± 1.0 ps. The first two agree well with the measured TFLS lifetimes, τ1,TFL < 0.18 ps and τ2,TFL = 0.50 ± 0.01 ps. τ1 is related to the relaxation of the initially excited Franck-Condon (FC) state of the pyridinecarboxamide, followed by the excited-state intramolecular proton transfer (ESIPT) step to the neighbouring pyridine. The subsequent return of the molecules to the electronic ground state takes place within τ2, mediated by a conical intersection (CI) at a twisted configuration of the pyridinecarboxamide moiety. The main components in all TEAS time profiles feature a rise with τ2 and a decay with τ4 and describe subsequent molecular transformations in the electronic ground state. τ3 is ascribed to vibrational cooling of the molecules. The final iminol exhibits a permanent UV absorption at λ = 247 nm, where its absorbance is stronger than that of the carboxamide reactant. The iminol structure is unambiguously identified by the TVA spectra, which show the build-up of corresponding vibrational bands with τ4,TVA = 23 ± 2 ps after the initial bleach of the reactant vibrational bands, in excellent agreement with the TEAS data. Its lifetime is >10 ns.


Using patient management as a surrogate for patient health outcomes in diagnostic test evaluation.
Before a new test is introduced in clinical practice, evidence is needed to demonstrate that its use will lead to improvements in patient health outcomes. Studies reporting test accuracy may not be sufficient, and clinical trials of tests that measure patient health outcomes are rarely feasible. Therefore, the consequences of testing on patient management are often investigated as an intermediate step in the pathway. There is a lack of guidance on the interpretation of this evidence, and patient management studies often neglect a discussion of the limitations of measuring patient management as a surrogate for health outcomes. We discuss the rationale for measuring patient management, describe the common study designs and provide guidance about how this evidence should be reported. Interpretation of patient management studies relies on the condition that patient management is a valid surrogate for downstream patient benefits. This condition presupposes two critical assumptions: the test improves diagnostic accuracy; and the measured changes in patient management improve patient health outcomes. The validity of this evidence depends on the certainty around these critical assumptions and the ability of the study design to minimise bias. Three common designs are test RCTs that measure patient management as a primary endpoint, diagnostic before-after studies that compare planned patient management before and after testing, and accuracy studies that are extended to report on the actual treatment or further tests received following a positive and negative test result. Patient management can be measured as a surrogate outcome for test evaluation if its limitations are recognised. The potential consequences of a positive and negative test result on patient management should be pre-specified and the potential patient benefits of these management changes clearly stated. Randomised comparisons will provide higher quality evidence about differences in patient management using the new test than observational studies. Regardless of the study design used, the critical assumption that patient management is a valid surrogate for downstream patient benefits or harms must be discussed in these studies.


Persistent high serum bicarbonate and the risk of heart failure in patients with chronic kidney disease (CKD): A report from the Chronic Renal Insufficiency Cohort (CRIC) study.
Serum bicarbonate varies over time in chronic kidney disease (CKD) patients, and this variability may portend poor cardiovascular outcomes. The aim of this study was to conduct a time-updated longitudinal analysis to evaluate the association of serum bicarbonate with long-term clinical outcomes: heart failure, atherosclerotic events, renal events (halving of estimated glomerular filtration rate [eGFR] or end-stage renal disease), and mortality. Serum bicarbonate was measured annually, in 3586 participants with CKD, enrolled in the Chronic Renal Insufficiency Cohort (CRIC) study. Marginal structural models were created to allow for integration of all available bicarbonate measurements and proper adjustment for time-dependent confounding. During the 6 years follow-up, 512 participants developed congestive heart failure (26/1000 person-years) and 749 developed renal events (37/1000 person-years). The risk of heart failure and death was significantly higher for participants who maintained serum bicarbonate >26 mmol/L for the entire duration of follow-up (hazard ratio [HR] 1.66; 95% confidence interval [CI], 1.23 to 2.23, and HR 1.36, 95% CI 1.02 to 1.82, respectively) compared with participants who kept their bicarbonate 22 to 26 mmol/L, after adjusting for demographics, co-morbidities, medications including diuretics, eGFR, and proteinuria. Participants who maintained serum bicarbonate <22 mmol/L had almost a 2-fold increased risk of renal disease progression (HR 1.97; 95% CI, 1.50 to 2.57) compared with participants with bicarbonate 22 to 26 mmol/L. In this large CKD cohort, persistent serum bicarbonate >26 mmol/L was associated with increased risk of heart failure events and mortality. Further studies are needed to determine the optimal range of serum bicarbonate in CKD to prevent adverse clinical outcomes.


Data on the configuration design of internet-connected home cooling systems by engineering students.
This experiment was carried out to record the step-by-step actions that humans take in solving a configuration design problem, either in small teams or individually. Specifically, study participants were tasked with configuring an internet-connected system of products to maintain temperature within a home, subject to cost constraints. Every participant was given access to a computer-based design interface that allowed them to construct and assess solutions. The interface was also used to record the data that is presented here. In total, data was collected for 68 participants, and each participant was allowed to perform 50 design actions in solving the configuration design problem. Major results based on the data presented here have been reported separately, including initial behavioral analysis (McComb et al.) [1], [2] and design pattern assessments via Markovian modeling (McComb et al., 2017; McComb et al., 2017) [3], [4].


Endosymbiont gene functions impaired and rescued by polymerase infidelity at poly(A) tracts.
Among host-dependent bacteria that have evolved by extreme reductive genome evolution, long-term bacterial endosymbionts of insects have the smallest (160-790 kb) and most A + T-rich (>70%) bacterial genomes known to date. These genomes are riddled with poly(A) tracts, and 5-50% of genes contain tracts of 10 As or more. Here, we demonstrate transcriptional slippage at poly(A) tracts within genes of Buchnera aphidicola associated with aphids and Blochmannia pennsylvanicus associated with ants. Several tracts contain single frameshift deletions; these apparent pseudogenes showed patterns of constraint consistent with purifying selection on the encoded proteins. Transcriptional slippage yielded a heterogeneous population of transcripts with variable numbers of As in the tract. Across several frameshifted genes, including B. aphidicola cell wall biosynthesis genes and a B. pennsylvanicus histidine biosynthesis gene, 12-50% of transcripts contained corrected reading frames that could potentially yield full-length proteins. In situ immunostaining confirmed the production of the cell wall biosynthetic enzyme UDP-N-acetylmuramyl pentapeptide synthase encoded by the frameshifted murF gene. Simulation studies indicated an overrepresentation of poly(A) tracts in endosymbiont genomes relative to other A + T-rich bacterial genomes. Polymerase infidelity at poly(A) tracts rescues the functionality of genes with frameshift mutations and, conversely, reduces the efficiency of expression for in-frame genes carrying poly(A) regions. These features of homopolymeric tracts could be exploited to manipulate gene expression in small synthetic genomes.


Naturally occurring circadian rhythm variation associated with clock gene loci in Swedish Arabidopsis accessions.
Circadian clocks have evolved to resonate with external day and night cycles. However, these entrainment signals are not consistent everywhere and vary with latitude, climate and seasonality. This leads to divergent selection for clocks which are locally adapted. To investigate the genetic basis for this circadian variation, we used a delayed fluorescence imaging assay to screen 191 naturally occurring Swedish Arabidopsis accessions for their circadian phenotypes. We demonstrate that the period length co-varies with both geography and population sub-structure. Several candidate loci linked to period, phase and relative amplitude error (RAE) were revealed by genome-wide association mapping and candidate genes were investigated using TDNA mutants. We show that natural variation in a single non-synonymous substitution within COR28 is associated with a long-period and late-flowering phenotype similar to that seen in TDNA knock-out mutants. COR28 is a known coordinator of flowering time, freezing tolerance and the circadian clock; all of which may form selective pressure gradients across Sweden. We demonstrate the effect of the COR28-58S SNP in increasing period length through a co-segregation analysis. Finally, we show that period phenotypic tails remain diverged under lower temperatures and follow a distinctive "arrow-shaped" trend indicative of selection for a cold-biased temperature compensation response.


A Systematic Review of the Effectiveness of Medical Cannabis for Psychiatric, Movement and Neurodegenerative Disorders.
The discovery of endocannabinoid's role within the central nervous system and its potential therapeutic benefits have brought forth rising interest in the use of cannabis for medical purposes. The present review aimed to synthesize and evaluate the available evidences on the efficacy of cannabis and its derivatives for psychiatric, neurodegenerative and movement disorders. A systematic search of randomized controlled trials of cannabis and its derivatives were conducted via databases (PubMed, Embase and the Cochrane Central Register of Controlled Trials). A total of 24 reports that evaluated the use of medical cannabis for Alzheimer's disease, anorexia nervosa, anxiety, dementia, dystonia, Huntington's disease, Parkinson's disease, post-traumatic stress disorder (PTSD), psychosis and Tourette syndrome were included in this review. Trial quality was assessed with the Cochrane risk of bias tool. There is a lack of evidence on the therapeutic effects of cannabinoids for amyotrophic lateral sclerosis and dystonia. Although trials with positive findings were identified for anorexia nervosa, anxiety, PTSD, psychotic symptoms, agitation in Alzheimer's disease and dementia, Huntington's disease, and Tourette syndrome, and dyskinesia in Parkinson's disease, definitive conclusion on its efficacy could not be drawn. Evaluation of these low-quality trials, as rated on the Cochrane risk of bias tools, was challenged by methodological issues such as inadequate description of allocation concealment, blinding and underpowered sample size. More adequately powered controlled trials that examine the long and short term efficacy, safety and tolerability of cannabis for medical use, and the mechanisms underpinning the therapeutic potential are warranted.


Retinal expression, regulation, and functional bioactivity of prostacyclin-stimulating factor.
Prostacyclin-stimulating factor (PSF) acts on vascular endothelial cells to stimulate the synthesis of the vasodilatory molecule prostacyclin (PGI2). We have examined the expression, regulation, and hemodynamic bioactivity of PSF both in whole retina and in cultured cells derived from this tissue. PSF was expressed in all retinal cell types examined in vitro, but immunohistochemical analysis revealed PSF mainly associated with retinal vessels. PSF expression was constitutive in retinal pericytes (RPCs) but could be modulated in bovine retinal capillary endothelial cells (RECs) by cell confluency, hypoxia, serum starvation, high glucose concentrations, or inversely by soluble factors present in early vs. late retinopathy, such as TGF-beta, VEGF, or bFGF. In addition, RPC-conditioned media dramatically increased REC PGI2 production, a response inhibited by blocking PSF with a specific antisense oligodeoxynucleotide (ODN). In vivo, PGI2 increased retinal blood flow (RBF) in control and diabetic animals. Furthermore, the early drop in RBF during the initial weeks after inducing diabetes in rats, as well as the later increase in RBF, both correlated with levels of retinal PSF. RBF also responded to treatment with RPC-conditioned media, and this effect could be partially blocked using the antisense PSF ODN. We conclude that PSF expressed by ocular cells can induce PGI2, retinal vascular dilation, and increased retinal blood flow, and that alterations in retinal PSF expression may explain the biphasic changes in RBF observed in diabetes.


Suitability of soccer training drills for endurance training.
Recent evidence suggests that certain soccer drills produce exercise intensities suitable for physical conditioning. However, it remains debatable whether soccer drills can provide a sufficiently unified exercise intensity among different players and on repetition of a drill, because movement patterns cannot be externally controlled during soccer drills. Good reliability and low variability of exercise intensity would enable all players to receive an appropriate training stimulus. The purpose of this study was to investigate intersubject variability and intrasubject reliability in exercise intensity during soccer drills. It was hypothesized that soccer drills that involve the highest exercise intensities would demonstrate the lowest intersubject variability and the highest intrasubject reliability. Heart rates of 23 professional soccer players were recorded during a range of soccer training drills. The drills consisted of 2 vs. 2 to 8 vs. 8 normal scoring games and 2 further possession games. Heart rate responses were examined for variability, reliability, and suitability for soccer endurance training. Coefficients of variation across players were less than 3% for all drills. Paired t-tests showed no significant differences in heart rate on repetition of the drills and 95% ratio limits of agreement were 1.8-3.8%. There were no significant correlations between exercise intensity and the statistical measures of variability and reliability. Several drills produced exercise intensities suitable for soccer endurance training with mean heart rate responses ranging from 87-91% HRmax. Soccer drills such as those used in the present study appear to be an adequate substitute for physical training without the ball and thus provide simultaneous skill and fitness training. The increase in training time spent developing technical ability and/or a reduction in total training time required may be useful for soccer teams.


Medical technology procurement in Europe: a cross-country comparison of current practice and policy.
Procurement policy can influence the diffusion of medical devices into national health systems, but limited comparative evidence exists on how countries procure such technologies. This paper discusses the procurement of select medical devices across five countries (England, France, Germany, Italy, and Spain) based on a review of published and grey literature and policy documents, as well as expert interviews. All countries have introduced various regulatory or policy measures that implicitly or explicitly influence device procurement, from lists of devices for purchase to changes in financing mechanisms. There has also been movement toward more centralized procurement with the introduction of purchasing groups or consortiums, notably in England, France, Germany, and Italy. While a number of stakeholder groups are involved in purchasing activities, a greater, more formalized role for physicians and governments is needed to ensure that technologies procured best meet patient needs and align with national health care priorities and other sectoral objectives. A general theme across all national procurement systems was a focus on cost-containment, but like other areas of technology policy (e.g., coverage), basing purchasing decisions on a broader range of criteria, such as quality and health outcomes, might better allow governments to achieve value for money and support patient access to beneficial innovations. More research is needed, however, to substantiate the role and influence of procurement on balancing the adoption and affordability of medical technologies.


Polymorphism of genes related to insulin sensitivity and the risk of biliary tract cancer and biliary stone: a population-based case-control study in Shanghai, China.
Biliary tract cancer, encompassing tumors of the gallbladder, extrahepatic bile ducts and ampulla of Vater, is a rare but highly fatal malignancy. Obesity and gallstones, both related to insulin resistance, are linked to an elevated risk of biliary cancer. The peroxisome proliferator-activated receptors (PPARs) and the retinoid X receptors (RXRs), expressed in adipose tissue, play a key role in the regulation of obesity-related insulin sensitivity, thus genetic variants of these two receptor genes may be related to biliary cancer and stones. We examined the associations of seven single-nucleotide polymorphisms in the PPAR-gamma, PPAR-delta, RXR-alpha, RXR-beta and INS genes with biliary cancer and stones in a population-based case-control study in Shanghai, China. We included 237 gallbladder, 127 extrahepatic bile duct and 47 ampulla of Vater cancer cases, 895 stone cases and 786 population controls. Relative to individuals with the RXR-beta C51T (rs2076310) CC genotype, those having the TT genotype had a 1.6-fold risk for bile duct cancer [odds ratio (OR) = 1.67; 95% confidence interval (CI) = 0.99-2.84], with a more pronounced association among men (OR = 2.30; 95% CI = 1.14-4.65; P interaction = 0.07). This marker was also associated with a higher risk of gallstones among subjects with a higher body mass index (BMI) (>or=23 kg/m(2)) (OR = 1.80; 95% CI = 1.09-2.94), although the interaction with BMI was not statistically significant (P interaction = 0.28). No association was found between other variants and biliary cancers and stones. Results from this population-based study suggest that certain genetic variants involved in the regulation of obesity-related insulin sensitivity may increase susceptibility to bile duct cancer and gallstones.


Toward passive BCI: asynchronous decoding of neural responses to direction- and angle-specific perturbations during a simulated cockpit scenario.
Neuroimaging studies have provided proof that loss of balance evokes specific neural transient wave complexes in electroencephalography (EEG), called perturbation evoked potentials (PEPs). Online decoding of balance perturbations from ongoing EEG signals can establish the possibility of implementing passive brain-computer interfaces (pBCIs) as a part of aviation/driving assistant systems. In this study, we investigated the feasibility of identifying the existence and expression of perturbations in four different conditions by using EEG signals. Fifteen healthy participants experienced four various postural changes while they sat in a glider cockpit. Sudden perturbations were exposed by a robot connected to a glider and moved to the right and left directions with tilting angles of 5 and 10 degrees. Perturbations occurred in an oddball paradigm in which participants were not aware of the time and expression of the perturbations. We employed a hierarchical approach to separate the perturbation and rest, and then discriminate the expression of perturbations. The performance of the BCI system was evaluated by using classification accuracy and F1 score. Asynchronously, we achieved average accuracies of 89.83 and 73.64% and average F1 scores of 0.93 and 0.60 for binary and multiclass classification, respectively. These results manifest the practicality of pBCI for the detection of balance disturbances in a realistic situation.


Antitumor effects of herbal mixture extract in the pancreatic adenocarcinoma cell line PANC1.
A recent study showned that complementary medicine is gradually gaining wide acceptance. In the present study, the herbal mixture extract (H3) composed of 3 oriental herbal plants was investigated for anticancer activity in vitro and in vivo. H3 inhibited PANC1 cell growth by promoting G0/G1 arrest (11% increase) and apoptotic cell death (9% increase). H3 also suppressed stem cell-like side population cells (4% decrease) and migration activity (24% decrease). In contrast, gemcitabine decreased side population cells and migration activity by 3 and 11%, respectively. These effects of H3 and gemcitabine were further studied by examining the expression of apoptosis-associated genes (CXCR4, JAK2 and XIAP) and stem cell-associated genes (ABCG2, POU5F1 and SOX2). We also found that H3 suppressed tumor growth by 46% in a PANC1‑xenograft model, while gemcitabine caused a 36% decrease. The antitumor effects of H3 were confirmed by western blot analysis for COX-2 and cytochrome c expression. Furthermore, necrotic cell death and erythrocyte-containing cavities were detected in tumor tissue by hematoxylin and eosin (H&E) staining. Notably, the combinatorial therapy (H3 and gemcitabine) increased tumor growth compared to that in the control. In conclusion, the present study shows that H3 has promise as a therapeutic agent against pancreatic cancer and its cancer stem cells.


Evaluation of a Fully Automatic Deep Learning-Based Method for the Measurement of Psoas Muscle Area.
Manual muscle mass assessment based on Computed Tomography (CT) scans is recognized as a good marker for malnutrition, sarcopenia, and adverse outcomes. However, manual muscle mass analysis is cumbersome and time consuming. An accurate fully automated method is needed. In this study, we evaluate if manual psoas annotation can be substituted by a fully automatic deep learning-based method. This study included a cohort of 583 patients with severe aortic valve stenosis planned to undergo Transcatheter Aortic Valve Replacement (TAVR). Psoas muscle area was annotated manually on the CT scan at the height of lumbar vertebra 3 (L3). The deep learning-based method mimics this approach by first determining the L3 level and subsequently segmenting the psoas at that level. The fully automatic approach was evaluated as well as segmentation and slice selection, using average bias 95% limits of agreement, Intraclass Correlation Coefficient (ICC) and within-subject Coefficient of Variation (CV). To evaluate performance of the slice selection visual inspection was performed. To evaluate segmentation Dice index was computed between the manual and automatic segmentations (0 = no overlap, 1 = perfect overlap). Included patients had a mean age of 81 ± 6 and 45% was female. The fully automatic method showed a bias and limits of agreement of -0.69 [-6.60 to 5.23] cm2, an ICC of 0.78 [95% CI: 0.74-0.82] and a within-subject CV of 11.2% [95% CI: 10.2-12.2]. For slice selection, 84% of the selections were on the same vertebra between methods, bias and limits of agreement was 3.4 [-24.5 to 31.4] mm. The Dice index for segmentation was 0.93 ± 0.04, bias and limits of agreement was -0.55 [1.71-2.80] cm2. Fully automatic assessment of psoas muscle area demonstrates accurate performance at the L3 level in CT images. It is a reliable tool that offers great opportunities for analysis in large scale studies and in clinical applications.


Combining Diffusion Tensor Imaging and Susceptibility Weighted Imaging on the Substantia Nigra of 1-Methyl-4-Phenyl-1, 2, 3, 6-Tetrahydropyridine (MPTP)-induced Rhesus Monkey Model of Parkinson's Disease.
The aim of this study was to evaluate whether combining diffusion tensor magnetic resonance imaging (DTI) and susceptibility weighted imaging (SWI) techniques would provide a sensitive method for differentiating between 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced rhesus monkey model of Parkinson's disease (PD) and wild-type controls. Seventeen rhesus monkeys were divided into two groups. A series of intramuscular injections of either saline (control group, n = 8) or MPTP (0.2 mg/kg body weight; PD group, n = 9) were given to the monkeys, twice a week. Then, SWI and DTI scans were obtained from the monkeys with Siemens Magnetom Verio 3.0T superconductive MRI system. Region of interest analysis was performed on substantia nigra pars compacta (SNc) and substantia nigra pars reticulata (SNr). In addition, immunohistochemical staining of tyrosine hydroxylase was applied to assess degeneration of SN dopaminergic neurons. Monkeys in the PD group displayed mild to moderate motor symptoms assessed using Kurlan's scale. With SWI scans, decreased width of SNc but increased width of SNr was found in PD group monkeys compared to controls. Calculation of the ratios of widths of SNc and SNr to the anterior and posterior mesencephalic diameter also reflected narrower SNc but wider SNr than controls. Decreased SWI signal intensity of SNc and SNr suggested iron deposition in both subregions of SN. The DTI scans showed lower fractional anisotropy (FA) values in SNc of the PD group monkeys, while no change of FA values in SNr was detected. Immunohistochemical test displayed generalized loss of dopaminergic neurons in SN of PD group monkeys. Combining the use of DTI and SWI can provide a sensitive method for differentiating between MPTP-induced rhesus monkey model of PD and wild-type controls. This effective imaging modality might provide additional information for characteristic identification of PD at early stages, thus enhancing the ability to make early diagnosis, and monitor progression of the natural history and treatment effects.


Multiple pathways regulate the induction of genes during zoosporogenesis in Phytophthora infestans.
Zoospores are a critical component of the disease cycles of most oomycete pathogens. To better understand this stage, genes induced during zoosporogenesis were identified from Phytophthora infestans, the potato late blight pathogen. Using cDNA arrays representing 2,600 genes expressed during zoosporogenesis, 69 genes showing >fourfold increases in mRNA levels were identified, of which 22 exhibited >100-fold induction. Included were putative protein kinases, transcription factors, ion channels, and other regulators. The expression of 15 genes was characterized in detail using zoosporogenesis time courses, other developmental stages, different temperature regimes, and tissue treated with signaling inhibitors. The latter were of interest because zoosporogenesis is known to be cold induced and inhibited by calcium channel blockers such as verapamil; moreover, in this study, inhibitors of phospholipase C (U-73122) and inositol trisphosphate receptor-gated calcium channels (2-aminoethoxydiphenyl borate) also were shown to block zoosporogenesis. The results indicated that the cytoplasmic and transcriptional changes occurring during zoosporogenesis are regulated by several pathways. For example, verapamil inhibited zoosporogenesis but not the up-regulation of most genes; the induction of some genes required while others were independent of calcium or phospholipid signaling; and, although most genes were induced in sporangia at 10 degrees C but not 24 degrees C, one was induced at both temperatures.


Hormonal regulation of skeletal muscle glycogen synthase through covalent phosphorylation.
Studies have been initiated to determine the hormonal regulation of glycogen synthase in rabbit skeletal muscle. It was found that glycogen synthase purified from control animals was quite highly phosphorylated (2.35 mol phosphate/mol synthase subunit) with 40% of the phosphate in the trypsin-sensitive or COOH-terminal domain, and 60% in the trypsin-insensitive or NH2-terminal domain. The phosphorylation state of synthase was elevated (3.9 mol/mol) by epinephrine injection and in the diabetic condition. With epinephrine, about 76% of the additional phosphate was incorporated in the trypsin-sensitive domain, which strongly supports the contention that this hormone acts through the cyclic AMP (cAMP)-dependent protein kinase. In the synthase purified from diabetic rabbits, 90% of the additional phosphate was in the trypsin-insensitive domain. Insulin treatment of the diabetics resulted in specific dephosphorylation of the trypsin-insensitive domain. These results indicate that in this system insulin is not acting by inhibition of the cAMP-dependent protein kinase.


Molecular processes underlying the floral transition in the soybean shoot apical meristem.
The transition to flowering is characterized by a shift of the shoot apical meristem (SAM) from leaf production to the initiation of a floral meristem. The flowering process is of vital importance for agriculture, but the associated events or regulatory pathways in the SAM are not well understood, especially at a system level. To address this issue, we have used a GeneChip containing 37 744 probe sets to generate a temporal profile of gene expression during the floral initiation process in the SAM of the crop legume, soybean (Glycine max). A total of 331 transcripts displayed significant changes in their expression profiles. The in silico and RT-PCR analysis on differentially regulated transcripts implies the intriguing involvement of sugar, auxin or abscisic acid (ABA) in events prior to the induction of floral homeotic transcripts. The novel involvement of ABA in the floral transition is further implicated by immunoassay, suggesting an increase in ABA levels in the SAM during this developmental transition. Furthermore, in situ localization, together with in silico data demonstrating a marked enhancement of abiotic stress-related transcripts, such as trehalose metabolism genes in SAMs, points to an overlap of abiotic stress and floral signalling pathways.


Towards a theoretical understanding of false positives in DNA motif finding.
Detection of false-positive motifs is one of the main causes of low performance in de novo DNA motif-finding methods. Despite the substantial algorithm development effort in this area, recent comprehensive benchmark studies revealed that the performance of DNA motif-finders leaves room for improvement in realistic scenarios. Using large-deviations theory, we derive a remarkably simple relationship that describes the dependence of false positives on dataset size for the one-occurrence per sequence motif-finding problem. As expected, we predict that false-positives can be reduced by decreasing the sequence length or by adding more sequences to the dataset. Interestingly, we find that the false-positive strength depends more strongly on the number of sequences in the dataset than it does on the sequence length, but that the dependence on the number of sequences diminishes, after which adding more sequences does not reduce the false-positive rate significantly. We compare our theoretical predictions by applying four popular motif-finding algorithms that solve the one-occurrence-per-sequence problem (MEME, the Gibbs Sampler, Weeder, and GIMSAN) to simulated data that contain no motifs. We find that the dependence of false positives detected by these softwares on the motif-finding parameters is similar to that predicted by our formula. We quantify the relationship between the sequence search space and motif-finding false-positives. Based on the simple formula we derive, we provide a number of intuitive rules of thumb that may be used to enhance motif-finding results in practice. Our results provide a theoretical advance in an important problem in computational biology.


Regulatory crackdown on stem cell therapy: what would the position be in South Africa?
This article attempts to answer the question of whether stem cell therapy falls within the current South African regulatory framework, using the Regenexx case in the USA as an example. The USA regulator, the Federal Drug Administration (FDA), is seeking to regulate the Regenexx autologous stem cell therapy as a 'drug' and a 'biological product'. The opposing position taken by the inventors of the Regenexx therapy is that re-implantation of one's own body parts can in principle not be a 'drug' and that the FDA is exceeding its mandate. In this article arguments are presented that the Regenexx therapy would qualify as a 'biological medicinal product' in the South African regulatory framework established in terms of the Medicines and Related Substances Control Act. As such, the Regenexx therapy would be subject to registration with the Medicines Control Council (MCC) as a legal precondition for its commercialisation. Furthermore, in order to convince the MCC of the safety, efficacy and quality of the Regenexx therapy, such therapy must - similar to any other new medicine - first be subjected to clinical trials. It is therefore concluded that stem cell therapy is indeed comprehensively regulated in South Africa, and that a recent opinion expressed in this journal that there exists a 'legislative vacuum' with relation to the regulation of stem cell therapy in South Africa is plainly incorrect.


Verification of cardiac mechanics software: benchmark problems and solutions for testing active and passive material behaviour.
Models of cardiac mechanics are increasingly used to investigate cardiac physiology. These models are characterized by a high level of complexity, including the particular anisotropic material properties of biological tissue and the actively contracting material. A large number of independent simulation codes have been developed, but a consistent way of verifying the accuracy and replicability of simulations is lacking. To aid in the verification of current and future cardiac mechanics solvers, this study provides three benchmark problems for cardiac mechanics. These benchmark problems test the ability to accurately simulate pressure-type forces that depend on the deformed objects geometry, anisotropic and spatially varying material properties similar to those seen in the left ventricle and active contractile forces. The benchmark was solved by 11 different groups to generate consensus solutions, with typical differences in higher-resolution solutions at approximately 0.5%, and consistent results between linear, quadratic and cubic finite elements as well as different approaches to simulating incompressible materials. Online tools and solutions are made available to allow these tests to be effectively used in verification of future cardiac mechanics software.


Effects of drought on the abundance and distribution of non-breeding shorebirds in central California, USA.
Conservation of migratory species requires anticipating the potential impacts of extreme climatic events, such as extreme drought. During drought, reduced habitat availability for shorebirds creates the potential for changes in their abundance and distribution, in part because many species are highly mobile and rely on networks of interior and coastal habitats. Understanding how shorebirds responded to a recent drought cycle that peaked from 2013 to 2015 in central California, USA, will help optimize management of wetlands and fresh water for wildlife. In the Central Valley, a vast interior region that is characterized by a mosaic of wetlands and agricultural lands, we found 22% and 29% decreases in the annual abundance of shorebirds during periods of 3-year drought (2013-2015) and 2-year extreme drought (2014-2015), respectively, when compared to non-drought years. Lower abundance of shorebirds coincided with significant decreases in the mean proportion flooded of survey units (7% and 9%, respectively) that were reliant on fresh water. Drought was associated with lower abundance within both the interior Central Valley and coastal San Francisco Bay for greater and lesser yellowlegs (Tringa melanoleuca and T. flavipes) and long- and short-billed dowitchers (Limnodromus scolopaceus and L. griseus). Only dunlins (Calidris alpina) had patterns of abundance that suggested substantial shifts in distribution between the Central Valley and coastal regions of San Francisco Bay and Point Reyes. Our results indicate that drought has the potential to reduce, at least temporally, shorebird populations and flooded habitat in the Central Valley, and the ability to respond to drought by taking advantage of nearby coastal habitats may limit the long-term effects of drought on some species. Successful conservation strategies must balance the impacts of reduced habitat availability at interior sites with the ability of some migratory shorebirds to adapt rapidly to shifting distributions of resources.


Variation in assembly stoichiometry in non-metazoan homologs of the hub domain of Ca2+ /calmodulin-dependent protein kinase II.
The multi-subunit Ca2+ /calmodulin-dependent protein kinase II (CaMKII) holoenzyme plays a critical role in animal learning and memory. The kinase domain of CaMKII is connected by a flexible linker to a C-terminal hub domain that assembles into a 12- or 14-subunit scaffold that displays the kinase domains around it. Studies on CaMKII suggest that the stoichiometry and dynamic assembly/disassembly of hub oligomers may be important for CaMKII regulation. Although CaMKII is a metazoan protein, genes encoding predicted CaMKII-like hub domains, without associated kinase domains, are found in the genomes of some green plants and bacteria. We show that the hub domains encoded by three related green algae, Chlamydomonas reinhardtii, Volvox carteri f. nagarensis, and Gonium pectoral, assemble into 16-, 18-, and 20-subunit oligomers, as assayed by native protein mass spectrometry. These are the largest known CaMKII hub domain assemblies. A crystal structure of the hub domain from C. reinhardtii reveals an 18-subunit organization. We identified four intra-subunit hydrogen bonds in the core of the fold that are present in the Chlamydomonas hub domain, but not in metazoan hubs. When six point mutations designed to recapitulate these hydrogen bonds were introduced into the human CaMKII-α hub domain, the mutant protein formed assemblies with 14 and 16 subunits, instead of the normal 12- and 14-subunit assemblies. Our results show that the stoichiometric balance of CaMKII hub assemblies can be shifted readily by small changes in sequence.


4-Hydroxynonenal, an endogenous aldehyde, causes pain and neurogenic inflammation through activation of the irritant receptor TRPA1.
TRPA1 is an excitatory ion channel expressed by a subpopulation of primary afferent somatosensory neurons that contain substance P and calcitonin gene-related peptide. Environmental irritants such as mustard oil, allicin, and acrolein activate TRPA1, causing acute pain, neuropeptide release, and neurogenic inflammation. Genetic studies indicate that TRPA1 is also activated downstream of one or more proalgesic agents that stimulate phospholipase C signaling pathways, thereby implicating this channel in peripheral mechanisms controlling pain hypersensitivity. However, it is not known whether tissue injury also produces endogenous proalgesic factors that activate TRPA1 directly to augment inflammatory pain. Here, we report that recombinant or native TRPA1 channels are activated by 4-hydroxy-2-nonenal (HNE), an endogenous alpha,beta-unsaturated aldehyde that is produced when reactive oxygen species peroxidate membrane phospholipids in response to tissue injury, inflammation, and oxidative stress. HNE provokes release of substance P and calcitonin gene-related peptide from central (spinal cord) and peripheral (esophagus) nerve endings, resulting in neurogenic plasma protein extravasation in peripheral tissues. Moreover, injection of HNE into the rodent hind paw elicits pain-related behaviors that are inhibited by TRPA1 antagonists and absent in animals lacking functional TRPA1 channels. These findings demonstrate that HNE activates TRPA1 on nociceptive neurons to promote acute pain, neuropeptide release, and neurogenic inflammation. Our results also provide a mechanism-based rationale for developing novel analgesic or anti-inflammatory agents that target HNE production or TRPA1 activation.


Comprehensive Biomarker Analyses in Patients with Advanced or Metastatic Non-Small Cell Lung Cancer Prospectively Treated with the Polo-Like Kinase 1 Inhibitor BI2536.
Polo like kinase 1 (PLK1) is frequently upregulated in tumors and is thus viewed as a promising therapeutic target in various cancers. Several PLK1 inhibitors have recently been developed and clinically tested in solid cancers, albeit with limited success. So far, no predictive biomarkers for PLK1 inhibitors have been established. To this end, we conducted a post-hoc biomarker analysis of tumor samples from non-small cell lung cancer (NSCLC) patients treated with the PLK1 inhibitor BI2536 in a phase II study. We analyzed formalin-fixed paraffin-embedded surplus tumor tissue from 47 study patients using immunohistochemistry (IHC) and DNA sequencing of KRAS, EGFR, BRAF, and PIK3CA. KRAS-mutated patients showed numerically prolonged progression-free survival, but statistical significance was not established. Interestingly, when pathways rather than single genes were analyzed, a positive correlation between IHC staining of activated ERK (p-ERK) and mutated KRAS was detected, whereas KRAS mutation status was found to be negatively correlated with activated AKT (p-AKT). With this hypothesis-generating study in BI2531-treated patients, we could not establish a correlation between KRAS mutations and relevant clinical endpoints. Future clinical trials with concomitant systematic biosampling and comprehensive molecular analyses are required to identify biomarkers predictive for response to PLK1 inhibitors.


16S rRNA sequences of uncultivated hot spring cyanobacterial mat inhabitants retrieved as randomly primed cDNA.
Cloning and analysis of cDNAs synthesized from rRNAs is one approach to assess the species composition of natural microbial communities. In some earlier attempts to synthesize cDNA from 16S rRNA (16S rcDNA) from the Octopus Spring cyanobacterial mat, a dominance of short 16S rcDNAs was observed, which appear to have originated only from certain organisms. Priming of cDNA synthesis from small ribosomal subunit RNA with random deoxyhexanucleotides can retrieve longer sequences, more suitable for phylogenetic analysis. Here we report the retrieval of 16S rRNA sequences from three formerly uncultured community members. One sequence type, which was retrieved three times from a total of five sequences analyzed, can be placed in the cyanobacterial phylum. A second sequence type is related to 16S rRNAs from green nonsulfur bacteria. The third sequence type may represent a novel phylogenetic type.


An unexpected role for platelets in blocking Th17 differentiation.
It is well known that platelets interact with cells of the innate immune system to promote tissue repair. In contrast, it is less clear whether these links extend to cells of the adaptive immune system, such as T cells. In this issue of the JCI, Morrell and colleagues provide compelling evidence that platelets are required to limit CD4+ Th17 differentiation through the actions of the chemokine platelet factor 4 (PF4). Absence of PF4 in the host leads to exaggerated Th17 differentiation after transplantation and rapid graft rejection. The authors' findings argue that platelets are not bit part players, but rather fully fledged, critical members of the adaptive immune system.


Breaking down the wall: fractionation of mycobacteria.
Mycobacterium spp. possess a complex cell envelope that consists of a plasma membrane, a peptidoglycan-arabinogalactan complex which in turn is esterified by mycolic acids that form with other non-bound lipids an asymmetric permeability barrier and an outer layer, also called a capsule in the case of pathogenic species. In order to investigate the functional roles of the cell envelope components, especially those of the major pathogens Mycobacterium tuberculosis and Mycobacterium leprae, it is necessary to fractionate the envelope by breaking the unusual wall that covers these bacteria. To this aim we first compared the efficiency of high pressure (cell disrupter/French press) with those of pathogen-compatible breakage methods such as sonication, bead beater and lysozyme treatment using the non-pathogenic Mycobacterium smegmatis. When the distribution of various specific markers of the cell envelope compartments, which include mycolic acids, arabinose, NADH oxidase activity, cell wall and cytosolic proteins, were determined sonication combined with lysozyme treatment was found to be the best option. The protocol of subcellular fractionation was then validated for pathogenic species by applying the method to Mycobacterium bovis BCG cells, an attenuated strain of the M. tuberculosis complex.


College programming for students in addiction recovery: A PRISMA-guided scoping review.
The health and well-being of students in recovery from substance use disorder are increasingly being recognized as a priority on college campuses. This scoping review maps the state of the existing literature evaluating collegiate recovery programming to highlight research gaps and inform policy. We conducted a systematic search of articles related to collegiate recovery programming published before August 2020. The 15 extracted study characteristics included publication type, study design, primary outcomes, reporting of behavioral addictions, mutual-help group attendance, sample demographic information, school size, ownership, and funding source. The PRISMA-guided search strategy identified 357 articles for abstract review; of 113 articles retained for full-text review, 54 studies met criteria for inclusion. Primary outcomes were coded into four domains: clinical, recovery experience, program characterization, and stigma. Most (57%) used quantitative observational designs and 41% employed qualitative research designs. Government or foundation grants funded 11% of the studies. The domains identified offer a framework for healthcare providers, college administrators, and researchers to understand and improve programs, thereby better serving this vulnerable student group.


Global epiproteomic signatures distinguish embryonic stem cells from differentiated cells.
Complex organisms contain a variety of distinct cell types but only a single genome. Therefore, cellular identity must be specified by the developmentally regulated expression of a subset of genes from an otherwise static genome. In mammals, genomic DNA is modified by cytosine methylation, resulting in a pattern that is distinctive for each cell type (the epigenome). Because nucleosomal histones are subject to a wide variety of post-translational modifications (PTMs), we reasoned that an analogous "epiproteome" might exist that could also be correlated with cellular identity. Here, we show that the quantitative evaluation of nucleosome PTMs yields epiproteomic signatures that are useful for the investigation of stem cell differentiation, chromatin function, cellular identity, and epigenetic responses to pharmacologic agents. We have developed a novel enzyme-linked immunosorbent assay-based method for the quantitative evaluation of the steady-state levels of PTMs and histone variants in preparations of native intact nucleosomes. We show that epiproteomic responses to the histone deacetylase inhibitor trichostatin A trigger changes in histone methylation as well as acetylation, and that the epiproteomic responses differ between mouse embryonic stem cells and mouse embryonic fibroblasts (MEFs). ESCs subjected to retinoic acid-induced differentiation contain reconfigured nucleosomes that include increased content of the histone variant macroH2A and other changes. Furthermore, ESCs can be distinguished from embryonal carcinoma cells and MEFs based purely on their epiproteomic signatures. These results indicate that epiproteomic nucleosomal signatures are useful for the investigation of stem cell identity and differentiation, nuclear reprogramming, epigenetic regulation, chromatin dynamics, and assays for compounds with epigenetic activities. Disclosure of potential conflicts of interest is found at the end of this article.


Wideband energy reflectance measurements: effects of negative middle ear pressure and application of a pressure compensation procedure.
The wideband energy reflectance (ER) technique has become popular as a tool for evaluating middle ear function. Negative middle ear pressure (MEP) is a prevalent form of middle ear dysfunction, which may impact application of ER measurements in differential diagnosis. A negative MEP may be countervailed by application of an equivalent negative ear canal pressure. The present study examined ER in the same ears under normal and experimentally induced negative MEP conditions. Thirty-five subjects produced at least one negative MEP each (-40 to -225 daPa). Negative MEP significantly altered ER in a frequency-specific manner that varied with MEP magnitude. ER increased for low- to mid-frequencies with the largest change (~0.20 to 0.40) occurring between 1 and 1.5 kHz. ER decreased for frequencies above 3 kHz with the largest change (~-0.10 to -0.25) observed between 4.5 and 5.5 kHz. Magnitude of changes increased as MEP became more negative, as did the frequencies at which maximum changes occurred, and the frequency at which enhancement transitioned to reduction. Ear canal pressure compensation restored ER to near baseline values. This suggests that the compensation procedure adequately mitigates the effects of negative MEP on ER. Theoretical issues and clinical implications are discussed.


Circannual changes in stress and feeding hormones and their effect on food-seeking behaviors.
Seasonal fluctuations in food availability show a tight association with seasonal variations in body weight and food intake. Seasonal variations in food intake, energy storage, and expenditure appear to be a widespread phenomenon suggesting they may have evolved in anticipation for changing environmental demands. These cycles appear to be driven by changes in external daylength acting on neuroendocrine pathways. A number of neuroendocrine pathways, two of which are the endocrine mechanisms underlying feeding and stress, appear to show seasonal changes in both their circulating levels and reactivity. As such, variation in the level or reactivity to these hormones may be crucial factors in the control of seasonal variations in food-seeking behaviors. The present review examines the relationship between feeding behavior and seasonal changes in circulating hormones. We hypothesize that seasonal changes in circulating levels of glucocorticoids and the feeding-related hormones ghrelin and leptin contribute to seasonal fluctuations in feeding-related behaviors. This review will focus on the seasonal circulating levels of these hormones as well as sensitivity to these hormones in the modulation of food-seeking behaviors.


Bioengineering Organs for Blood Detoxification.
For patients with severe kidney or liver failure the best solution is currently organ transplantation. However, not all patients are eligible for transplantation and due to limited organ availability, most patients are currently treated with therapies using artificial kidney and artificial liver devices. These therapies, despite their relative success in preserving the patients' life, have important limitations since they can only replace part of the natural kidney or liver functions. As blood detoxification (and other functions) in these highly perfused organs is achieved by specialized cells, it seems relevant to review the approaches leading to bioengineered organs fulfilling most of the native organ functions. There, the culture of cells of specific phenotypes on adapted scaffolds that can be perfused takes place. In this review paper, first the functions of kidney and liver organs are briefly described. Then artificial kidney/liver devices, bioartificial kidney devices, and bioartificial liver devices are focused on, as well as biohybrid constructs obtained by decellularization and recellularization of animal organs. For all organs, a thorough overview of the literature is given and the perspectives for their application in the clinic are discussed.