Target-specific mechanics of phagocytosis: protrusive neutrophil response to zymosan differs from the uptake of antibody-tagged pathogens.
The physical mechanisms that control target-specific responses of human neutrophils to distinct immune threats are poorly understood. Using dual-micropipette manipulation, we have quantified and compared the time courses of neutrophil phagocytosis of two different targets: zymosan (a prominent model of fungal infection), and antibody-coated (Fc) particles. Our single-live-cell/single-target approach exposes surprising differences between these two forms of phagocytosis. Unlike the efficient uptake of 3-μm Fc targets (within ~66 seconds), the engulfment of similarly sized zymosan is slow (~167 seconds), mainly due to the formation of a characteristic pedestal that initially pushes the particle outwards by ~1 μm. Despite a roughly twofold difference in maximum cortical tensions, the top 'pull-in' speeds of zymosan and Fc targets are indistinguishable at ~33 nm/second. Drug inhibition shows that both actin as well as myosin II partake in the regulation of neutrophil cortical tension and cytoplasmic viscosity; other than that, myosin II appears to play a minor role in both forms of phagocytosis. Remarkably, an intact actin cytoskeleton is required to suppress, in antibody-mediated phagocytosis, the initially protrusive deformation that distinguishes the neutrophil response to zymosan.


Age at gluten introduction and risk of celiac disease.
The goal of this study was to determine whether age at introduction to gluten was associated with risk for celiac disease (CD) in genetically predisposed children. TEDDY (The Environmental Determinants of Diabetes in the Young) is a prospective birth cohort study. Newborn infants (N = 6436) screened for high-risk HLA-genotypes for CD were followed up in Finland, Germany, Sweden, and the United States. Information about infant feeding was collected at clinical visits every third month. The first outcome was persistent positive for tissue transglutaminase autoantibodies (tTGA), the marker for CD. The second outcome was CD, defined as either a diagnosis based on intestinal biopsy results or on persistently high levels of tTGA. Swedish children were introduced to gluten earlier (median: 21.7 weeks) compared with children from Finland (median: 26.1 weeks), Germany, and the United States (both median: 30.4 weeks) (P < .0001). During a median follow-up of 5.0 years (range: 1.7-8.8 years), 773 (12%) children developed tTGA and 307 (5%) developed CD. Swedish children were at increased risk for tTGA (hazard ratio: 1.74 [95% CI: 1.47-2.06]) and CD (hazard ratio: 1.76 [95% CI: 1.34-2.24]) compared with US children, respectively (P < .0001).Gluten introduction before 17 weeks or later than 26 weeks was not associated with increased risk for tTGA or CD, adjusted for country, HLA, gender, and family history of CD, neither in the overall analysis nor on a country-level comparison. In TEDDY, the time to first introduction to gluten introduction was not an independent risk factor for developing CD.


The Role of Active Engagement of Peer Observation in the Acquisition of Surgical Skills in Virtual Reality Tasks for Novices.
Peer-assisted learning has been regarded as an adjunct to teaching modalities. It remains inconclusive regarding the benefits of peer observation in skills learning. Hence, we investigated whether the active engagement (AE) of peer observation in addition to expert demonstration would facilitate the performance in the virtual reality (VR) tasks. The programs involved 4 VR tasks including basic (camera targeting), intermediate (energy dissection and energy switching), and advanced (suture sponge) tasks in the da Vinci Skills Simulators, which were set up in the operating room at Taipei Medical University Hospital. Fifty medical students participated in the study. The AE of the participants was defined as the total number of peer observations in addition to expert observation before their performance. We assessed the correlations between AE and surgical task performance using Pearson correlation and the concept of learning analytics. Medical students (sixth-year students in Taiwan, equivalent to fourth-year students in the US system) from Taipei Medical University were recruited. AE was correlated with the energy dissection task (r = 0.329, p = 0.02) and marginally associated with the energy switching task (r = 0.271, p = 0.057). However, AE was not correlated with either task scores for camera targeting (r = 0.096, p = 0.509) or task scores for suture sponge (r = -0.091, p = 0.529). Our findings suggest that AE of peer observation may facilitate learning energy dissection task, which is an intermediate-level task, but not in other basic or advanced tasks in a VR context. The study highlights the potential effect of AE of peer observation on surgical learning based on a distinct level of tasks. Tasks that fit the learners' level are recommended. Nevertheless, the effectiveness of peer observation on surgical training still has to be explored to ensure favorable results and optimal learning outcomes.


Generating multiplex gradients of biomolecules for controlling cellular adhesion in parallel microfluidic channels.
Here we present a microfluidic platform to generate multiplex gradients of biomolecules within parallel microfluidic channels, in which a range of multiplex concentration gradients with different profile shapes are simultaneously produced. Nonlinear polynomial gradients were also generated using this device. The gradient generation principle is based on implementing parrallel channels with each providing a different hydrodynamic resistance. The generated biomolecule gradients were then covalently functionalized onto the microchannel surfaces. Surface gradients along the channel width were a result of covalent attachments of biomolecules to the surface, which remained functional under high shear stresses (50 dyn/cm(2)). An IgG antibody conjugated to three different fluorescence dyes (FITC, Cy5 and Cy3) was used to demonstrate the resulting multiplex concentration gradients of biomolecules. The device enabled generation of gradients with up to three different biomolecules in each channel with varying concentration profiles. We were also able to produce 2-dimensional gradients in which biomolecules were distributed along the length and width of the channel. To demonstrate the applicability of the developed design, three different multiplex concentration gradients of REDV and KRSR peptides were patterned along the width of three parallel channels and adhesion of primary human umbilical vein endothelial cell (HUVEC) in each channel was subsequently investigated using a single chip.


The teleost anatomy ontology: anatomical representation for the genomics age.
The rich knowledge of morphological variation among organisms reported in the systematic literature has remained in free-text format, impractical for use in large-scale synthetic phylogenetic work. This noncomputable format has also precluded linkage to the large knowledgebase of genomic, genetic, developmental, and phenotype data in model organism databases. We have undertaken an effort to prototype a curated, ontology-based evolutionary morphology database that maps to these genetic databases (http://kb.phenoscape.org) to facilitate investigation into the mechanistic basis and evolution of phenotypic diversity. Among the first requirements in establishing this database was the development of a multispecies anatomy ontology with the goal of capturing anatomical data in a systematic and computable manner. An ontology is a formal representation of a set of concepts with defined relationships between those concepts. Multispecies anatomy ontologies in particular are an efficient way to represent the diversity of morphological structures in a clade of organisms, but they present challenges in their development relative to single-species anatomy ontologies. Here, we describe the Teleost Anatomy Ontology (TAO), a multispecies anatomy ontology for teleost fishes derived from the Zebrafish Anatomical Ontology (ZFA) for the purpose of annotating varying morphological features across species. To facilitate interoperability with other anatomy ontologies, TAO uses the Common Anatomy Reference Ontology as a template for its upper level nodes, and TAO and ZFA are synchronized, with zebrafish terms specified as subtypes of teleost terms. We found that the details of ontology architecture have ramifications for querying, and we present general challenges in developing a multispecies anatomy ontology, including refinement of definitions, taxon-specific relationships among terms, and representation of taxonomically variable developmental pathways.


Patterns of Care for Stage IA Cervical Cancer: Use of Definitive Radiation Therapy Versus Hysterectomy.
The standard of care for clinical IA cervical cancer is surgery, but nonoperative cases may receive definitive radiation therapy (RT). Herein, we investigated national practice patterns associated with the administration of definitive RT as compared with hysterectomy-based surgery (HYS) as well as delivery of adjuvant RT after HYS. The National Cancer Data Base (NCDB) was queried for clinical IA primary cervical cancer cases (2004-2013) receiving definitive RT or HYS with or without adjuvant RT. Patients with unknown RT or surgery status were excluded, as were benign histologies and receipt of non-HYS such as fertility-sparing surgery. Patient, tumor, and treatment parameters were extracted. Univariable and multivariable logistic regression determined variables associated with receipt of RT and HYS. In total, 3816 patients were analyzed (n = 3514 [92.1%] HYS alone, n = 100 [2.6%] RT alone, n = 202 [5.3%] combination). On multivariable analysis of HYS versus definitive RT, RT was more likely to be given to patients who were older (P < 0.001) and with Medicare (P = 0.011), Medicaid/other government insurance (P = 0.011), or uninsured/unknown status (P = 0.003). In addition, treatment with surgery alone was associated with patients in the 2 highest income quartiles (P = 0.013, P = 0.054). On multivariable analysis of patients receiving RT in addition to HYS, adjuvant RT was added most commonly for positive margins (P < 0.001) and increasing age (P < 0.001). This is the largest analysis to date evaluating definitive RT for IA cervical cancer. Younger age and higher socioeconomic status are associated with receipt of HYS instead of definitive RT, and positive margins are most associated with the addition of adjuvant RT. Although these data must be further validated with better defined patient selection and do not imply causation, several socioeconomic findings discovered herein need to be addressed to ensure the highest quality cancer care to all patients.


Generation and propagation of partially coherent beams with nonconventional correlation functions: a review [invited].
Partially coherent beams with nonconventional correlation functions have displayed many extraordinary properties, such as self-focusing and self-splitting, which are totally different from those of partially coherent beams with conventional Gaussian correlated Schell-model functions and are useful in many applications, such as optical trapping, free-space optical communications, and material thermal processing. In this paper, we present a review of recent developments on generation and propagation of partially coherent beams with nonconventional correlation functions.


Importance of cumulative exposure to elevated cholesterol and blood pressure in development of atherosclerotic coronary artery disease in systemic lupus erythematosus: a prospective proof-of-concept cohort study.
Previous studies have shown that traditional risk factors such as hypercholesterolemia and hypertension account for only a small proportion of the dramatically increased risk of atherosclerotic coronary artery disease (CAD) in systemic lupus erythematosus (SLE). However, in these studies, exposure to risk factors was measured only at baseline. In this study, our objective was to compare measures of cumulative exposure with remote and recent values for each of total cholesterol (TC), systolic (SBP), and diastolic (DBP) blood pressure in terms of ability to quantify risk of atherosclerotic CAD in patients with SLE. Patients in the Toronto lupus cohort had TC and BP measured at each clinic visit and were followed up prospectively for the occurrence of CAD. For each patient, arithmetic mean, time-adjusted mean (AM) and area-under-the-curve (AUC) were calculated for serial TC, SBP, and DBP measurements. Proportional hazards regression models were used to compare these summary measures with recent and first-available ("remote") measurements in terms of ability to quantify risk of CAD events, defined as myocardial infarction, angina, or sudden cardiac death. The 991 patients had a mean ± SD of 19 ± 19 TC measurements per patient. Over a follow-up of 6.7 ± 6.4 years, 86 CAD events occurred; although remote TC was not significantly predictive of CAD, mean and AM TC were more strongly predictive (hazard ratio (HR) 2.07; P = 0.003) than recent TC (HR 1.86, P = 0.001). AUC TC was not predictive of CAD. A similar pattern was seen for DBP and SBP. Older age, male sex, higher baseline and recent disease activity score, and corticosteroid use also increased CAD risk, whereas antimalarials were protective. In contrast to the population-based Framingham model, first-available TC and BP are not predictive of CAD among patients with SLE, in whom measures reflecting cumulative exposure over time are better able to quantify CAD risk. This is an important consideration in future studies of dynamic risk factors for CAD in a chronic relapsing-remitting disease such as SLE. Our findings also underpin the importance of adequate control of SLE disease activity while minimizing corticosteroid use, and highlight the cardioprotective effect of antimalarials.


Laboratory animal allergy in a pharmaceutical company.
A cross sectional survey was carried out on 138 workers exposed to laboratory animals. Sixty (44%) had symptoms in a self completed questionnaire that were consistent with laboratory animal allergy (LAA) of whom 15 (11%) had chest symptoms. There was a positive skin prick test to one or more animal urine extracts (rat, mouse, guinea pig, rabbit) in 13% and 38% had a positive radioallergosorbent test to urine extract. LAA chest symptoms were almost five times more common in atopic than non-atopic subjects (who were distinguished by skin test response to common, non-animal aeroallergens). A positive skin test to animal urine was associated with LAA chest symptoms and with atopy. Nose, eye, or skin symptoms without chest symptoms were not associated with atopy. There was an inverse relation between duration of employment at the firm and LAA chest symptoms, suggesting selection of affected people out of employment with animals.


Genome-Wide DNA Methylation Patterns in Persistent Attention-Deficit/Hyperactivity Disorder and in Association With Impulsive and Callous Traits.
Attention-deficit/hyperactivity disorder (ADHD) is a neurodevelopmental disorder that often persists into adulthood. ADHD and related personality traits, such as impulsivity and callousness, are caused by genetic and environmental factors and their interplay. Epigenetic modifications of DNA, including methylation, are thought to mediate between such factors and behavior and may behave as biomarkers for disorders. Here, we set out to study DNA methylation in persistent ADHD and related traits. We performed epigenome-wide association studies (EWASs) on peripheral whole blood from participants in the NeuroIMAGE study (age range 12-23 years). We compared participants with persistent ADHD (n = 35) with healthy controls (n = 19) and with participants with remittent ADHD (n = 19). Additionally, we performed EWASs of impulsive and callous traits derived from the Conners Parent Rating Scale and the Callous-Unemotional Inventory, respectively, across all participants. For every EWAS, the linear regression model analyzed included covariates for age, sex, smoking scores, and surrogate variables reflecting blood cell type composition and genetic background. We observed no epigenome-wide significant differences in single CpG site methylation between participants with persistent ADHD and healthy controls or participants with remittent ADHD. However, epigenome-wide analysis of differentially methylated regions provided significant findings showing that hypermethylated regions in the APOB and LPAR5 genes were associated with ADHD persistence compared to ADHD remittance (p = 1.68 * 10-24 and p = 9.06 * 10-7, respectively); both genes are involved in cholesterol signaling. Both findings appeared to be linked to genetic variation in cis. We found neither significant epigenome-wide single CpG sites nor regions associated with impulsive and callous traits; the top-hits from these analyses were annotated to genes involved in neurotransmitter release and the regulation of the biological clock. No link to genetic variation was observed for these findings, which thus might reflect environmental influences. In conclusion, in this pilot study with a small sample size, we observed several DNA-methylation-disorder/trait associations of potential significance for ADHD and the related behavioral traits. Although we do not wish to draw conclusions before replication in larger, independent samples, cholesterol signaling and metabolism may be of relevance for the onset and/or persistence of ADHD.


Light enhanced amino acid uptake by dominant bacterioplankton groups in surface waters of the Atlantic Ocean.
(35)S-Methionine and (3)H-leucine bioassay tracer experiments were conducted on two meridional transatlantic cruises to assess whether dominant planktonic microorganisms use visible sunlight to enhance uptake of these organic molecules at ambient concentrations. The two numerically dominant groups of oceanic bacterioplankton were Prochlorococcus cyanobacteria and bacteria with low nucleic acid (LNA) content, comprising 60% SAR11-related cells. The results of flow cytometric sorting of labelled bacterioplankton cells showed that when incubated in the light, Prochlorococcus and LNA bacteria increased their uptake of amino acids on average by 50% and 23%, respectively, compared with those incubated in the dark. Amino acid uptake of Synechococcus cyanobacteria was also enhanced by visible light, but bacteria with high nucleic acid content showed no light stimulation. Additionally, differential uptake of the two amino acids by the Prochlorococcus and LNA cells was observed. The populations of these two types of cells on average completely accounted for the determined 22% light enhancement of amino acid uptake by the total bacterioplankton community, suggesting a plausible way of harnessing light energy for selectively transporting scarce nutrients that could explain the numerical dominance of these groups in situ.


The Structural Stability of Graphene Anticorrosion Coating Materials is Compromised at Low Potentials.
Corrosion of engineered structures is a major problem causing an estimated economic loss of more than 2 trillion US dollars annually worldwide. Graphene has recently emerged as highly promising, low-cost, and transparent anticorrosion coating material. Herein, it is shown that a multilayer graphene film grown on Ni by chemical vapor deposition undergoes abrupt stability failure under galvanic-corrosion conditions. The multilayer graphene coating was examined by optical microscopy, SEM, energy dispersive X-ray spectroscopy, Raman spectroscopy, and cyclic voltammetry after exposure to potentials between 600 and 1300 mV in alkaline solution. A fast and simple electrochemical method is proposed to sensitively quantify the damage caused by the applied potential bias. It is based on quantification of the oxidation signals generated by the underlying Ni-metal catalyst that is exposed by damage to the graphene film. It is shown that film damage can start at potentials as low as 900 mV and that macroscopic and extensive damage can be caused at potentials above 1000 mV. In addition, once the graphene film has been damaged, the corrosion rate of the underlying metal is significantly increased. These findings are of great importance for potential applications of multilayer graphene films in coating metal structures with huge industrial and economic implications.


Nonprocessive methylation by Dot1 leads to functional redundancy of histone H3K79 methylation states.
Whereas mono-, di- and trimethylation states of lysines on histones typically have specific functions, no specific functions have been attributed so far to the different methylation states of histone H3 Lysine 79 (H3K79) generated by Dot1. Here we show that Dot1, in contrast to other known histone methyltransferases, introduces multiple methyl groups via a nonprocessive mechanism. The kinetic mechanism implies that the H3K79 methylation states cannot be generated independently, suggesting functional redundancy. Indeed, gene silencing in yeast, which is dependent on Dot1, relied on global H3K79 methylation levels and not on one specific methylation state. Furthermore, our findings suggest that histone H2B ubiquitination affects H3K79 trimethylation by enhancing synthesis of all H3K79 methylation states. Our results suggest that multiple methylation of H3K79 leads to a binary code, which is expected to limit the possibilities for regulation by putative demethylases or binding proteins.


Training in Neurovascular Intervention Usefulness of in-Vitro Model and Clinical Practice.
We introduce our training tools and system of neurovascular intervention. An in vitro cerebral vascular model was used for the young residents to understand the basic interventional techniques and devices. The model included several vascular lesions such as cerebral aneurysm, dural arterio-venous fistula, or carotid artery stenosis. Endovascular procedures in the model were performed under fluoroscopic or direct visual control, and consecutive haemodynamic changes were visualized by using digital subtraction angiography and direct observation. Thus, traineess could have an easy understanding of clinical conditions. New medical devices, such as platinum coils, were successfully implanted in the model under stable conditions. After the initial training using vascular model, the residents had started clinical experiences under the control of senior surgeons. Although it is difficult to describe usefulness of our clinical training, we believe that we provide enough good quality and quantity of clinical cases to the residents. Because our endovascular team has recently had150-200 interventional procedures every year, one resident can have experienced more than 100 cases per year. The qualification of a Board Certified Specialist of the Japanese Society of Intravascular Neurosurgery (JSIN) requires that the applicant must have experienced more than 100 cases for four years. So our residents can have enough case materials to qualify the board examination.


Medication-Related Osteonecrosis of the Jaw: MASCC/ISOO/ASCO Clinical Practice Guideline.
To provide guidance regarding best practices in the prevention and management of medication-related osteonecrosis of the jaw (MRONJ) in patients with cancer. Multinational Association of Supportive Care in Cancer/International Society of Oral Oncology (MASCC/ISOO) and ASCO convened a multidisciplinary Expert Panel to evaluate the evidence and formulate recommendations. Guideline development involved a systematic review of the literature and a formal consensus process. PubMed and EMBASE were searched for studies of the prevention and management of MRONJ related to bone-modifying agents (BMAs) for oncologic indications published between January 2009 and December 2017. Results from an earlier systematic review (2003 to 2008) were also included. The systematic review identified 132 publications, only 10 of which were randomized controlled trials. Recommendations underwent two rounds of consensus voting. Currently, MRONJ is defined by (1) current or previous treatment with a BMA or angiogenic inhibitor, (2) exposed bone or bone that can be probed through an intraoral or extraoral fistula in the maxillofacial region and that has persisted for longer than 8 weeks, and (3) no history of radiation therapy to the jaws or metastatic disease to the jaws. In patients who initiate a BMA, preventive care includes comprehensive dental assessments, discussion of modifiable risk factors, and avoidance of elective dentoalveolar surgery (ie, surgery that involves the teeth or contiguous alveolar bone) during BMA treatment. It remains uncertain whether BMAs should be discontinued before dentoalveolar surgery. Staging of MRONJ should be performed by a clinician with experience in the management of MRONJ. Conservative measures comprise the initial approach to MRONJ treatment. Ongoing collaboration among the dentist, dental specialist, and oncologist is essential to optimal patient care.


ZEBrA: Zebra finch Expression Brain Atlas-A resource for comparative molecular neuroanatomy and brain evolution studies.
An in-depth understanding of the genetics and evolution of brain function and behavior requires a detailed mapping of gene expression in functional brain circuits across major vertebrate clades. Here we present the Zebra finch Expression Brain Atlas (ZEBrA; www.zebrafinchatlas.org, RRID: SCR_012988), a web-based resource that maps the expression of genes linked to a broad range of functions onto the brain of zebra finches. ZEBrA is a first of its kind gene expression brain atlas for a bird species and a first for any sauropsid. ZEBrA's >3,200 high-resolution digital images of in situ hybridized sections for ~650 genes (as of June 2019) are presented in alignment with an annotated histological atlas and can be browsed down to cellular resolution. An extensive relational database connects expression patterns to information about gene function, mouse expression patterns and phenotypes, and gene involvement in human diseases and communication disorders. By enabling brain-wide gene expression assessments in a bird, ZEBrA provides important substrates for comparative neuroanatomy and molecular brain evolution studies. ZEBrA also provides unique opportunities for linking genetic pathways to vocal learning and motor control circuits, as well as for novel insights into the molecular basis of sex steroids actions, brain dimorphisms, reproductive and social behaviors, sleep function, and adult neurogenesis, among many fundamental themes.


Enhancement of morphine antinociception by a CCKB antagonist in mice is mediated via opioid delta receptors.
This study investigated the possible involvement of opioid delta receptors in the modulation of morphine antinociceptive potency produced by L365,260 a CCKB antagonist. Intracerebroventricular (i.c.v.), intratheoal (i.th.) or subcutaneous (s.c.) L365,260 alone did not produce any antinociceptive actions in the mouse warm-water tail-nick test. Treatment with L365,260 by any of these routes produced a leftward shift of the corresponding morphine dose-effect curve that was blocked by pretreatment with a receptor selective dose of s.c. naltrindole, an opioid delta receptor antagonist. Pretreatment with i.c.v. antisera to [Leu5]enkephalin also blocked the leftward displacement of the i.c.v. morphine dose-effect curve resulting from L365,260 but did not directly alter the i.c.v. morphine dose-effect curve; antisera to [Met5]enkephalin did not alter the effects of morphine or the modulation of morphine antinociception produced by L365,260. Repeated pretreatment with L365,260 resulted in a progressive decrease in the magnitude of the morphine modulatory action (i.e., L365,260 "tolerance"). In these "L365,260-tolerant" mice, the dose-effect curve for i.c.v. [D-Ala2, Glu4]deltrophin (a selective delta agonist) was displaced to the right by approximately 8.2-fold. The i.c.v. administration of [Leu5]enkephalin produced a leftward displacement of the i.c.v. morphine dose-effect curve that diminished after repeated administration (i.e., [Leu5]enkephalin "tolerance"). In "[Leu5]enkephalin-tolerant" mice, L365,260 failed to produce the leftward shift of the morphine dose-effect curve seen in control animals. That is, two-way antinociceptive cross-tolerance was observed between an opioid delta agonist and a CCKB receptor antagonist. Intracerebroventricular thiorphan, a peptidase inhibitor, did not elicit antinociception directly. Co-administration of thiorphan with L365,260 elicited significant antinociception that was blocked by naltrindole or antisera to [Leu5]enkephalin; antisera to [Met5]enkephalin had no effect. Repeated administration of i.c.v. [D-Ala2, Glu4]deltorphin resulted in a progressively decreasing antinociceptive effect (i.e., [D-Ala2, Glu4]deltorphin "tolerance"). In "[D-Ala2, Glu4]deltorphin-tolerant" mice, the thiorphan/L365,260 antinociceptive effect was inhibited. Collectively, these data suggest that CCK interacts at the CCKB receptor to inhibit tonically the release and/or availability of an endogenous substance acting at opioid delta receptors. The subsequent enhancement of morphine antinociceptive potency may reflect the well-known modulation of morphine antinociception produced by opioid delta receptor agonists. In this case, the latter may be [Leu5]enkephalin or a [Leu5]enkephalin-like substance.


Population genetic analysis infers migration pathways of Phytophthora ramorum in US nurseries.
Recently introduced, exotic plant pathogens may exhibit low genetic diversity and be limited to clonal reproduction. However, rapidly mutating molecular markers such as microsatellites can reveal genetic variation within these populations and be used to model putative migration patterns. Phytophthora ramorum is the exotic pathogen, discovered in the late 1990s, that is responsible for sudden oak death in California forests and ramorum blight of common ornamentals. The nursery trade has moved this pathogen from source populations on the West Coast to locations across the United States, thus risking introduction to other native forests. We examined the genetic diversity of P. ramorum in United States nurseries by microsatellite genotyping 279 isolates collected from 19 states between 2004 and 2007. Of the three known P. ramorum clonal lineages, the most common and genetically diverse lineage in the sample was NA1. Two eastward migration pathways were revealed in the clustering of NA1 isolates into two groups, one containing isolates from Connecticut, Oregon, and Washington and the other isolates from California and the remaining states. This finding is consistent with trace forward analyses conducted by the US Department of Agriculture's Animal and Plant Health Inspection Service. At the same time, genetic diversities in several states equaled those observed in California, Oregon, and Washington and two-thirds of multilocus genotypes exhibited limited geographic distributions, indicating that mutation was common during or subsequent to migration. Together, these data suggest that migration, rapid mutation, and genetic drift all play a role in structuring the genetic diversity of P. ramorum in US nurseries. This work demonstrates that fast-evolving genetic markers can be used to examine the evolutionary processes acting on recently introduced pathogens and to infer their putative migration patterns, thus showing promise for the application of forensics to plant pathogens.


Combining Optogenetics and Electrophysiology to Analyze Projection Neuron Circuits.
A set of methods is described for channelrhodopsin-2 (ChR2)-based synaptic circuit analysis that combines photostimulation of virally transfected presynaptic neurons' axons with whole-cell electrophysiological recordings from retrogradely labeled postsynaptic neurons. The approach exploits the preserved photoexcitability of ChR2-expressing axons in brain slices and can be used to assess either local or long-range functional connections. Stereotaxic injections are used both to express ChR2 selectively in presynaptic axons of interest (using rabies virus [RV] or adeno-associated virus [AAV]) and to label two types of postsynaptic projection neurons of interest with fluorescent retrograde tracers. In brain slices, tracer-labeled postsynaptic neurons are targeted for whole-cell electrophysiological recordings, and synaptic connections are assessed by sampling voltage or current responses to light-emitting diode (LED) photostimulation of ChR2-expressing axons. The data are analyzed to estimate the relative amplitude of synaptic input and other connectivity parameters. Pharmacological and electrophysiological manipulations extend the versatility of the basic approach, allowing the dissection of monosynaptic versus disynaptic responses, excitatory versus inhibitory responses, and more. The method enables rapid, quantitative characterization of synaptic connectivity between defined pre- and postsynaptic classes of neurons.


BMP1 and TLL1 Are Required for Maintaining Periodontal Homeostasis.
Mutations in bone morphogenetic protein 1 (BMP1) in humans or deletion of BMP1 and related protease tolloid like 1 (TLL1) in mice lead to osteogenesis imperfecta (OI). Here, we show progressive periodontal defects in mice in which both BMP1 and TLL1 have been conditionally ablated, including malformed periodontal ligament (PDL) (recently shown to play key roles in normal alveolar bone formation), significant loss in alveolar bone mass ( P < 0.01), and a sharp reduction in cellular cementum. Molecular mechanism studies revealed a dramatic increase in the uncleaved precursor of type I collagen (procollagen I) and a reduction in dentin matrix protein 1 (DMP1), which is partially responsible for defects in extracellular matrix (ECM) formation and mineralization. We also showed a marked increase in the expression of matrix metallopeptidase 13 (MMP13) and tartrate-resistant acid phosphatase (TRAP), leading to an acceleration in periodontal breakdown. Finally, we demonstrated that systemic application of antibiotics significantly improved the alveolar bone and PDL damage of the knockdown phenotype, which are thus shown to be partially secondary to pathogen-induced inflammation. Together, identification of the novel roles of BMP1 and TLL1 in maintaining homeostasis of periodontal formation, partly via biosynthetic processing of procollagen I and DMP1, provides novel insights into key contributions of the extracellular matrix environment to periodontal homeostasis and contributes toward understanding of the pathology of periodontitis.


Comparison of conventional vs self-etching adhesive bonds to caries-affected dentin.
The mechanism of resin adhesion to caries-affected dentin is still unclear. This study evaluated the interfacial morphology of two bonding systems to caries-affected dentin, coupled with the measurement of microtensile bond strengths (mu TBS). Carious human molars were prepared as previously described in Nakajima and others (1995) and were bonded with Single Bond (SB) or FluoroBond (FB) according to the manufacturer's instructions, followed by creation of AP-X composite buildups. After one day of storage in 37 degrees C water, the teeth were serially sectioned vertically into 0.8 mm slabs, trimmed to yield a 1 mm2 test area, and tested to failure in a Bencor device used in an Instron machine operated at 1 mm/min. Resin-dentin interfaces were observed with SEM before or after acid/base challenge. Bonding to normal dentin with the two bonding systems (SB and FB) showed tensile bond strengths significantly higher than those to caries-affected dentin. The moist bonding technique significantly increased bond strength of SB to normal and caries-affected dentin. SEM examination revealed that typical hybrid layer and resin tags could not be formed to caries-affected dentin. The results suggested that resin penetration may be prevented by occlusion of dentinal tubules by mineral deposits that may also impart acid-resistance to the intertubular matrix of caries-affected dentin.


Sex differences in emotional and psychophysiological responses to musical stimuli.
Although it is known that men and women differ in their music preferences and emotional reactions to music, little is known about sex differences in physiological reactions to music. In our study, we therefore set out to examine the differential reactivity to two musical stimuli that elicit distinct psychological and physiological reaction patterns. Fifty-three healthy subjects (mean age: 26.13, SD: 3.97; 26 males, 27 females) were examined. Heart rate, electrodermal activity, skin temperature, salivary cortisol, salivary alpha-amylase, and psychological variables were assessed during the course of the whole study. Following baseline assessment, two musical stimuli, which were carefully selected and rated in a pre-study as relaxing and pleasant (renaissance music) and arousing and unpleasant (heavy metal), respectively, were introduced. They were presented on two different days in a randomized order. Whereas psychological variables did not differ between men and women, results of electrophysiological measures indicate significantly different reactivity patterns between men and women. Women displayed elevated response curves to the arousing and unpleasant stimulus, whereas men did not. However, no differences were found with regards to endocrine measures in saliva. Our results demonstrate sex differences in reactivity patterns to musical stimuli in psychophysiological measures. In our study, we were able to show that women tend to show hypersensitivity to aversive musical stimuli. This finding is in accordance with previous literature on sex differences in emotion research. Furthermore, our study indicates that the confounding effects of sex differences have to be considered when using musical stimuli for emotion induction.


Prevalence and predictors of self-medication in a selected urban and rural district of Sri Lanka.
Self-medication is widely practised in many developing countries. The determinants of self-medication need to be understood to design adequate medicine information policies and patient-dispenser education strategies. Hence, the prevalence of medicine use and predictors of self-medication were determined in Sri Lanka. In a community-based cross- sectional study, data were collected from 1800 adults selected from Gampaha and Polonnaruwa districts respectively. Study participants were sampled using a multistage cluster sampling technique. Trained public health midwives administered the questionnaire. Two Likert scales provided information on access to medical care and satisfaction with available pharmacy services. About 95% of the sampled population participated in the study. Overall, prevalence of medication use (allopathic, traditional, home remedies) in urban and rural population was 33.9% and 35.3%, respectively. Self-medication prevalence of allopathic drugs in the urban sector (12.2%) was significantly higher than in the rural (7.9%) sector(p<0.05). In the urban sector, small household size and preference to have medicines from outside the pharmacies predisposed to self-medication. The higher acceptability of medical services and regularity of medical care decreased the likelihood of self-medication. In the rural sector, lower satisfaction about the healthcare providers' concern for clients, lower satisfaction about affordability of medical care and higher satisfaction with technical competence of the pharmacy staff increased the likelihood of self-medication. In both urban and rural sectors, when symptom count increased, tendency to self-medicate decreased. Self-medication prevalence was higher in urban compared to rural areas in Sri Lanka. Some aspects of access to medical care, satisfaction with pharmacy services and perceived severity of the disease were found to be important determinants of self-medication.


Increasing incidence of hepatocellular carcinoma possibly associated with non-A, non-B hepatitis in Japan, disclosed by hepatitis B virus DNA analysis of surgically resected cases.
At the National Cancer Center Hospital in Japan, the total number of surgically treated hepatocellular carcinomas (HCCs) has been increasing steadily and rapidly over the last 10 years, whereas the number of cases positive for hepatitis B surface antigen in sera (HBsAg) has remained almost stable. Thus, the relative percentage of HCC cases with serum HBsAg has shown a marked decrease. In order to examine whether this increased proportion of HBsAg-seronegative patients carries hepatitis B virus (HBV) DNA in the liver, we extracted DNA from the formalin-fixed and paraffin-embedded cancerous and noncancerous liver tissues of 79 patients with HCC. The HCCs examined included 49 specimens resected during a period from 1970 to 1980 and 30 resected in 1986 and 1987. We were able to detect reliably the presence of HBV DNA by dot-blot hybridization. The presence of HBV DNA in liver tissues showed a good correlation with positivity for serum HBsAg in both examined groups. In total, HBV DNA was detected in 81% (21 of 26) of HBsAg-seropositive cases and in only 8% (4 of 53) of HBsAg-seronegative cases, indicating that the increased number of HBsAg-seronegative cases had no HBV involvement. Among these HBsAg-seronegative HCC patients, 89.7% showed a histology of cirrhosis or chronic active hepatitis in the noncancerous liver and 29.1% had a history of blood transfusion. These results suggest an increasing incidence of non-A, non-B hepatitis-associated HCCs in Japan and the possible transmission of factors by means other than blood transfusion.


Conversion of thyroxine to triiodothyronine in normal human subjects.
The conversion of thyroxine to triiodothyronine, previously demonstrated in athyreotic human subjects, has been investigated in normal subjects who were given intravenous injections of purified thyroxine labeled with carbon-14 in ring A and in the alanine side chain. Evidence for the conversion of T4 to T3 was provided by the finding of carbon-14 in the T3 fraction isolated from serums. It is estimated that an appreciable fraction of T4 may be transformed to T3 in normal man.


The Medical ANtiarrhythmic Treatment or Radiofrequency Ablation in Paroxysmal Atrial Fibrillation (MANTRA-PAF) trial: clinical rationale, study design, and implementation.
No large randomized multicentre trial has evaluated the efficacy of radiofrequency ablation (RFA) vs. anti-arrhythmic drug (AAD) therapy as a first-line treatment of paroxysmal atrial fibrillation (AF). The Medical ANtiarrhythmic Treatment or Radiofrequency Ablation (MANTRA-PAF) trial is a randomized, controlled, parallel group, multicentre study designed to test whether catheter-based RFA is superior to optimized AAD therapy in suppressing relapse within 24 months of symptomatic and/or asymptomatic AF in patients with paroxysmal AF without prior AAD therapy. The primary endpoint is cumulative AF burden on repeated 7 days Holter monitoring. Secondary endpoints are: thromboembolic events, hospitalization due to arrhythmia, pro-arrhythmic events, procedure/treatment-related side effects, health economics, quality of life, and change in left ventricular function. Ten centres in Scandinavia and Germany are participating in the study. Enrolment was started in 2005 and as of November 2008, 260 patients have been enrolled into the study. It is expected that enrolment will end by March 2009, when 300 patients have been included. The MANTRA-PAF trial will determine whether catheter-based RFA is superior to optimized AAD therapy as a first-line treatment in suppressing long-term relapse of symptomatic and/or asymptomatic AF.


Dispersed rat parotid acinar cells. III. Characterization of cholinergic receptors.
The in vitro characterization of cholinergic receptors in dispersed rat parotid acinar cells was accomplished through investigations of the net transmembrane fluxes of K in response to exposure of the cells to selected cholinergic agonists and antagonists. Interaction of acetylcholine bromide (ACh) and carbamylcholine (carbachol) with the cholinergic receptors resulted in rapid net efflux of K from the cells. This cellular response was demonstrable in concentrations of carbachol as low as 10(-8) M. With gradual increase in the concentrations of the agonist an increase in the K efflux was observed up to 10(-5) M. At higher concentrations of this cholinergic agonist no further increases in the net K efflux were observed. The response of the cells to cholinergic agonists was inhibited by atropine but not by the adrenergic antagonists phentolamine or propranolol, suggesting cholinergic agonist-antagonist interactions at the receptor site. The dispersed rat parotid acinar cells appear to have functionally intact cholinergic receptors and could be used as valuable experimental tools for the study of receptor physiology and pharmacology as well as of other aspects of secretory function at the cellular level.


Do circulating plasma AVT and/or cortisol levels control pulsatile urea excretion in the gulf toadfish (Opsanus beta)?
Previous work has shown that pulsatile urea excretion at the gills of the gulf toadfish is due to periodic activation of a facilitated diffusion transport system with molecular and pharmacological similarity to the UT-A transport system of the mammalian kidney. In mammals, AVP and glucocorticoids are two important endocrine regulators of this system. The present study focused on the potential role of circulating AVT (the teleost homologue of AVP) and cortisol levels as possible triggers for urea pulses. Long-term (34-84 h) monitoring of plasma levels by repetitive sampling at 2-h intervals from chronic cannulae in individual toadfish demonstrated that circulating AVT concentrations are low (10(-12)-10(-11) M), and show no relationship to the occurrence of natural urea pulses. In contrast, plasma cortisol levels decline greatly prior to natural pulses and rise rapidly thereafter. AVT injections into the caudal artery or ventral aorta elicited pulse events, but these were extremely small (1-10%) relative to natural pulses, and occurred only at unphysiological dose levels (10(-9) M in the plasma). AVP was a partial agonist, but isotocin, insulin-like growth factor-1, and atrial natriuretic peptide were without effect at the same concentration. Artificially raising plasma cortisol levels by cortisol injection tended to reduce responsiveness to AVT. Pharmacological reduction of plasma cortisol levels by metyrapone injection elicited small pulses similar to those caused by AVT. Following such pulse events, AVT was ineffective in inducing pulses. We conclude that decreases in circulating cortisol play an important permissive role in urea pulsing, but that circulating AVT levels are not involved.


The role of site accessibility in microRNA target recognition.
MicroRNAs are key regulators of gene expression, but the precise mechanisms underlying their interaction with their mRNA targets are still poorly understood. Here, we systematically investigate the role of target-site accessibility, as determined by base-pairing interactions within the mRNA, in microRNA target recognition. We experimentally show that mutations diminishing target accessibility substantially reduce microRNA-mediated translational repression, with effects comparable to those of mutations that disrupt sequence complementarity. We devise a parameter-free model for microRNA-target interaction that computes the difference between the free energy gained from the formation of the microRNA-target duplex and the energetic cost of unpairing the target to make it accessible to the microRNA. This model explains the variability in our experiments, predicts validated targets more accurately than existing algorithms, and shows that genomes accommodate site accessibility by preferentially positioning targets in highly accessible regions. Our study thus demonstrates that target accessibility is a critical factor in microRNA function.


Urinary creatinine concentrations in the U.S. population: implications for urinary biologic monitoring measurements.
Biologic monitoring (i.e., biomonitoring) is used to assess human exposures to environmental and workplace chemicals. Urinary biomonitoring data typically are adjusted to a constant creatinine concentration to correct for variable dilutions among spot samples. Traditionally, this approach has been used in population groups without much diversity. The inclusion of multiple demographic groups in studies using biomonitoring for exposure assessment has increased the variability in the urinary creatinine levels in these study populations. Our objectives were to document the normal range of urinary creatinine concentrations among various demographic groups, evaluate the impact that variations in creatinine concentrations can have on classifying exposure status of individuals in epidemiologic studies, and recommend an approach using multiple regression to adjust for variations in creatinine in multivariate analyses. We performed a weighted multivariate analysis of urinary creatinine concentrations in 22,245 participants of the Third National Health and Nutrition Examination Survey (1988-1994) and established reference ranges (10th-90th percentiles) for each demographic and age category. Significant predictors of urinary creatinine concentration included age group, sex, race/ethnicity, body mass index, and fat-free mass. Time of day that urine samples were collected made a small but statistically significant difference in creatinine concentrations. For an individual, the creatinine-adjusted concentration of an analyte should be compared with a "reference" range derived from persons in a similar demographic group (e.g., children with children, adults with adults). For multiple regression analysis of population groups, we recommend that the analyte concentration (unadjusted for creatinine) should be included in the analysis with urinary creatinine added as a separate independent variable. This approach allows the urinary analyte concentration to be appropriately adjusted for urinary creatinine and the statistical significance of other variables in the model to be independent of effects of creatinine concentration.


Fatty acid facts, Part III: Cardiovascular disease, or, a fish diet is not fishy.
Preclinical and clinical studies have demonstrated that omega-3 polyunsaturated fatty acids (n-3 PUFAs) play a significant role in the prevention of cardiovascular disease. These fatty acids are called essential fatty acids as they fulfil essential functions and the mammalian cell cannot synthesize them de novo. Dietary sources of n-3 PUFAs include fish oils rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The clinical relevance of these molecules is derived from the incorporation of EPA and DHA into cell membranes. The presence of EPA/DHA alters the physical characteristics of the membrane. Both these altered physicochemical membrane properties and the presence of n-3 PUFAs released by the action of phospholipid lipases (resulting in antiinflammatory eicosanoids) improve biological functions such as signal transduction, ion channelling and ligand binding to nuclear receptors. EPA/DHA also reduce or quench gene expression of cyclooxygenase-2 and other enzymes, thereby diminishing the formation of proinflammatory molecules. Increased EPA/DHA concentration also gives rise to antiinflammatory lipid mediators, called lipoxins, resolvins and protectins. Another important function of n-3 PUFAs is scavenging of free radicals, which diminishes inflammatory response and oxidation of lipoprotein particles, notably low density lipoproteins. The interplay of these molecular processes has distinct cardioprotective effects, which involve actions on lipid metabolism, lipoprotein particle size, blood pressure, vascular function, coagulation potential, inflammatory response, atheroma formation and antiarrhythmic. In view of these actions, fish oil preparations and/or intake of oily fish are recommended as primary and secondary prevention of cardiovascular disease and sudden cardiac death. Large, ongoing trials will further elucidate the presumed favorable effects of EPA/DHA in heart failure and diabetes. This review provides a summary of the physiological mechanisms of the action of EPA and DHA and highlights the epidemiological evidence for a reduction in cardiac events and mortality.


Antimicrobial susceptibility testing using high surface-to-volume ratio microchannels.
This study reports the use of microfluidics, which intrinsically has a large surface-to-volume ratio, toward rapid antimicrobial susceptibility testing at the point of care. By observing the growth of uropathogenic Escherichia coli in gas permeable polymeric microchannels with different dimensions, we demonstrate that the large surface-to-volume ratio of microfluidic systems facilitates rapid growth of bacteria. For microchannels with 250 microm or less in depth, the effective oxygenation can sustain the growth of E. coli to over 10(9) cfu/mL without external agitation or oxygenation, which eliminates the requirement of bulky instrumentation and facilitates rapid bacterial growth for antimicrobial susceptibility testing at the point of care. The applicability of microfluidic rapid antimicrobial susceptibility testing is demonstrated in culture media and in urine with clinical bacterial isolates that have different antimicrobial resistance profiles. The antimicrobial resistance pattern can be determined as rapidly as 2 h compared to days in standard clinical procedures facilitating diagnostics at the point of care.


Antiobesogenic effects of central GIPR antagonism.
Developing effective treatments for obesity and related metabolic disease remains a challenge. One logical strategy targets the appetite-regulating actions of gut hormones such as incretins. One of these incretins, glucose-dependent insulinotropic polypeptide (GIP), has garnered much attention as a potential target: however, whether it is beneficial to boost or block the action of GIP to promote weight loss remains an unresolved question. In this issue of the JCI, Kaneko and colleagues show that antagonizing GIP signaling in the CNS enhances the weight-reducing effects of leptin in rodents with diet-induced obesity. The authors posit that an increase in circulating intestinally derived GIP, as a consequence of overnutrition, acts in the brain to impair hypothalamic leptin action, resulting in increased food intake and body weight gain. This research advances the idea that multiple GIP signaling pathways and mechanisms exist in the obese state and offers intriguing new insights into the antiobesogenic consequences of antagonizing brain GIP action.


Hypoglycemic effect and mechanism of honokiol on type 2 diabetic mice.
Honokiol is one of the main bioactive constituents of the traditional Chinese herbal drug Magnolia bark (Cortex Magnoliae officinalis, Hou Po). The aim of this study was to probe its anti-type 2 diabetes mellitus effects and the underlying mechanism. Type 2 diabetic mouse model was established by intraperitoneally injecting with streptozotocin. Fasting blood glucose, body weight, and lipid profile were measured. The subcutaneous adipose tissue, skeletal muscle, and liver were isolated as well as homogenized. The phospho-insulin receptor β-subunit (IRβ), IRβ, phospho-AKT, AKT, phospho-ERK1/2, ERK1/2, phosphotyrosine, and actin were examined by Western blot assay. Cell viability or cytotoxicity was analyzed by using MTT method. The inhibitory potencies of honokiol on the protein tyrosine phosphatase 1B (PTP1B) activity were performed in reaction buffer. Molecular docking and dynamic simulation were also analyzed. In in vivo studies, oral treatment with 200 mg/kg honokiol for 8 weeks significantly decreases the fasting blood glucose in type 2 diabetes mellitus mice. The phosphorylations of the IRβ and the downstream insulin signaling factors including AKT and ERK1/2 significantly increase in adipose, skeletal muscle, and liver tissue of the honokiol-treated mice. Moreover, honokiol enhanced the insulin-stimulated phosphorylations of IRβ, AKT, and ERK1/2 in a dose-dependent manner in C2C12 myotube cells. Meanwhile, honokiol enhanced insulin-stimulated GLUT4 translocation. Importantly, honokiol exhibited reversible competitive inhibitory activity against PTP1B with good selectivity in vitro and in vivo. Furthermore, using molecular docking and dynamic simulation approaches, we determined the potential binding mode of honokiol to PTP1B at an atomic level. These findings indicated the hypoglycemic effects of honokiol and its mechanism that honokiol improved the insulin sensitivity by targeting PTP1B. Therefore, our study may highlight honokiol as a promising insulin sensitizer for the therapy of type 2 diabetes.


Population Structure in the Roundtail Chub (Gila robusta Complex) of the Gila River Basin as Determined by Microsatellites: Evolutionary and Conservation Implications.
Ten microsatellite loci were characterized for 34 locations from roundtail chub (Gila robusta complex) to better resolve patterns of genetic variation among local populations in the lower Colorado River basin. This group has had a complex taxonomic history and previous molecular analyses failed to identify species diagnostic molecular markers. Our results supported previous molecular studies based on allozymes and DNA sequences, which found that most genetic variance was explained by differences among local populations. Samples from most localities were so divergent species-level diagnostic markers were not found. Some geographic samples were discordant with current taxonomy due to admixture or misidentification; therefore, additional morphological studies are necessary. Differences in spatial genetic structure were consistent with differences in connectivity of stream habitats, with the typically mainstem species, G. robusta, exhibiting greater genetic connectedness within the Gila River drainage. No species exhibited strong isolation by distance over the entire stream network, but the two species typically found in headwaters, G. nigra and G. intermedia, exhibited greater than expected genetic similarity between geographically proximate populations, and usually clustered with individuals from the same geographic location and/or sub-basin. These results highlight the significance of microevolutionary processes and importance of maintaining local populations to maximize evolutionary potential for this complex. Augmentation stocking as a conservation management strategy should only occur under extreme circumstances, and potential source populations should be geographically proximate stocks of the same species, especially for the headwater forms.


Role of miR-34c in ketamine-induced neurotoxicity in neonatal mice hippocampus.
Ketamine is a commonly used pediatric anesthetic, but it might affect development, or even induce neurotoxicity in the neonatal brain. We have used an in vivo neonatal mouse model to induce ketamine-related neurotoxicity in the hippocampus, and found that miR-34c, a microRNA associated with pathogenesis of Alzheimer's disease, was significantly upregulated during ketamine-induced hippocampal neurodegeneration. Functional assay of silencing miR-34c demonstrated that downregulation of miR-34c activated PKC-ERK pathway, upregulated anti-apoptotic protein BCL2, and ameliorated ketamine-induced apoptosis in the hippocampus. Cognitive examination with the Morris water maze test showed that ketamine-induced memory impairment was significantly improved by miR-34c downregulation. Thus, miR-34c is important in regulating ketamine-induced neurotoxicity in hippocampus.


Impact of malaria related messages on insecticide-treated net (ITN) use for malaria prevention in Ghana.
Media messages have been used in Ghana to promote insecticide-treated net (ITN)/bed net usage in an effort to impact on malaria prevention. The aim of this study was to assess the effect of such malaria-related messages delivered through electronic/print media and by volunteers/health workers on the use of ITNs by children living in a household. Data was collected from September to November of 2008 using a structured, interviewer-administered questionnaire by the Ghana Statistical Service as part of a national demographic and health survey (DHS). Secondary data analysis was performed on the collected data using multivariate logistic regression for both individual messages and a composite (any of) message variable. From the 11,788 households surveyed, 45% had at least one net. Households with male heads were more likely to have a child sleeping under a bed net the previous night (p = 0.0001). Individual Messages delivered by a health worker or a dedicated radio programme, had the highest effect for one or more children sleeping under a net the night before (OR(adjusted) = 1.65; 95% CI = 1.44 to 1.88 and OR(adjusted) = 1.26; 95% CI =1.12 to 1.42 respectively) while hearing any of the eight messages (composite score) resulted in the highest odds for one or more children (OR(adjusted) = 3.06; 95% CI = 2.27 to 4.12) sleeping under a bed net. Efforts to relate ITN messages to the public are very useful in increasing use of bed nets and having multiple ways of reaching the public increases their effect, with the biggest effect seen when health workers and volunteers were used to deliver malaria-related messages to the public.


Two-thirds of primary care physicians accepted new Medicaid patients in 2011-12: a baseline to measure future acceptance rates.
As part of the Affordable Care Act, primary care physicians providing services to patients insured through Medicaid in some states will receive higher payments in 2013 and 2014 than in the past. Payments for some services will increase to match Medicare rates. This change may lead to wider acceptance of new Medicaid patients among primary care providers. Using data from the 2011-12 National Ambulatory Medical Care Survey Electronic Medical Records Supplement, I summarize baseline rates of acceptance of new Medicaid patients among office-based physicians by specialty and practice type. I also report state-level acceptance rates for both primary care and other physicians. About 33 percent of primary care physicians (those in general and family medicine, internal medicine, or pediatrics) did not accept new Medicaid patients in 2011-12, ranging from a low of 8.9 percent in Minnesota to a high of 54.0 percent in New Jersey. Primary care physicians in New Jersey, California, Alabama, and Missouri were less likely than the national average to accept new Medicaid patients in 2011-12. The data presented here provide a baseline for comparison of new Medicaid acceptance rates in 2013-14.


APBP-1, a DNA/RNA-binding protein, interacts with the chick aggrecan regulatory region.
Expression of the extracellular proteoglycan aggrecan is both cell-specific and developmentally regulated. Previous studies identified six functionally defined cis elements in the aggrecan promoter region which were shown to repress aggrecan gene expression (1). Using competition electrophoretic mobility shift assays (EMSAs) we have now identified in nuclear extracts a functional repressor cis element, (T/C)TCCCCT(A/C)RRC, which occurs at multiple locations within the chick aggrecan regulatory region. We purified the factor that binds to this cis element and established that it, APBP-1 (aggrecan promoter-binding protein-1), is a 19-kDa protein that has significant homology to CIRP (cold inducible RNA-binding protein). Recombinantly expressed APBP-1 mimics the native cis element-trans factor interaction in EMSAs. In situ hybridization demonstrates that aggrecan and APBP-1 RNA expression are restricted to complementary tissues in the developing limb, and Northern blot analysis of chick limb bud mRNA shows that APBP-1 mRNA expression is inversely correlated with aggrecan mRNA expression. Functional analyses by transient transfections and Northern blot analyses suggest APBP-1 has the capacity to repress aggrecan expression, indicating that this factor may be important regulator of aggrecan gene expression.


Family Quality of Life of South African Families Raising Children with Autism Spectrum Disorder.
This article describes the family quality of life among families who are raising a young child with autism spectrum disorder. Survey research was conducted with 180 families of children with autism spectrum disorder who were receiving disability-related services in the Gauteng province of South Africa. The principle measure used was the Beach Center Family Quality of Life Scale to assess five subdomains: family interaction, parenting, emotional well-being, material/physical well-being, and disability-related support. Results indicated that families felt the most satisfied with disability-related support and the least satisfied with the family's emotional well-being. Family income, family type, and the severity level of autism were significantly associated with how satisfied families felt about their quality of life.


Copper modulates the phenotypic response of activated BV2 microglia through the release of nitric oxide.
Microglia are resident immune cells of the central nervous system. Their persistent activation in neurodegenerative diseases, traditionally attributed to neuronal dysfunction, may be due to a microglial failure to modulate the release of cytotoxic mediators such as nitric oxide (NO). The persistent activation of microglia with the subsequent release of NO vis-á-vis the accumulation of redox transition metals such as copper (Cu) in neurodegenerative diseases, prompted the hypothesis that copper would alter NO signaling by changing the redox environment of the cell and that, by altering the fate of NO, microglia would adopt a different phenotype. We have used the microglial cell model, BV2, to examine the effects of Cu(I) on NO production and activation as they have been shown to be phenotypically plastic. Our results show that cell viability is not affected by Cu(I) in BV2 microglia and that it has no effect on iNOS mRNA, protein expression and nitrite release. However, when LPS is added to Cu(I)-treated medium, nitrite release is abrogated while iNOS expression is not significantly altered. This effect is Cu(I)-specific and it is not observed with other non-redox metals, suggesting that Cu(I) modulates NO reactivity. Immunofluorescence analysis shows that the M1 (inflammatory) phenotype of BV2 microglia observed in response to LPS, is shifted to an M2 (adaptive) phenotype when Cu(I) is administered in combination with LPS. This same shift is not observed when iNOS function is inhibited by 1400W. In the present study we show that Cu(I) modulates the release of NO to the media, without altering iNOS expression, and produces phenotypic changes in BV2 microglia.


Should doctors intentionally do less than the best?
The papers of Burley and Harris, and Draper and Chadwick, in this issue, raise a problem: what should doctors do when patients request an option which is not the best available? This commentary argues that doctors have a duty to offer that option which will result in the individual affected by that choice enjoying the highest level of wellbeing. Doctors can deviate from this duty and submaximise--bring about an outcome that is less than the best--only if there are good reasons to do so. The desire to have a child which is genetically related provides little, if any, reason to submaximise. The implication for cloning, preimplantation diagnosis and embryo transfer is that doctors should only produce a clone or transfer embryos expected to enjoy a level of wellbeing which is less than that enjoyed by other children the couple could have, if there is a good reason to employ that technology. This paper sketches what might constitute a good reason to submaximise.


European Federation of Neurological Societies/Peripheral Nerve Society guideline on management of multifocal motor neuropathy.
Several diagnostic criteria for multifocal motor neuropathy have been proposed in recent years and a beneficial effect of intravenous immunoglobulin (IVIg) and various other immunomodulatory drugs has been suggested in several trials and uncontrolled studies. The objectives were to prepare consensus guidelines on the definition, investigation and treatment of multifocal motor neuropathy. Disease experts and a patient representative considered references retrieved from MEDLINE and the Cochrane Library in July 2004 and prepared statements which were agreed in an iterative fashion. The Task Force agreed good practice points to define clinical and electrophysiological diagnostic criteria for multifocal motor neuropathy and investigations to be considered. The principal recommendations and good practice points were: (i) IVIg (2 g/kg given over 2-5 days) should be considered as the first line treatment (level A recommendation) when disability is sufficiently severe to warrant treatment. (ii) Corticosteroids are not recommended (good practice point). (iii) If initial treatment with IVIg is effective, repeated IVIg treatment should be considered (level C recommendation). The frequency of IVIg maintenance therapy should be guided by the individual response (good practice point). Typical treatment regimens are 1 g/kg every 2-4 weeks or 2 g/kg every 4-8 weeks (good practice point). (iv) If IVIg is not or not sufficiently effective then immunosuppressive treatment may be considered. Cyclophosphamide, ciclosporin, azathioprine, interferon beta1a, or rituximab are possible agents (good practice point). (v) Toxicity makes cyclophosphamide a less desirable option (good practice point).


Cost effectiveness of denosumab compared with oral bisphosphonates in the treatment of post-menopausal osteoporotic women in Belgium.
Denosumab has recently been shown to be well tolerated, to increase bone mineral density (BMD) and to significantly reduce the risk of hip, vertebral and non-vertebral fractures in the FREEDOM (Fracture REduction Evaluation of Denosumab in Osteoporosis every 6 Months) trial. It is becoming increasingly important to evaluate not only the therapeutic value of a new drug but also the cost effectiveness compared with the most relevant treatment alternatives. The objective of this study was to estimate the cost effectiveness of denosumab compared with oral bisphosphonates (branded and generic drugs) in the treatment of post-menopausal osteoporotic women in Belgium. Cost effectiveness of 3 years of treatment with denosumab was compared with branded risedronate and branded and generic alendronate using an updated version of a previously validated Markov microsimulation model. The model was populated with relevant cost, adherence and epidemiological data for Belgium from a payer perspective and the results were presented as costs per QALY gained (&U20AC;, year 2009 values). Analyses were performed in populations (aged ≥60 years) in which osteoporosis medications are currently reimbursed in many European countries, i.e. those with BMD T-score of -2.5 or less or prevalent vertebral fracture. Patients receiving denosumab were assumed to have a 46% lower risk of discontinuation than those receiving oral bisphosphonates, and the effect of denosumab after treatment cessation was assumed to decline linearly to zero over a maximum of 1 year. Denosumab was cost effective compared with all other therapies, assuming a willingness to pay of &U20AC;40 000 per QALY gained. In particular, denosumab was found to be cost effective compared with branded alendronate and risedronate at a threshold value of &U20AC;30 000 per QALY and denosumab was dominant (i.e. lower cost and greater effectiveness) compared with risedronate from the age of 70 years in women with a T-score of -2.5 or less and no prior fractures. The cost effectiveness of denosumab compared with generic alendronate was estimated at &U20AC;38 514, &U20AC;22 220 and &U20AC;27 862 per QALY for women aged 60, 70 and 80 years, respectively, with T-scores of -2.5 or less. The equivalent values were &U20AC;37 167, &U20AC;19 718 and &U20AC;19 638 per QALY for women with prevalent vertebral fractures. This study suggests, on the basis of currently available data, that denosumab is a cost-effective strategy compared with oral bisphosphonates (including generic alendronate) for the treatment of post-menopausal osteoporotic women, aged ≥60 years in Belgium. Denosumab therefore appears to have the potential to become a first-line treatment for post-menopausal women with osteoporosis. However, further studies would be required to evaluate the long-term safety and adherence of denosumab in real-world clinical practice as well as head-to-head effectiveness compared with oral bisphosphonates.


Protective role of p450 epoxyeicosanoids in subarachnoid hemorrhage.
Patients recovering from aneurysmal subarachnoid hemorrhage (SAH) are at risk for developing delayed cerebral ischemia (DCI). Experimental and human studies implicate the vasoconstrictor P450 eicosanoid 20-hydroxyeicosatetraenoic acid (20-HETE) in the pathogenesis of DCI. To date, no studies have evaluated the role of vasodilator epoxyeicosatrienoic acids (EETs) in DCI. Using mass spectrometry, we measured P450 eicosanoids in cerebrospinal fluid (CSF) from 34 SAH patients from 1 to 14 days after admission. CSF eicosanoid levels were compared in patients who experienced DCI versus those who did not. We then studied the effect of EETs in a model of SAH using mice lacking the enzyme soluble epoxide hydrolase (sEH), which catabolizes EETs into their inactive diol. To assess changes in vessel morphology and cortical perfusion in the mouse brain, we used optical microangiography, a non-invasive coherence-based imaging technique. Along with increases in 20-HETE, we found that CSF levels of 14,15-EET were elevated in SAH patients compared to control CSF, and levels were significantly higher in patients who experienced DCI compared to those who did not. Mice lacking sEH had elevated 14,15-EET and were protected from the delayed decrease in microvascular cortical perfusion after SAH, compared to wild type mice. Our findings suggest that P450 eicosanoids play an important role in the pathogenesis of DCI. While 20-HETE may contribute to the development of DCI, 14,15-EET may afford protection against DCI. Strategies to enhance 14,15-EET, including sEH inhibition, should be considered as part of a comprehensive approach to prevent DCI.


Model-Based Generation of Large Databases of Cardiac Images: Synthesis of Pathological Cine MR Sequences From Real Healthy Cases.
Collecting large databases of annotated medical images is crucial for the validation and testing of feature extraction, statistical analysis, and machine learning algorithms. Recent advances in cardiac electromechanical modeling and image synthesis provided a framework to generate synthetic images based on realistic mesh simulations. Nonetheless, their potential to augment an existing database with large amounts of synthetic cases requires further investigation. We build upon these works and propose a revised scheme for synthesizing pathological cardiac sequences from real healthy sequences. Our new pipeline notably involves a much easier registration problem to reduce potential artifacts, and takes advantage of mesh correspondences to generate new data from a given case without additional registration. The output sequences are thoroughly examined in terms of quality and usability on a given application: the assessment of myocardial viability, via the generation of 465 synthetic cine MR sequences (15 healthy and 450 with pathological tissue viability [random location, extent, and grade, up to myocardial infarct]). We demonstrate that: 1) our methodology improves the state-of-the-art algorithms in terms of realism and accuracy of the simulated images and 2) our methodology is well-suited for the generation of large databases at small computational cost.


Surgical treatment of venous malformations in Klippel-Trénaunay syndrome.
Klippel-Trénaunay syndrome (KTS) is a complex congenital anomaly, characterized by varicosities and venous malformations (VMs) of one or more limbs, port-wine stains, and soft tissue and bone hypertrophy. Venous drainage is frequently abnormal because of embryonic veins, agenesis, hypoplasia, valvular incompetence, or aneurysms of deep veins. We previously reported on the surgical management of KTS. In this article, we update our experience. Twenty patients with KTS underwent surgical treatment for VMs between July 1, 1987, and January 1, 2000. This group represented 6.9% of 290 patients with KTS who were seen at our institution during this 12.5-year study period. Surgical indications, venous anatomy (determined with duplex scan, contrast phlebography, magnetic resonance imaging or magnetic resonance phlebography), operative procedures, and complications were reviewed, and outcomes were recorded. Twelve male and eight female patients (mean age, 23.4 years; range, 7.7-40.6 years) underwent 30 vascular surgical procedures in 21 lower limbs. All 20 patients (100%) had varicose veins or VMs, 13 (65%) had port-wine stains, and 18 (90%) had limb hypertrophy. Pain was the most common complaint, which was present in 16 patients (80%), followed by swelling in 15 (75%), bleeding in 8 (40%), and superficial thrombophlebitis and cellulitis in 3 (15%). Imaging confirmed patent deep veins in 18 patients, hypoplastic femoral vein in 1, and entrapped popliteal veins bilaterally in 1. Four patients (20%) had large persistent sciatic veins (PSVs). The CEAP clinical classification was C-3 for 17 patients (85%), C-4 for 1 patient (5%), and C-6 for 2 patients (10%). Stripping of large lateral veins, avulsion, and excision of varicosities or VMs were performed on all limbs. Three patients required staged resections. The release of entrapped popliteal veins was performed in both limbs of one patient; another underwent a popliteal-saphenous bypass graft. One patient underwent excision of a PSV. Open and endoscopic perforator vein ligation was performed in one patient each. Two patients (12%) had hematomas that required evacuation. No patients had caval filter placement; none had postoperative deep venous thrombosis or pulmonary embolus. The mean follow-up was 63.6 months (range, 0-138 months). All patients reported initial improvement, but some varicosities recurred in 10 patients (50%), an ulcer did not heal in one, and a new ulcer developed in one, 8 years after surgery. Three patients underwent reoperation for recurrent varicosities. Follow-up CEAP scores were C-2 in 10 patients (50%), C-3 in 6 patients (30%), C-4 and C-5 in 1 patient each (5%), and C-6 in 2 patients (10%). Clinical scores improved from 4.3 +/- 2.2 to 3.1 +/- 2.3. (P =.03). The management of patients with KTS continues to be primarily nonoperative, but those patients with patent deep veins can be considered for excision of symptomatic varicose veins and VMs. Although the recurrence rate is high, clinical improvement is significant, and reoperations can be performed if needed. Occasionally, deep vein reconstruction, excision of PSVs, or subfascial endoscopic perforator surgery is indicated. Because KTS is rare, patients should receive multidisciplinary care in qualified vascular centers.


How culture shaped the human genome: bringing genetics and the human sciences together.
Researchers from diverse backgrounds are converging on the view that human evolution has been shaped by gene-culture interactions. Theoretical biologists have used population genetic models to demonstrate that cultural processes can have a profound effect on human evolution, and anthropologists are investigating cultural practices that modify current selection. These findings are supported by recent analyses of human genetic variation, which reveal that hundreds of genes have been subject to recent positive selection, often in response to human activities. Here, we collate these data, highlighting the considerable potential for cross-disciplinary exchange to provide novel insights into how culture has shaped the human genome.


Do-not-resuscitate orders among trauma patients.
Do-not-resuscitate (DNR) orders are an important means to communicate end-of-life wishes. Previous studies have demonstrated variable prevalence of DNR orders among hospitalized trauma patients. This study was conducted to identify the prevalence and type of DNR orders among trauma patients and to identify associations of DNR orders with injury severity, length of stay, and whether CPR was performed in cases of cardiac arrest. In this retrospective study, medical records were reviewed for 263 trauma patients at Miami Valley Hospital in Dayton, Ohio, in 2014 with a DNR order. Among 3394 trauma patients in 2014, 263 (8%) patients had a DNR order. Participants were 43% male and 57% female. The mean age was 76 (range, 16-90+) years. The most common mechanisms of injury included fall (n = 214, 81.4%) and motor vehicle collision (n = 16, 6.1%). Most DNR orders in this patient population were instituted during the hospitalization (n = 176, 67%). The most common types of advance directives included DNR order (n = 224, 85.2%), living will (n = 124, 47.2%), and durable power of health care attorney (n = 126, 47.9%). A minority of patients died during hospitalization (n = 100, 38.0%). Among patients who were deceased, 14 (14.0%) had CPR performed. Among trauma patients with DNR orders, most DNR orders were instituted during the hospital admission. Most deceased patients with DNR orders did not have CPR performed during the hospital stay.


[Sporotrichosis: development and challenges of an epidemic].
In the state of Rio de Janeiro, Brazil, sporotrichosis reached epidemic levels, involving humans and cats. Preliminary data indicate that approximately 2200 human cases were diagnosed between 1998 and December of 2009, and 3244 cats were treated. The geographic distribution of cases reveals a concentration in the City of Rio de Janeiro metropolitan area. The disease affects mostly women of a low socioeconomic status, aged 40 to 55 years, who work as housekeepers. Itraconazole has been the drug of choice for treatment. Although sporotrichosis does not usually affect organs other than the skin, mucosa, and subcutaneous tissue, it has an indirect social impact resulting from absenteeism, pain, and discomfort during the active disease stage, and the unpleasant appearance of the scars. In turn, systemic involvement is frequent in cats, leading to serious and difficult to treat forms of the disease and death. Considering that treatment time in animals is longer than in human beings, treating cats with sporotrichosis has been the greatest obstacle and the most important challenge for the control of this epidemic infection.


Detection of abnormal left ventricular geometry in patients without cardiovascular disease through machine learning: An ECG-based approach.
Cardiac remodeling is recognized as an important aspect of cardiovascular disease (CVD) progression. Machine learning (ML) techniques were applied to basic clinical parameters and electrocardiographic features, in order to detect abnormal left ventricular geometry (LVG) even before the onset of left ventricular hypertrophy (LVH), in a population without established CVD. The authors enrolled 528 patients with and without essential hypertension, but no other indications of CVD. All patients underwent a full echocardiographic evaluation and were classified into 3 groups; normal geometry (NG), concentric remodeling without LVH (CR), and LVH. Abnormal LVG was identified as increased relative wall thickness (RWT) and/or left ventricular mass index (LVMi). The authors trained supervised ML models to classify patients with abnormal LVG and calculated SHAP values to perform feature importance and interaction analysis. Hypertension, age, body mass index over the Sokolow-Lyon voltage, QRS-T angle, and QTc duration were some of the most important features. Our model was able to distinguish NG from CR+LVH combined, with 87% accuracy on an unseen test set, 75% specificity, 97% sensitivity, and area under the receiver operating curve (AUC/ROC) equal to 0.91. The authors also trained our model to classify NG and CR (NG + CR) against those with LVH, with 89% test set accuracy, 93% specificity, 67% sensitivity, and an AUC/ROC value of 0.89, for a 0.4 decision threshold. Our ML algorithm effectively detects abnormal LVG even at early stages. Innovative solutions are needed to improve risk stratification of patients without established CVD, and ML may enable progress in this direction.


Effect of two different training programs with the same workload on soccer overhead throwing velocity.
The purpose of this study was to determine whether two throwing programs, based upon velocity or resistance with the same workload, would enhance soccer overhead throwing velocity. Sports science students (n = 64, age 21.1 +/- 2.1 y, mass 71.1 +/- 11 kg, height 1.75 +/- 0.09 m; mean +/- SD) divided into two groups matched on performance, participated in the study. The resistance-training group trained overhead throwing with a 5-kg medicine ball for two sets of 8 reps per session, whereas a velocity training group threw four sets of 16 reps with a regular soccer ball. These training programs were matched on workload. Throwing performance with a soccer ball and a 5-kg medicine ball were tested before and after a training period of 6 wk with two sessions per week. Both groups significantly increased the throwing velocity with the soccer ball (resistance-training group: 3.2% [1.0-5.5%]; P = .003 and velocity-training group: 5.1% [2.6-7.7%]; P < .001), whereas no substantial changes were found for throwing with the 5-kg medicine ball after the training period. No substantial differences between the groups were found, which indicates that both forms of training increased the throwing velocity. It is concluded that both velocity and resistance throwing training programs after a short period of training with the same workload can increase throwing velocity and that workload is of importance in designing training programs and comparing them with each other.


Improved statistical methods enable greater sensitivity in rhythm detection for genome-wide data.
Robust methods for identifying patterns of expression in genome-wide data are important for generating hypotheses regarding gene function. To this end, several analytic methods have been developed for detecting periodic patterns. We improve one such method, JTK_CYCLE, by explicitly calculating the null distribution such that it accounts for multiple hypothesis testing and by including non-sinusoidal reference waveforms. We term this method empirical JTK_CYCLE with asymmetry search, and we compare its performance to JTK_CYCLE with Bonferroni and Benjamini-Hochberg multiple hypothesis testing correction, as well as to five other methods: cyclohedron test, address reduction, stable persistence, ANOVA, and F24. We find that ANOVA, F24, and JTK_CYCLE consistently outperform the other three methods when data are limited and noisy; empirical JTK_CYCLE with asymmetry search gives the greatest sensitivity while controlling for the false discovery rate. Our analysis also provides insight into experimental design and we find that, for a fixed number of samples, better sensitivity and specificity are achieved with higher numbers of replicates than with higher sampling density. Application of the methods to detecting circadian rhythms in a metadataset of microarrays that quantify time-dependent gene expression in whole heads of Drosophila melanogaster reveals annotations that are enriched among genes with highly asymmetric waveforms. These include a wide range of oxidation reduction and metabolic genes, as well as genes with transcripts that have multiple splice forms.


Potential benefits of cattle vaccination as a supplementary control for bovine tuberculosis.
Vaccination for the control of bovine tuberculosis (bTB) in cattle is not currently used within any international control program, and is illegal within the EU. Candidate vaccines, based upon Mycobacterium bovis bacillus Calmette-Guérin (BCG) all interfere with the action of the tuberculin skin test, which is used to determine if animals, herds and countries are officially bTB-free. New diagnostic tests that Differentiate Infected from Vaccinated Animals (DIVA) offer the potential to introduce vaccination within existing eradication programs. We use within-herd transmission models estimated from historical data from Great Britain (GB) to explore the feasibility of such supplemental use of vaccination. The economic impact of bovine Tuberculosis for farmers is dominated by the costs associated with testing, and associated restrictions on animal movements. Farmers' willingness to adopt vaccination will require vaccination to not only reduce the burden of infection, but also the risk of restrictions being imposed. We find that, under the intensive sequence of testing in GB, it is the specificity of the DIVA test, rather than the sensitivity, that is the greatest barrier to see a herd level benefit of vaccination. The potential negative effects of vaccination could be mitigated through relaxation of testing. However, this could potentially increase the hidden burden of infection within Officially TB Free herds. Using our models, we explore the range of the DIVA test characteristics necessary to see a protective herd level benefit of vaccination. We estimate that a DIVA specificity of at least 99.85% and sensitivity of >40% is required to see a protective benefit of vaccination with no increase in the risk of missed infection. Data from experimentally infected animals suggest that this target specificity could be achieved in vaccinates using a cocktail of three DIVA antigens while maintaining a sensitivity of 73.3% (95%CI: 61.9, 82.9%) relative to post-mortem detection.


Inhibitory effects of topical thymoquinone on corneal neovascularization.
Thymoquinone, one of the biologically active components of black seed oil, has anti-inflammatory and antioxidant properties. We aimed to study the effect of thymoquinone on corneal neovascularization in rats and to compare its efficacy with that of triamcinolone acetonide. Chemical cauterization of the cornea was performed with silver nitrate/potassium nitrate sticks in 40 eyes in 40 rats. An examiner blinded to the experiments scored the intensity of the cauterization. Topical instillation of thymoquinone 0.1%, thymoquinone 0.4%, and triamcinolone acetonide was continued for 7 days. The inhibitory effects of the drugs on corneal neovascularization were tested and compared with each other and with controls with a computer program that evaluates percent areas of cornea covered by neovascularization. The means of percent area of corneal neovascularization in the thymoquinone 0.1%, thymoquinone 0.4%, triamcinolone acetonide, and control groups were 60.1%, 45%, 46%, and 72%, respectively. The inhibitory effect of thymoquinone 0.4% was found to be equal to that of triamcinolone acetonide (P = 0.87). The thymoquinone 0.4% and triamcinolone groups were different from the thymoquinone 0.1% and control groups (P < 0.05). There was also a significant difference between the percent area of corneal neovascularization in the thymoquinone 0.1% group and that of the controls (P < 0.05). The mean burn stimulus intensities were not different among the groups (P = 0.54). Thymoquinone was shown to have an inhibitory effect, comparable with that of triamcinolone, on corneal neovascularization in this rat model. However, thymoquinone decreased corneal neovascularization in a dose-dependent manner.


Origin of the low-temperature endotherm of acid-doped ice VI: new hydrogen-ordered phase of ice or deep glassy states?
On the basis of a low-temperature endotherm, it has recently been argued that cooling acid-doped ice VI at high pressures leads to a new hydrogen-ordered phase. We show that the endotherms are in fact caused by the glass transitions of deep glassy states related to ice VI. As expected for such endothermic overshoot effects, they display a characteristic dependence on pressure and cooling rate, they can be produced by sub-T g annealing at ambient pressure, and they can be made to appear or disappear depending on the heating rate and the initial extent of relaxation. It is stressed that the existence of a new crystalline phase, as it has been suggested, cannot depend on the heating rate at which it is heated. X-ray diffraction shows that samples for which the low-temperature endotherm is present, weak or absent, as observed at a heating rate of 5 K min-1, are structurally very similar. Furthermore, we show that the reported shifts of the (102) Bragg peak upon heating are fully consistent with our scenario and also with our earlier neutron diffraction study. Deuterated acid-doped ice VI cooled at high pressure also displays a low-temperature endotherm and its neutron diffraction pattern is consistent with deep glassy ice VI. Accessing deep glassy states of ice with the help of acid doping opens up a fascinating new chapter in ice research. Compared to pure ice VI, the glass transition temperature is lowered by more than 30 K by the acid dopant. Future work should focus on the deep glassy states related to all the other hydrogen-disordered ices including the 'ordinary' ice Ih.


Predictive analytics and machine learning in stroke and neurovascular medicine.
Advances in predictive analytics and machine learning supported by an ever-increasing wealth of data and processing power are transforming almost every industry. Accuracy and precision of predictive analytics have significantly increased over the past few years and are evolving at an exponential pace. There have been significant breakthroughs in using Predictive Analytics in healthcare where it is held as the foundation of precision medicine. Yet, although the research in the field is expanding with the profuse volume of papers applying machine learning algorithms to medical data, very few have contributed meaningfully to clinical care. This lack of impact stands in stark contrast to the enormous relevance of machine learning to many other industries. Regardless of the status of its current contribution, the field of predictive analytics is expected to fundamentally change the way we diagnose and treat diseases, as well as the conduct of biomedical science research. In this review, we describe the main tools and techniques in predictive analytics and will analyze the trends in application of these techniques over the recent years. We will also provide examples of its application in medicine and more specifically in stroke and neurovascular research and outline current limitations.


Determinants of Health Literacy and Its Associations With Health-Related Behaviors, Depression Among the Older People With and Without Suspected COVID-19 Symptoms: A Multi-Institutional Study.
Purpose: We examined factors associated with health literacy among elders with and without suspected COVID-19 symptoms (S-COVID-19-S). Methods: A cross-sectional study was conducted at outpatient departments of nine hospitals and health centers 14 February-2 March 2020. Self-administered questionnaires were used to assess patient characteristics, health literacy, clinical information, health-related behaviors, and depression. A sample of 928 participants aged 60-85 years were analyzed. Results: The proportion of people with S-COVID-19-S and depression were 48.3 and 13.4%, respectively. The determinants of health literacy in groups with and without S-COVID-19-S were age, gender, education, ability to pay for medication, and social status. In people with S-COVID-19-S, one-score increment of health literacy was associated with 8% higher healthy eating likelihood (odds ratio, OR, 1.08; 95% confidence interval, 95%CI, 1.04, 1.13; p < 0.001), 4% higher physical activity likelihood (OR, 1.04; 95%CI, 1.01, 1.08, p = 0.023), and 9% lower depression likelihood (OR, 0.90; 95%CI, 0.87, 0.94; p < 0.001). These associations were not found in people without S-COVID-19-S. Conclusions: The older people with higher health literacy were less likely to have depression and had healthier behaviors in the group with S-COVD-19-S. Potential health literacy interventions are suggested to promote healthy behaviors and improve mental health outcomes to lessen the pandemic's damage in this age group.


Role of oxidative stress in transition of hypertrophy to heart failure.
In an attempt to define the role of increased oxidative stress in the transition from compensatory hypertrophy to heart failure, this study examined the effects of long-term vitamin E therapy on the occurrence of heart failure subsequent to chronic pressure overload in guinea pigs. Hyperfunctional heart hypertrophy has been shown to be accompanied by an increase in the endogenous antioxidant reserve, whereas congestive heart failure is accompanied by a decrease in this reserve. The effects of vitamin E, a naturally occurring antioxidant, on the development of heart failure from a hypertrophic stage were examined. The ascending aorta in guinea pigs was coarcted. For vitamin treatment, slow-release pellets were implanted at the time of the operation. The animals were assessed at 10 and 20 weeks for hemodynamic function, myocardial structure, antioxidant agents and oxidative stress. Banding of the ascending aorta in guinea pigs resulted in hyperfunctional hypertrophy at 10 weeks, which was followed by congestive heart failure at 20 weeks. Hypertrophied hearts showed decreased oxidative stress, as evidenced by a higher oxidation-reduction (redox) state and less lipid peroxidation, whereas the failure stage was characterized by increased oxidative stress. Supplementation of animals with timed-release vitamin E tablets resulted in an increased myocardial content of the vitamin, and the banded animals did not develop any signs of heart failure at 20 weeks. Hemodynamic function at 20 weeks in these vitamin E-treated animals was also better maintained. The myocardial reduced glutathione/oxidized glutathione ratio of vitamin E-treated animals at 20 weeks was higher and lipid peroxidation was less compared with the untreated animals. Ultrastructural abnormalities were significantly less in the vitamin E-treated hearts compared with the untreated failing hearts at 20 weeks. An improved myocardial redox state with vitamin E therapy, coupled with the modulation of the development of heart failure, may indicate a pathophysiologic role for increased oxidative stress in the pathogenesis of heart failure. This study suggests the potential therapeutic value of long-term antioxidant treatment in modulating or preventing the pathogenesis of heart failure.