Local causal states and discrete coherent structures.
Coherent structures form spontaneously in nonlinear spatiotemporal systems and are found at all spatial scales in natural phenomena from laboratory hydrodynamic flows and chemical reactions to ocean, atmosphere, and planetary climate dynamics. Phenomenologically, they appear as key components that organize the macroscopic behaviors in such systems. Despite a century of effort, they have eluded rigorous analysis and empirical prediction, with progress being made only recently. As a step in this, we present a formal theory of coherent structures in fully discrete dynamical field theories. It builds on the notion of structure introduced by computational mechanics, generalizing it to a local spatiotemporal setting. The analysis' main tool employs the local causal states, which are used to uncover a system's hidden spatiotemporal symmetries and which identify coherent structures as spatially localized deviations from those symmetries. The approach is behavior-driven in the sense that it does not rely on directly analyzing spatiotemporal equations of motion, rather it considers only the spatiotemporal fields a system generates. As such, it offers an unsupervised approach to discover and describe coherent structures. We illustrate the approach by analyzing coherent structures generated by elementary cellular automata, comparing the results with an earlier, dynamic-invariant-set approach that decomposes fields into domains, particles, and particle interactions.


alpha-Glucosidase and N-acetylglucosamine-6-sulphatase are the major mannose-6-phosphate glycoproteins in human urine.
Most newly synthesized lysosomal enzymes contain a transient carbohydrate modification, mannose 6-phosphate (Man-6-P), which signals their vesicular transport from the Golgi to the lysosome via Man-6-P receptors (MPRs). We have examined Man-6-P glycoproteins in human urine by using a purified soluble fragment of the soluble cation-independent MPR (sCI-MPR) as a preparative and analytical affinity reagent. In a survey of urine samples from seven healthy subjects, the pattern of Man-6-P glycoproteins detected with iodinated sCI-MPR as a probe in a blotting assay was essentially identical in each, regardless of sex or age. Two bands of approx. 100 and 110 kDa were particularly prominent. Man-6-P glycoproteins in human urine were purified by affinity chromatography on immobilized sCI-MPR. Seven distinct bands revealed by SDS/PAGE and Coomassie Blue staining were subjected to N-terminal sequence analysis. The prominent 100 and 110 kDa Man-6-P glycoproteins were identified as N-acetylglucosamine-6-sulphatase and alpha-glucosidase respectively. This identification was confirmed by molecular mass determinations on the two major bands after deglycosylation. Sequence analysis revealed arylsulphatase A and several previously unidentified proteins as minor species. Man-6-P glycoproteins were also purified on an analytical scale to determine the proportion of a number of lysosomal enzyme activities represented by the mannose-6-phosphorylated forms. The lysosomal enzymes in urine containing the highest proportion of mannose-6-phosphorylated form were beta-mannosidase (82%), hexosaminidase (27%) and alpha-glucosidase (24%). The profiles of Man-6-P glycoproteins detected by blotting in urine and plasma were not similar, suggesting that the urinary species are not derived from the bloodstream.


Photoperiodic regulation of prolactin secretion: changes in intra-pituitary signalling and lactotroph heterogeneity.
Many mammalian species utilise day-length (photoperiod) to adapt their physiology to seasonal changes in environmental conditions, via secretion of pineal melatonin. Photoperiodic regulation of prolactin secretion is believed to occur via melatonin-mediated changes in the secretion of a putative prolactin secretagogue, tuberalin, from the pituitary pars tuberalis. Despite the in vivo and in vitro evidence in support of this intra-pituitary signalling mechanism, the identity of tuberalin has yet to be elucidated. This paper reviews recent advances in the characterisation of tuberalin and the regulation of its secretion. Furthermore, the hypothesis that pituitary lactotroph cells display heterogeneity in their response to changing photoperiod and tuberalin secretion is examined.


Global access to affordable direct oral anticoagulants.
Poor control of cardiovascular disease accounts for a substantial proportion of the disease burden in developing countries, but often essential anticoagulant medicines for preventing strokes and embolisms are not widely available. In 2019, direct oral anticoagulants were added to the World Health Organization's WHO Model list of essential medicines. The aims of this paper are to summarize the benefits of direct oral anticoagulants for patients with cardiovascular disease and to discuss ways of increasing their usage internationally. Although the cost of direct oral anticoagulants has provoked debate, the affordability of introducing these drugs into clinical practice could be increased by: price negotiation; pooled procurement; competitive tendering; the use of patent pools; and expanded use of generics. In 2017, only 14 of 137 countries that had adopted national essential medicines lists included a direct oral anticoagulant on their lists. This number could increase rapidly if problems with availability and affordability can be tackled. Once the types of patient likely to benefit from direct oral anticoagulants have been clearly defined in clinical practice guidelines, coverage can be more accurately determined and associated costs can be better managed. Government action is required to ensure that direct oral anticoagulants are covered by national budgets because the absence of reimbursement remains an impediment to achieving universal coverage. Tackling cardiovascular disease with the aid of direct oral anticoagulants is an essential component of efforts to achieve the World Health Organization's target of reducing premature deaths due to noncommunicable disease by 25% by 2025.


Strain, disinfectant, concentration, and contact time quantitatively impact disinfectant efficacy.
Transmission of healthcare-associated infections caused by antibiotic- and multi-drug resistant (MDR) pathogens (e.g. Methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa) are a major concern in patient care facilities. Disinfectant usage is critical to control and prevent pathogen transmission, yet the relationships among strain, disinfectant type, contact time, and concentration are not well-characterized. We hypothesized that there would be significant differences in disinfectant efficacy among clinically relevant strains under off-label disinfectant conditions, but there would be less no differences among at registered label use concentrations and contact times. The purpose of this study was to quantify the effect of disinfectant concentration and contact time on the bactericidal efficacy of clinically relevant strains of S. aureus and P. aeruginosa. Accelerated hydrogen peroxide (AHP), quaternary ammonium compounds (Quat), and sodium hypochlorite were tested at label and reduced contact times and concentrations against four MDR P. aeruginosa strains and four MRSA strains. Quantitative EPA method MB-25-02 was used to measure disinfectant efficacy reported as log10 reduction. Both off-label disinfectant concentrations and contact times significantly affected efficacy of all disinfectants tested. Bactericidal efficacy varied among MRSA and P. aeruginosa strains. The quantitative disinfectant efficacy method used highlights the inter-strain variability that exists within a bacterial species. It also underscores the need for a disinfectant validation method that takes these variances into account.


Methodological issues in studies of air pollution and daily counts of deaths or hospital admissions.
To review the issues and methodologies in epidemiologic time series studies of daily counts of mortality and hospital admissions and illustrate some of the methodologies. This is a review paper with an example drawn from hospital admissions of the elderly in Cleveland, Ohio, USA. The central issue is control for seasonality. Both over and under control are possible, and the use of diagnostics, including plots, is necessary. Weather dependence is probably non-linear, and adequate methods are necessary to adjust for this. In Cleveland, the use of categorical variables for weather and sinusoidal terms for filtering season are illustrated. After control for season, weather, and day of the week effects, hospital admission of persons aged 65 and older in Cleveland for respiratory illness was associated with ozone (RR = 1.09, 95% CI 1.02, 1.16) and particulates (PM10 (RR = 1.12, 95% CI 1.01, 1.24), and marginally associated with sulphur dioxide (SO2) (RR = 1.03, 95% CI = 0.99, 1.06). All of the relative risks are for a 100 micrograms/m3 increase in the pollutant. Several adequate methods exist to control for weather and seasonality while examining the associations between air pollution and daily counts of mortality and morbidity. In each case, care and judgement are required.


Anti-cancer properties and mechanisms of action of thymoquinone, the major active ingredient of Nigella sativa.
Over the past two decades, studies have documented the wide-range anti-cancer effects of Nigella sativa, known as black seed or black cumin. Thymoquinone (TQ), its major active ingredient, has also been extensively studied and reported to possess potent anti-cancer properties. Herein, we provide a comprehensive review of the findings related to the anti-cancer activity of TQ. The review focuses on analyzing experimental studies performed using different in vitro and in vivo models to identify the anti-proliferative, pro-apoptotic, anti-oxidant, cytotoxic, anti-metastatic, and NK-dependent cytotoxic effects exerted by TQ. In addition, we pinpoint the molecular mechanisms underlying these effects and the signal transduction pathways implicated by TQ. Our analysis show that p53, NF-κB, PPARγ, STAT3, MAPK, and PI3K/AKT signaling pathways are among the most significant pathways through which TQ mediates its anti-cancer activity. Experimental findings and recent advances in the field highlight TQ as an effective therapeutic agent for the suppression of tumor development, growth and metastasis for a wide range of tumors.


Optimal timing of one-shot interventions for epidemic control.
The interventions and outcomes in the ongoing COVID-19 pandemic are highly varied. The disease and the interventions both impose costs and harm on society. Some interventions with particularly high costs may only be implemented briefly. The design of optimal policy requires consideration of many intervention scenarios. In this paper we investigate the optimal timing of interventions that are not sustainable for a long period. Specifically, we look at at the impact of a single short-term non-repeated intervention (a "one-shot intervention") on an epidemic and consider the impact of the intervention's timing. To minimize the total number infected, the intervention should start close to the peak so that there is minimal rebound once the intervention is stopped. To minimise the peak prevalence, it should start earlier, leading to initial reduction and then having a rebound to the same prevalence as the pre-intervention peak rather than one very large peak. To delay infections as much as possible (as might be appropriate if we expect improved interventions or treatments to be developed), earlier interventions have clear benefit. In populations with distinct subgroups, synchronized interventions are less effective than targeting the interventions in each subcommunity separately.


Characterization of attaching and effacing Escherichia coli (AEEC) isolated from pigs and sheep.
Attaching and effacing Escherichia coli (AEEC) are characterized by their ability to cause attaching-and-effacing (A/E) lesions in the gut mucosa of human and animal hosts leading to diarrhoea. The genetic determinants for the production of A/E lesions are located on the locus of enterocyte effacement (LEE), a pathogenicity island that also contains the genes encoding intimin (eae). This study reports data on the occurrence of eae positive E. coli carried by healthy pigs and sheep at the point of slaughter, and on serotypes, intimin variants, and further virulence factors of isolated AEEC strains. Faecal samples from 198 finished pigs and 279 sheep were examined at slaughter. The proportion of eae positive samples was 89% for pigs and 55% for sheep. By colony dot-blot hybridization, AEEC were isolated from 50 and 53 randomly selected porcine and ovine samples and further characterized. Strains of the serotypes O2:H40, O3:H8 and O26:H11 were found in both pigs and sheep. In pigs O2:H40, O2:H49, O108:H9, O145:H28 and in sheep O2:H40, O26:H11, O70:H40, O146:H21 were the most prevalent serotypes among typable strains. Eleven different intimin types were detected, whereas gamma2/theta was the most frequent, followed by beta1, epsilon and gamma1. All but two ovine strains tested negative for the genes encoding Shiga toxins. All strains tested negative for the bfpA gene and the EAF plasmid. EAST1 (astA) was present in 18 of the isolated strains. Our data show that pigs and sheep are a source of serologically and genetically diverse intimin-harbouring E. coli strains. Most of the strains show characteristics of atypical enteropathogenic E. coli. Nevertheless, there are stx-negative AEEC strains belonging to serotypes and intimin types that are associated with classical enterohaemorrhagic E. coli strains (O26:H11, beta1; O145:H28, gamma1).


The effect of a unique propolis compound (Propoelix™) on clinical outcomes in patients with dengue hemorrhagic fever.
Dengue fever is a mosquito-borne virus belonging to the family Flaviviridae. It is an old virus that has re-emerged globally over the past 20 years and now causes a global burden of 50 million infections per year across approximately 100 countries. Despite this, there is no safe vaccine available, and therapy is largely supportive. Its pathogenesis is multifaceted and currently still poorly understood, leading to a lack of disease-specific therapy. Propolis is a natural antiviral and anti-inflammatory product derived from the saps of plants and mixed with the saliva of honeybees. Propoelix™ is a uniquely potent and water-soluble extract of propolis containing high concentrations of anti-inflammatory compounds like caffeic acid phenethyl ester. The primary objective is to determine the effectiveness of a unique propolis extract (Propoelix™) on the clinical course of patients with dengue hemorrhagic fever (DHF). The secondary objective is to examine the effect of Propoelix™ on tumor necrosis factor-α (TNF-α) levels in patients with DHF. A double-blind, randomized, placebo-controlled trial was conducted at the Department of Internal Medicine, Gatot Soebroto Central Army Hospital in Jakarta, Indonesia, from May 2012 to July 2013. Sixty-three patients who met the inclusion criteria were enrolled in the trial. Patients were randomized to receive either two capsules of Propoelix™ 200 mg three times a day or placebo daily for 7 days. Clinical and laboratory variables of both groups, including the anti-inflammatory marker TNF-α, were investigated. Patients were deemed technically fit for discharge if their platelet counts had recovered and exceeded 100,000/μL but were all observed as inpatients for 7 days. There were 31 patients in the Propoelix™ treatment group and 32 patients in the placebo group. Platelet counts in the Propoelix™-treated group showed a trend toward a faster recovery by day 3 of admission and became statistically significant by day 6 (101.42±48.79 vs 80.78±43.35 [10(3)/mL], P=0.042) and day 7 (146.67±64.68 vs 107.84±57.22 [10(3)/mL], P=0.006). Patients treated with Propoelix™ had a significantly greater decline in TNF-α levels on day 7 of therapy compared with patients in the placebo group (P=0.018). They also had a significantly shorter length of hospitalization compared with those in the placebo group (4.69±0.78 days vs 5.46±1.16 days, P=0.012). Propoelix™ appears to hasten the improvement in platelet counts and TNF-α levels and shortens the duration of hospitalization in patients with DHF.


A max-flow-based approach to the identification of protein complexes using protein interaction and microarray data.
The emergence of high-throughput technologies leads to abundant protein-protein interaction (PPI) data and microarray gene expression profiles, and provides a great opportunity for the identification of novel protein complexes using computational methods. By combining these two types of data, we propose a novel Graph Fragmentation Algorithm (GFA) for protein complex identification. Adapted from a classical max-flow algorithm for finding the (weighted) densest subgraphs, GFA first finds large (weighted) dense subgraphs in a protein-protein interaction network, and then, breaks each such subgraph into fragments iteratively by weighting its nodes appropriately in terms of their corresponding log-fold changes in the microarray data, until the fragment subgraphs are sufficiently small. Our tests on three widely used protein-protein interaction data sets and comparisons with several latest methods for protein complex identification demonstrate the strong performance of our method in predicting novel protein complexes in terms of its specificity and efficiency. Given the high specificity (or precision) that our method has achieved, we conjecture that our prediction results imply more than 200 novel protein complexes.


Text data extraction for a prospective, research-focused data mart: implementation and validation.
Translational research typically requires data abstracted from medical records as well as data collected specifically for research. Unfortunately, many data within electronic health records are represented as text that is not amenable to aggregation for analyses. We present a scalable open source SQL Server Integration Services package, called Regextractor, for including regular expression parsers into a classic extract, transform, and load workflow. We have used Regextractor to abstract discrete data from textual reports from a number of 'machine generated' sources. To validate this package, we created a pulmonary function test data mart and analyzed the quality of the data mart versus manual chart review. Eleven variables from pulmonary function tests performed closest to the initial clinical evaluation date were studied for 100 randomly selected subjects with scleroderma. One research assistant manually reviewed, abstracted, and entered relevant data into a database. Correlation with data obtained from the automated pulmonary function test data mart within the Northwestern Medical Enterprise Data Warehouse was determined. There was a near perfect (99.5%) agreement between results generated from the Regextractor package and those obtained via manual chart abstraction. The pulmonary function test data mart has been used subsequently to monitor disease progression of patients in the Northwestern Scleroderma Registry. In addition to the pulmonary function test example presented in this manuscript, the Regextractor package has been used to create cardiac catheterization and echocardiography data marts. The Regextractor package was released as open source software in October 2009 and has been downloaded 552 times as of 6/1/2012. Collaboration between clinical researchers and biomedical informatics experts enabled the development and validation of a tool (Regextractor) to parse, abstract and assemble structured data from text data contained in the electronic health record. Regextractor has been successfully used to create additional data marts in other medical domains and is available to the public.


Bonding of a self-etching primer to non-carious cervical sclerotic dentin: interfacial ultrastructure and microtensile bond strength evaluation.
The objectives of this study were 1) to examine the ultrastructural features of the resin-sclerotic dentin interface following the application of Clearfil Liner Bond II sigma to natural cervical wedge-shaped lesions, and 2) to evaluate the regional tensile bond strength of this self-etching primer at different locations on natural and artificially-created cervical lesions. Deep cervical natural lesions were bonded using the self-etching primer. Micromorphology of the bonded interface at different locations within the lesions were examined using scanning electron microscopy (SEM), transmission electron microscopy (TEM) and scanning transmission electron microscopy/energy dispersive x-ray analysis (STEM/EDX). Ultrastructural features were further compared with the use of the same self-etching primer on artificial lesions created in sound cervical dentin. A nontrimming technique was used to evaluate the regional tensile bond strength from the occlusal, gingival, and the deepest central part of both natural and artificial cervical lesions. Beams with a mean area of 0.46 +/- 0.03 mm2 were prepared and were pulled to failure using a Bencor Multi-T testing device attached to an Instron universal tester. Bond strength results were evaluated using a two-way ANOVA design. A hypermineralized layer devoid of intact, banded collagen was invariably present on the surface of the natural lesions. Depending upon its thickness at different locations of the lesion, the action of a self-etching primer may be limited to this surface layer alone, producing a hybridized hypermineralized surface layer. Penetration of the self-etching primer into the underlying sclerotic dentin produced a hybridized complex containing a hybridized hypermineralized surface layer as well as a subsurface layer of hybridized intertubular dentin. Bacterial colonization of the lesion surface resulted in the formation of an additional zone of hybridized intermicrobial matrix over the surface of the lesions. Dentinal tubules remained blocked with sclerotic casts, and resin tags were rarely observed. Regional tensile bond strength results showed that the overall bond strength to natural sclerotic dentin was about 20% lower than sound cervical dentin, but was independent of the different locations within the lesions from which bond strength was evaluated. There were four factors that may have influenced the overall decrease in bond strength in natural cervical sclerotic lesions: a) the presence of a hybridized intermicrobial matrix together with entrapped bacteria may have weakened the bonds, b) inability of a self-etching primer to etch through a thick, hypermineralized surface layer, c) presence of a layer of possibly remineralized, denatured collagen at the base of the hypermineralized surface layer, and d) retention of acid-resistant sclerotic casts that obliterate the tubular lumina and prevent effective resin tag formation.


3-Methyleneoxindole: an affinity label of glutathione S-transferase pi which targets tryptophan 38.
The compound 3-methyleneoxindole (MOI), a photooxidation product of the plant auxin indole-3-acetic acid, functions as an affinity label of the dimeric pi class glutathione S-transferase (GST) isolated from pig lung. MOI inactivates the enzyme to a limit of 14% activity. The k for inactivation by MOI is decreased 20-fold by S-hexylglutathione but only 2-fold by S-methylglutathione, suggesting that MOI does not react entirely within the glutathione site. The striking protection against inactivation provided by S-(hydroxyethyl)ethacrynic acid indicates that MOI reacts in the active site region involving both the glutathione and the xenobiotic substrate sites. Incorporation of [(3)H]MOI up to approximately 1 mol/mol of enzyme dimer concomitant with maximum inactivation suggests that there are interactions between subunits. Fractionation of the proteolytic digest of [(3)H]MOI-modified GST pi yielded Trp38 as the only labeled amino acid. The crystal structure of the human GST pi-ethacrynic acid complex (2GSS) shows that the indole of Trp38 is less than 4 A from ethacrynic acid. Similarly, MOI may bind in this substrate site. In contrast to its effect on the pi class GST, MOI inactivates much less rapidly and extensively alpha and mu class GSTs isolated from the rat. These results show that MOI reacts preferentially with GST pi. Such a compound may be useful in novel combination chemotherapy to enhance the efficacy of alkylating cancer drugs while minimizing toxic side effects.


Cognitive skills, student achievement tests, and schools.
Cognitive skills predict academic performance, so schools that improve academic performance might also improve cognitive skills. To investigate the impact schools have on both academic performance and cognitive skills, we related standardized achievement-test scores to measures of cognitive skills in a large sample (N = 1,367) of eighth-grade students attending traditional, exam, and charter public schools. Test scores and gains in test scores over time correlated with measures of cognitive skills. Despite wide variation in test scores across schools, differences in cognitive skills across schools were negligible after we controlled for fourth-grade test scores. Random offers of enrollment to oversubscribed charter schools resulted in positive impacts of such school attendance on math achievement but had no impact on cognitive skills. These findings suggest that schools that improve standardized achievement-test scores do so primarily through channels other than improving cognitive skills.


TORC2, a coactivator of cAMP-response element-binding protein, promotes Epstein-Barr virus reactivation from latency through interaction with viral BZLF1 protein.
Reactivation of the Epstein-Barr virus from latency is dependent on expression of the viral BZLF1 protein. The BZLF1 promoter (Zp) normally exhibits only low basal activity but is activated in response to chemical inducers such as 12-O-tetradecanoylphorbol-13-acetate and calcium ionophore. We found here that Transducer of Regulated cAMP-response Element-binding Protein (CREB) (TORC) 2 enhances Zp activity 10-fold and more than 100-fold with co-expression of the BZLF1 protein. Mutational analysis of Zp revealed that the activation by TORC is dependent on ZII and ZIII cis elements, binding sites for CREB family transcriptional factors and the BZLF1 protein, respectively. Immunoprecipitation, chromatin immunoprecipitation, and reporter assay using Gal4-luc and Gal4BD-BZLF1 fusion protein indicate that TORC2 interacts with BZLF1, and that the complex is efficiently recruited onto Zp. These observations clearly indicate that TORC2 activates the promoter through interaction with the BZLF1 protein as well as CREB family transcriptional factors. Induction of the lytic replication resulted in the translocation of TORC2 from cytoplasm to viral replication compartments in nuclei, and furthermore, activation of Zp by TORC2 was augmented by calcium-regulated phosphatase, calcineurin. Silencing of endogenous TORC2 gene expression by RNA interference decreased the levels of the BZLF1 protein in response to 12-O-tetradecanoylphorbol-13-acetate/ionophore. Based on these results, we conclude that Epstein-Barr virus exploits the calcineurin-TORC signaling pathway through interactions between TORC and the BZLF1 protein in reactivation from latency.


Cerebral venous thrombosis and hepatic venous thrombosis during pregnancy.
Cerebral venous thrombosis (CVT) and Budd-Chiari syndrome (BCS) are rarely encountered thrombotic diseases of two different vascular systems. A 20-year-old woman in the 14th week of pregnancy was brought to the emergency department with complaints of sudden headache and loss of vision. Thrombosis in the left transverse and sigmoid sinus was demonstrated at MR angiography. She was diagnosed with BCS by using hepatic Doppler ultrasonography after an elevation of liver enzymes. Although CVT and BCS are rarely seen during pregnancy, they need emergency treatment because of high mortality.


Perindopril changes the mesenteric pressure profile of conscious hypertensive and normotensive rats.
Information about how antihypertensive therapy affects the arterial blood pressure profile in conscious animals is at present not available. Here we report measurements of part of the pressure profile in conscious spontaneously hypertensive rats (SHR, n = 7) and Wistar-Kyoto (WKY, n = 7) rats before and after treatment with the angiotensin-converting enzyme inhibitor perindopril. The previously developed technique that we used, provided simultaneous measurements of the undisturbed arterial blood pressure at the base of mesenteric arcades (P(arc); diameter, approximately 100 microns) and systemic mean blood pressure (MBP). The ratio P(arc)/MBP was 63 +/- 2% (mean +/- SEM) in SHR and 64 +/- 3% in WKY rats. When a bolus of perindopril (0.8 mg/kg) was injected into the aorta, P(arc)/MBP fell within 2 minutes to 51 +/- 2% (P < .05) for SHR and 56 +/- 2% (P < .05) for WKY rats, and these levels were maintained for the next hour. In contrast, MBP did not change for approximately 5 minutes in either strain, whereas after 1 hour MBP still had not changed significantly in WKY rats, but MBP had fallen by 16 +/- 2% (P < .05) in SHR.(ABSTRACT TRUNCATED AT 250 WORDS)


Inhibitory centers in sexual behavior in the male rat.
Small lesions placed near the diencephalic, mesencephalic junction, in either the lateral or medial mammillary region, resulted in an increase of copulatory behavior. This increase was expressed both by increased numbers of copulation plugs formed per 14-day interval and by increased percentage of days on which copulation occurred. Inhibitory structures thus form an essential part of the circuitry involved in mediation of sex behavior in the male.


Comparative study of unsupervised dimension reduction techniques for the visualization of microarray gene expression data.
Visualization of DNA microarray data in two or three dimensional spaces is an important exploratory analysis step in order to detect quality issues or to generate new hypotheses. Principal Component Analysis (PCA) is a widely used linear method to define the mapping between the high-dimensional data and its low-dimensional representation. During the last decade, many new nonlinear methods for dimension reduction have been proposed, but it is still unclear how well these methods capture the underlying structure of microarray gene expression data. In this study, we assessed the performance of the PCA approach and of six nonlinear dimension reduction methods, namely Kernel PCA, Locally Linear Embedding, Isomap, Diffusion Maps, Laplacian Eigenmaps and Maximum Variance Unfolding, in terms of visualization of microarray data. A systematic benchmark, consisting of Support Vector Machine classification, cluster validation and noise evaluations was applied to ten microarray and several simulated datasets. Significant differences between PCA and most of the nonlinear methods were observed in two and three dimensional target spaces. With an increasing number of dimensions and an increasing number of differentially expressed genes, all methods showed similar performance. PCA and Diffusion Maps responded less sensitive to noise than the other nonlinear methods. Locally Linear Embedding and Isomap showed a superior performance on all datasets. In very low-dimensional representations and with few differentially expressed genes, these two methods preserve more of the underlying structure of the data than PCA, and thus are favorable alternatives for the visualization of microarray data.


Detecting Antigen-Specific T Cell Responses: From Bulk Populations to Single Cells.
A new generation of sensitive T cell-based assays facilitates the direct quantitation and characterization of antigen-specific T cell responses. Single-cell analyses have focused on measuring the quality and breadth of a response. Accumulating data from these studies demonstrate that there is considerable, previously-unrecognized, heterogeneity. Standard assays, such as the ICS, are often insufficient for characterization of rare subsets of cells. Enhanced flow cytometry with imaging capabilities enables the determination of cell morphology, as well as the spatial localization of the protein molecules within a single cell. Advances in both microfluidics and digital PCR have improved the efficiency of single-cell sorting and allowed multiplexed gene detection at the single-cell level. Delving further into the transcriptome of single-cells using RNA-seq is likely to reveal the fine-specificity of cellular events such as alternative splicing (i.e., splice variants) and allele-specific expression, and will also define the roles of new genes. Finally, detailed analysis of clonally related antigen-specific T cells using single-cell TCR RNA-seq will provide information on pathways of differentiation of memory T cells. With these state of the art technologies the transcriptomics and genomics of Ag-specific T cells can be more definitively elucidated.


A modelled comparison of the effects of using different ways to compensate for missed treatment days in radiotherapy.
There is much evidence for the detrimental effect on tumour control of missed treatment days during radiotherapy, amounting for example to approximately a 1.6% absolute decrease in local control probability per day of treatment prolongation in the case of head and neck squamous cell cancer. Various methods to compensate for missed treatment days are compared quantitatively in this article, using the linear-quadratic formalism. The overall time and fraction size can be maintained by either treating on weekend days (the preferred way (Method 1a), although with unsocial hours and at extra cost) or using two fractions per day to "catch up' (Method 1b). The latter might incur a small loss of tolerance regarding late reactions, when intervals of 6-8 h are used rather than 24 h, and there may be logistical/scheduling difficulties with larger numbers of patients in some centres when using this method. A second type of strategy retains overall treatment time, and also one fraction per day, but the size of the dose per fraction is increased. For example, this may be done for the same number of "post-gap' days as gap days (Method 2). However, with this method, calculated isoeffect doses regarding late reactions indicate a probable decrease in tumour control rate (Method 2a). Otherwise, isoeffective doses regarding tumour control result in an increase in late reactions (Method 2b). In addition, this method is unsuitable for short regimens already using high doses per fraction. To reduce this problem, overall treatment time can also be retained by using fewer fractions, all of greater size in the case of planned gaps (statutory holidays), or larger remaining fractions after unplanned gaps (Method 2c). The problem also with this method is that equivalence for tumour control gives an increase in late reactions. The least satisfactory strategy (Method 3) is to accept the protraction caused by the missed treatment days, and give either the same prescribed number of (slightly larger) fractions or the planned treatment followed by one (or more) extra fraction to compensate for the gap. This would retain the expected local control rate, but there would be an increase in late reactions. An example of this, using average parameter values, is that a 3-day gap (necessitating four extra days to complete treatment with one fraction of 2.4 Gy) might maintain a 70% local control rate for glottic carcinoma, but severe reactions might rise from 1% to 4% and minor/moderate reactions from 37% to 50%. In this example, the inclusion of an extra weekend would increase the required extra dose and hence may further increase the morbidity rates. A final point is that the effect of treatment interruptions for an individual patient is expected to be greater than that for a group of patients because of interpatient heterogeneity tending to flatten dose-response curves. Calculations show that the above value of 1.6% loss of local control per day for a group of patients may reflect values for individual patients that range around a median value of as much as 5% per day, so stressing further the importance of gaps in treatment. It is concluded that, wherever possible, treatment days should not be missed. If they are missed, it is important to compensate for them, preferably by one of the first of the above methods (1a or 1b), in order to keep as close as possible to the original/standard prescription in terms of total dose, dose per fraction and overall time.


EMLA application exceeding two hours improves pediatric emergency department venipuncture success.
The purpose of this study was to determine whether placing Eutectic Mixture of Local Anesthetics (EMLA) at emergency department (ED) triage improves venipuncture success. Emergency department triage nurses prospectively identified patients aged 0-18 years assessed to have 50% or greater chance of needing venipuncture while in the emergency department. Identified patients received EMLA or no intervention according to randomized 24-h blocks. Data were collected on need for venipuncture, venipuncture success (defined as obtaining access in one attempt), and duration and success of EMLA application (defined as EMLA occluded between 1 and 5 h). Parents' satisfaction and perception of pain were assessed with a 5-item scale. Consent was obtained from 267 out of 287 patients, in whom 111 of 154 venipunctures were successful (72%). EMLA (n = 100) and no-intervention (n = 167) groups did not differ by age or dehydration-related illnesses. Nurses flagged patients more often on days when EMLA was not applied (χ(2) = 37.8, df2, p < 0.0001), but with lower specificity of needing venipuncture (48.5% no-intervention venipuncture rate vs. 73% venipuncture rate with EMLA (χ(2) = 14.4, df2, p = 0.0001). Of the 73 EMLA patients undergoing venipuncture, 2 removed EMLA at unknown times; 2 had application duration longer than 5 h, and nurses chose non-EMLA venipuncture sites for 8. Punctures through EMLA sites with appropriate duration were successful in 51/61 (84%) versus 58/89 (65%) in untreated skin (p = 0.01, odds ratio (OR) = 2.8; 95% confidence interval (CI) [1.3, 6.2]). Success in intention-to-treat groups did not differ (OR = 1.8, 95% CI [0.87, 3.7]). Improved venipuncture success was seen linearly with increased application duration (p = 0.018). Parents perceived less pain with punctures through EMLA sites (p = 0.006). Venipuncture through an EMLA site reduced pain and improved success. Rates improved with increasing application duration. Barriers to triage EMLA placement should be evaluated. Application longer than 2 h should be studied further as a means to improve success. EMLA was supplied as a part of the Investigator-sponsored study program of AstraZeneca.


Graphene Quantum Capacitors for High Frequency Tunable Analog Applications.
Graphene quantum capacitors (GQC) are demonstrated to be enablers of radio-frequency (RF) functions through voltage-tuning of their capacitance. We show that GQC complements MEMS and MOSFETs in terms of performance for high frequency analog applications and tunability. We propose a CMOS compatible fabrication process and report the first experimental assessment of their performance at microwaves frequencies (up to 10 GHz), demonstrating experimental GQCs in the pF range with a tuning ratio of 1.34:1 within 1.25 V, and Q-factors up to 12 at 1 GHz. The figures of merit of graphene variable capacitors are studied in detail from 150 to 350 K. Furthermore, we describe a systematic, graphene specific approach to optimize their performance and predict the figures of merit achieved if such a methodology is applied.


SMS texts on corruption help Ugandan voters hold elected councillors accountable at the polls.
Many politicians manipulate information to prevent voters from holding them accountable; however, mobile text messages may make it easier for nongovernmental organizations to credibly share information on official corruption that is difficult for politicians to counter directly. We test the potential for texts on budget management to improve democratic accountability by conducting a large (n = 16,083) randomized controlled trial during the 2016 Ugandan district elections. In cooperation with a local partner, we compiled, simplified, and text-messaged official information on irregularities in local government budgets. Verified recipients of messages that described more irregularities than expected reported voting for incumbent councillors 6% less often; verified recipients of messages conveying fewer irregularities than expected reported voting for incumbent councillors 5% more often. The messages had no observable effect on votes for incumbent council chairs, potentially due to voters' greater reliance on other sources of information for higher profile elections. These mixed results suggest that text messages on budget corruption help voters hold some politicians accountable in settings where elections are not free and fair.


National TB program shortages as potential factor for poor-quality TB care cascade: Healthcare workers' perspective from Beira, Mozambique.
Mozambique is one of the countries with the deadly implementation gaps in the tuberculosis (TB) care and services delivery. In-hospital delays in TB diagnosis and treatment, transmission and mortality still persist, in part, due to poor-quality of TB care cascade. We aimed to assess, from the healthcare workers' (HCW) perspective, factors associated with poor-quality TB care cascade and explore local sustainable suggestions to improve in-hospital TB management. In-depth interviews and focus group discussions were conducted with different categories of HCW. Audio-recording and written notes were taken, and content analysis was performed through atlas.ti7. Bottlenecks within hospital TB care cascade, lack of TB staff and task shifting, centralized and limited time of TB laboratory services, and fear of healthcare workers getting infected by TB were mentioned to be the main factors associated with implementation gaps. Interviewees believe that task shifting from nurses to hospital auxiliary workers, and from higher and well-trained to lower HCW are accepted and feasible. The expansion and use of molecular TB diagnostic tools are seen by the interviewees as a proper way to fight effectively against both sensitive and MDR TB. Ensuring provision of N95 respiratory masks is believed to be an essential requirement for effective engagement of the HCW on high-quality in-hospital TB care. For monitoring and evaluation, TB quality improvement teams in each health facility are considered to be an added value. Shortage of resources within the national TB control programme is one of the potential factors for poor-quality of the TB care cascade. Task shifting of TB care and services delivery, decentralization of the molecular TB diagnostic tools, and regular provision of N95 respiratory masks should contribute not just to reduce the impact of resource scarceness, but also to ensure proper TB diagnosis and treatment to both sensitive and MDR TB.


Energy spectra considerations for synchrotron radiotherapy trials on the ID17 bio-medical beamline at the European Synchrotron Radiation Facility.
The aim of this study was to validate the kilovoltage X-ray energy spectrum on the ID17 beamline at the European Synchrotron Radiation Facility (ESRF). The purpose of such validation was to provide an accurate energy spectrum as the input to a computerized treatment planning system, which will be used in synchrotron microbeam radiotherapy trials at the ESRF. Calculated and measured energy spectra on ID17 have been reported previously but recent additions and safety modifications to the beamline for veterinary trials warranted a fresh investigation. The authors used an established methodology to compare X-ray attenuation measurements in copper sheets (referred to as half value layer measurements in the radiotherapy field) with the predictions of a theoretical model. A cylindrical ionization chamber in air was used to record the relative attenuation of the X-ray beam intensity by increasing thicknesses of high-purity copper sheets. The authors measured the half value layers in copper for two beamline configurations, which corresponded to differing spectral conditions. The authors obtained good agreement between the measured and predicted half value layers for the two beamline configurations. The measured first half value layer was 1.754 ± 0.035 mm Cu and 1.962 ± 0.039 mm Cu for the two spectral conditions, compared with theoretical predictions of 1.763 ± 0.039 mm Cu and 1.984 ± 0.044 mm Cu, respectively. The calculated mean energies for the two conditions were 105 keV and 110 keV and there was not a substantial difference in the calculated percentage depth dose curves in water between the different spectral conditions. The authors observed a difference between their calculated energy spectra and the spectra previously reported by other authors, particularly at energies greater than 100 keV. The validation of the beam spectrum by the copper half value layer measurements means the authors can provide an accurate spectrum as an input to a treatment planning system for the forthcoming veterinary trials of microbeam radiotherapy to spontaneous tumours in cats and dogs.


Graphene-Assisted Synthesis of 2D Polyglycerols as Innovative Platforms for Multivalent Virus Interactions.
2D nanomaterials have garnered widespread attention in biomedicine and bioengineering due to their unique physicochemical properties. However, poor functionality, low solubility, intrinsic toxicity, and nonspecific interactions at biointerfaces have hampered their application in vivo. Here, biocompatible polyglycerol units are crosslinked in two dimensions using a graphene-assisted strategy leading to highly functional and water-soluble polyglycerols nanosheets with 263 ± 53 nm and 2.7 ± 0.2 nm average lateral size and thickness, respectively. A single-layer hyperbranched polyglycerol containing azide functional groups is covalently conjugated to the surface of a functional graphene template through pH-sensitive linkers. Then, lateral crosslinking of polyglycerol units is carried out by loading tripropargylamine on the surface of graphene followed by lifting off this reagent for an on-face click reaction. Subsequently, the polyglycerol nanosheets are detached from the surface of graphene by slight acidification and centrifugation and is sulfated to mimic heparin sulfate proteoglycans. To highlight the impact of the two-dimensionality of the synthesized polyglycerol sulfate nanosheets at nanobiointerfaces, their efficiency with respect to herpes simplex virus type 1 and severe acute respiratory syndrome corona virus 2 inhibition is compared to their 3D nanogel analogs. Four times stronger in virus inhibition suggests that 2D polyglycerols are superior to their current 3D counterparts.


Role of plasma elongation on turbulent transport in magnetically confined plasmas.
The theoretical study of plasma turbulence is of central importance to fusion research. Experimental evidence indicates that the confinement time results mainly from the turbulent transport of energy, the magnitude of which depends on the turbulent state resulting from nonlinear saturation mechanisms, in particular, the self-generation of coherent macroscopic structures and large scale flows. Plasma geometry has a strong impact on the structure and magnitude of these flows and also modifies the mode linear growth rates. Nonlinear global gyrokinetic simulations in realistic tokamak magnetohydrodynamic equilibria show how plasma shape can control the turbulent transport. Results are best described in terms of an effective temperature gradient. With increasing plasma elongation, the nonlinear critical effective gradient is not modified while the stiffness of transport is decreasing.


Self-regulation in self-propelled nematic fluids.
We consider the hydrodynamic theory of an active fluid of self-propelled particles with nematic aligning interactions. This class of materials has polar symmetry at the microscopic level, but forms macrostates of nematic symmetry. We highlight three key features of the dynamics. First, as in polar active fluids, the control parameter for the order-disorder transition, namely the density, is dynamically convected by the order parameter via active currents. The resulting dynamical self-regulation of the order parameter is a generic property of active fluids and destabilizes the uniform nematic state near the mean-field transition. Secondly, curvature-driven currents render the system unstable deep in the nematic state, as found previously. Finally, and unique to self-propelled nematics, nematic order induces local polar order that in turn leads to the growth of density fluctuations. We propose this as a possible mechanism for the smectic order of polar clusters seen in numerical simulations.


Epidemiology of Chagas disease in Ecuador. A brief review.
Chagas disease is a complex public health problem that has been underestimated in Ecuador. Here we review the relevant published information, and present unpublished and new data that help to understand the current Chagas disease epidemiological situation and its evolution in the country. Three main characteristics have been identified: (i) persistence of Trypanosoma cruzi transmission in already known foci; (ii) a marked endemicity in some urban areas of Guayaquil; and (iii) the transformation of new Amazon foci into truly endemic areas. The situation in other suspect areas remains uncertain. Five Triatominae species have been implicated in the transmission of T. cruzi to people in Ecuador (Triatoma dimidiata, Rhodnius ecuadoriensis, R. pictipes, R. robustus and Panstrongylus geniculatus), but some others may also play a role in some areas (P. rufotuberculatus, P. howardi, T. carrioni and P. chinai). Other Triatominae reported seem to have little or no epidemiological relevance (T. venosa, T. dispar, Eratyrus mucronatus, E. cuspidatus, P. lignarius and Cavernicola pilosa). High frequency of acute cases and severe chronic disease has been observed. Although cardiomyopathy is more frequent, serious digestive disease is also present. It is estimated that around 120,000-200,000 people may be infected. 2.2 to 3.8 million people are estimated to live under transmission risk conditions.


Czech Teratology Information Service: comparison of treatments by psychotropic and antiepileptic drugs.
Care, treatment and follow-up in psychiatric and epileptic pregnant women were compared with women inquiring Czech Teratology Information Service (CZTIS) due to other exposure to drugs during pregnancy. Data were collected by CZTIS, member of European Network of Teratology Information Services from 1996. Exposed groups were compared with pregnant women exposed to drugs which were not classified as major teratogens or hyperthermia. Groups do not vary in age, reproductive history and other parameters. We observed higher frequency of miscarriage and voluntary termination of pregnancy in the group of psychiatric patients. The number of malformation in prospective follow-up cases was lower than in control group. Chronic diseases as epilepsy or psychiatric disorders have to be treated during pregnancy. Women should obtain accurate information about possible risk before pregnancy. Co-operation is needed in these cases. Physicians should keep in mind that appropriate information is to be given to the patient according to her disease, education and comprehension of the problem. If there is any doubt they should organize help for their patients.


Open TG-GATEs: a large-scale toxicogenomics database.
Toxicogenomics focuses on assessing the safety of compounds using gene expression profiles. Gene expression signatures from large toxicogenomics databases are expected to perform better than small databases in identifying biomarkers for the prediction and evaluation of drug safety based on a compound's toxicological mechanisms in animal target organs. Over the past 10 years, the Japanese Toxicogenomics Project consortium (TGP) has been developing a large-scale toxicogenomics database consisting of data from 170 compounds (mostly drugs) with the aim of improving and enhancing drug safety assessment. Most of the data generated by the project (e.g. gene expression, pathology, lot number) are freely available to the public via Open TG-GATEs (Toxicogenomics Project-Genomics Assisted Toxicity Evaluation System). Here, we provide a comprehensive overview of the database, including both gene expression data and metadata, with a description of experimental conditions and procedures used to generate the database. Open TG-GATEs is available from http://toxico.nibio.go.jp/english/index.html.


Healthcare interventions for depression in low socioeconomic status populations: A systematic review and meta-analysis.
The prevalence and impact of depressive disorders in developed countries are associated with certain population characteristics, including socioeconomic status. The aim of this systematic review and meta-analysis was to identify, characterize and analyze the short- and long-term effectiveness of healthcare interventions for depressive disorders in low socioeconomic status populations. The main biomedical databases were searched and fifteen articles assessing seventeen interventions were included in the review. Most interventions were implemented in the US (n=11) and culturally adapted (n=11). We conducted two meta-analyses for assessing both the short- (n=11) and long-term effectiveness (n=12) of interventions. There was a statistically significant reduction in overall depressive symptoms (-0.58, 95% CI [-0.74, -0.41]) at short-term (up to three months after the intervention), especially for combined and psychotherapeutic interventions. The overall effect slightly decreased at long-term (-0.42, 95% CI [-0.63, -0.21]). Those interventions including culturally specific training for providers and booster sessions seemed to be more effective in reducing depressive disorders at short and long term, respectively. In conclusion, healthcare interventions are effective in decreasing clinically significant depressive disorders in low socioeconomic status populations. Future interventions should take into account the key characteristics identified in this review.


Anti-obesity efficacy of nanoemulsion oleoresin capsicum in obese rats fed a high-fat diet.
This study determined the effects of oleoresin capsicum (OC) and nanoemulsion OC (NOC) on obesity in obese rats fed a high-fat diet. THE RATS WERE RANDOMLY SEPARATED INTO THREE GROUPS: a high-fat (HF) diet group, HF + OC diet group, and HF + NOC diet group. All groups were fed the diet and water ad libitum for 14 weeks. NOC reduced the body weight and adipose tissue mass, whereas OC did not. OC and NOC reduced mRNA levels of adipogenic genes, including peroxisome proliferator-activated receptor (PPAR)-γ, sterol regulatory element-binding protein-1c, and fatty acid-binding protein in white adipose tissue. The mRNA levels of genes related to β-oxidation or thermogenesis including PPAR-α, palmitoyltransferase-1α, and uncoupling protein-2 were increased by the OC and NOC relative to the HF group. Both OC and NOC clearly stimulated AMP-activated protein kinase (AMPK) activity. In particular, PPAR-α, palmitoyltransferase-1α, uncoupling protein-2 expression, and AMPK activity were significantly increased in the NOC group compared to in the OC group. NOC decreased glycerol-3-phosphate dehydrogenase activity whereas OC did not. From these results, NOC could be suggested as a potential anti-obesity agent in obese rats fed a HF diet. The effects of the NOC on obesity were associated with changes of multiple gene expression, activation of AMPK, and inhibition of glycerol-3-phosphate dehydrogenase in white adipose tissue.


Effect of mutation type and location on clinical outcome in 1,013 probands with Marfan syndrome or related phenotypes and FBN1 mutations: an international study.
Mutations in the fibrillin-1 (FBN1) gene cause Marfan syndrome (MFS) and have been associated with a wide range of overlapping phenotypes. Clinical care is complicated by variable age at onset and the wide range of severity of aortic features. The factors that modulate phenotypical severity, both among and within families, remain to be determined. The availability of international FBN1 mutation Universal Mutation Database (UMD-FBN1) has allowed us to perform the largest collaborative study ever reported, to investigate the correlation between the FBN1 genotype and the nature and severity of the clinical phenotype. A range of qualitative and quantitative clinical parameters (skeletal, cardiovascular, ophthalmologic, skin, pulmonary, and dural) was compared for different classes of mutation (types and locations) in 1,013 probands with a pathogenic FBN1 mutation. A higher probability of ectopia lentis was found for patients with a missense mutation substituting or producing a cysteine, when compared with other missense mutations. Patients with an FBN1 premature termination codon had a more severe skeletal and skin phenotype than did patients with an inframe mutation. Mutations in exons 24-32 were associated with a more severe and complete phenotype, including younger age at diagnosis of type I fibrillinopathy and higher probability of developing ectopia lentis, ascending aortic dilatation, aortic surgery, mitral valve abnormalities, scoliosis, and shorter survival; the majority of these results were replicated even when cases of neonatal MFS were excluded. These correlations, found between different mutation types and clinical manifestations, might be explained by different underlying genetic mechanisms (dominant negative versus haploinsufficiency) and by consideration of the two main physiological functions of fibrillin-1 (structural versus mediator of TGF beta signalling). Exon 24-32 mutations define a high-risk group for cardiac manifestations associated with severe prognosis at all ages.


Pressure reentrant assembly: direct simulation of volumes of micellization.
Surfactants exhibit maxima in their critical micelle concentrations upon application of hydrostatic pressure, which is attributable to changes in their volumes of micellization from positive to negative values with increasing pressure. We present a direct molecular simulation analysis of the volumes of micellization of an anionic, cationic, and nonionic surfactant in aqueous solution at pressures up to 2500 bar. Excellent agreement with experiment is observed. A Kirkwood-Buff theory analysis based on proximal solvent distributions permits the breakdown of the volumes of micellization into constituent surfactant headgroup and tailgroup contributions. Although the micellization volume crossover is analogous to the transfer of an alkane from water to its pure liquid, significant differences are observed, including lower compressibilities of micelle volumes compared to that of the alkane liquid, negative partial compressibilites for anionic sulfated surfactant monomers, and large nonionic ethoxy headgroup contributions to the micellization volume.


Sequence diversity within a subgroup of mouse immunoglobulin kappa chains controlled by the IgK-Ef2 locus.
We previously showed that a chromosome 6 locus, IgK-Ef2, controls a pair of prominent bands in normal mouse light-chain isoelectric focusing profiles. Screening of myeloma light chains derived from BALB/c mice (an IgK-EF2 alpha strain) led to the identification of seven light chains cofocusing with the polymorphic bands controlled by IgK-Ef2. Complete sequencing of the variable (V) regions of four of the light chains indicates that they are all members of the same subgroup (Vk-1A) and they differ from one another by 1--3 substitutions. One of the protein differs from the prototype V-region sequence only in the deletion of a single residue at position 95 immediately preceding of J region. The other two differ from the protype V region by 3 (two framework [fr], one complementarity-determined [cdr]) and one (fr) residues, respectively. Complete V-region sequences of two closely related light chains derived from NZB mice (an IgK-Ef2b strain) indicate the NZB proteins are derived from a distinct Vk gene (Vk-1B), differing by four substitutions from the Vk-1A sequence. The results suggest that the IgK-Ef2 polymorphism may be a result of, at least in part, the loss of the gene(s) coding for the Vk-1A subgroups in IgK-Ef2b strains of mice. The nature of the sequence diversity found in the Vk-1A subgroup indicates that either it is coded by a repeated series of virtually identical genes or that somatic mutation of a single Vk-1A gene may give rise to substitutions in framework as well as cdr regions.


Submicron Size Particles of a Murine Monoclonal Antibody Are More Immunogenic Than Soluble Oligomers or Micron Size Particles Upon Subcutaneous Administration in Mice.
Protein aggregates are one of the several risk factors for undesired immunogenicity of biopharmaceuticals. However, it remains unclear which features determine whether aggregates will trigger an unwanted immune response. The aim of this study was to determine the effect of aggregates' size on their relative immunogenicity. A monoclonal murine IgG1 was stressed by exposure to low pH and elevated temperature followed by stirring to obtain aggregates widely differing in size. Aggregate fractions enriched in soluble oligomers, submicron size particles and micron size particles were isolated via centrifugation or size-exclusion chromatography and characterized physicochemically. The secondary and tertiary structures of aggregates were altered in a similar way for all the fractions, while no substantial chemical degradation was observed. Development of anti-drug antibodies was measured after subcutaneous administration of each enriched fraction to BALB/c mice. Among all tested fractions, the most immunogenic was the one highly enriched in submicron size particles (∼100-1000 nm). Fractions composed of micron size (>1-100 μm) particles or soluble oligomers (<100 nm) were not immunogenic under the dosing regimen studied in this work. These results show that aggregate size is an important factor for protein immunogenicity.


Towards a theoretical foundation for morphological computation with compliant bodies.
The control of compliant robots is, due to their often nonlinear and complex dynamics, inherently difficult.The vision of morphological computation proposes to view these aspects not only as problems, but rather also as parts of the solution. Non-rigid body parts are not seen anymore as imperfect realizations of rigid body parts, but rather as potential computational resources. The applicability of this vision has already been demonstrated for a variety of complex robot control problems. Nevertheless, a theoretical basis for understanding the capabilities and limitations of morphological computation has been missing so far. We present a model for morphological computation with compliant bodies, where a precise mathematical characterization oft he potential computational contribution of a complex physical body is feasible. The theory suggests that complexity and nonlinearity, typically unwanted properties of robots, are desired features in order to provide computational power. We demonstrate that simple generic models of physical bodies,based on mass-spring systems, can be used to implement complex nonlinear operators. By adding a simple readout(which is static and linear) to the morphology, such devices are able to emulate complex mappings of input to output streams in continuous time. Hence, by outsourcing parts of the computation to the physical body, the difficult problem of learning to control a complex body, could be reduced to a simple and perspicuous learning task, which can not get stuck in local minima of an error function.


Increased expression of PAD2 after repeated intracerebroventricular infusions of soluble Abeta(25-35) in the Alzheimer's disease model rat brain: effect of memantine.
Peptidylarginine deiminases (PADs) convert the arginine residues in proteins into citrulline residues in a Ca(2+)-dependent manner. We previously showed that a bilateral injection of ibotenic acid into the rat nucleus basalis magnocellularis elevated the PAD2 activity in the hippocampus and striatum. In this study, we examined whether repeated intracerebroventricular infusions of soluble Abeta25-35 would affect the PAD2 expression in any regions of the rat brain. We also assessed the protective effect of memantine on Abeta-induced PAD2 alterations. The infusion of Abeta(25-35) increased the activity and protein level of PAD2 in the hippocampus, and co-treatment with memantine suppressed these changes. An immunohistochemical analysis showed that an increased level of PAD2 was coincident with GFAP-positive astrocytes and CD11b-positive microglia. In addition, immunofluoresecence staining revealed that citrullinepostive immunoreactivity coincided with the occurrence of GFAP-positive astrocytes. Co-treatment with memantine reversed the activation of the astrocytes and microglia, thus attenuating the PAD2 increment. These biochemical and immunohistochemical results suggest that PAD2 might play an important role in the pathology of early Alzheimer's disease, and may correlate with the changes in glial cells that are recovered by memantine treatment.


Transplant immunosuppression increases and prolongs transgene expression following adenoviral-mediated transfection of rat lungs.
Gene therapy provides the potential to modify donor organs to better withstand transplantation, but this has yet to be realized. In vivo gene transfer using adenoviral vectors has had limited success because of host immune response that induces inflammation and limits the amount and duration of transgene expression. We hypothesize that transplantation immunosuppression can attenuate the post-transfection host-immune response to allow for improved gene transfer following adenoviral-mediated transfection. We intratracheally transfected with adenovirus containing the beta-galactosidase gene and randomized the rats to either the immunosuppression group, receiving daily cyclosporine, azathioprine, and methylprednisolone, or the control group, receiving no immunosuppression. We evaluated transgene expression and post-transfection inflammation at time points ranging from 1 day to 5 weeks. Following transfection, control rats showed relatively low levels of transgene expression, which rapidly decreased to non-detectable levels. In contrast, immunosuppressed rats demonstrated significantly higher levels of transgene expression overall (p < 0.00005), peaking at almost 3 times that of the control group (p < 0.02), and showing prolonged and elevated transgene expression at 5 weeks (p < 0.02). On histologic sections of the lungs, immunosuppressed rats exhibited overall lesser grades of post-transfection inflammation. Transplant immunosuppression provides the means to attenuate the severe immune response to adenoviral-mediated gene transfection and thereby increase and prolong transgene expression.


Classification of failure of limb salvage after reconstructive surgery for bone tumours : a modified system Including biological and expandable reconstructions.
Previous classification systems of failure of limb salvage focused primarily on endoprosthetic failures and lacked sufficient depth for the effective study of the causes of failure. In order to address these inadequacies, the International Society of Limb Salvage (ISOLS) formed a committee to recommend revisions of the previous systems. The purpose of this study was to report on their recommendations. The modifications were prepared using an earlier, evidence-based model with subclassification based on the existing medical literature. Subclassification for all five primary types of failure of limb salvage following endoprosthetic reconstruction were formulated and a complementary system was derived for the failure of biological reconstruction. An additional classification of failure in paediatric patients was also described. Limb salvage surgery presents a complex array of potential mechanisms of failure, and a complete and precise classification of types of failure is required. Earlier classification systems lacked specificity, and the evidence-based system outlined here is designed to correct these weaknesses and to provide a means of reporting failures of limb salvage in order to allow the interpretation of outcome following reconstructive surgery.


The dialectics of universality: The heterodox critical social theory of Robert Fine.
Robert Fine was among the most original social theorists in Britain of the past 30 years, and the aim of this paper is to offer a first systematic assessment of his intellectual contribution. There are sound intellectual reasons to explore Fine's scholarship. He maintained a problematic relation with mainstream sociology and, against the reduction of sociology to questions of method, culture, or class, he argued that sociologists must continue to ask difficult normative questions as part of the social world they ought to explain. And there are also pressing political concerns that justify a reconsideration of his writings. Global politics is currently marked by a populist wave that decries the very ideas and values that were central to Fine's social theory: the need to uphold the rule of law at home and abroad, the politics of cosmopolitan solidarity, and the significance of antisemitism and its relationships with different forms of authoritarian politics. My main argument is that there is a dialectics of universality that drives forward Fine's intellectual project. By this, I mean that a universalistic idea of humanity-an all-inclusive conception of all human beings-is the most important normative intuition of modern times. This idea of humanity moves forward in history through a dual process of emancipation and domination: successful forms of social, legal, and political inclusion help make visible previous dynamics of exclusion but may also create or recreate discriminatory practices. Building on the work of French historian Michael Löwy on heterodox Jewish thinkers, I explain the three main tenets of Fine's work: (a) his reconstruction of critical social theory; (b) the notion of cosmopolitan solidarity; and (c) the significance and main features of modern antisemitism.


Structural characterizations of fly ash-based geopolymer after adsorption of various metal ions.
Geopolymer, an amorphous substance, has been viewed as good adsorbent or catalyst and attracted much attentions from all over the world. In order to achieve the better applications of geopolymer in these fields, a deep understanding of the microstructure of geopolymer would be strongly required. In the present study, geopolymer was synthesized from coal fly ash, and the structural analysis of geopolymer after adsorption of various metal ions (Li+, Cs+, Sr2+ and Co2+) was studied using XRD, SEM-EDX, FTIR, UV-VIS DRS, TG-DTA as well as surface area and pore distribution analysis. Pair distribution function preferably illustrated that geopolymer was successfully prepared from calcined fly ash. Geopolymer possesses different affinities towards various metal ions. After exchanging with other metal ions, the main structure of geopolymer was maintained. Metal ions with a large radius would have greater effect on the existing state and amount of water molecules in geopolymer. Moreover, the specific surface area of geopolymer after exchanging with metal ions decreased as a function of the radius of them. The spectra corresponding to d-d transitions indicated that the Co2+ could be incorporated into the deformed six-member rings or eight-membered rings. It could be deduced that the sites for ion exchange could be different rings or even cavities distributed on the surface layer of geopolymer. Furthermore, the rings distributed in the geopolymer structure were predominant in the 6-, 8-, 10- or even 12-member rings to maintain the structure stability and charge balance with the cations.


Effect of air-flow on the evaluation of refractive surgery ablation patterns.
An Allegretto Eye-Q laser platform (Wavelight GmbH, Erlangen, Germany) was used to study the effect of air-flow speed on the ablation of artificial polymer corneas used for testing refractive surgery patterns. Flat samples of two materials (PMMA and Filofocon A) were ablated at four different air flow conditions. The shape and profile of the ablated surfaces were measured with a precise non-contact optical surface profilometer. Significant asymmetries in the measured profiles were found when the ablation was performed with the clinical air aspiration system, and also without air flow. Increasing air-flow produced deeper ablations, improved symmetry, and increased the repeatability of the ablation pattern. Shielding of the laser pulse by the plume of smoke during the ablation of plastic samples reduced the central ablation depth by more than 40% with no-air flow, 30% with clinical air aspiration, and 5% with 1.15 m/s air flow. A simple model based on non-inertial dragging of the particles by air flow predicts no central shielding with 2.3 m/s air flow, and accurately predicts (within 2 μm) the decrease of central ablation depth by shielding. The shielding effects for PMMA and Filofocon A were similar despite the differences in the ablation properties of the materials and the different full-shielding transmission coefficient, which is related to the number of particles ejected and their associated optical behavior. Air flow is a key factor in the evaluation of ablation patterns in refractive surgery using plastic models, as significant shielding effects are found with typical air-flow levels used under clinical conditions. Shielding effects can be avoided by tuning the air flow to the laser repetition rate.


Board Review Course as Intervention: Impact on PANCE Performance.
To analyze the effectiveness of an independent commercial 3-day Physician Assistant National Certifying Examination (PANCE)-Physician Assistant National Recertifying Examination (PANRE) board review course to improve first-time pass rates on the PANCE. Data were extracted from academic files at 2 universities over 2 years. In the first year (2014), the universities had not implemented a requirement for participation in a commercial board review course. In the second year (2015), both universities required participation in the course. There were 116 students at Idaho State University and 85 students at the University of Utah. Multiple regression analyses with PANCE score as the outcome variable were conducted with number of days to PANCE, number of practice tests, location of review course, Physician Assistant Clinical Knowledge Rating and Assessment Tool (PACKRAT) II performance, first-time PANCE pass rates, and first-time PANCE scores. After controlling for demographics, PACKRAT II scores, and students' home university, the study found that participation in the board certification course did not seem to affect student performance on the PANCE (R change = 0.000, p = .728). Post hoc analysis revealed a significant main effect for number of tests (F(3,94) = 3.35, p = .022, (Equation is included in full-text article.)= 0.097). Although not statistically significant, differences between on-site and off-site attendance may be of practical significance. There were no significant differences in PANCE performance of PA students who participated in a program-sponsored board review course and the performance of those who did not participate the previous year. We offer suggestions for future research to maximize the utility of a board review course.


Measuring inequalities in the distribution of health workers: the case of Tanzania.
The overall human resource shortages and the distributional inequalities in the health workforce in many developing countries are well acknowledged. However, little has been done to measure the degree of inequality systematically. Moreover, few attempts have been made to analyse the implications of using alternative measures of health care needs in the measurement of health workforce distributional inequalities. Most studies have implicitly relied on population levels as the only criterion for measuring health care needs. This paper attempts to achieve two objectives. First, it describes and measures health worker distributional inequalities in Tanzania on a per capita basis; second, it suggests and applies additional health care needs indicators in the measurement of distributional inequalities. We plotted Lorenz and concentration curves to illustrate graphically the distribution of the total health workforce and the cadre-specific (skill mix) distributions. Alternative indicators of health care needs were illustrated by concentration curves. Inequalities were measured by calculating Gini and concentration indices. There are significant inequalities in the distribution of health workers per capita. Overall, the population quintile with the fewest health workers per capita accounts for only 8% of all health workers, while the quintile with the most health workers accounts for 46%. Inequality is perceptible across both urban and rural districts. Skill mix inequalities are also large. Districts with a small share of the health workforce (relative to their population levels have an even smaller share of highly trained medical personnel. A small share of highly trained personnel is compensated by a larger share of clinical officers (a middle-level cadre) but not by a larger share of untrained health workers. Clinical officers are relatively equally distributed. Distributional inequalities tend to be more pronounced when under-five deaths are used as an indicator of health care needs. Conversely, if health care needs are measured by HIV prevalence, the distributional inequalities appear to decline. The measure of inequality in the distribution of the health workforce may depend strongly on the underlying measure of health care needs. In cases of a non-uniform distribution of health care needs across geographical areas, other measures of health care needs than population levels may have to be developed in order to ensure a more meaningful measurement of distributional inequalities of the health workforce.


Immunoglobulins inhibit pathophysiological effects of anti-GQ1b-positive sera at motor nerve terminals through inhibition of antibody binding.
High-dose intravenous immunoglobulin (IVIg) is an effective treatment for many antibody-mediated neuromuscular diseases, suggesting that IVIg directly interferes with the pathways through which the pathogenic antibodies exert their effects. However, the precise mechanisms of action are unclear. Serum anti-GQ1b antibodies are strongly associated with ophthalmoplegia in patients with Miller Fisher syndrome (MFS) and Guillain-Barré syndrome (GBS). They induce complement-mediated alpha-latrotoxin-like effects on mouse neuromuscular junctions (NMJs) ex vivo, comprising transient muscle fibre twitching, due to a dramatic increase in the frequency of miniature end-plate potentials (spontaneous quantal acetylcholine release), followed by transmission block. To clarify the mechanisms by which IVIg may act in MFS and GBS, we investigated its effects on the interaction of anti-GQ1b antibodies with GQ1b in vitro and on anti-GQ1b antibody-mediated NMJ injury ex vivo, using anti-GQ1b-positive serum samples from MFS/GBS patients. We show that IVIg inhibits the binding of anti-GQ1b antibodies to GQ1b, thereby preventing complement activation and subsequent pathophysiological effects in our ex vivo mouse NMJ model. These results provide further support for the hypothesis that anti-ganglioside antibodies are the pathogenic factors in MFS/GBS and show that this NMJ model provides a suitable system for investigating the therapeutic effects of IVIg in antibody-mediated neuromuscular diseases.


Synergism of antioxidant action of vitamins E, C and quercetin is related to formation of molecular associations in biomembranes.
Vitamins E, C and polyphenols (flavonoids and non-flavonoids) are major natural antioxidants capable of preventing damage generated by oxidative stress. Here we show the capacity of these antioxidants to form non-covalent association within lipid bilayers close to the membrane/cytosol interface. Antioxidant regeneration is significantly enhanced in these complexes.


Determination of toxic and essential metals in rock and sea salts using pulsed nanosecond laser-induced breakdown spectroscopy.
A spectrometer based on pulsed nanosecond laser-induced breakdown spectroscopy (LIBS) was employed for the quantitative determination of heavy and essential metals in salts from various sources available in Pakistan. Six salt samples were collected from sea salt and rock salt. Toxic metals (Cu, Cd, and Ni) and other microessentials (Fe, Ca, Co, Mg, Mn, S, and Zn) were investigated from the recorded spectra. The detection system was calibrated using a parametric dependence study. The quantitative analyses were accomplished under the assumption of local thermodynamic equilibrium and optically thin plasma. The results by the LIBS technique were in agreement with the outcomes of the same samples studied using a more standard approach like inductively coupled plasma-atomic emission spectroscopy (ICP-AES). When the concentrations of heavy and essential metals were calculated using a calibration-free LIBS method that does not need a standard salt specimen and dilution, both LIBS and ICP-AES were also in good agreement. The limit of detection of the experimental set up was determined for the observed heavy metals in the studied samples.


The impact of COVID-19 on the wellbeing of the UK nursing and midwifery workforce during the first pandemic wave: A longitudinal survey study.
The specific challenges experienced by the nursing and midwifery workforce in previous pandemics have exacerbated pre-existing professional and personal challenges, and triggered new issues. We aimed to determine the psychological impact of the COVID-19 pandemic on the UK nursing and midwifery workforce and identify potential factors associated with signs of post-traumatic stress disorder. A United Kingdom national online survey was conducted at three time-points during the first wave of the COVID-19 pandemic between April and August 2020 (T1 and T2 during initial wave; T3 at three-months following the first wave). All members of the UK registered and unregistered nursing and midwifery workforce were eligible to participate. The survey was promoted via social media and through organisational email and newsletters. The primary outcome was an Impact of Events Scale-Revised score indicative of a post-traumatic stress disorder diagnosis (defined using the cut-off score ≥33). Multivariable logistic regression modelling was used to assess the association between explanatory variables and post-traumatic stress disorder. We received 7840 eligible responses (T1- 2040; T2- 3638; T3- 2162). Overall, 91.6% participants were female, 77.2% were adult registered nurses, and 28.7% were redeployed during the pandemic. An Impact of Events Scale-Revised score ≥33 (probable post-traumatic stress disorder) was observed in 44.6%, 37.1%, and 29.3% participants at T1, T2, and T3 respectively. At all three time-points, both personal and workplace factors were associated with probable post-traumatic stress disorder, although some specific associations changed over the course of the pandemic. Increased age was associated with reduced probable post-traumatic stress disorder at T1 and T2 (e.g. 41-50 years at T1 odds ratio (OR) 0.60, 95% confidence interval (CI) 0.42-0.86), but not at T3. Similarly, redeployment with inadequate/ no training was associated with increased probable post-traumatic stress disorder at T1 and T2, but not at T3 (T1 OR 1.37, 95% CI 1.06-1.77; T3 OR 1.17, 95% CI 0.89-1.55). A lack of confidence in infection prevention and control training was associated with increased probable post-traumatic stress disorder at all three time-points (e.g. T1 OR 1.48, 95% CI 1.11-1.97). A negative psychological impact was evident 3-months following the first wave of the pandemic. Both personal and workplace are associated with adverse psychological effects linked to the COVID-19 pandemic. These findings will inform how healthcare organisations should respond to staff wellbeing needs both during the current pandemic, and in planning for future pandemics.


Human intestinal folate conjugase: adaptation after jejunoileal bypass.
During intestinal absorption, dietary polyglutamyl folates are hydrolyzed to monoglutamyl folates by pteroylpolyglutamate hydrolase (folate conjugase). This enzyme is present in the brush border and intracellular fractions of human jejunal mucosa. We compared the activities of brush border and intracellular folate conjugase (BBFC and ICFC), and other mucosal enzymes in the jejunum in continuity and bypassed loop in nine patients who underwent intestinal reconstitution operations 3 to 9 yr after jejunoileal bypass. Control jejunum was obtained from seven obese patients undergoing gastric bypass surgery. Jejunal morphometry showed mucosal hyperplasia with taller villi, larger crypts, and no change in cell size in the jejunum in continuity as compared to control or bypass jejunum. In brush border fractions, specific activities of sucrase and BBFC were significantly greater in the jejunum in continuity than in the control or bypass jejunum. In contrast, the specific activities of the other brush border and intracellular enzymes, including ICFC, were similar in all three segments. This is the first evidence that BBFC, like sucrase, adapts to luminal nutrition. The difference in response of BBFC and ICFC to diet and/or pancreato-biliary secretions provides evidence that these are distinct enzymes which are regulated by different mechanisms.


Aerococcus Viridans: Case Report, Microbiology, and Literature Review.
BACKGROUND Aerococcus viridans are Gram-positive, catalase and oxidase-negative, microaerophilic, and non-motile bacteria species that are rarely associated with human infections such as arthritis, bacteremia, endocarditis, and meningitis. The bacteria are also fastidious (i.e., have complex nutritional requirements) and often confused with Streptococci species or treated as a contaminant. CASE REPORT We report a case of Aerococcus septicemia in an 85-year-old female patient, who transferred from a nursing home to an acute care hospital in Washington DC, USA. She had a 2-day history of worsening mental status, fever of 38.9°C (102°F), and tachycardia. Urinalysis revealed numerous white blood cells and bacteria. Laboratory tests revealed a white blood cell count of 14 000 cells/mL (85% neutrophils, 8% lymphocytes, 5% bands, and 2% monocytes), hemoglobin of 12.6 g/dL, and serum creatinine of 0.8 mg/dL. Blood and urine cultures obtained during admission grew penicillin-resistant A. viridans, identified via matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) on day 3 of admission. The patient received empiric vancomycin with piperacillin-tazobactam, and we deescalated to vancomycin monotherapy to complete a 14-day treatment course. CONCLUSIONS This case report highlights the role of MALDI-TOF for identifying fastidious organisms, and we were able to form a better clinical correlation between patient symptoms and causative organisms. We believe that antimicrobial therapy (in accordance with susceptibility results) should be initiated in symptomatic patients who have A. viridans isolated in significant amounts in urine or from a sterile site.


Alterations to metabolically active bacteria in the mucosa of the small intestine predict anti-obesity and anti-diabetic activities of grape seed extract in mice.
Epidemiological and clinical studies suggest that grapes and grape-derived products may reduce the risk for chronic disease. Grape seed extract specifically has been gaining interest due to its reported ability to prevent weight gain, moderate hyperglycemia, and reduce inflammation. The purpose of this study was to examine the long-term effects of two doses of grape seed extract (10 and 100 mg kg-1 body wt per d in mice) on markers of metabolic syndrome in the context of a moderately high-fat diet. After 12 weeks, the lower dose of grape seed extract was more effective at inhibiting fat gain and improving glucose tolerance and insulin sensitivity. Neither the high fat diet nor grape seed extract altered skeletal muscle substrate metabolism. Most interestingly, when examining the profile of metabolically active microbiota in the mucosa of the small intestine, cecum, and colonic tissue, grape seed extract seemed to have the most dramatic effect on small intestinal tissue, where the population of Firmicutes was lower compared to control groups. This effect was not observed in the cecal or colonic tissues, suggesting that the main alterations to gut microbiota due to flavan-3-ol supplementation occur in the small intestine, which has not been reported previously. These findings suggest that grape seed extract can prevent early changes in glucose tolerance and alter small intestinal gut microbiota, prior to the onset of skeletal muscle metabolic derangements, when grape seed extract is consumed at a low dose in the context of a moderately high fat diet.


The social accountability of medical schools and its indicators.
There is growing interest worldwide in social accountability for medical and other health professional schools. Attempts have been made to apply the concept primarily to educational reform initiatives with limited concern towards transforming an entire institution to commit and assess its education, research and service delivery missions to better meet priority health needs in society for an efficient, equitable an sustainable health system. In this paper, we clarify the concept of social accountability in relation to responsibility and responsiveness by providing practical examples of its application; and we expand on a previously described conceptual model of social accountability (the CPU model), by further delineating the parameters composing the model and providing examples on how to translate them into meaningful indicators. The clarification of concepts of social responsibility, responsiveness and accountability and the examples provided in designing indicators may help medical schools and other health professional schools in crafting their own benchmarks to assess progress towards social accountability within the context of their particular environment.


Mechanism of contractile responses to bradykinin in canine basilar arteries.
This study was undertaken to investigate the role of endothelium in the modulation of vascular responses to bradykinin and to elucidate the receptor types and mechanism of action of bradykinin in isolated basilar artery. The results showed a contractile response to bradykinin in basilar artery. This contractile response to bradykinin was partially modulated by endothelium in a dose-dependent manner. In addition, both des-Arg9-[Leu8]bradykinin and [d-Arg0,Hyp3,Thi5,8,D-Phe7]bradykinin significantly antagonized the responses to bradykinin. However, the blocking effect and the apparent affinity of [d-Arg0,Hyp3,Thi5,8,D-Phe7]bradykinin (pA2 = 9.6 +/- 0.4) were greater than those with des-Arg9-[Leu8]bradykinin (pA.2 = 7.8 +/- 0.3). These results suggest that two apparently distinct types of BK receptors may exist in basilar arteries. Furthermore, the contractile response to bradykinin in basilar artery was significantly inhibited by 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate (10(-5) M), H-7 (10(-5) M) and TMB-8 (10(-5) M), but not by indomethacin (10(-5) M) or nordihydroquariaretic acid (10(-5) M). On the other hand, nifedipine, Ca(2+)-free medium, EGTA and Ca(2+)-free medium/EGTA significantly reduced the bradykinin-induced contraction, indicating that part of the contractile response of basilar artery to bradykinin is dependent on extracellular Ca2+. In conclusion, the mechanism of the contractile responses to bradykinin in basilar artery may involve increased intracellular Ca2+ levels acting on the BK1 and BK2 receptor, followed by activation of the phosphoinositide pathway and receptor-mediated Ca2+ channel.


Bone morphogenetic protein signaling is required for maintenance of differentiated phenotype, control of proliferation, and hypertrophy in chondrocytes.
To examine the role of bone morphogenetic protein (BMP) signaling in chondrocytes during endochondral ossification, the dominant negative (DN) forms of BMP receptors were introduced into immature and mature chondrocytes isolated from lower and upper portions of chick embryo sternum, respectively. We found that control sternal chondrocyte populations expressed type IA, IB, and II BMP receptors as well as BMP-4 and -7. Expression of a DN-type II BMP receptor (termed DN-BMPR-II) in immature lower sternal (LS) chondrocytes led to a loss of differentiated functions; compared with control cells, the DN-BMPR- II-expressing LS chondrocytes proliferated more rapidly, acquired a fibroblastic morphology, showed little expression of type II collagen and aggrecan genes, and upregulated type I collagen gene expression. Expression of DN-BMPR-II in mature hypertrophic upper sternal (US) chondrocytes caused similar effects. In addition, the DN-BMPR-II-expressing US cells exhibited little alkaline phosphatase activity and type X collagen gene expression, while the control US cells produced both alkaline phosphatase and type X collagen. Both DN-BMPR-II-expressing US and LS chondrocytes failed to respond to treatment with BMP-2 . When we examined the effects of DN forms of types IA and IB BMP receptors, we found that DN-BMPR-IA had little effect, while DN-BMPR-IB had similar but weaker effects compared with those of DN-BMPR-II. We conclude that BMP signaling, particularly that mediated by the type II BMP receptor, is required for maintenance of the differentiated phenotype, control of cell proliferation, and expression of hypertrophic phenotype.


Deer responses to repellent stimuli.
Four repellents representing different modes of action (neophobia, irritation, conditioned aversion, and flavor modification) were tested with captive white-tailed deer in a series of two-choice tests. Two diets differing significantly in energy content were employed in choice tests so that incentive to consume repellent-treated diets varied according to which diet was treated. When the high-energy diet was treated with repellents, only blood (flavor modification) and capsaicin (irritation) proved highly effective. Rapid habituation to the odor of meat and bone meal (neophobia) presented in a sachet limited its effectiveness as a repellent under conditions with a high feeding motivation. Thiram, a stimulus used to condition aversions, was not strongly avoided in these trials, that included only limited exposures to the repellent. These data support previous studies indicating that habituation to odor limits the effectiveness of repellents that are not applied directly to food, while topically-applied irritants and animal-based products produce significant avoidance.


Binding of six chimeric analogs of omega-conotoxin MVIIA and MVIIC to N- and P/Q-type calcium channels.
Replacement of the N-terminal half of omega-conotoxin MVIIC, a peptide blocker of P/Q-type calcium channels, with that of omega-conotoxin MVIIA significantly increased the affinity for N-type calcium channels. To identify the residues essential for subtype selectivity, we examined single reverse mutations from MVIIA-type to MVIIC-type in this chimeric analog. A reverse mutation from Lys(7) to Pro(7) decreased the affinity for both P/Q- and N-type channels, whereas that from Leu(11) to Thr(11) increased the affinity for P/Q-type channels and decreased the affinity for N-type channels. The roles of these two residues were confirmed by synthesizing two MVIIC analogs in which Pro(7) and Thr(11) were replaced with Lys(7) and Leu(11), respectively.


Psychiatric problems and quality of life in a clinical sample of adolescents: The role of peer relations.
The quality of peer relations is linked to mental health in childhood and adolescence, but few studies have investigated its clinical relevance. In particular, the potential mediating role of peer functioning in the associations between different dimensions of symptoms and quality of life (QoL) has not been sufficiently examined. In a clinical sample of adolescents, we examined peer relations in light of psychiatric diagnoses, as well as QoL and symptoms of mental health problems, with particular focus on symptoms of anxiety, depression, and ADHD. We also examined the potential mediating role of peer problems in the relationship between such symptoms and QoL. The sample consisted of 603 adolescents (ages 13-18) referred to clinical assessment. Psychiatric diagnoses according to the criteria of the International Statistical Classification of Diseases and Related Health Problems, 10th revision, were collected from participants' clinical charts. Symptoms of disorders, QoL, and quality of peer relations were measured by self-report questionnaires. Adolescents diagnosed with anxiety/depressive disorder reported more peer problems and lower QoL than adolescents with attention deficit/hyperactivity disorder. These findings were supported with symptom ratings. A path model with bootstrapping was used to assess the potential mediating role of peer problems in the association between symptoms and QoL, showing that peer problems partly mediated the relationship between emotional symptoms and QoL, but not the relationship between ADHD-symptoms and QoL. Improvement of peer relations may be a fruitful path for enhancing QoL among adolescents with symptoms of anxiety and depression.