CDC5 inhibits the hyperphosphorylation of the checkpoint kinase Rad53, leading to checkpoint adaptation.
The Saccharomyces cerevisiae polo-like kinase Cdc5 promotes adaptation to the DNA damage checkpoint, in addition to its numerous roles in mitotic progression. The process of adaptation occurs when cells are presented with persistent or irreparable DNA damage and escape the cell-cycle arrest imposed by the DNA damage checkpoint. However, the precise mechanism of adaptation remains unknown. We report here that CDC5 is dose-dependent for adaptation and that its overexpression promotes faster adaptation, indicating that high levels of Cdc5 modulate the ability of the checkpoint to inhibit the downstream cell-cycle machinery. To pinpoint the step in the checkpoint pathway at which Cdc5 acts, we overexpressed CDC5 from the GAL1 promoter in damaged cells and examined key steps in checkpoint activation individually. Cdc5 overproduction appeared to have little effect on the early steps leading to Rad53 activation. The checkpoint sensors, Ddc1 (a member of the 9-1-1 complex) and Ddc2 (a member of the Ddc2/Mec1 complex), properly localized to damage sites. Mec1 appeared to be active, since the Rad9 adaptor retained its Mec1 phosphorylation. Moreover, the damage-induced interaction between phosphorylated Rad9 and Rad53 remained intact. In contrast, Rad53 hyperphosphorylation was significantly reduced, consistent with the observation that cell-cycle arrest is lost during adaptation. Thus, we conclude Cdc5 acts to attenuate the DNA damage checkpoint through loss of Rad53 hyperphosphorylation to allow cells to adapt to DNA damage. Polo-like kinase homologs have been shown to inhibit the ability of Claspin to facilitate the activation of downstream checkpoint kinases, suggesting that this function is conserved in vertebrates.


Fracture of the anconeal process in two cats.
Two adult Domestic Shorthaired cats were presented with acute forelimb lameness. In one case, there was a history of trauma. A fracture of the anconeal process was diagnosed on the flexed mediolateral radiographs of the elbow in both cats. The fracture was accompanied by a bony avulsion of the tricipital tendon in one animal. Both cats underwent surgical removal of the fractured anconeal process. Follow-up radiographic evaluation was available for one cat six months after treatment; radiographs showed evidence of osteoarthritis. The long-term clinical outcome, based on owner telephone follow-up conducted four and nine years postoperatively, was considered to be satisfactory.


Applications of nanotechnology in cancer.
Modern cancer therapy is more individualized to specific cancer subtypes, in an attempt to treat those patients who are likely to obtain greater benefit and avoid treatment induced side effects in those who will not. Nanotechnology heralds an era whereby cancer could be diagnosed by a single agent, treated simultaneously while the diagnosis is being made, and its response to treatment monitored. Whilst nanotechnology is still mostly in the research stage, several applications are ready for translation from the bench to the bedside, in particular in the field of breast cancer. This is exciting new area of research where science fiction may become a reality.


MERS-CoV spillover at the camel-human interface.
Middle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic virus from camels causing significant mortality and morbidity in humans in the Arabian Peninsula. The epidemiology of the virus remains poorly understood, and while case-based and seroepidemiological studies have been employed extensively throughout the epidemic, viral sequence data have not been utilised to their full potential. Here, we use existing MERS-CoV sequence data to explore its phylodynamics in two of its known major hosts, humans and camels. We employ structured coalescent models to show that long-term MERS-CoV evolution occurs exclusively in camels, whereas humans act as a transient, and ultimately terminal host. By analysing the distribution of human outbreak cluster sizes and zoonotic introduction times, we show that human outbreaks in the Arabian peninsula are driven by seasonally varying zoonotic transfer of viruses from camels. Without heretofore unseen evolution of host tropism, MERS-CoV is unlikely to become endemic in humans.


A close examination of the structure and dynamics of HC(NH2)2PbI3 by MD simulations and group theory.
The formamidinium lead iodide hybrid perovskite is studied using first principles molecular dynamics simulations and further analyzed using group theory. The simulations are performed on large supercells containing 768 atoms under isothermal and fully anisotropic isobaric conditions. Two trajectories, one at 300 K and another at 450 K, were extended for over 50 ps in order to perform a detailed assessment of the rotational dynamics of organic cations. The characteristic rotations of the cation are analyzed by defining two rotation axes. It is found that the formamidinium molecules rotate preferentially around the direction parallel to the line connecting the two nitrogen atoms. The rotational dynamics shows some characteristics already observed in methylammonium lead iodide, like the heterogeneous dynamics at room temperature that disappears at 450 K. The orientational probability of the molecules is explored in terms of an expansion in cubic harmonics up to the 12th order. It reveals a strong directionality at room temperature that relaxes when increasing the temperature. These findings are further rationalized using Landau and group theories suggesting a mixed displacive/order-disorder structural instability at lower temperatures.


Modeling-Guided Design of Paper Microfluidic Networks: A Case Study of Sequential Fluid Delivery.
Paper-based microfluidic devices are popular for their ability to automate multistep assays for chemical or biological sensing at a low cost, but the design of paper microfluidic networks has largely relied on experimental trial and error. A few mathematical models of flow through paper microfluidic devices have been developed and have succeeded in explaining experimental flow behavior. However, the reverse engineering problem of designing complex paper networks guided by appropriate mathematical models is largely unsolved. In this article, we demonstrate that a two-dimensional paper network (2DPN) designed to sequentially deliver three fluids to a test zone on the device can be computationally designed and experimentally implemented without experimental trial and error. This was accomplished by three new developments in modeling flow through paper networks: (i) coupling of the Richards equation of flow through porous media to the species transport equation, (ii) modeling flow through assemblies of multiple paper materials (test membrane and wicking pad), and (iii) incorporating limited-volume fluid sources. We demonstrate the application of this model in the optimal design of a paper-based signal-enhanced immunoassay for a malaria protein, PfHRP2. This work lays the foundation for the development of a computational design toolbox to aid in the design of paper microfluidic networks.


Proximal Femoral Shortening After Cephalomedullary Nail Insertion for Intertrochanteric Fractures.
To assess the incidence of proximal femoral shortening (PFS) and its effect on the patient outcomes when intertrochanteric fractures were treated with a cephalomedullary nail (CMN). Retrospective cohort study. Level II trauma center. Forty-eight consecutive patients with OTA/AO 31-A intertrochanteric fractures. All patients were treated with a Gamma3 CMN (Stryker, Kalamazoo, MI). PFS was assessed for abductor lever arm (x vector), femoral height (y vector), and overall shortening (z vector) on anteroposterior radiographs. Fixation success and retained ambulatory capacity were noted. Shortening of >5 mm of the x, y, and z vectors was evident in 18, 20, and 29 patients, respectively. Shortening of >10 mm of the x, y, and z vectors was measured in 5, 6, and 8 patients, respectively. Mean shortening of the x, y, and z vectors was 4.5, 5.5, and 7 mm, respectively. Greater PFS was found to be associated with fixation failure and inability to retain ambulatory capacity, independently (P ≤ 0.05 and P ≤ 0.025, respectively). Of note, an unstable fracture pattern was not found to be associated with greater PFS. PFS is a common phenomenon after CMN of intertrochanteric fractures with a Gamma CMN. In addition, greater PFS seems to be associated with fixation failure and inability to retain ambulatory capacity postoperatively. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


In vivo efficacy of apramycin in murine infection models.
Apramycin is a unique aminoglycoside with a dissociation of antibacterial activity and ototoxicity. We assessed the antibacterial efficacy of apramycin in two murine models of infection, Mycobacterium tuberculosis aerosol infection and Staphylococcus aureus septicemia. In both infection models, the efficacy of apramycin was comparable to that of amikacin. These results suggest that apramycin has the potential to become a clinically useful agent against drug-resistant pathogens and support further development of this promising unique aminoglycoside.


Focal ductal branch dilatation on magnetic resonance cholangiopancreatography: a hint for early diagnosis of pancreatic carcinoma.
A 63-year-old man with a combination of early pancreatic carcinoma and an intraductal papillary adenoma was reported. A pancreatic cyst was detected by chance at the head of the pancreas by computed tomography for a follow-up study of early rectal carcinoma previously operated. Detailed studies by endoscopic retrograde pancreatography (ERP) showed irregular narrowing of the main pancreatic duct at the pancreatic body and magnetic resonance cholangiopancreatography (MRCP) revealed dilatation of ductal branches draining there. Brushing cytology of the pancreatic duct demonstrated cancer cells and total pancreatectomy was performed. Stepwise histo-pathological examinations of the specimen showed two foci of invasive carcinoma in the neck and body and multiple foci of severe dysplasia, some of which contained carcinoma in situ, in the body of the pancreas. The cystic tumor in the head of the pancreas was an intraductal papillary adenoma. In this case, the scrutiny of a pancreatic cyst including MRCP and ERP led to an early diagnosis of pancreatic cancer. Dilatation of ductal branches depicted by MRCP might be a new hint for early diagnosis of pancreatic carcinoma.


Structural elucidation and molecular characterization of Marinobacter sp. α-amylase.
Halophiles have been perceived as potential source of novel enzymes in recent years. The interest emanates from their ability to catalyze efficiently under high salt and organic solvents. Marinobacter sp. EMB8 α-amylase was found to be active and stable in salt and organic solvents. A study was carried out using circular dichroism (CD), fluorescence spectroscopy, and bioinformatics analysis of similar protein sequence to ascertain molecular basis of salt and solvent adaptability of α-amylase. Structural changes recorded in the presence of varying amounts of NaCl exhibited an increase in negative ellipticity as a function of salt, confirming that salt stabilizes the protein and increases the secondary structure, making it catalytically functional. The data of intrinsic and extrinsic fluorescence (using 1-anilinonaphthalene 8-sulfonate [ANS] as probe) further confirmed the role of salt. The α-amylase was active in the presence of nonpolar solvents, namely, hexane and decane, but inactivated by ethanol. The decrease in the activity was correlated with the loss of tertiary structure in the presence of ethanol. Guanidine hydrochloride and pH denaturation indicated the molten globule state at pH 4.0. Partial N-terminal amino acid sequence of the purified α-amylase revealed the relatedness to Pseudoalteromonas sp. α-amylase. "FVHLFEW" was found as the N-terminal signature sequence. Bioinformatics analysis was done using M. algicola α-amylase protein having the same N-terminal signature sequence. The three-dimensional structure of Marinobacter α-amylase was deduced using the I-TASSER server, which reflected the enrichment of acidic amino acids on the surface, imparting the stability in the presence of salt. Our study clearly indicate that salt is necessary for maintaining the secondary and tertiary structure of halophilic protein, which is a necessary prerequisite for catalysis.


Visual Abstraction and Exploration of Multi-class Scatterplots.
Scatterplots are widely used to visualize scatter dataset for exploring outliers, clusters, local trends, and correlations. Depicting multi-class scattered points within a single scatterplot view, however, may suffer from heavy overdraw, making it inefficient for data analysis. This paper presents a new visual abstraction scheme that employs a hierarchical multi-class sampling technique to show a feature-preserving simplification. To enhance the density contrast, the colors of multiple classes are optimized by taking the multi-class point distributions into account. We design a visual exploration system that supports visual inspection and quantitative analysis from different perspectives. We have applied our system to several challenging datasets, and the results demonstrate the efficiency of our approach.


Corneal Endothelial Cell Segmentation by Classifier-Driven Merging of Oversegmented Images.
Corneal endothelium images obtained by in vivo specular microscopy provide important information to assess the health status of the cornea. Estimation of clinical parameters, such as cell density, polymegethism, and pleomorphism, requires accurate cell segmentation. State-of-the-art techniques to automatically segment the endothelium are error-prone when applied to images with low contrast and/or large variation in cell size. Here, we propose an automatic method to segment the endothelium. Starting with an oversegmented image comprised of superpixels obtained from a stochastic watershed segmentation, the proposed method uses intensity and shape information of the superpixels to identify and merge those that constitute a cell, using support vector machines. We evaluated the automatic segmentation on a data set of in vivo specular microscopy images (Topcon SP-1P), obtaining 95.8% correctly merged cells and 2.0% undersegmented cells. We also evaluated the parameter estimation against the results of the vendor's built-in software, obtaining a statistically significant better precision in all parameters and a similar or better accuracy. The parameter estimation was also evaluated on three other data sets from different imaging modalities (confocal microscopy, phase-contrast microscopy, and fluorescence confocal microscopy) and tissue types (ex vivo corneal endothelium and retinal pigment epithelium). In comparison with the estimates of the data sets' authors, we achieved statistically significant better accuracy and precision in all parameters except pleomorphism, where a similar accuracy and precision were obtained.


Matrix metalloproteinases in plants: a brief overview.
Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases belonging to the metzincin clan. MMPs have been characterized in detail in mammals, and they have been shown to play key roles in many physiological and pathological processes. Plant MMP-like proteases exist, but relatively few have been characterized. It has been speculated that plant MMPs are involved in remodeling of the plant extracellular matrix during growth, development and stress response. However, the precise functions and physiological substrates in higher plants remain to be determined. In this brief overview, we summarize the current knowledge of MMPs in higher plants and algae.


Folding of guanine quadruplex molecules-funnel-like mechanism or kinetic partitioning? An overview from MD simulation studies.
Guanine quadruplexes (GQs) play vital roles in many cellular processes and are of much interest as drug targets. In contrast to the availability of many structural studies, there is still limited knowledge on GQ folding. We review recent molecular dynamics (MD) simulation studies of the folding of GQs, with an emphasis paid to the human telomeric DNA GQ. We explain the basic principles and limitations of all types of MD methods used to study unfolding and folding in a way accessible to non-specialists. We discuss the potential role of G-hairpin, G-triplex and alternative GQ intermediates in the folding process. We argue that, in general, folding of GQs is fundamentally different from funneled folding of small fast-folding proteins, and can be best described by a kinetic partitioning (KP) mechanism. KP is a competition between at least two (but often many) well-separated and structurally different conformational ensembles. The KP mechanism is the only plausible way to explain experiments reporting long time-scales of GQ folding and the existence of long-lived sub-states. A significant part of the natural partitioning of the free energy landscape of GQs comes from the ability of the GQ-forming sequences to populate a large number of syn-anti patterns in their G-tracts. The extreme complexity of the KP of GQs typically prevents an appropriate description of the folding landscape using just a few order parameters or collective variables. We reconcile available computational and experimental studies of GQ folding and formulate basic principles characterizing GQ folding landscapes. This article is part of a Special Issue entitled "G-quadruplex" Guest Editor: Dr. Concetta Giancola and Dr. Daniela Montesarchio.


Impact of Directive 2013/11/EU on Consumer ADR Quality: Evidence from France and the UK.
One of the objectives of Directive 2013/11/EU was to promote high-quality consumer alternative dispute resolution (ADR) schemes in the European Union (EU) through the creation of certification processes and regular monitoring by Member States. To obtain and keep certification, ADR bodies must continuously comply with several binding requirements set down in the Directive testifying-among other things-of their impartiality, expertise, transparency, accessibility, as well as of the fairness, timeliness and effectiveness of their procedures. The objective of this regulatory architecture was to trigger some long-term effects on the procedural design and functioning of ADR bodies and to enhance their credibility and legitimacy vis-à-vis consumers and traders. As such, the new rules have aimed to respond to the criticisms sometimes expressed about the way ADR providers operate, in particular concerns regarding schemes' lack of independence, limited accountability and possible effects on due process. Yet, doubts have been expressed about the ability of the Directive to secure a consistent approach fully supporting high-quality ADR in the EU. This paper intends to test these doubts against facts and evidence. Based notably on replies to a questionnaire sent to Competent Authorities, it zooms in on experiences in two Member States, namely France and the United Kingdom (UK) (more specifically for the latter in the civil aviation and non-regulated sectors). It highlights how the binding quality criteria have been working in practice and the impacts that the Directive has had on ADR bodies in those Member States and sectors. It sheds light on several persisting issues, and makes some policy recommendations, which may be relevant for policymakers not only in France and the UK, but also in other Member States and at the EU level when further developing a sustainable framework for high-quality ADR. In 2019, the European Commission is expected to publish a report on the implementation of the Consumer ADR Directive in all Member States. This contribution may be viewed as a first small step in that direction.


A ghrelin gene variant may predict crossover rate from restricting-type anorexia nervosa to other phenotypes of eating disorders: a retrospective survival analysis.
Patients with anorexia nervosa restricting type (AN-R) often develop bulimic symptoms and crossover to AN-binge eating/purging type (AN-BP), or to bulimia nervosa (BN). We have reported earlier that genetic variants of an orexigenic peptide ghrelin are associated with BN. Here, the relationship between a ghrelin gene variant and the rate of change from AN-R to other phenotypes of eating disorders (EDs) was investigated. Participants were 165 patients with ED, initially diagnosed as AN-R. The dates of their AN-R onset and changes in diagnosis to other subtypes of ED were investigated retrospectively. Ghrelin gene 3056 T-->C SNP (single nucleotide polymorphism) was genotyped. Probability and hazard ratios were analyzed using life table analysis and Cox's proportional hazard regression model, in which the starting point was the time of AN-R onset and the outcome events were the time of (i) onset of binge eating, that is, when patients changed to binge eating AN and BN and (ii) recovery of normal weight, that is, when patients changed to BN or remission. Patients with the TT genotype at 3056 T-->C had a higher probability and hazard ratio for recovery of normal weight. The ghrelin SNP was not related with the onset of binge eating. The 3056 T-->C SNP of the ghrelin gene is related to the probability and the rate of recovery of normal body weight from restricting-type AN.


Beyond viral suppression of HIV - the new quality of life frontier.
In 2016, the World Health Organization (WHO) adopted a new Global Health Sector Strategy on HIV for 2016-2021. It establishes 15 ambitious targets, including the '90-90-90' target calling on health systems to reduce under-diagnosis of HIV, treat a greater number of those diagnosed, and ensure that those being treated achieve viral suppression. The WHO strategy calls for person-centered chronic care for people living with HIV (PLHIV), implicitly acknowledging that viral suppression is not the ultimate goal of treatment. However, it stops short of providing an explicit target for health-related quality of life. It thus fails to take into account the needs of PLHIV who have achieved viral suppression but still must contend with other intense challenges such as serious non-communicable diseases, depression, anxiety, financial stress, and experiences of or apprehension about HIV-related discrimination. We propose adding a 'fourth 90' to the testing and treatment target: ensure that 90 % of people with viral load suppression have good health-related quality of life. The new target would expand the continuum-of-services paradigm beyond the existing endpoint of viral suppression. Good health-related quality of life for PLHIV entails attention to two domains: comorbidities and self-perceived quality of life. Health systems everywhere need to become more integrated and more people-centered to successfully meet the needs of virally suppressed PLHIV. By doing so, these systems can better meet the needs of all of their constituents - regardless of HIV status - in an era when many populations worldwide are living much longer with multiple comorbidities.


The effects of a hot drink on nasal airflow and symptoms of common cold and flu.
Hot drinks are a common treatment for common cold and flu but there are no studies reported in the scientific and clinical literature on this mode of treatment. This study investigated the effects of a hot fruit drink on objective and subjective measures of nasal airflow, and on subjective scores for common cold/flu symptoms in 30 subjects suffering from common cold/flu. The results demonstrate that the hot drink had no effect on objective measurement of nasal airflow but it did cause a significant improvement in subjective measures of nasal airflow. The hot drink provided immediate and sustained relief from symptoms of runny nose, cough, sneezing, sore throat, chilliness and tiredness, whereas the same drink at room temperature only provided relief from symptoms of runny nose, cough and sneezing. The effects of the drinks are discussed in terms of a placebo effect and physiological effects on salivation and airway secretions. In conclusion the results support the folklore that a hot tasty drink is a beneficial treatment for relief of most symptoms of common cold and flu.


[Epidemiology and drug susceptibility of Pseudomonas aeruginosa strains isolated in the Chugoku region of Japan. Infection Forum in the Chugoku Region].
Recently, there have been reports concerning an increased frequency of isolation of multi-drug resistant Pseudomonas aeruginosa (MDRP) strains in hospitals and other clinical settings as well as the associated risk of their hospital-acquired infections; in such a situation, it has been a major challenge to establish methods of managing and treating the infections. In order to investigate the trend of P. aeruginosa, the Infection Forum in the Chugoku Region has conducted to a multi-center collaborative study to isolate P. aeruginosa strains from sputum and urine samples collected between October 2006 and September 2008, analyzed the drug susceptibility and the pulsed-field gel electrophoresis (PFGE) patterns of each strain, and assessed epidemiologic characteristics. Of the 738 P. aeruginosa strains collected in this study, 152 (20.6%), 179 (24.3%), 47 (6.4%), and 39 (5.3%) were found to be ciprofloxacin-resistant, imipenem-resistant, amikacin-resistant, and MDRP, respectively. Among the various antimicrobial agents tested, arbekacin (ABK) revealed the strongest inhibitory effects on each drug-resistant and MDRP strain; therefore, ABK was considered as a potential candidate for future treatment of diseases caused by P. aeruginosa. The study also showed that the detection rates of MDRP varied a lot from hospital to hospital. In addition, PFGE-based cluster analyses revealed several strains isolated in the same hospital exhibited a similar PFGE pattern and the same drug susceptibility, suggesting the presence of "unique" hospital-specific strains.


The Crl protein activates cryptic genes for curli formation and fibronectin binding in Escherichia coli HB101.
Curli are thin, coiled, temperature-regulated fibres on fibronectin-binding Escherichia coli. The subunit protein of curli was highly homologous at its amino terminus to SEF-17, the subunit protein of thin, aggregative fimbriae of Salmonella enteritidis 27655 strain 3b, suggesting that these fibres form a novel class of surface organelles on enterobacteria. E. coli HB101 is non-curliated and unable to bind soluble, iodinated fibronectin. The phenotypically cryptic curlin subunit gene, csgA, in HB101 is transcriptionally activated by expressing the cytoplasmic Crl on a multicopy plasmid. Transcriptional activation of csgA by Crl was observed after growth at 26 degrees C but not at 37 degrees C, even though crl transcription was not thermoregulated. A deletion of the 39 carboxy-terminal residues abolished Crl activity, whereas a deletion of 10 residues at the C-terminus did not, implying that a region between residue 93 and 122 in the 132-amino-acid-residue large Crl protein is required for activating curli expression in E. coli HB101. crl is a normal housekeeping gene in E. coli and it is suggested that its gene product may either be a DNA-binding protein affecting chromatin structure as has been suggested for histone-like protein H1 or interact with specific regulatory protein(s) controlling transcription of genes required for curli formation and fibronectin binding.


Engineering a stable future for DNA-origami as a biomaterial.
DNA as a biomaterial has evoked great interest as a potential platform for therapeutics and diagnostics and as hydrogel scaffolds due to the relative ease of programming its robust and uniform shape, site-specific functionality and controlled responsive behavior. However, for a stable self-assembled product, a relatively high cation concentration is required to prevent denaturation. Physiological and cell-culture conditions do not match these concentrations and present additional nucleases that cause a serious threat to the integrity of DNA-based materials. For the translation of this promising technology towards bioengineering challenges, stability needs to be guaranteed. Over the past years, various methods have been developed addressing the stability-related weaknesses of DNA-origami. This mini-review explains the common stability issues and compares the stabilization strategies recently developed. We present a detailed overview of each method in order to ease the selection process on which method to use for future users of DNA-origami as a biomaterial.


Development of ruthenium antitumor drugs that overcome multidrug resistance mechanisms.
Organometallic ruthenium(II) complexes of the general formula [Ru(eta6-p-cymene)Cl2(L)] and [Ru(eta6-p-cymene)Cl(L)2][BPh4] with modified phenoxazine- and anthracene-based multidrug resistance (MDR) modulator ligands (L) have been synthesized, spectroscopically characterized, and evaluated in vitro for their cytotoxic and MDR reverting properties in comparison with the free ligands. For an anthracene-based ligand, coordination to a ruthenium(II) arene fragment led to significant improvement of cytotoxicity as well as Pgp inhibition activity. A similar, but weaker effect was also observed when using a benzimidazole-phenoxazine derivative as Pgp inhibitor. The most active compound in terms of both Pgp inhibition and cytotoxicity is [Ru(eta6-p-cymene)Cl2(L)], where L is an anthracene-based ligand. Studies show that it induces cell death via inhibition of DNA synthesis. Moreover, because the complex is fluorescent, its uptake in cells was studied, and relative to the free anthracene-based ligand, uptake of the complex is accelerated and accumulation of the complex in the cell nucleus is observed.


Family-focused physical activity, diet and obesity interventions in African-American girls: a systematic review.
Obesity interventions that involve family members may be effective with racial/ethnic minority youth. This review assessed the nature and effectiveness of family involvement in obesity interventions among African-American girls aged 5-18 years, a population group with high rates of obesity. Twenty-six databases were searched between January 2011 and March 2012, yielding 27 obesity pilot or full-length prevention or treatment studies with some degree of family involvement and data specific to African-American girls. Interventions varied in type and level of family involvement, cultural adaptation, delivery format and behaviour change intervention strategies; most targeted parent-child dyads. Some similarities in approach based on family involvement were identified. The use of theoretical perspectives specific to African-American family dynamics was absent. Across all studies, effects on weight-related behaviours were generally promising but often non-significant. Similar conclusions were drawn for weight-related outcomes among the full-length randomized controlled trials. Many strategies appeared promising on face value, but available data did not permit inferences about whether or how best to involve family members in obesity prevention and treatment interventions with African-American girls. Study designs that directly compare different types and levels of family involvement and incorporate relevant theoretical elements may be an important next step.


Validation of the charm 3 SL3 beta-lactam test for screening raw milk in compliance with the U.S. pasteurized milk ordinance. Performance Tested Method 071002.
The Charm 3 SL3 beta-Lactam Test is a 3 min receptor-based lateral-flow Rapid One-Step Assay (ROSA) that detects the six beta-lactam drugs of concern approved for dairy cattle in the United States. The method is a biochemical formulation change of the SL3 beta-Lactam Test evaluated and approved in 2007. The Charm 3 SL3 was evaluated under the AOAC Research Institute Performance Tested Method (PTM) program following the protocol of the U.S. Food and Drug Administration, Center for Veterinary Medicine. The method was approved as PTM 071002 on May 8, 2009. The following drugs were detected in three combined lots: penicillin G at 3.8 ppb, ampicillin at 8.0 ppb, amoxicillin at 8.4 ppb, cephapirin at 20.0 ppb, ceftiofur (total metabolites) at 79 ppb, and cloxacillin at 8.6 ppb > or = 90% of the time with 95% confidence. These detection levels are lower than, but within 75% of, the U.S. Safe Level/Tolerances. Lot-to-lot repeatability was typically within 20% of these determined levels. The test kit was found to be suitable for testing thawed frozen samples. It was also found to respond with equal or better sensitivity to samples that contained incurred analytes, i.e., both the microbiologically active parent drug and its active metabolites. There were no interferences from somatic cells at 1.1 million/mL, bacterial cells at 300 000 CFU/mL, or 32 other non-beta-lactam drugs at 100 ppb. Ruggedness experiments indicated that the test procedure is robust. These results meet the fit-for-purpose approval criteria for inclusion in the National Conference for Interstate Milk Shipments milk testing program.


S-layer, surface-accessible, and concanavalin A binding proteins of Methanosarcina acetivorans and Methanosarcina mazei.
The outermost cell envelope structure of many archaea and bacteria contains a proteinaceous lattice termed the surface layer or S-layer. It is typically composed of only one or two abundant, often posttranslationally modified proteins that self-assemble to form the highly organized arrays. Surprisingly, over 100 proteins were annotated to be S-layer components in the archaeal species Methanosarcina acetivorans C2A and Methanosarcina mazei Gö1, reflecting limitations of current predictions. An in vivo biotinylation methodology was devised to affinity tag surface-exposed proteins while overcoming unique challenges in working with these fragile organisms. Cells were adapted to growth under N2 fixing conditions, thus, minimizing free amines reactive to the NHS-label, and high pH media compatible with the acylation chemistry was used. A 3-phase separation procedure was employed to isolate intact, labeled cells from lysed-cell derived proteins. Streptavidin affinity enrichment followed by stringent wash conditions removed nonspecifically bound proteins. This methodology revealed S-layer proteins in M. acetivorans C2A and M. mazei Gö1 to be MA0829 and MM1976, respectively. Each was demonstrated to exist as multiple glycosylated forms using SDS-PAGE coupled with glycoprotein-specific staining, and by interaction with the lectin, Concanavalin A. A number of additional surface-exposed proteins and glycoproteins were identified and included all three subunits of the thermosome: the latter suggests that the chaperonin complex is both surface- and cytoplasmically localized. This approach provides an alternative strategy to study surface proteins in the archaea.


Solid lipid nanodispersions containing mixed lipid core and a polar heterolipid: characterization.
This paper describes the characterization of solid lipid nanodispersions (SLN) prepared with a 1:1 mixture of theobroma oil and goat fat as the main lipid matrix and Phospholipon 90G (P90G) as a stabilizer heterolipid, using polysorbate 80 as the mobile surfactant, with a view to applying the SLN in drug delivery. The 1:1 lipid mixture and P90G constituting the lipid matrix was first homogeneously prepared by fusion. Thereafter, the SLN were formulated with a gradient of polysorbate 80 and constant lipid matrix concentration by melt-high pressure homogenisation. The SLN were characterized by time-resolved particle size analysis, zeta potential and osmotic pressure measurements, differential scanning calorimetry (DSC) and wide angle X-ray diffraction (WAXD). Transmission electron microscopy (TEM) and isothermal heat conduction microcalorimetry (IMC) which monitors the in situ crystallization were also carried out on the SLN containing P90G and 1.0 % w/w of polysorbate 80. The results obtained in these studies were compared with SLN prepared with theobroma oil with and without phospholipid. Particle size analysis of SLN indicated reduction in size with increase in concentration of mobile surfactant and was in the lower nanometer range after 3 months except SLN prepared without P90G or polysorbate 80. The lipid nanoparticles had negative potentials after 3 months. WAXD and DSC studies revealed low crystalline SLN after 3 months of storage except in WAXD of SLN formulated with 1.0 % w/w polysorbate 80. TEM micrograph of the SLN containing 1.0 % w/w polysorbate 80 revealed discrete particles whose sizes were in consonance with the static light scattering measurement. In situ crystallization studies in IMC revealed delayed crystallization of the SLN with 1.0 % w/w polysorbate 80. Results indicate lipid mixtures produced SLN with lower crystallinity and higher particle sizes compared with SLN prepared with theobroma oil alone with or without P90G, and would lead to higher drug incorporation efficiency when used in formulation of actives. Mixtures of theobroma oil and goat fat would be suitable for the preparation of nanostructured lipid carriers. SLN of theobroma oil containing phospholipid could prove to be a good ocular or parenteral drug delivery system considering the low particle size, particle size stability and in vivo tolerability of the component lipids. SLN prepared with lipid admixture, which had higher increase in d(90%) on storage are suitable for preparation of topical and transdermal products.


Injury analyses in rural children: comparison of Old-Order Anabaptists and non-Anabaptists.
Southwestern Ontario largely comprises rural farming districts and is home to numerous Old-Order Anabaptist settlements. Our objective was to describe the injuries sustained by rural children, both Old-Order Anabaptist and non-Anabaptist, to better target injury prevention programs. We retrospectively examined injury data of rural children in Southwestern Ontario with injury severity scores ≥ 12 obtained from hospital and trauma databases (1997-2007). A total of 422 rural children were included in this study: 7.8% Anabaptist (n = 33) and 92.2% non-Anabaptist (n = 389). The age of injured Anabaptist children (median, 7 years; interquartile range = 10) was younger than non-Anabaptist children (median, 14 years; interquartile range = 7; p < 0.001). Anabaptist children were most frequently injured on their property (48.5%; n = 16 of 33; p < 0.001). Non-Anabaptist children were mostly injured on roads (56.8%; n = 221 of 389; p < 0.05) and by motor vehicle collisions (MVCs; 40.1%; n = 156 of 389; p = 0.02). Frequent causes of injury among Anabaptist children were falls (24.2%; n = 8 of 33; p = 0.02), animals (15.2%; n = 5 of 33; p = 0.004), and buggies (9.1%; n = 3 of 33). Approximately half of both groups injured in MVCs did not use seat belts. There were no significant differences between cohorts in sex, injury severity scores, hospitalization days, rates of complications, interventions, comorbidities, or mortality rates. Injuries to Anabaptist children occur at a young age, primarily on their property, and exhibit a unique spectrum of mechanisms. In contrast, injuries to non-Anabaptist children occur at an older age, primarily on roads, and in MVCs. The use of protective devices was low among all rural children. Development of collaborative injury prevention programs targeted to distinct rural communities, including Anabaptist and non-Anabaptist, are needed for reducing injuries among rural children.


Combining in Vitro Folding with Cell Free Protein Synthesis for Membrane Protein Expression.
Cell free protein synthesis (CFPS) has emerged as a promising methodology for protein expression. While polypeptide production is very reliable and efficient using CFPS, the correct cotranslational folding of membrane proteins during CFPS is still a challenge. In this contribution, we describe a two-step protocol in which the integral membrane protein is initially expressed by CFPS as a precipitate followed by an in vitro folding procedure using lipid vesicles for converting the protein precipitate to the correctly folded protein. We demonstrate the feasibility of using this approach for the K(+) channels KcsA and MVP and the amino acid transporter LeuT. We determine the crystal structure of the KcsA channel obtained by CFPS and in vitro folding to show the structural similarity to the cellular expressed KcsA channel and to establish the feasibility of using this two-step approach for membrane protein production for structural studies. Our studies show that the correct folding of these membrane proteins with complex topologies can take place in vitro without the involvement of the cellular machinery for membrane protein biogenesis. This indicates that the folding instructions for these complex membrane proteins are contained entirely within the protein sequence.


Barriers to the Administration of Epinephrine in Schools.
Anaphylaxis is a serious and growing concern in the school setting as the prevalence of food allergies and food-induced severe allergic reactions continues to increase. A cross-sectional, web-based survey was conducted regarding anaphylactic events that occurred during the 2014-2015 school year. Eligible schools were enrolled in the EPIPEN4SCHOOLS® program (Mylan Specialty L.P., Canonsburg, PA), which provides free epinephrine auto-injectors to qualifying US schools. Participating schools completed a 29-item survey on anaphylactic event occurrence and treatment, epinephrine stock, school policies regarding anaphylaxis, school staff training, and school nursing coverage. Responses were provided by 12,275 schools. Epinephrine was administered on school property for 63.7% of reported anaphylactic events (1272/1998). In 38.5% (235/610) of events for which epinephrine was not used, antihistamines were cited as the reason. Only 59.4% of schools cited epinephrine as their standard first-line therapy for anaphylaxis. School nurses were most likely to be trained in anaphylaxis recognition and permitted to administer epinephrine; however, just 53.6% of schools had a full-time nurse on staff. Process-related barriers to the appropriate use of epinephrine go beyond access to medication. Widespread staff training and review of school policies are needed to ensure that anaphylaxis is appropriately managed in schools.


More Than Words: The Use of Video in Ethnographic Research With People With Intellectual Disabilities.
There is a tendency to exclude people with intellectual disabilities from participating in research about their own lives. While the use of participatory research approaches is increasing, the methods used for engaging people with intellectual disabilities in research are generally limited to interviews and focus groups. Yet a focus on the spoken or written word can present a challenge for those who may prefer to use alternative forms of communication. The purpose of this article is to share the methodological insights gained from a visual ethnographic study that sought to explore the experiences of people with intellectual disabilities engaged in nature based (or "green care") therapeutic interventions for health and well-being. If used within carefully negotiated relationships, we suggest that video can be an empowering visual medium for doing research that can help to elicit the experiences of people with intellectual disabilities firsthand, without having to rely on the views and perspectives of other people.


Evidence of Pasteurella haemolytica linked immune complex disease in natural and experimental models.
The pathogenesis of bovine pneumonic pasteurellosis is not completely understood, and studies have not established that Pasteurella haemolytica A1 (Ph1) virulence is exclusively responsible for the development of acute pulmonary lesions. The purpose of this investigation was to determine if immune complex disease is involved in the pathogenesis of bovine pneumonic pasteurellosis. A retrospective immunohistologic study of lung tissue from natural cases of bovine pneumonic pasteurellosis (44) as performed, and immune complexes were observed in alveloar spaces and walls in 88% of these cases. To study this pathologic mechanism experimentally, groups of mice were immunized with purified Ph1 outer membranes (OMs) or sham immunized on days 0 and 14. Mice were challenged intratracheally on day 24 with either live Ph1 or Ph1 OMs, and pulmonary lesions were assessed 24 h after challenge. Placebo immunized mice developed focal infiltrates of neutrophils and macrophages centered around large caliber bronchi. Mice immunized with Ph1 OMs and challenged with live Ph1 or OMs developed severe bronchointerstitial pneumonia with diffuse neutrophilic infiltration, focal necrosis, hemorrhage and edema, that is histologically similar to bovine pneumonic pasteurellosis. Immunohistology revealed flocculent aggregates of IgG and complement positive material within alveolar spaces and walls from mice challenged with live Ph1, and fine granular deposits of IgG and complement positive material were observed lining the alveolar walls from mice challenged with Ph1 OMs. Immunized mice exhibited high serum IgG antibody titers to Ph1 outer membrane proteins (OMPs). Results of this study suggest that immune complex disease plays a role in the pathogenesis of bovine pneumonic pasteurellosis.


Estimated US Infection- and Vaccine-Induced SARS-CoV-2 Seroprevalence Based on Blood Donations, July 2020-May 2021.
People who have been infected with or vaccinated against SARS-CoV-2 have reduced risk of subsequent infection, but the proportion of people in the US with SARS-CoV-2 antibodies from infection or vaccination is uncertain. To estimate trends in SARS-CoV-2 seroprevalence related to infection and vaccination in the US population. In a repeated cross-sectional study conducted each month during July 2020 through May 2021, 17 blood collection organizations with blood donations from all 50 US states; Washington, DC; and Puerto Rico were organized into 66 study-specific regions, representing a catchment of 74% of the US population. For each study region, specimens from a median of approximately 2000 blood donors were selected and tested each month; a total of 1 594 363 specimens were initially selected and tested. The final date of blood donation collection was May 31, 2021. Calendar time. Proportion of persons with detectable SARS-CoV-2 spike and nucleocapsid antibodies. Seroprevalence was weighted for demographic differences between the blood donor sample and general population. Infection-induced seroprevalence was defined as the prevalence of the population with both spike and nucleocapsid antibodies. Combined infection- and vaccination-induced seroprevalence was defined as the prevalence of the population with spike antibodies. The seroprevalence estimates were compared with cumulative COVID-19 case report incidence rates. Among 1 443 519 specimens included, 733 052 (50.8%) were from women, 174 842 (12.1%) were from persons aged 16 to 29 years, 292 258 (20.2%) were from persons aged 65 years and older, 36 654 (2.5%) were from non-Hispanic Black persons, and 88 773 (6.1%) were from Hispanic persons. The overall infection-induced SARS-CoV-2 seroprevalence estimate increased from 3.5% (95% CI, 3.2%-3.8%) in July 2020 to 20.2% (95% CI, 19.9%-20.6%) in May 2021; the combined infection- and vaccination-induced seroprevalence estimate in May 2021 was 83.3% (95% CI, 82.9%-83.7%). By May 2021, 2.1 SARS-CoV-2 infections (95% CI, 2.0-2.1) per reported COVID-19 case were estimated to have occurred. Based on a sample of blood donations in the US from July 2020 through May 2021, vaccine- and infection-induced SARS-CoV-2 seroprevalence increased over time and varied by age, race and ethnicity, and geographic region. Despite weighting to adjust for demographic differences, these findings from a national sample of blood donors may not be representative of the entire US population.


Orthotopic lower urinary tract reconstruction in women using the Kock ileal neobladder: updated experience in 34 patients.
Orthotopic lower urinary tract reconstruction has revolutionized urinary diversion following cystectomy. Initially performed solely in male patients, orthotopic diversion has now become a viable option in women. Currently, the orthotopic neobladder is the diversion of choice for women requiring lower urinary tract reconstruction at our institution. We evaluate and update our clinical and functional experience with orthotopic reconstruction in female patients. Since June 1990, 34 women 31 to 86 years old (median age 67) have undergone orthotopic lower urinary tract reconstruction following cystectomy. Indications for cystectomy included transitional cell carcinoma in 29 patients, urachal adenocarcinoma in 1, mesenchymal tumor of endometrial origin in 1, cervical carcinoma in 1 and a fibrotic radiated bladder in 1. In addition, 1 woman underwent undiversion to the native urethra following a previous simple cystectomy and cutaneous diversion for eosinophilic cystitis. Data were analyzed according to postoperative early and late complications, survival, tumor recurrence, pathological evaluation of the cystectomy specimen, continence status, voiding pattern and patient satisfaction. The median followup in this group of patients was 30 months (range 17 to 70). There were no perioperative deaths, and 4 early (11%) and 3 (9%) late complications. Four patients died, none with a urethral recurrence, including 3 of metastatic bladder cancer and 1 of unrelated causes. In another patient with an extensive mesenchymal tumor of the uterus a sigmoid tumor recurred requiring conversion of the orthotopic reservoir to a cutaneous diversion. All of the remaining 29 patients are alive without evidence of disease. Intraoperative frozen section of the distal surgical margin (proximal urethra) accurately evaluated (confirmed by permanent section) the proximal urethra prospectively for tumor in all 29 specimens removed for transitional cell carcinoma, including 28 specimens (97%) without evidence of tumor and 1 specimen with carcinoma in situ. Complete daytime and nighttime continence was reported by 29 (88%) and 27 (82%) of 33 evaluable patients, respectively. A total of 28 patients (85%) void to completion, while 5 (15%) require some form of intermittent catheterization to empty the neobladder. Patient satisfaction is overwhelming. The excellent clinical and functional results demonstrated with further followup confirm our initial experience with orthotopic diversion in women. Careful selection of appropriate female candidates for orthotopic diversion is critical, and includes preoperative evaluation of the bladder neck and intraoperative frozen section analysis of the distal cystectomy margin. Furthermore, close monitoring of the retained urethra is mandatory in all women undergoing orthotopic diversion. We believe that the orthotopic neobladder is the urinary diversion of choice in women following cystectomy.


Differential contributions of motilin receptor extracellular domains for peptide and non-peptidyl agonist binding and activity.
The family of G protein-coupled receptors that includes receptors for motilin, ghrelin, and growth hormone secretagogue has substantial potential importance as drug targets. Understanding of the molecular basis of hormone binding and receptor activation should provide insights that are helpful in the development of such drugs. We previously examined the unique second extracellular loop domain of the motilin receptor, identifying key epitopes in perimembranous locations at each end of this long loop (Matsuura, B., Dong, M., and Miller, L. J. (2002) J. Biol. Chem. 277, 9834-9839). Here, we have extended that work, examining the other predicted extracellular domains of the motilin receptor by using sequential deletions of segments ranging from one to six amino acid residues and site-directed alanine replacement mutagenesis approaches. Each construct was transiently expressed in COS cells, and characterized for motilin- and erythromycin-stimulated intracellular calcium responses and motilin radioligand binding. Only those receptor segments that included key Cys residues in positions 25, 30, and 111 or perimembranous regions at the ends of the amino terminus and the first and third extracellular loops disrupted motilin biological activity. Each of these Cys deletions also disrupted action of erythromycin. Alanine replacements for each of the potentially important amino acid residues in the perimembranous segments revealed that residues Gly36, Pro103, Leu109, and Phe332 were responsible for the selective negative impact on motilin biological activity, while responding normally to erythromycin. These results support the presence of functionally important disulfide bonds in the motilin receptor ectodomain and demonstrate that the structural determinants for binding and biological activity of peptide and non-peptidyl agonist ligands are distinct, with a broad extracellular perimembranous base contributing to normal motilin binding.


Extracellular vesicles and vesicle-free secretome of the protozoa Acanthamoeba castellanii under homeostasis and nutritional stress and their damaging potential to host cells.
Acanthamoeba castellanii (Ac) are ubiquitously distributed in nature, and by contaminating medical devices such as heart valves and contact lenses, they cause a broad range of clinical presentations to humans. Although several molecules have been described to play a role in Ac pathogenesis, including parasite host-tissue invasion and escaping of host-defense, little information is available on their mechanisms of secretion. Herein, we describe the molecular components secreted by Ac, under different protein availability conditions to simulate host niches. Ac extracellular vesicles (EVs) were morphologically and biochemically characterized. Dynamic light scattering analysis of Ac EVs identified polydisperse populations, which correlated to electron microscopy measurements. High-performance thin liquid chromatography of Ac EVs identified phospholipids, steryl-esters, sterol and free-fatty acid, the last two also characterized by GC-MS. Secretome composition (EVs and EVs-free supernatants) was also determined and proteins biological functions classified. In peptone-yeast-glucose (PYG) medium, a total of 179 proteins were identified (21 common proteins, 89 exclusive of EVs and 69 in EVs-free supernatant). In glucose alone, 205 proteins were identified (134 in EVs, 14 common and 57 proteins in EVs-free supernatant). From those, stress response, oxidative and protein and amino acid metabolism proteins prevailed. Qualitative differences were observed on carbohydrate metabolism enzymes from Krebs cycle and pentose phosphate shunt. Serine proteases and metalloproteinases predominated. Analysis of the cytotoxicity of Ac EVs (upon uptake) and EVs-free supernatant to epithelial and glioblastoma cells revealed a dose-dependent effect. Therefore, the Ac secretome differs depending on nutrient conditions, and is also likely to vary during infection.


Addressing gaps on risk and resilience factors for alcohol use outcomes in sexual and gender minority populations.
In 2011, the Institute of Medicine released a report that constituted the first comprehensive effort by a federal body to understand the current state of science pertinent to the health needs of sexual and gender minority populations. This mini-review summarises recent empirical, methodological and theoretical advances in alcohol-related research among to lesbian, gay, bisexual and transgender (LGBT) populations and highlights progress towards addressing gaps, with a particular interest in those identified by the Institute of Medicine report. Articles published since 2011 were identified from PsycINFO and PubMed database searches, using various combinations of keyword identifiers (alcohol, alcohol abuse, substance abuse, LGBT, lesbian, gay, bisexual, transgender). Reference sections of included articles were also examined for additional citations. Recent empirical work has contributed to a greater understanding of sub-group differences within this diverse population. Evidence has supported theorised influences that can account for alcohol-related disparities, yet important gaps remain. Studies that examine the role of gender identity and its intersection with sexual identity within transgender and gender non-conforming sub-populations are lacking. Methodological advances in this literature have begun to allow for examinations of how minority-specific and general risk factors of alcohol misuse may contribute to patterns of alcohol involvement over time and within social-relational contexts The recommendations made in the current mini-review are meant to facilitate future collaborative efforts, scale development, thoughtful methodological design and analysis and theoretically driven nuanced hypotheses to better understand, and ultimately address, alcohol-related disparities among sexual and gender minority populations. [Talley AE, Gilbert PA, Mitchell J, Goldbach J, Marshall BDL, Kaysen D. Addressing gaps on risk and resilience factors for alcohol use outcomes in sexual and gender minority populations. Drug Alcohol Rev 2016;35:484-493].


NTS neurons with carotid chemoreceptor inputs arborize in the rostral ventrolateral medulla.
Neurons that were excited by hypoxic stimulation of carotid chemoreceptors were recorded in the caudal portion of the nucleus of the solitary tract (cNTS) of urethan-anesthetized, vagotomized, aortic-deafferented, artificially ventilated rats (n = 23). The focus of the study is on 26 chemosensitive neurons (classified as early- and late-response cells) that were tonically activated by chemoreceptor stimulation and never fired in bursts synchronized with the phrenic nerve discharge (PND) cycle. The discharge of early-response cells (n = 14) started up to 2.5 s before the onset of PND activation, whereas the discharge of late-response cells (n = 14) started 1.5-5 s after onset of PND response. Four early-response cells were antidromically activated from the rostral ventrolateral medulla (RVLM; latencies: 7-13 ms), and two had axenal collaterals in the region of the nucleus ambiguus. Four late-response neurons were antidromically activated from the RVLM (latencies: 6-12 ms), but no collateral was found in this area. The basal discharge of early- and late-response cells ranged from 0 to 10 and 0 to 30 spikes/s, respectively, but most of them had a very low spontaneous firing rate (median: 0.2 and 0.6 spikes/s, respectively). Neither type was excited by baroreceptor stimulation. The cNTS also contained neurons that were firing in bursts synchronized with the PND cycle. These cells were activated by chemoreceptor stimulation and were not antidromically activated from the RVLM. Chemosensitive neurons made up 33% of cNTS neurons antidromically activated from the RVLM (8/24). In conclusion, a population of cNTS chemosensitive neurons devoid of respiratory modulation projects through the RVLM and arborizes in this region.


Residual allergenicity of amino acid-based and extensively hydrolysed cow's milk formulas.
Criteria for labelling infant feeds as suitable for the dietary management of cow's milk protein allergy (CMPA) rely on proving the hypoallergenicity of such feeds or clinical studies showing that the feeds are tolerated by 90% of children with proven CMPA. South African (SA) labelling legislation does not indicate what testing is necessary to prove hypoallergenicity. To evaluate all extensively hydrolysed cow's milk formulas and amino acid-based formulas available in SA for residual allergen content, protein size and amino-acid content. All amino-acid and extensively hydrolysed formulas were found to be similar in composition, with no residual cow's milk allergens detectable by enzyme-linked immunosorbent assay. Furthermore, proteins were absent and only small molecules in the size range of amino acids and possibly of very small oligopeptides were detected. These findings indicate that the formulas are extremely likely to be compliant with the definition of hypoallergenicity as tolerance in 90% of proven sufferers from cow's milk allergy. The formulas may therefore be labelled as suitable for the dietary management of infants with CMPA.


Unraveling additive from nonadditive effects using genomic relationship matrices.
The application of quantitative genetics in plant and animal breeding has largely focused on additive models, which may also capture dominance and epistatic effects. Partitioning genetic variance into its additive and nonadditive components using pedigree-based models (P-genomic best linear unbiased predictor) (P-BLUP) is difficult with most commonly available family structures. However, the availability of dense panels of molecular markers makes possible the use of additive- and dominance-realized genomic relationships for the estimation of variance components and the prediction of genetic values (G-BLUP). We evaluated height data from a multifamily population of the tree species Pinus taeda with a systematic series of models accounting for additive, dominance, and first-order epistatic interactions (additive by additive, dominance by dominance, and additive by dominance), using either pedigree- or marker-based information. We show that, compared with the pedigree, use of realized genomic relationships in marker-based models yields a substantially more precise separation of additive and nonadditive components of genetic variance. We conclude that the marker-based relationship matrices in a model including additive and nonadditive effects performed better, improving breeding value prediction. Moreover, our results suggest that, for tree height in this population, the additive and nonadditive components of genetic variance are similar in magnitude. This novel result improves our current understanding of the genetic control and architecture of a quantitative trait and should be considered when developing breeding strategies.


The association between pulse wave velocity and metabolic syndrome and adiponectin in patients with impaired fasting glucose: cardiovascular risks and adiponectin in IFG.
We aimed to assess how metabolic profiles, surrogate markers of insulin resistance, and subclinical atherosclerosis are interrelated in subjects with impaired fasting glucose (IFG) and investigate whether the diagnosis of metabolic syndrome (MetS) further increases the risk of cardiovascular disease among subjects already at risk. We analyzed 1739 Korean subjects with IFG. The parameters of MetS, plasma adiponectin level, and pulse wave velocity (PWV) were assessed. Subjects with MetS had unfavorable metabolic parameters, lower adiponectin level, and higher peripheral PWV compared to those without MetS. Adiponectin correlated with fasting glucose, waist circumference, triglyceride, HDL-cholesterol, BMI, HOMA-IR, and the number of MetS components. In addition to blood pressure, peripheral PWV was associated with triglyceride, waist circumference, and the number of MetS components while aortic PWV correlated positively with fasting plasma glucose. Multiple linear regression analysis revealed that adiponectin correlated with HDL-cholesterol, HOMA-IR, fasting glucose, waist circumference, and triglyceride, peripheral PWV with blood pressure, body mass index, waist circumference, and the number of MetS components, and aortic PWV with fasting plasma glucose. In subjects with IFG, concurrent MetS increases PWV and has an unfavorable effect on cardiovascular risks, and these risks were further increased by additional MetS components.


Epigenetic mechanisms in the regulation of the maize Suppressor-mutator transposon.
Transcription and transposition of the maize Suppressor-mutator (Spm) transposon are epigenetically controlled. Methylation of specific element sequences prevents transcription and transposition in a heritable manner. Reactivation and demethylation occur in the presence of an active element, implying the existence of an element-encoded epigenetic activator. The methylation target sequences are the 0.2 kb promoter and an 0.35 kb GC-rich downstream sequence. Two Spm-encoded proteins, TnpA and TnpD, participate in transposition. In addition, TnpA has positive and negative regulatory activities. TnpA represses and activates the unmethylated and methylated Spm promoters, respectively, and it participates in the transient and heritable demethylation of the promoter and GC-rich region. There is evidence that TnpA-mediated repressor and epigenetic activator functions occur by different molecular mechanisms.


Utilizing temporal information in fMRI decoding: classifier using kernel regression methods.
This paper describes a general kernel regression approach to predict experimental conditions from activity patterns acquired with functional magnetic resonance image (fMRI). The standard approach is to use classifiers that predict conditions from activity patterns. Our approach involves training different regression machines for each experimental condition, so that a predicted temporal profile is computed for each condition. A decision function is then used to classify the responses from the testing volumes into the corresponding category, by comparing the predicted temporal profile elicited by each event, against a canonical hemodynamic response function. This approach utilizes the temporal information in the fMRI signal and maintains more training samples in order to improve the classification accuracy over an existing strategy. This paper also introduces efficient techniques of temporal compaction, which operate directly on kernel matrices for kernel classification algorithms such as the support vector machine (SVM). Temporal compacting can convert the kernel computed from each fMRI volume directly into the kernel computed from beta-maps, average of volumes or spatial-temporal kernel. The proposed method was applied to three different datasets. The first one is a block-design experiment with three conditions of image stimuli. The method outperformed the SVM classifiers of three different types of temporal compaction in single-subject leave-one-block-out cross-validation. Our method achieved 100% classification accuracy for six of the subjects and an average of 94% accuracy across all 16 subjects, exceeding the best SVM classification result, which was 83% accuracy (p=0.008). The second dataset is also a block-design experiment with two conditions of visual attention (left or right). Our method yielded 96% accuracy and SVM yielded 92% (p=0.005). The third dataset is from a fast event-related experiment with two categories of visual objects. Our method achieved 77% accuracy, compared with 72% using SVM (p=0.0006).


Treatment and management of chronic fatigue syndrome/myalgic encephalomyelitis: all roads lead to Rome.
This review explores the current evidence on benefits and harms of therapeutic interventions in chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) and makes recommendations. CFS/ME is a complex, multi-system, chronic medical condition whose pathophysiology remains unknown. No established diagnostic tests exist nor are any FDA-approved drugs available for treatment. Because of the range of symptoms of CFS/ME, treatment approaches vary widely. Studies undertaken have heterogeneous designs and are limited by sample size, length of follow-up, applicability and methodological quality. The use of rintatolimod and rituximab as well as counselling, behavioural and rehabilitation therapy programs may be of benefit for CFS/ME, but the evidence of their effectiveness is still limited. Similarly, adaptive pacing appears to offer some benefits, but the results are debatable: so is the use of nutritional supplements, which may be of value to CFS/ME patients with biochemically proven deficiencies. To summarize, the recommended treatment strategies should include proper administration of nutritional supplements in CFS/ME patients with demonstrated deficiencies and personalized pacing programs to relieve symptoms and improve performance of daily activities, but a larger randomized controlled trial (RCT) evaluation is required to confirm these preliminary observations. At present, no firm conclusions can be drawn because the few RCTs undertaken to date have been small-scale, with a high risk of bias, and have used different case definitions. Further, RCTs are now urgently needed with rigorous experimental designs and appropriate data analysis, focusing particularly on the comparison of outcomes measures according to clinical presentation, patient characteristics, case criteria and degree of disability (i.e. severely ill ME cases or bedridden).


Absence of detectable melatonin and preservation of cortisol and thyrotropin rhythms in tetraplegia.
The human circadian timing system regulates the temporal organization of several endocrine functions, including the production of melatonin (via a neural pathway that includes the spinal cord), TSH, and cortisol. In traumatic spinal cord injury, afferent and efferent circuits that influence the basal production of these hormones may be disrupted. We studied five subjects with chronic spinal cord injury (three tetraplegic and two paraplegic, all neurologically complete injuries) under stringent conditions in which the underlying circadian rhythmicity of these hormones could be examined. Melatonin production was absent in the three tetraplegic subjects with injury to their lower cervical spinal cord and was of normal amplitude and timing in the two paraplegic subjects with injury to their upper thoracic spinal cord. The amplitude and the timing of TSH and cortisol rhythms were robust in the paraplegics and in the tetraplegics. Our results indicate that neurologically complete cervical spinal injury results in the complete loss of pineal melatonin production and that neither the loss of melatonin nor the loss of spinal afferent information disrupts the rhythmicity of cortisol or TSH secretion.


Low interbasin connectivity in a facultatively diadromous fish: evidence from genetics and otolith chemistry.
Southern smelts (Retropinna spp.) in coastal rivers of Australia are facultatively diadromous, with populations potentially containing individuals with diadromous or wholly freshwater life histories. The presence of diadromous individuals is expected to reduce genetic structuring between river basins due to larval dispersal via the sea. We use otolith chemistry to distinguish between diadromous and nondiadromous life histories and population genetics to examine interbasin connectivity resulting from diadromy. Otolith strontium isotope ((87) Sr:(86) Sr) transects identified three main life history patterns: amphidromy, freshwater residency and estuarine/marine residency. Despite the potential for interbasin connectivity via larval mixing in the marine environment, we found unprecedented levels of genetic structure for an amphidromous species. Strong hierarchical structure along putative taxonomic boundaries was detected, along with highly structured populations within groups using microsatellites (FST = 0.046-0.181), and mtDNA (ΦST = 0.498-0.816). The presence of strong genetic subdivision, despite the fact that many individuals reside in saline water during their early life history, appears incongruous. However, analysis of multielemental signatures in the otolith cores of diadromous fish revealed strong discrimination between river basins, suggesting that diadromous fish spend their early lives within chemically distinct estuaries rather than the more homogenous marine environment, thus avoiding dispersal and maintaining genetic structure.


Duodenal Microbiota in Stunted Undernourished Children with Enteropathy.
Environmental enteric dysfunction (EED) is an enigmatic disorder of the small intestine that is postulated to play a role in childhood undernutrition, a pressing global health problem. Defining the incidence of this disorder, its pathophysiological features, and its contribution to impaired linear and ponderal growth has been hampered by the difficulty in directly sampling the small intestinal mucosa and microbial community (microbiota). In this study, among 110 young children (mean age, 18 months) with linear growth stunting who were living in an urban slum in Dhaka, Bangladesh, and had not benefited from a nutritional intervention, we performed endoscopy in 80 children who had biopsy-confirmed EED and available plasma and duodenal samples. We quantified the levels of 4077 plasma proteins and 2619 proteins in duodenal biopsy samples obtained from these children. The levels of bacterial strains in microbiota recovered from duodenal aspirate from each child were determined with the use of culture-independent methods. In addition, we obtained 21 plasma samples and 27 fecal samples from age-matched healthy children living in the same area. Young germ-free mice that had been fed a Bangladeshi diet were colonized with bacterial strains cultured from the duodenal aspirates. Of the bacterial strains that were obtained from the children, the absolute levels of a shared group of 14 taxa (which are not typically classified as enteropathogens) were negatively correlated with linear growth (length-for-age z score, r = -0.49; P = 0.003) and positively correlated with duodenal proteins involved in immunoinflammatory responses. The representation of these 14 duodenal taxa in fecal microbiota was significantly different from that in samples obtained from healthy children (P<0.001 by permutational multivariate analysis of variance). Enteropathy of the small intestine developed in gnotobiotic mice that had been colonized with cultured duodenal strains obtained from children with EED. These results provide support for a causal relationship between growth stunting and components of the small intestinal microbiota and enteropathy and offer a rationale for developing therapies that target these microbial contributions to EED. (Funded by the Bill and Melinda Gates Foundation and others; ClinicalTrials.gov number, NCT02812615.).


Impaired skin microvascular function in children, adolescents, and young adults with type 1 diabetes.
Vascular disease in type 1 diabetes is a complex and multifactorial process, which probably begins in childhood in association with the onset of diabetes. To determine the possible factors involved, we measured microvascular responses to endothelium-dependent (acetylcholine) and endothelium-independent (sodium nitroprusside) vasodilators in 56 patients with type 1 diabetes (aged 9-22 years) and 22 control subjects. Skin perfusion was measured at the dorsum of the foot using laser Doppler flowmetry during low-current iontophoresis of acetylcholine and sodium nitroprusside. Maximum vasodilator function was measured during local 44 degrees C skin heating. Vascular responses were significantly reduced in patients with type 1 diabetes compared with responses in control subjects: acetylcholine (P<0.01, analysis of variance [ANOVA]), sodium nitroprusside (P<0.01, ANOVA), and local heating (P<0.02. Mann-Whitney U test). Endothelium-dependent responses were related to duration of diabetes (r = -0.38, P<0.01) and to glycemic control (r = 0.37, P<0.01). Significant correlations were found in the patient group between responses to acetylcholine and sodium nitroprusside (r = 0.28, P<0.05) but not to heating, suggesting that a common factor (e.g., nitric oxide activity) may be responsible for the abnormal vascular responses to these chemicals. Early changes in microvascular function are present in young patients with type 1 diabetes, long before the initial clinical presentation. These abnormalities may be related to complex interactions between structural abnormalities and functional changes in the endothelium, smooth muscle, and nitric oxide activity.


The consequences of malnutrition following discharge from rehabilitation to the community: a systematic review of current evidence in older adults.
The prevalence of malnutrition in the rehabilitation setting is estimated to be 30-50%, with older adults at higher nutritional risk. Malnutrition also exists in the community setting, where 10-30% of adults are malnourished; however, the relationship between the two settings has been little explored. The present study aimed to determine the association between malnutrition in older adults admitted for rehabilitation and nutrition status, functional status, quality of life, institutionalisation, acute care admissions and mortality once discharged to the community. Six electronic databases were searched for relevant publications (1990-2013) using controlled vocabulary. Longitudinal papers were included in which older adults (≥65 years) were admitted for rehabilitation if nutrition assessment was performed during admission with relevant outcomes measured following discharge to the community. Five observational studies were eligible for review which had similar populations. The five reviews comprised 1020 participants in total and, once discharged, follow-up ranged from immediate to 26 months. Malnutrition during rehabilitation was negatively associated with physical function and quality of life, and positively associated with risk of institutionalisation, hospitalisation and mortality. Although these studies were of high quality and strength, the overall contribution to the evidence is limited as a result of the small number of heterogenic studies. No intervention studies were identified. Malnutrition in older adults admitted for rehabilitation has a negative effect on functional recovery and quality of life following discharge to the community. This review highlights an evidence gap along the continuum of care for malnourished older adults, where further observational and intervention research is needed following discharge from rehabilitation to the community.


Proline betaine and its biotransformation products in fasting urine samples are potential biomarkers of habitual citrus fruit consumption.
The lack of robust measures of dietary exposure hinders a quantitative understanding of causal relationships between diet and health. Non-targeted metabolite fingerprinting was used to explore the relationships between citrus exposure in free-living human subjects, estimated by a FFQ, and the chemical content of urine. Volunteers (study 1, n 12; study 2, n 11) were classified into high-, medium- and low-frequency citrus consumption groups. Overnight and spot fasting urine samples were obtained after exposure to a standardised citrus-free evening meal. The urine samples were analysed by flow injection electrospray-ionisation MS followed by supervised multivariate data classification analysis to discover discriminatory features associated with the level of citrus exposure. Good separation of high and low citrus consumption classes was achieved. Deeper exploration of high-ranked explanatory mass signals revealed several correlated signals derived from proline betaine. Targeted analysis of the relative levels of proline betaine in both fasting and overnight urine samples demonstrated good correlation with FFQ exposure data. Acute exposure of volunteers to orange juice resulted in the appearance of proline betaine and several biotransformed products in postprandial urine samples. Biomarker validation showed sensitivities of 80·8-92·2 % and specificities of 74·2-94·1 % (false discovery rate-adjusted P values < 0·05) for elevated proline betaine in participants who reported high citrus consumption. Proline betaine biotransformation products displayed weaker quantitative relationships with habitual citrus exposure. Targeted screening for the presence of biotransformation products of hesperidin and narirutin, known to be abundant in oranges, revealed that they were relatively poor indicators of citrus exposure.


Unconventional medical practices among Ghanaian students: A university-based survey.
Research on unconventional medical practices among students has proliferated lately in the global space, hitherto, little is known explicitly in Ghana. This paper teases out insights for recent utilisation patterns of traditional medical therapies at Kwame Nkrumah University of Science and Technology (KNUST), Ghana. A sample of 754, randomly selected undergraduates were involved in a retrospective cross-sectional survey. Data were analysed using multivariate logistic regression and Pearson's χ2 test with p < 0.05 as significant. Overall prevalence of traditional therapies consumption was 89.1% in the last 12 months. Herbal-based products (67%), prayer healing (15%) and body-mind therapies (11%) were principally used and, accessed through purchases from pharmacy shops (29%) and encounter with faith healers (26%). Although students' knowledge on traditional therapies was acquired through family members (50%) and media (23%), literary materials remained significant information routes for Science related students compared to the Non-science related counterparts (p < 0.001). Pursuing Non-science-related programme [odds ratio (OR) 6.154 (95% confidence interval (CI) 3.745-10.111; p < 0.001)] and having Christian faith [OR 2.450 (95% CI 1.359-4.415; p = 0.003)] were strongly associated with students' traditional therapies use. Although students exhibited positive attitude towards unconventional therapies, there is an urgent need to validate the quality of traditional therapies through randomised clinical trials and regulatory practices to ensure quality control. Health forces should intensify efforts towards intercultural health care system in Ghana.


Classification of support staff in a consortium medical library: a case study.
A representative committee of Houston Academy of Medicine-Texas Medical Center Library staff and faculty, under the direction of the library administration, successfully redesigned a job classification system for the library's nonprofessional staff. In the new system all nonprofessionals are assigned to one of five grade levels, each with a corresponding salary range. To determine its appropriate grade level each job is analyzed and assigned a numerical value using a point system based on a set of five factors, each of which is assigned a relative number of points. The factors used to measure jobs are: education and experience, complexity of work, administrative accountability, manual skill, and contact with users. Each factor is described according to degrees, so that a job can be given partial credit for a factor. An advisory staff classification committee now participates in the ongoing administration of the classification system.


The lifetime risk of total hip and knee arthroplasty: results from the UK general practice research database.
To estimate the lifetime risk of undergoing primary total hip (THR) or knee (TKR) replacement in the UK. A Population-based cohort study of 25,845 patients who had undergone a THR and 23,260 patients who had undergone a TKR between 1991 and 2006, using data from the UK General Practice Research Database. The estimated mortality-adjusted lifetime risk of THR at age 50 for the year 2005 was 11.6% (95% CI: 11.1, 12.1) for women and 7.1% (95% CI: 6.7, 7.5) for men. For TKR the risks were 10.8% (95% CI: 10.3, 11.3) for women and 8.1% (95% CI: 7.6, 8.5) for men. Between 1991 and 2006, the lifetime risk of THR at age 50 rose from 4.0% (95% CI: 3.5, 4.4) to 11.1% (95% CI: 10.6, 11.6) for women and for men from 2.2% (95% CI: 1.8, 2.5) to 6.6% (95% CI: 6.2, 7.0). Over the same period, for TKR the risk for women increased from 2.9% (95% CI: 2.6, 3.3) to 10.6% (95% CI: 10.1, 11.1) and for men from 1.8% (95% CI: 1.5, 2.2) to 7.7% (95% CI: 7.3, 8.2). The lifetime risk of undergoing THR or TKR is estimated to be substantially less than the risk of developing symptomatic hip or knee osteoarthritis. For the knee, the difference between these risk estimates is particularly wide. The reasons for the size of these differences are not clear, and further work is needed to quantify the extent of latent demand for these cost-effective and established interventions among the population with symptomatic osteoarthritis of the hip or knee.


Parkinson's disease in Africa: A systematic review of epidemiologic and genetic studies.
Parkinson's disease (PD) occurs worldwide, but little is known about PD in Africa. We systematically reviewed publications on PD in Africa, with emphasis on epidemiologic and genetic studies. Articles published between 1944 and December 2004 were identified using several strategies. The studies emanated from 13 African countries (Kenya, Uganda, Tanzania, Ethiopia, Nigeria, Senegal, Ghana, Togo, Libya, Tunisia, Algeria, Zimbabwe, and South Africa). The publications fell into four categories: clinical series (n = 17), prevalence studies (n = 7), incidence studies (n = 1), and genetic studies (n = 3). The clinical series documented the occurrence of PD in Africa and described its clinical characteristics. The prevalence studies suggested some intracontinental geographic variation in PD prevalence. Overall, the prevalence figures and the incidence rates of PD in Africa appeared lower than those reported for European and North American populations. Few genetic studies of PD have been reported from Africa, and none in blacks. There are no case-control or cohort studies of PD reported from Africa. This review provides a summary of PD research in Africa over the past 60 years and highlights the information gaps and potential areas for future research.


Long-term depression of synaptic inhibition is expressed postsynaptically in the developing auditory system.
Inhibitory transmission is critically involved in the functional maturation of neural circuits within the brain. However, the mechanisms involved in its plasticity and development remain poorly understood. At an inhibitory synapse of the developing auditory brain stem, we used whole cell recordings to determine the site of induction and expression of long-term depression (LTD), a robust activity-dependent phenomenon that decreases inhibitory synaptic gain and is postulated to underlie synapse elimination. Recordings were obtained from lateral superior olivary (LSO) neurons, and hyperpolarizing inhibitory potentials were evoked by stimulation of the medial nucleus of the trapezoid body (MNTB). Both postsynaptic glycine and GABAA receptors could independently display LTD when isolated pharmacologically. Focal application of GABA, but not glycine, on the postsynaptic LSO neuron was sufficient to induce depression of the amino acid-evoked response, or MNTB-evoked inhibitory postsynaptic potentials. This GABA-mediated depression, in the absence of MNTB stimulation, was blocked by a GABAB receptor antagonist. To assess whether a change in neurotransmitter release is associated with the LTD, the polyvalent cation, ruthenium red, was used to increase the frequency of miniature inhibitory synaptic events. Consistent with a postsynaptic locus of expression, we found that the mean amplitude of miniature events decreased after LTD with no change in their frequency of occurrence. Furthermore, there was no change in the paired-pulse ratio or release kinetics of evoked inhibitory responses. Together, these results provide direct evidence that activity-dependent LTD of inhibition has a postsynaptic locus of induction and alteration, and that GABA but not glycine plays a pivotal role.


Characterization of branched ultrahigh molar mass polymers by asymmetrical flow field-flow fractionation and size exclusion chromatography.
The molar mass distribution (MMD) of synthetic polymers is frequently analyzed by size exclusion chromatography (SEC) coupled to multi angle light scattering (MALS) detection. For ultrahigh molar mass (UHM) or branched polymers this method is not sufficient, because shear degradation and abnormal elution effects falsify the calculated molar mass distribution and information on branching. High temperatures above 130 °C have to be applied for dissolution and separation of semi-crystalline materials like polyolefins which requires special hardware setups. Asymmetrical flow field-flow fractionation (AF4) offers the possibility to overcome some of the main problems of SEC due to the absence of an obstructing porous stationary phase. The SEC-separation mainly depends on the pore size distribution of the used column set. The analyte molecules can enter the pores of the stationary phase in dependence on their hydrodynamic volume. The archived separation is a result of the retention time of the analyte species inside SEC-column which depends on the accessibility of the pores, the residence time inside the pores and the diffusion ability of the analyte molecules. The elution order in SEC is typically from low to high hydrodynamic volume. On the contrary AF4 separates according to the diffusion coefficient of the analyte molecules as long as the chosen conditions support the normal FFF-separation mechanism. The separation takes place in an empty channel and is caused by a cross-flow field perpendicular to the solvent flow. The analyte molecules will arrange in different channel heights depending on the diffusion coefficients. The parabolic-shaped flow profile inside the channel leads to different elution velocities. The species with low hydrodynamic volume will elute first while the species with high hydrodynamic volume elute later. The AF4 can be performed at ambient or high temperature (AT-/HT-AF4). We have analyzed one low molar mass polyethylene sample and a number of narrow distributed polystyrene standards as reference materials with known structure by AT/HT-SEC and AT/HT-AF4. Low density polyethylenes as well as polypropylene and polybutadiene, containing high degrees of branching and high molar masses, have been analyzed with both methods. As in SEC the relationship between the radius of gyration (R(g)) or the molar mass and the elution volume is curved up towards high elution volumes, a correct calculation of the MMD and the molar mass average or branching ratio is not possible using the data from the SEC measurements. In contrast to SEC, AF4 allows the precise determination of the MMD, the molar mass averages as well as the degree of branching because the molar mass vs. elution volume curve and the conformation plot is not falsified in this technique. In addition, higher molar masses can be detected using HT-AF4 due to the absence of significant shear degradation in the channel. As a result the average molar masses obtained from AF4 are higher compared to SEC. The analysis time in AF4 is comparable to that of SEC but the adjustable cross-flow program allows the user to influence the separation efficiency which is not possible in SEC without a costly change of the whole column combination.


CD11b⁺, Ly6G⁺ cells produce type I interferon and exhibit tissue protective properties following peripheral virus infection.
The goal of the innate immune system is containment of a pathogen at the site of infection prior to the initiation of an effective adaptive immune response. However, effector mechanisms must be kept in check to combat the pathogen while simultaneously limiting undesirable destruction of tissue resulting from these actions. Here we demonstrate that innate immune effector cells contain a peripheral poxvirus infection, preventing systemic spread of the virus. These innate immune effector cells are comprised primarily of CD11b⁺Ly6C⁺Ly6G⁻ monocytes that accumulate initially at the site of infection, and are then supplemented and eventually replaced by CD11b⁺Ly6C⁺Ly6G⁺ cells. The phenotype of the CD11b⁺Ly6C⁺Ly6G⁺ cells resembles neutrophils, but the infiltration of neutrophils typically occurs prior to, rather than following, accumulation of monocytes. Indeed, it appears that the CD11b⁺Ly6C⁺Ly6G⁺ cells that infiltrated the site of VACV infection in the ear are phenotypically distinct from the classical description of both neutrophils and monocyte/macrophages. We found that CD11b⁺Ly6C⁺Ly6G⁺ cells produce Type I interferons and large quantities of reactive oxygen species. We also observed that depletion of Ly6G⁺ cells results in a dramatic increase in tissue damage at the site of infection. Tissue damage is also increased in the absence of reactive oxygen species, although reactive oxygen species are typically thought to be damaging to tissue rather than protective. These data indicate the existence of a specialized population of CD11b⁺Ly6C⁺Ly6G⁺ cells that infiltrates a site of virus infection late and protects the infected tissue from immune-mediated damage via production of reactive oxygen species. Regulation of the action of this population of cells may provide an intervention to prevent innate immune-mediated tissue destruction.


Evaluation of a Web-Based Training Program for Professional Carers Working With People With Learning Disabilities and Challenging Behavior: A Pilot Study with SSED-Design.
The interaction between people with intellectual disabilities and professional carers is often influenced by communicative difficulties contributing challenging behaviours. The aims of this study were to evaluate to a web-based training program aimed at improving carers' abilities to interact with people with learning disabilities who exhibit challenging behaviours and to explore carers' experiences of participating in such a program. A single-subject experimental design and mixed methods were used to integrate qualitative and quantitative data. Triangulation of questionnaires, interviews with carers, and assessments of one woman's behaviour was performed. The participants were professional carers aged 20 to 55 years. The web-based training program increased carers' abilities to handle challenging behaviours and decreased challenging behaviours in daily care. The program improved the opportunities to offer training to carers who work in community-based accommodations with limited time to receive training.


Experimental Determination of the Molar Absorption Coefficient of Cesium Lead Halide Perovskite Quantum Dots.
Lead halide perovskite (CsPbX3, where X = Cl, Br, or I) quantum dots (QDs), with tunable optical and electronic properties, have attracted attention because of their promising applications in solar cells and next-generation optoelectronic devices. Hence, it is crucial to investigate in detail the fundamental size-dependent properties of these perovskite QDs to obtain high-quality nanocrystals for practical use. We propose a direct method for determining the concentration of solution-processed CsPbX3 QDs by means of spectrophotometry, in which the molar absorption coefficient (ε) is obtained using absorption and the Beer-Lambert law. By tuning the size of CsPbX3 QDs, we obtain their corresponding ε leading to a calibration curve for calculating the nanocrystal concentrations. The ε at the band edge for CsPbX3 (X = Cl, Br, or I) nanocrystals was found to be strongly dependent on the bandgap of the nanocrystals. We also obtained a reliable size dependence of the bandgap calibration curves to estimate the size of QDs from the absorption spectra.


Genetic features of Mongolian ethnic groups revealed by Y-chromosomal analysis.
About 20 ethnic groups reside in Mongolia. On the basis of genetic and anthropological studies, it is believed that Mongolians have played a pivotal role in the peopling of Central and East Asia. However, the genetic relationships among these ethnic groups have remained obscure, as have their detailed relationships with adjacent populations. We analyzed 16 binary and 17 STR polymorphisms of human Y chromosome in 669 individuals from nine populations, including four indigenous ethnic groups in Mongolia (Khalkh, Uriankhai, Zakhchin, and Khoton). Among these four Mongolian populations, the Khalkh, Uriankhai, and Zakhchin populations showed relatively close genetic affinities to each other and to Siberian populations, while the Khoton population showed a closer relationship to Central Asian populations than to even the other Mongolian populations. These findings suggest that the major Mongolian ethnic groups have a close genetic affinity to populations in northern East Asia, although the genetic link between Mongolia and Central Asia is not negligible.


Psychotherapy for Depression in Older Veterans Via Telemedicine: Effect on Quality of Life, Satisfaction, Treatment Credibility, and Service Delivery Perception.
To analyze the impact of telepsychology and same-room care on functioning, satisfaction, and perception of care based on a noninferiority trial of psychotherapy delivered via telemedicine or same-room care to elderly patients with depression. 241 elderly patients with depression (meeting DSM-IV diagnostic criteria) were randomly assigned to either telemedicine (n = 120) or same-room treatment (n = 121) between April 1, 2007, and July 31, 2011. The primary outcomes included quality of life (36-item Short Form Survey [SF-36]), satisfaction (Charleston Psychiatric Outpatient Satisfaction Scale), treatment credibility, and service delivery perception scores obtained at 4 weeks, 8 weeks, 3 months, and 12 months. Comparisons of intervention means were carried out at each time point using independent sample t tests and SAS Procedure MIANALYZE to combine results across the multiply imputed complete data sets. If significant differences were detected for a given outcome within a domain, a Bonferroni correction was applied to determine if significance was maintained. None of the SF-36 scores showed a significant difference between the 2 treatment groups by the end of the study period, with little significance shown throughout the intermediate time points. Similarly, over all time points, there was no statistically significant difference in patient satisfaction or treatment credibility. This study found that telemedicine is a viable alternative modality for providing evidence-based psychotherapy for elderly patients with depression. Results provide evidence that quality of life and satisfaction with care are not adversely influenced by the decision to use a telehealth modality instead of in-person treatment, and, as a result, resources can be devoted to offering services in patients' homes through telemedicine. ClinicalTrials.gov identifier: NCT00324701.