Cell Behaviors during Closure of the Choroid Fissure in the Developing Eye.
Coloboma is a defect in the morphogenesis of the eye that is a consequence of failure of choroid fissure fusion. It is among the most common congenital defects in humans and can significantly impact vision. However, very little is known about the cellular mechanisms that regulate choroid fissure closure. Using high-resolution confocal imaging of the zebrafish optic cup, we find that apico-basal polarity is re-modeled in cells lining the fissure in proximal to distal and inner to outer gradients during fusion. This process is accompanied by cell proliferation, displacement of vasculature, and contact between cells lining the choroid fissure and periocular mesenchyme (POM). To investigate the role of POM cells in closure of the fissure, we transplanted optic vesicles onto the yolk, allowing them to develop in a situation where they are depleted of POM. The choroid fissure forms normally in ectopic eyes but fusion fails in this condition, despite timely apposition of the nasal and temporal lips of the retina. This study resolves some of the cell behaviors underlying choroid fissure fusion and supports a role for POM in choroid fissure fusion.


Quantitative trait variation in ASD probands and toddler sibling outcomes at 24 months.
Younger siblings of children with autism spectrum disorder (ASD) are at increased likelihood of receiving an ASD diagnosis and exhibiting other developmental concerns. It is unknown how quantitative variation in ASD traits and broader developmental domains in older siblings with ASD (probands) may inform outcomes in their younger siblings. Participants included 385 pairs of toddler siblings and probands from the Infant Brain Imaging Study. ASD probands (mean age 5.5 years, range 1.7 to 15.5 years) were phenotyped using the Autism Diagnostic Interview-Revised (ADI-R), the Social Communication Questionnaire (SCQ), and the Vineland Adaptive Behavior Scales, Second Edition (VABS-II). Siblings were assessed using the ADI-R, VABS-II, Mullen Scales of Early Learning (MSEL), and Autism Diagnostic Observation Schedule (ADOS) and received a clinical best estimate diagnosis at 24 months using DSM-IV-TR criteria (n = 89 concordant for ASD; n = 296 discordant). We addressed two aims: (1) to determine whether proband characteristics are predictive of recurrence in siblings and (2) to assess associations between proband traits and sibling dimensional outcomes at 24 months. Regarding recurrence risk, proband SCQ scores were found to significantly predict sibling 24-month diagnostic outcome (OR for a 1-point increase in SCQ = 1.06; 95% CI = 1.01, 1.12). Regarding quantitative trait associations, we found no significant correlations in ASD traits among proband-sibling pairs. However, quantitative variation in proband adaptive behavior, communication, and expressive and receptive language was significantly associated with sibling outcomes in the same domains; proband scores explained 9-18% of the variation in cognition and behavior in siblings with ASD. Receptive language was particularly strongly associated in concordant pairs (ICC = 0.50, p < 0.001). Proband ASD symptomology, indexed by the SCQ, is a predictor of familial ASD recurrence risk. While quantitative variation in social communication and restricted and repetitive behavior were not associated among sibling pairs, standardized ratings of proband language and communication explained significant variation in the same domains in the sibling at 24 months, especially among toddlers with an ASD diagnosis. These data suggest that proband characteristics can alert clinicians to areas of developmental concern for young children with familial risk for ASD.


Qualitative evaluation of a diabetes electronic decision support tool: views of users.
Quality care of type 2 diabetes is complex and requires systematic use of clinical data to monitor care processes and outcomes. An electronic decision support (EDS) tool for the management of type 2 diabetes in primary care was developed by the Australian Pharmaceutical Alliance. The aim of this qualitative study was to evaluate the uptake and use of the EDS tool as well as to describe the impact of the EDS tool on the primary care consultation for diabetes from the perspectives of general practitioners and practice nurses. This was a qualitative study of telephone interviews. General Practitioners and Practice Nurses from four Divisions of General Practice who had used the EDS tool for a minimum of six weeks were invited to participate. Semi-structured interviews were conducted and the interview transcripts were coded and thematically analysed using NVivo 8 software. In total 15 General Practitioners and 2 Practice Nurses completed the interviews. The most commonly used feature of the EDS tool was the summary side bar; its major function was to provide an overview of clinical information and a prompt or reminder to diabetes care. It also assisted communication and served an educational role as a visual aide in the consultation. Some participants thought the tool resulted in longer consultations. There were a range of barriers to use related to the design and functionality of the tool and to the primary care context. The EDS tool shows promise as a way of summarising information about patients' diabetes state, reminder of required diabetes care and an aide to patient education.


An outbreak of waterborne giardiasis associated with heavy water runoff due to warm weather and volcanic ashfall.
From mid-June through early August 1980, an outbreak of gastrointestinal illness in Red Lodge, Montana affected approximately 780 persons, as estimated from attack rates of 33 per cent and 15 per cent in urban and rural residents, respectively. Giardia lamblia was identified in stool specimens from 51 per cent of 47 persons with a history of untreated gastrointestinal illness and in 13 per cent of 24 specimens from asymptomatic persons (p = .00045, Fisher's Exact Test). The epidemic curve was bimodal with peaks in mid-June and mid-July. Each peak occurred about three weeks after an episode of very heavy water runoff resulting from warm sunny weather and snow darkened by ashfall from the Mt. St. Helens volcanic eruption of May 18, 1980. Unfiltered and inadequately chlorinated surface water was supplied by the city water system, which was implicated as the vehicle of transmission in the outbreak. Water systems providing unfiltered surface water are more likely to become contaminated during periods of heavy water runoff.


Detecting recombination in evolving nucleotide sequences.
Genetic recombination can produce heterogeneous phylogenetic histories within a set of homologous genes. These recombination events can be obscured by subsequent residue substitutions, which consequently complicate their detection. While there are many algorithms for the identification of recombination events, little is known about the effects of subsequent substitutions on the accuracy of available recombination-detection approaches. We assessed the effect of subsequent substitutions on the detection of simulated recombination events within sets of four nucleotide sequences under a homogeneous evolutionary model. The amount of subsequent substitutions per site, prior evolutionary history of the sequences, and reciprocality or non-reciprocality of the recombination event all affected the accuracy of the recombination-detecting programs examined. Bayesian phylogenetic-based approaches showed high accuracy in detecting evidence of recombination event and in identifying recombination breakpoints. These approaches were less sensitive to parameter settings than other methods we tested, making them easier to apply to various data sets in a consistent manner. Post-recombination substitutions tend to diminish the predictive accuracy of recombination-detecting programs. The best method for detecting recombined regions is not necessarily the most accurate in identifying recombination breakpoints. For difficult detection problems involving highly divergent sequences or large data sets, different types of approach can be run in succession to increase efficiency, and can potentially yield better predictive accuracy than any single method used in isolation.


Guanidinoacetic acid is efficacious in improving growth performance and muscle energy homeostasis in broiler chicks fed arginine-deficient or arginine-adequate diets.
Two studies were conducted to test the efficacy of guanidinoacetic acid (GAA) to spare Arg and serve as a precursor of creatine (Cr) by evaluating growth performance and muscle cellular energy homeostasis in broiler chicks. In both studies, 12 replicate pens of 6 chicks received dietary treatments beginning at day 2 post-hatch. At conclusion of each study, muscle biopsy samples were collected within 60 s of euthanasia for analysis of Cr-related energy metabolites. In study 1, Arg-deficient starter and grower basal diets were supplemented with 0 (negative control, NC), 0.06, 0.12, or 0.18% GAA, or supplemental Arg (positive control, PC; 0.37 and 0.32% L-Arg in starter and grower phases, respectively). Dietary GAA elicited graded improvements, with final BW, overall BW gain, and overall G:F being increased (P < 0.05) by 0.12% GAA compared with the NC diet with no difference to PC diet. Increases (P < 0.001) of phosphocreatine (PCr), total Cr (tCr), and glycogen concentrations, as well as the PCr-to-adenosine triphosphate (ATP) and glycogen:ATP ratios, were observed with supplementation of 0.12% GAA compared with the NC diet, even exceeding responses to the PC diet. In study 2, Arg-adequate starter and grower basal diets were supplemented with 0 (negative control, NC), 0.06, or 0.12% GAA, 0.12% Cr monohydrate (PC1), or salmon protein (PC2; containing total Arg concentrations equal to those of the NC diet in each phase and containing similar Cr as in PC1). Overall G:F was increased (P < 0.05) by PC1, but not by PC2, compared with the NC, while GAA supplementation elicited a response intermediate to NC and PC1 diets. However, GAA supplementation increased (P < 0.01) concentrations of tCr and glycogen, as well as the PCr:ATP and glycogen:ATP ratios, when compared with the NC (Arg-adequate) diet. Collectively, these data indicate that GAA can be used to replace Arg in practical, Arg-deficient diets and improve muscle energy homeostasis in broiler chicks receiving either Arg-deficient or Arg-adequate practical diets.


Vulnerability of agriculture to climate change increases the risk of child malnutrition: Evidence from a large-scale observational study in India.
The impact of climate change on agriculture and food security has been examined quite thoroughly by researchers globally as well as in India. While existing studies provide evidence on how climate variability affects the food security and nutrition, research examining the extent of effect vulnerability of agriculture to climate change can have on nutrition in India are scarce. This study examined a) the association between the degree of vulnerability in agriculture to climate change and child nutrition at the micro-level b) spatial effect of climate vulnerability on child nutrition, and c) the geographical hotspots of both vulnerability in agriculture to climate change and child malnutrition. The study used an index on vulnerability of agriculture to climate change and linked it to child malnutrition indicators (stunting, wasting, underweight and anaemia) from the National Family Health Survey 4 (2015-16). Mixed-effect and spatial autoregressive models were fitted to assess the direction and strength of the relationship between vulnerability and child malnutrition at macro and micro level. Spatial analyses examined the within-district and across-district spill-over effects of climate change vulnerability on child malnutrition. Both mixed-effect and spatial autoregressive models found that the degree of vulnerability was positively associated with malnutrition among children. Children residing in districts with a very high degree of vulnerability were more like to have malnutrition than those residing in districts with very low vulnerability. The analyses found that the odds of a child suffering from stunting increased by 32%, wasting by 42%, underweight by 45%, and anaemia by 63% if the child belonged to a district categorised as very highly vulnerable when compared to those categorised as very low. The spatial analysis also suggested a high level of clustering in the spatial distribution of vulnerability and malnutrition. Hotspots of child malnutrition and degree of vulnerability were mostly found to be clustered around western-central part of India. Study highlights the consequences that vulnerability of agriculture to climate change can have on child nutrition. Strategies should be developed to mitigate the effect of climate change on areas where there is a clustering of vulnerability and child malnutrition.


Health-related quality of life among children with mental health problems: a population-based approach.
Children with mental health problems have been neglected in health-related quality of life (HRQOL) studies. Therefore, the aims of the current study were 1) to assess the influence of the presence of mental or physical health problems on HRQOL; and 2) to analyze the effects of item overlap between mental health problems and HRQOL-measurements. Proxy- and self-rated HRQOL (KIDSCREEN-27) of children 9-14 years old was assessed across children with mental health problems (n = 535), children with physical health problems (n = 327), and healthy controls (n = 744). Multiple linear regression analyses were conducted with health status, severity of symptoms, status of medication use, gender and nationality as independent, and HRQOL scores as dependent variables. The effects of item overlap were analyzed by repeating regression analyses while excluding those HRQOL items that contextually overlapped the most frequently-occurring mental health problem (attention deficits). Severity of symptoms was the strongest predictor of reduced HRQOL. However, all other predictors (except for the status of medication use) also contributed to the prediction of some HRQOL scores. Controlling for item overlap did not meaningfully alter the results. When children with different health constraints are compared, the severity of their particular health problems should be considered. Furthermore, item overlap seems not to be a major problem when the HRQOL of children with mental health problems is studied. Hence, HRQOL assessments are useful to gather information that goes beyond the clinical symptoms of a health problem. This information can, for instance, be used to improve clinical practice.


Naturally-occurring modification restricts the anticodon domain conformational space of tRNA(Phe).
Post-transcriptional modifications contribute chemistry and structure to RNAs. Modifications of tRNA at nucleoside 37, 3'-adjacent to the anticodon, are particularly interesting because they facilitate codon recognition and negate translational frame-shifting. To assess if the functional contribution of a position 37-modified nucleoside defines a specific structure or restricts conformational flexibility, structures of the yeast tRNA(Phe) anticodon stem and loop (ASL(Phe)) with naturally occurring modified nucleosides differing only at position 37, ASL(Phe)-(Cm(32),Gm(34),m(5)C(40)), and ASL(Phe)-(Cm(32),Gm(34),m(1)G(37),m(5)C(40)), were determined by NMR spectroscopy and restrained molecular dynamics. The ASL structures had similarly resolved stems (RMSD approximately 0.6A) of five canonical base-pairs in standard A-form RNA. The "NOE walk" was evident on the 5' and 3' sides of the stems of both RNAs, and extended to the adjacent loop nucleosides. The NOESY cross-peaks involving U(33) H2' and characteristic of tRNA's anticodon domain U-turn were present but weak, whereas those involving the U(33) H1' proton were absent from the spectra of both ASLs. However, ASL(Phe)-(Cm(32),Gm(34),m(1)G(37),m(5)C(40)) exhibited the downfield shifted 31P resonance of U(33)pGm(34) indicative of U-turns; ASL(Phe)-(Cm(32),Gm(34),m(5)C(40)) did not. An unusual "backwards" NOE between Gm(34) and A(35) (Gm(34)/H8 to A(35)/H1') was observed in both molecules. The RNAs exhibited a protonated A(+)(38) resulting in the final structures having C(32).A(+)(38) intra-loop base-pairs, with that of ASL(Phe)-(Cm(32),Gm(34),m(1)G(37),m(5)C(40)) being especially well defined. A single family of low-energy structures of ASL(Phe)-(Cm(32),Gm(34), m(1)G(37),m(5)C(40)) (loop RMSD 0.98A) exhibited a significantly restricted conformational space for the anticodon loop in comparison to that of ASL(Phe)-(Cm(32),Gm(34),m(5)C(40)) (loop RMSD 2.58A). In addition, the ASL(Phe)-(Cm(32),Gm(34),m(1)G(37),m(5)C(40)) average structure had a greater degree of similarity to that of the yeast tRNA(Phe) crystal structure. A comparison of the resulting structures indicates that modification of position 37 affects the accuracy of decoding and the maintenance of the mRNA reading frame by restricting anticodon loop conformational space.


Childhood cancer registries in Ontario, Canada: lessons learned from a comparison of two registries.
Two childhood cancer registries exist in Ontario. One (POGO) accrues by active registration by pediatric cancer centers, utilizing a histologically based classification system. The other accrues by passive linkage within a larger adult oriented cancer registry (OCR) using a topographically based classification. A reportedly high incidence of childhood cancer based on the latter registry prompted a comparison of the content of the two registries over a 2-year period with the hypothesis that there would be systematic accrual errors. All registrations in both registries for the specified period were reviewed systematically and validated by pathology reports. A small number (2.6%) of registrations in the passive registry were not incident cases, while 2 particular pathologic diagnoses were included in the histologically based registry and not the topographically based registry. These were low grade gliomas and Langerhans cell histiocytosis (LCH). The validated annual incident rate (15.6 per 100000 children 0-14 years of age, excluding LCH) is slightly higher than that reported in other industrialized countries. Ninety-six percent of children aged 0-14 were treated in pediatric oncology centers, while only 46% of adolescents aged 15-17 were treated in such centers. Of the remaining adolescents, more than one-third had lymphomas. This maldistribution of care provided for the young adolescent population may compromise their survival prospects. The results of this study should prompt revision of health care policy and patterns of service delivery.


Dynamic regulation of nuclear architecture and mechanics-a rheostatic role for the nucleus in tailoring cellular mechanosensitivity.
Nuclear architecture, a function of both chromatin and nucleoskeleton structure, is known to change with stem cell differentiation and differs between various somatic cell types. These changes in nuclear architecture are associated with the regulation of gene expression and genome function in a cell-type specific manner. Biophysical stimuli are known effectors of differentiation and also elicit stimuli-specific changes in nuclear architecture. This occurs via the process of mechanotransduction whereby extracellular mechanical forces activate several well characterized signaling cascades of cytoplasmic origin, and potentially some recently elucidated signaling cascades originating in the nucleus. Recent work has demonstrated changes in nuclear mechanics both with pluripotency state in embryonic stem cells, and with differentiation progression in adult mesenchymal stem cells. This review explores the interplay between cytoplasmic and nuclear mechanosensitivity, highlighting a role for the nucleus as a rheostat in tuning the cellular mechano-response.


Development of a 3D pharmacophore for nonspecific ligand recognition of alpha1, alpha2, alpha3, alpha5, and alpha6 containing GABA(A)/benzodiazepine receptors.
Transfected cells containing GABA(A) benzodiazepine receptors (BDZRs) have been utilized to systematically determine the affinity of ligands at alpha1, alpha2, alpha3, alpha5 and alpha6 subtypes in combination with beta2 and gamma2. All but a few of the ligands thus far studied have relatively high affinities for each of these alpha subtype receptors. Thus, these ligands must contain common stereochemical properties favorable for recognition by each of the subtype combinations. In the present work, such a common three-dimensional (3D) pharmacophore for recognition of alpha1, alpha2, alpha3, alpha5 and alpha6 containing GABA(A)/BDZRs types of receptors has been developed and assessed, using as a database receptor affinities measured in transfected cells for 27 diverse compounds. The 3D-recognition pharmacophore developed consists of three proton accepting groups, a hydrophobic group, and the centroid of an aromatic ring found in a common geometric arrangement in the 19 nonselective ligands used. Three tests were made to assess this pharmacophore: (i) Four low affinity compounds were used as negative controls, (ii) Four high affinity compounds, excluded from the pharmacophore development, were used as compounds for pharmacophore validation, (iii) The 3D pharmacophore was used to search 3D databases. The results of each of these types of assessments provided robust validation of the 3D pharmacophore. This 3D pharmacophore can now be used to discover novel nonselective ligands that could be activation selective at different behavioral end points. Additionally, it may serve as a guide in the design of more selective ligands, by determining if candidate ligands proposed for synthesis conform to this pharmacophore and selecting those that do not for further experimental assessment.


Formaldehyde as a basis for residential ventilation rates.
Traditionally, houses in the US have been ventilated by passive infiltration in combination with active window opening. However in recent years, the construction quality of residential building envelopes has been improved to reduce infiltration, and the use of windows for ventilation also may have decreased due to a number of factors. Thus, there has been increased interest in engineered ventilation systems for residences. The amount of ventilation provided by an engineered system should be set to protect occupants from unhealthy or objectionable exposures to indoor pollutants, while minimizing energy costs for conditioning incoming air. Determining the correct ventilation rate is a complex task, as there are numerous pollutants of potential concern, each having poorly characterized emission rates, and poorly defined acceptable levels of exposure. One ubiquitous pollutant in residences is formaldehyde. The sources of formaldehyde in new houses are reasonably understood, and there is a large body of literature on human health effects. This report examines the use of formaldehyde as a means of determining ventilation rates and uses existing data on emission rates of formaldehyde in new houses to derive recommended levels. Based on current, widely accepted concentration guidelines for formaldehyde, the minimum and guideline ventilation rates for most new houses are 0.28 and 0.5 air changes per hour, respectively.


Opposite effects of 3,4-methylenedioxymethamphetamine (MDMA) on sensorimotor gating in rats versus healthy humans.
Prepulse inhibition of acoustic startle refers to the reduction in the startle response when the startling stimulus is preceded by a weak prepulse stimulus. This phenomenon provides an operational measure of sensorimotor gating that has been found to be reduced in patients with schizophrenia and rats treated with serotonin agonists or serotonin releasers. In this study, we compared the effects of a serotonin releaser, MDMA, on prepulse inhibition in laboratory rats and healthy human volunteers. In particular, we investigated whether MDMA disrupts PPI in humans as observed in animal studies. Rats were tested after placebo and MDMA in a counterbalanced order at an interval of 1 week, with separate groups of rats being used for each dose of MDMA (1.7, 5.4 and 17.0 mg/kg). On each test day, rats were first tested after no injections and retested 2 h later, 10 min after a subcutaneous injection of placebo or MDMA. For the human study, a placebo-controlled within-subject design and double-blind procedures were used. Subjects were examined twice at a 2 to 4 week interval after either placebo or drug administration (order being counterbalanced). On each test day, subjects underwent baseline testing including psychological and PPI measures. Ninety minutes later, subjects received placebo or MDMA (1.7 mg/kg PO) and were retested after 75 min during the peak of behavioral effects of MDMA. As expected, MDMA decreased prepulse inhibition in a dose-related fashion in rats. In contrast, a typical recreational dose of MDMA (1.7 mg/kg, orally) increased prepulse inhibition in subjects experiencing robust psychological effects. This surprising disparity between the effects of the drug in rats and humans may reflect a species-specific difference in the mechanism of action of MDMA or in the behavioral expression of a similar pharmacological effect, or both.


Management of gastroesophageal reflux disease.
The primary treatment goals in patients with gastroesophageal reflux disease are relief of symptoms, prevention of symptom relapse, healing of erosive esophagitis, and prevention of complications of esophagitis. In patients with reflux esophagitis, treatment is directed at acid suppression through the use of lifestyle modifications (e.g., elevating the head of the bed, modifying the size and composition of meals) and pharmacologic agents (a histamine H2-receptor antagonist [H2RA] taken on demand or a proton pump inhibitor IPPI] taken 30 to 60 minutes before the first meal of the day). The preferred empiric approach is step-up therapy (treat initially with an H2RA for eight weeks; if symptoms do not improve, change to a PPI) or step-down therapy (treat initially with a PPI; then titrate to the lowest effective medication type and dosage). In patients with erosive esophagitis identified on endoscopy, a PPI is the initial treatment of choice. Diagnostic testing should be reserved for patients who exhibit warning signs (i.e., weight loss, dysphagia, gastrointestinal bleeding) and patients who are at risk for complications of esophagitis (i.e., esophageal stricture formation, Barrett's esophagus, adenocarcinoma). Antireflux surgery, including open and laparoscopic versions of Nissen fundoplication, is an alternative treatment in patients who have chronic reflux with recalcitrant symptoms. Newer endoscopic modalities, including the Stretta and endocinch procedures, are less invasive and have fewer complications than antireflux surgery, but response rates are lower.


Discourses of depression of Australian general practitioners working with gay men.
The data for this article are from a primary health care project on HIV and depression, in which the prevalence, nature, clinical management, and self-management of depression among homosexually active men attending high-HIV-caseload general practice clinics were investigated. One of the qualitative arms consisted of in-depth interviews with general practitioners (GPs) with high caseloads of gay men. The approach to discourse analysis was informed by Halliday's systemic functional linguistics. GPs constructed three discourses of depression: engaging with psychiatric discourse, engaging with the patient's world, and engaging with social structures. When GPs drew on the discourse of psychiatry, this discourse was positioned as only one possible construction of depression. This discourse was also contextualized in the social lives of gay men, and it was explicitly challenged and rejected. Engaging with their patients' social world was considered vital for recognizing depression in gay men. Finally, the GPs' construction of depression was inextricably linked to social disadvantage and marginalization. Depression is highly heterogeneous and constructed in terms of social relationships rather than as an independent entity that resides in the individual. There is a synergy between GPs' constructions of depression and men's experiences of depression, which differs from conventional medical views, and which enables GPs to be highly effective in dealing with the mental health issues of their gay patients.


Completely Automated CNN Architecture Design Based on Blocks.
The performance of convolutional neural networks (CNNs) highly relies on their architectures. In order to design a CNN with promising performance, extensive expertise in both CNNs and the investigated problem domain is required, which is not necessarily available to every interested user. To address this problem, we propose to automatically evolve CNN architectures by using a genetic algorithm (GA) based on ResNet and DenseNet blocks. The proposed algorithm is completely automatic in designing CNN architectures. In particular, neither preprocessing before it starts nor postprocessing in terms of CNNs is needed. Furthermore, the proposed algorithm does not require users with domain knowledge on CNNs, the investigated problem, or even GAs. The proposed algorithm is evaluated on the CIFAR10 and CIFAR100 benchmark data sets against 18 state-of-the-art peer competitors. Experimental results show that the proposed algorithm outperforms the state-of-the-art CNNs hand-crafted and the CNNs designed by automatic peer competitors in terms of the classification performance and achieves a competitive classification accuracy against semiautomatic peer competitors. In addition, the proposed algorithm consumes much less computational resource than most peer competitors in finding the best CNN architectures.


The sea lamprey germline genome provides insights into programmed genome rearrangement and vertebrate evolution.
The sea lamprey (Petromyzon marinus) serves as a comparative model for reconstructing vertebrate evolution. To enable more informed analyses, we developed a new assembly of the lamprey germline genome that integrates several complementary data sets. Analysis of this highly contiguous (chromosome-scale) assembly shows that both chromosomal and whole-genome duplications have played significant roles in the evolution of ancestral vertebrate and lamprey genomes, including chromosomes that carry the six lamprey HOX clusters. The assembly also contains several hundred genes that are reproducibly eliminated from somatic cells during early development in lamprey. Comparative analyses show that gnathostome (mouse) homologs of these genes are frequently marked by polycomb repressive complexes (PRCs) in embryonic stem cells, suggesting overlaps in the regulatory logic of somatic DNA elimination and bivalent states that are regulated by early embryonic PRCs. This new assembly will enhance diverse studies that are informed by lampreys' unique biology and evolutionary/comparative perspective.


Comparative effect of coffee robusta and coffee arabica (Qahwa) on memory and attention.
The comparative effects of coffee robusta and coffee arabica (Qahwa) on different attention and memory related assignments were measured in a double-blind study of 300 healthy young adult women who were randomly assigned to one of three different drinks: Group I (coffee robusta sachet dissolved in 100 ml of hot water): Group II (coffee arabica): and group III (100 ml water only). Cognitive function was assessed by standardized tests. Several monitoring cognitive tests and tasks were specifically chosen and performed to investigate the comparative effects of coffee robusta (CR) and coffee arabica (Qahwa; AC) on sleepiness (sleep and clear headed scale), attention (trail A & B, symbol digit, letter cancellation), general cognitive ability (stroop test) and memory (card test). Data was interpreted by analysis of variance (ANOVA). The present study revealed that coffee robusta has beneficial effects on attention, general cognitive ability and memory. Higher though non-significant cognitive scores were associated with coffee robusta consumption. Although, consumption of coffee arabica (Qahwa) has significant effects (P < 0.05) on sleepiness, attention, general cognitive ability and memory and it significantly improve reaction time and correct responses. Since different tasks were related to the sustained attention and working memory processes, results would suggest that coffee arabica (qahwa) could increase the memory and efficiency of the attentional system might be due to the presence of chlorogenic acids (CGA) which are found in less quantity in coffee robusta. However, more studies using larger samples and different tasks are necessary to better understand the effects of coffee robusta and arabica (Qahwa) on attention and memory.


Brief periods of auditory perceptual training can determine the sensory targets of speech motor learning.
The perception of speech is notably malleable in adults, yet alterations in perception seem to have little impact on speech production. However, we hypothesized that speech perceptual training might immediately influence speech motor learning. To test this, we paired a speech perceptual-training task with a speech motor-learning task. Subjects performed a series of perceptual tests designed to measure and then manipulate the perceptual distinction between the words head and had. Subjects then produced head with the sound of the vowel altered in real time so that they heard themselves through headphones producing a word that sounded more like had. In support of our hypothesis, the amount of motor learning in response to the voice alterations depended on the perceptual boundary acquired through perceptual training. The studies show that plasticity in adults' speech perception can have immediate consequences for speech production in the context of speech learning.


Phospholipase C is required for the control of stomatal aperture by ABA.
The calcium-releasing second messenger inositol 1,4,5-trisphosphate is involved in the regulation of stomatal aperture by ABA. In other signalling pathways, inositol 1,4,5-trisphosphate is generated by the action of phospholipase C. We have studied the importance of phospholipase C in guard cell ABA-signalling pathways. Immunolocalisation of a calcium-activated phospholipase C confirmed the presence of phospholipase C in tobacco guard cells. Transgenic tobacco plants with considerably reduced levels of phospholipase C in their guard cells were only partially able to regulate their stomatal apertures in response to ABA. These results suggest that phospholipase C is involved in the amplification of the calcium signal responsible for reductions in stomatal aperture in response to ABA. As full ABA-induced inhibition of stomatal opening was not observed, our results support a role for the action of other calcium-releasing second messengers in the guard cell ABA-signalling pathway. It is not known whether these different calcium-releasing second messengers act in the same or parallel ABA-signalling pathways.


The rewards of nicotine: regulation by tissue plasminogen activator-plasmin system through protease activated receptor-1.
Nicotine, a primary component of tobacco, is one of the most abused drugs worldwide. Approximately four million people die each year because of diseases associated with tobacco smoking. Mesolimbic dopaminergic neurons mediate the rewarding effects of abused drugs, including nicotine. Here we show that the tissue plasminogen activator (tPA)-plasmin system regulates nicotine-induced reward and dopamine release by activating protease activated receptor-1 (PAR1). In vivo microdialysis revealed that microinjection of either tPA or plasmin into the nucleus accumbens (NAc) significantly potentiated whereas plasminogen activator inhibitor-1 reduced the nicotine-induced dopamine release in the NAc in a dose-dependent manner. Nicotine-induced dopamine release was markedly diminished in tPA-deficient (tPA-/-) mice, and the defect of dopamine release in tPA-/- mice was restored by microinjection of either exogenous tPA or plasmin into the NAc. Nicotine increased tPA protein levels and promoted the release of tPA into the extracellular space in the NAc. Immunohistochemistry revealed that PAR1 immunoreactivity was localized to the nerve terminals positive for tyrosine hydroxylase in the NAc. Furthermore, we demonstrated that plasmin activated PAR1 and that nicotine-induced place preference and dopamine release were diminished in PAR1-deficient (PAR1-/-) mice. Targeting the tPA-plasmin-PAR1 system would provide new therapeutic approaches to the treatment of nicotine dependence.


Prospects and problems in the use of recombinant DNA for the production of a malaria vaccine.
Recent advances in malaria immunology and the discovery of an in vitro culture method for Plasmodium falciparum have opened the way for the feasibility of formulating a vaccine as a control measure for this most virulent species of human malaria. The successful use of recombinant DNA technology for the production of scarce cellular constituents has made available a great option for mass synthesis of malaria antigens, which otherwise could be prohibitive if it were to depend on the use of human serum and cells. The translation of feasibility to reality of production of a practical human malaria vaccine will depend on the definition of a safe and effective antigen.


Agent-based computational model of the prevalence of gonococcal infections after the implementation of HIV pre-exposure prophylaxis guidelines.
Recently, the first comprehensive guidelines were published for pre-exposure prophylaxis (PrEP) for the prevention of HIV infection in populations with substantial risk of infection. Guidelines include a daily regimen of emtricitabine/tenofovir disoproxil fumarate (TDF/FTC) as well as condom usage during sexual activity. The relationship between the TDF/FTC intake regimen and condom usage is not yet fully understood. If men who have sex with men (MSM,) engage in high-risk sexual activities without using condoms when prescribed TDF/FTC they might be at an increased risk for other sexually transmitted diseases (STD). Our study focuses on the possible occurrence of behavioral changes among MSM in the United States over time with regard to condom usage. In particular, we were interested in creating a model of how increased uptake of TDF/FTC might cause a decline in condom usage, causing significant increases in non-HIV STD incidence, using gonococcal infection incidence as a biological endpoint. We used the agent-based modeling software NetLogo, building upon an existing model of HIV infection. We found no significant evidence for increased gonorrhea prevalence due to increased PrEP usage at any level of sample-wide usage, with a range of 0-90% PrEP usage. However, we did find significant evidence for decreased prevalence of HIV, with a maximal effect being reached when 5% to 10% of the MSM population used PrEP. Our findings appear to indicate that attitudes of aversion, within the medical community, toward the promotion of PrEP due to the potential risk of increased STD transmission are unfounded.


Principles of effective consultation: an update for the 21st-century consultant.
Little information in the literature exists to guide consult interactions between different medical specialties. A total of 323 general internists, family medicine physicians, general surgeons, orthopedic surgeons, and obstetricians/gynecologists (OB/GYNs) from 3 academic medical centers completed a survey addressing their ideal relationship with consultants. Differences between surgeons and nonsurgeons were calculated using logistic regression, adjusting for location and trainee status. Differences between different specialties of surgeons were calculated using analysis of variance with Scheffe post hoc analysis There was a 72% response rate. About half of respondents were surgeons and the rest were general internists and family medicine physicians. More nonsurgeons (69%) desired the consultant to focus on a narrow question than did surgeons (41%). Over half (59%) of family medicine physicians and internists preferred to retain order-writing authority on their patients compared with 37% of surgeons (P<.001). Of the surgeons preferring to retain authority, 70% believed it was appropriate for consultants to write orders after a verbal discussion. Orthopedic surgeons desired consultants to write orders and co-manage patients significantly more compared with general surgeons and OB/GYNs (P<.001). Only 29% of physicians thought literature references were useful in consultations. Most physicians (75%) desired direct verbal communication with the specialist providing the consultation. Most family physicians (78%) believed there was little need for general internal medicine input, preferring to consult medicine subspecialists directly. Specialty-dependent differences exist in consult preferences of physicians. These differences vary from the extremes of orthopedic surgeons desiring a comprehensive co-management approach with the consultant to general internists and family medicine physicians desiring to retain control over order writing and have a more focused consultant approach.


Gallamine exerts biphasic allosteric effects at muscarinic receptors.
Although gallamine and a number of other compounds have been reported to slow the rate of dissociation of labeled ligands, especially [3H]N-methylscopolamine (NMS), from muscarinic receptors of heart and brain, there has been some dispute as to whether the dissociation of [3H]quinuclidinyl benzilate (QNB) is subject to such allosteric regulation. The present studies were intended to determine whether past discrepancies might be due to differences between tissues. We have found that gallamine modulates the dissociation of [3H]QNB from muscarinic receptors of the heart in a biphasic manner. Low concentrations (micromolar) accelerate the rate of dissociation, whereas higher concentrations (millimolar) slow it; at about 0.1 mM, the two effects cancel each other. Similar results were obtained with muscarinic receptors from the brainstem, but gallamine had only marginal effects on the dissociation of [3H]QNB in the forebrain. On the other hand, verapamil exerts only monophasic effects (slowing) on the dissociation of both [3H]NMS and [3H]QNB from heart receptors and gallamine slows the dissociation of [3H]NMS to a similar extent in all three tissues. Thus, it appears that past discrepancies in the literature can be attributed to the tissues and concentrations of gallamine that were used. Furthermore, the biphasic effects of gallamine suggest that there are multiple allosteric regulatory sites associated with muscarinic receptors.


The role of computed tomography in assessing pleural malignancy prior to thoracoscopy.
Computed tomography (CT) scanning is part of the routine diagnostic work up of patients with suspected pleural malignancy but has a wide variation in the reported sensitivity and specificity. This review was to appraise the recent literature on the utility of CT scanning. When investigating patients for suspected pleural malignancy, the sensitivity of a malignant CT report may be higher than previously reported (68%), but the specificity seems significantly lower (78%). The predictive value of CT scanning (on all patients with pleural effusions) may be increased using a CT scoring system. Recent meta-analyses of the utility of PET give differing opinions on the value of this imaging modality. Further work needs to be done to define its place in the diagnostic pathway. CT scoring systems may allow further risk stratification. However, a low negative predictive value of a 'negative' CT scan could lead to false reassurance and missed malignancy. PET/CT does not currently appear to add additional diagnostic value. Pulmonary emboli should be considered in all patients being investigated for clinically suspected malignant pleural disease. Respiratory physicians should be mindful of rare or unusual presentations.


Archaea in Yellowstone Lake.
The Yellowstone geothermal complex has yielded foundational discoveries that have significantly enhanced our understanding of the Archaea. This study continues on this theme, examining Yellowstone Lake and its lake floor hydrothermal vents. Significant Archaea novelty and diversity were found associated with two near-surface photic zone environments and two vents that varied in their depth, temperature and geochemical profile. Phylogenetic diversity was assessed using 454-FLX sequencing (~51,000 pyrosequencing reads; V1 and V2 regions) and Sanger sequencing of 200 near-full-length polymerase chain reaction (PCR) clones. Automated classifiers (Ribosomal Database Project (RDP) and Greengenes) were problematic for the 454-FLX reads (wrong domain or phylum), although BLAST analysis of the 454-FLX reads against the phylogenetically placed full-length Sanger sequenced PCR clones proved reliable. Most of the archaeal diversity was associated with vents, and as expected there were differences between the vents and the near-surface photic zone samples. Thaumarchaeota dominated all samples: vent-associated organisms corresponded to the largely uncharacterized Marine Group I, and in surface waters, ~69-84% of the 454-FLX reads matched archaeal clones representing organisms that are Nitrosopumilus maritimus-like (96-97% identity). Importance of the lake nitrogen cycling was also suggested by >5% of the alkaline vent phylotypes being closely related to the nitrifier Candidatus Nitrosocaldus yellowstonii. The Euryarchaeota were primarily related to the uncharacterized environmental clones that make up the Deep Sea Euryarchaeal Group or Deep Sea Hydrothermal Vent Group-6. The phylogenetic parallels of Yellowstone Lake archaea to marine microorganisms provide opportunities to examine interesting evolutionary tracks between freshwater and marine lineages.


Determination of trace 2,4-dinitrophenol in surface water samples based on hydrophilic molecularly imprinted polymers/nickel fiber electrode.
A rapid, sensitive and selective electrochemical method was proposed for the determination of 2,4-dinitrophenol (2,4-DNP) in surface water samples, using hydrophilic molecular imprinted polymers (MIPs) as the recognition element and nickel (Ni) fiber as the catalytic element. Hydrophilic MIPs were synthesized using 2,4-DNP as the template, acrylamide as the monomer, glycidilmethacrylate as the pro-hydrophilic co-monomer and acetonitrile as the solvent. Hydrophilic modification could enhance the accessibility of 2,4-DNP to the imprinted cavities and improve the selective recognition properties of traditional MIPs in water medium. Subsequently, hydrophilic MIPs/Ni fiber electrode was prepared to determine trace 2,4-DNP by cyclic voltammetry. The parameters affecting the analytical performance were investigated. Under optimized conditions, the linear range was 0.7-30 μg L⁻¹ and the detection limit was 0.1 μg L⁻¹. Finally, the proposed method was applied to measure 2,4-DNP in surface water samples. The spiked recoveries were changed from 91.3% to 102.6% and the RSD was not higher than 5.1%. There was no statistically significant difference between the results obtained by the proposed method and the traditional chromatographic method.


Mapping cell behavior across a wide range of vertical silicon nanocolumn densities.
Over the past decade, vertical nanostructures have provided novel approaches for biomedical applications such as intracellular delivery/detection, specific cell capture, membrane potential measurement, and cellular activity regulation. Although the feasibility of the vertical nanostructures as a new biological tool has been thoroughly demonstrated, a better understanding of cell behavior on vertical nanostructures, in particular the effects of geometry, is essential for advanced applications. To investigate the cell behavior according to the variation of the spacing between vertical nanostructures, we have interfaced fibroblasts (NIH3T3) with density-controlled vertical silicon nanocolumn arrays (vSNAs). Over a wide range of vSNA densities, we observe three distinct cell settling regimes and investigate both overall cell behavior (adhesions, morphology, and mobility) and detailed biomacromolecule variance (F-actin and focal adhesion) across these regimes. We expect that these systematic observations could serve as a guide for improved nanostructure array design for the desired cell manipulation.


In vitro functional response of human tendon cells to different dosages of low-frequency pulsed electromagnetic field.
Chronic tendinopathy is a degenerative process causing pain and disability. Current treatments include biophysical therapies, such as pulsed electromagnetic fields (PEMF). The aim of this study was to compare, for the first time, the functional in vitro response of human tendon cells to different dosages of PEMF, varying in field intensity and duration and number of exposures. Tendon cells, isolated from human semitendinosus and gracilis tendons (hTCs; n = 6), were exposed to different PEMF treatments (1.5 or 3 mT for 8 or 12 h, single or repeated treatments). Scleraxis (SCX), COL1A1, COL3A1 and vascular endothelial growth factor-A (VEGF-A) expression and cytokine production were assessed. None of the different dosages provoked apoptotic events. Proliferation of hTCs was enhanced by all treatments, whereas only 3 mT-PEMF treatment increased cell viability. However, the single 1.5 mT-PEMF treatment elicited the highest up-regulation of SCX, VEGF-A and COL1A1 expression, and it significantly reduced COL3A1 expression with respect to untreated cells. The treated hTCs showed a significantly higher release of IL-1β, IL-6, IL-10 and TGF-β. Interestingly, the repeated 1.5 mT-PEMF significantly further increased IL-10 production. 1.5 mT-PEMF treatment was able to give the best results in in vitro healthy human tendon cell culture. Although the clinical relevance is not direct, this investigation should be considered an attempt to clarify the effect of different PEMF protocols on tendon cells, in particular focusing on the potential applicability of this cell source for regenerative medicine purpose, both in surgical and in conservative treatment for tendon disorders.


Indirect comparison in evaluating relative efficacy illustrated by antimicrobial prophylaxis in colorectal surgery.
This paper aims to explore the potential usefulness and limitations of indirect comparisons in evaluating the relative efficacy of interventions. From a systematic review of antimicrobial prophylaxis in colorectal surgery, we identified 11 sets of randomized trials that can be used to compare antibiotics both directly and indirectly. The discrepancy between the direct and the indirect comparison is defined as the absolute value of difference in log odds ratio. The adjusted indirect comparison has the advantages that the prognostic factors of participants in different trials can be partially taken into account and more uncertainty be incorporated into its result by providing a wider confidence interval. However, considerable discrepancies exist between the direct and the adjusted indirect comparisons. When there is no direct comparison, the adjusted indirect method may be used to obtain some evidence about the relative efficacy of competing interventions, although such indirect results should be interpreted with great caution. Further empirical and methodologic research is needed to explore the validity and generalizability of the adjusted indirect comparison for evaluating different interventions.


Qualitative study of the consequences of vaccine-induced immune thrombocytopenia and thrombosis: the experiences of family members.
To explore the experiences of family members of patients who died or survived following a diagnosis of vaccine-induced immune thrombocytopenia and thrombosis (VITT). A semistructured qualitative study, conducted via Zoom. Participants discussed their experiences during hospitalisation and following discharge. Sixteen family members of patients with VITT (survivors=11; bereaved=5), recruited via a Facebook support group and advertising on Twitter. Analysis identified two themes common to both groups of participants: the stress of hospitalisation and the experience of multiple losses. A third theme, living with VITT, was unique to the survivor group and a fourth, battling against the system, was predominantly reported by bereaved participants. This is a significantly challenged group of people, with multiple emotional, financial, social and psychological losses. These losses have been compounded by experiences of limited governmental and societal recognition of the problems they face.


Incorporating fuzzy membership functions into the perceptron algorithm.
The perceptron algorithm, one of the class of gradient descent techniques, has been widely used in pattern recognition to determine linear decision boundaries. While this algorithm is guaranteed to converge to a separating hyperplane if the data are linearly separable, it exhibits erratic behavior if the data are not linearly separable. Fuzzy set theory is introduced into the perceptron algorithm to produce a ``fuzzy algorithm'' which ameliorates the convergence problem in the nonseparable case. It is shown that the fuzzy perceptron, like its crisp counterpart, converges in the separable case. A method of generating membership functions is developed, and experimental results comparing the crisp to the fuzzy perceptron are presented.


Core Body and Skin Temperature in Type 1 Narcolepsy in Daily Life; Effects of Sodium Oxybate and Prediction of Sleep Attacks.
Previous laboratory studies in narcolepsy patients showed altered core body and skin temperatures, which are hypothesised to be related to a disturbed sleep wake regulation. In this ambulatory study we assessed temperature profiles in normal daily life, and whether sleep attacks are heralded by changes in skin temperature. Furthermore, the effects of three months of treatment with sodium oxybate (SXB) were investigated. Twenty-five narcolepsy patients and 15 healthy controls were included. Core body, proximal and distal skin temperatures, and sleep-wake state were measured simultaneously for 24 hours in ambulatory patients. This procedure was repeated in 16 narcolepsy patients after at least 3 months of stable treatment with SXB. Increases in distal skin temperature and distal-to-proximal temperature gradient (DPG) strongly predicted daytime sleep attacks (P < 0.001). As compared to controls, patients had a higher proximal and distal skin temperature in the morning, and a lower distal skin temperature during the night (all P < 0.05). Furthermore, they had a higher core body temperature during the first part of the night (P < 0.05), which SXB decreased (F = 4.99, df = 1, P = 0.03) to a level similar to controls. SXB did not affect skin temperature. This ambulatory study demonstrates that daytime sleep attacks were preceded by clear changes in distal skin temperature and DPG. Furthermore, changes in core body and skin temperature in narcolepsy, previously only studied in laboratory settings, were partially confirmed. Treatment with SXB resulted in a normalisation of the core body temperature profile. Future studies should explore whether predictive temperature changes can be used to signal or even prevent sleep attacks.


Heart Rate Variability Biofeedback Improves Emotional and Physical Health and Performance: A Systematic Review and Meta Analysis.
We performed a systematic and meta analytic review of heart rate variability biofeedback (HRVB) for various symptoms and human functioning. We analyzed all problems addressed by HRVB and all outcome measures in all studies, whether or not relevant to the studied population, among randomly controlled studies. Targets included various biological and psychological problems and issues with athletic, cognitive, and artistic performance. Our initial review yielded 1868 papers, from which 58 met inclusion criteria. A significant small to moderate effect size was found favoring HRVB, which does not differ from that of other effective treatments. With a small number of studies for each, HRVB has the largest effect sizes for anxiety, depression, anger and athletic/artistic performance and the smallest effect sizes on PTSD, sleep and quality of life. We found no significant differences for number of treatment sessions or weeks between pretest and post-test, whether the outcome measure was targeted to the population, or year of publication. Effect sizes are larger in comparison to inactive than active control conditions although significant for both. HRVB improves symptoms and functioning in many areas, both in the normal and pathological ranges. It appears useful as a complementary treatment. Further research is needed to confirm its efficacy for particular applications.


Sequence data and association statistics from 12,940 type 2 diabetes cases and controls.
To investigate the genetic basis of type 2 diabetes (T2D) to high resolution, the GoT2D and T2D-GENES consortia catalogued variation from whole-genome sequencing of 2,657 European individuals and exome sequencing of 12,940 individuals of multiple ancestries. Over 27M SNPs, indels, and structural variants were identified, including 99% of low-frequency (minor allele frequency [MAF] 0.1-5%) non-coding variants in the whole-genome sequenced individuals and 99.7% of low-frequency coding variants in the whole-exome sequenced individuals. Each variant was tested for association with T2D in the sequenced individuals, and, to increase power, most were tested in larger numbers of individuals (>80% of low-frequency coding variants in ~82 K Europeans via the exome chip, and ~90% of low-frequency non-coding variants in ~44 K Europeans via genotype imputation). The variants, genotypes, and association statistics from these analyses provide the largest reference to date of human genetic information relevant to T2D, for use in activities such as T2D-focused genotype imputation, functional characterization of variants or genes, and other novel analyses to detect associations between sequence variation and T2D.


Robust but delayed thalamocortical activation of dendritic-targeting inhibitory interneurons.
GABA-releasing cortical interneurons are crucial for the neural transformations underlying sensory perception, providing "feedforward" inhibition that constrains the temporal window for synaptic integration. To mediate feedforward inhibition, inhibitory interneurons need to fire in response to ascending thalamocortical inputs, and most previous studies concluded that ascending inputs activate mainly or solely proximally targeting, parvalbumin-containing "fast-spiking" interneurons. However, when thalamocortical axons fire at frequencies that are likely to occur during natural exploratory behavior, activation of fast-spiking interneurons is rapidly and strongly depressed, implying the paradoxical conclusion that feedforward inhibition is absent when it is most needed. To address this issue, we took advantage of lines of transgenic mice in which either parvalbumin- or somatostatin-containing interneurons express GFP and recorded the responses of interneurons from both subtypes to thalamocortical stimulation in vitro. We report that during thalamocortical activation at behaviorally expected frequencies, fast-spiking interneurons were indeed activated only transiently because of rapid depression of their thalamocortical inputs, but a subset of layer 5 somatostatin-containing interneurons were robustly and persistently activated after a delay, due to the facilitation and temporal summation of their thalamocortical excitatory postsynaptic potentials. Somatostatin-containing interneurons are considered distally targeting. Thus, they are likely to provide delayed dendritic inhibition during exploratory behavior, contributing to the maintenance of a balance between cortical excitation and inhibition while leaving a wide temporal window open for synaptic integration and plasticity in distal dendrites.


Case-reporting of acute hepatitis B and C among injection drug users.
Although public health surveillance system data are widely used to describe the epidemiology of communicable disease, occurrence of hepatitis B and C virus (HBV and HCV, respectively) infections may be misrepresented by under-reporting in injection drug users (IDUs). This study was carried out to examine the relationship between HBV and HCV incidence and case-reporting of hepatitis B and C in Seattle IDUs. Names of participants in a Seattle IDU cohort study who acquired HBV or HCV infection over a 12-month follow-up period were compared to a database of persons with acute hepatitis B and C reported to the health department surveillance unit over the same period. Of 2,208 IDUs enrolled in the cohort who completed a follow-up visit, 63/759 acquired HBV infection, 53/317 acquired HCV infection, and 3 subjects acquired both HBV and HCV. Of 113 cohort subjects who acquired HBV or HCV, only 2 (1.5%) cases were reported; both had acute hepatitis B. The upper 95% confidence limit for case-reporting of hepatitis C in the cohort was 5.7%, and for hepatitis B, it was 7.5%. In this study, reporting of acute hepatitis in IDUs was extremely low, raising questions regarding the use of community surveillance data to estimate underlying incidence in that population group.


New insight into the mechanism of accumulation and intraerythrocytic compartmentation of albitiazolium, a new type of antimalarial.
Bis-thiazolium salts constitute a new class of antihematozoan drugs that inhibit parasite phosphatidylcholine biosynthesis. They specifically accumulate in Plasmodium- and Babesia-infected red blood cells (IRBC). Here, we provide new insight into the choline analogue albitiazolium, which is currently being clinically tested against severe malaria. Concentration-dependent accumulation in P. falciparum-infected erythrocytes reached steady state after 90 to 120 min and was massive throughout the blood cycle, with cellular accumulation ratios of up to 1,000. This could not occur through a lysosomotropic effect, and the extent did not depend on the food vacuole pH, which was the case for the weak base chloroquine. Analysis of albitiazolium accumulation in P. falciparum IRBC revealed a high-affinity component that was restricted to mature stages and suppressed by pepstatin A treatment, and thus likely related to drug accumulation in the parasite food vacuole. Albitiazolium also accumulated in a second high-capacity component present throughout the blood cycle that was likely not related to the food vacuole and also observed with Babesia divergens-infected erythrocytes. Accumulation was strictly glucose dependent, drastically inhibited by H+/K+ and Na+ ionophores upon collapse of ionic gradients, and appeared to be energized by the proton-motive force across the erythrocyte plasma membrane, indicating the importance of transport steps for this permanently charged new type of antimalarial agent. This specific, massive, and irreversible accumulation allows albitiazolium to restrict its toxicity to hematozoa-infected erythrocytes. The intraparasitic compartmentation of albitiazolium corroborates a dual mechanism of action, which could make this new type of antimalarial agent resistant to parasite resistance.


THYMECTOMY: PROLONGATION OF IMMUNOLOGICAL TOLERANCE IN THE ADULT MOUSE.
The loss of acquired immunological tolerance of mice to bovine gamma globulin depended on the presence of the thymus. Mice were repeatedly injected with bovine gamma-globulin from birth until the age of 5 to 10 weeks and then thymectomized or sham operated. After 130 to 160 days without antigen, an accelerated (immune) disappearance of I(125) bovine gamma-globulin could be uniformly induced in controls while thymectomized mice remained tolerant. Adult thymectomized mice made tolerant by a single injection of bovine gamma-globulin lost tolerance more slowly than sham-operated controls.


Effects of improvement in periodontal inflammation by toothbrushing on serum lipopolysaccharide concentration and liver injury in rats.
Periodontitis increases the serum lipopolysaccharide level, contributing to liver injury. Toothbrushing improves periodontitis and may also affect serum lipopolysaccharide concentration and periodontitis-induced liver injury. The purpose of the present study was to examine whether the improvement in periodontal inflammation by toothbrushing clinically affects the serum lipopolysaccharide level and hepatic pathological changes in rat periodontitis. Thirty male Wistar rats were divided into 5 groups, 2 groups receiving topical application of pyrogen-free water to the gingival sulcus for 4 or 8 weeks. The next 2 groups received topical application of lipopolysaccharide and proteases for 4 or 8 weeks. The last group received topical application of lipopolysaccharide and proteases for 8 weeks, and the palatal gingiva was brushed with a powered toothbrush once a day for 4 weeks prior to the end of the experimental period. Topical application of lipopolysaccharide and proteases induced not only periodontal inflammation but also an elevation in the serum lipopolysaccharide concentration, with increasing hepatic inflammation, steatosis and 8-hydroxydeoxyguanosine levels in a time-dependent manner. The rats that received gingival stimulation showed decreased polymorphonuclear leukocyte infiltration and collagen loss levels in the periodontal lesions. Furthermore, this group also showed a decrease in serum lipopolysaccharide concentration and hepatic inflammation, steatosis and 8-hydroxydeoxyguanosine levels, compared with the group receiving no treatment. Toothbrushing promoted healing of periodontal lesions, decreased serum lipopolysaccharide concentration and suppressed liver injury in a rat periodontitis model.


The Association between Periodontal Status, Serum Lipid Levels, Lipoprotein Associated Phosholipase A2 (Lp-PLA2) in Chronic Periodontitis Subjects and Healthy Controls.
Lipoprotein associated phospholipase A2 (Lp-PLA2) has been explored as a potential biomarker for Cardiovascular Disease (CVD). The human Lp-PLA2 is a serine-dependent, Ca2+- independent enzyme. It gives way to oxidised free fatty acids and lysophosphatidyl choline by hydrolysing oxidised phospholipids, that leads to smooth muscle cell apoptosis, inflammatory cell chemotaxis and endothelial cell dysfunction in CVD. Owing to its role in inflammation, it may influence the development and progression of periodontitis as well. To compare the demographic variables, Gingival Index(GI), Bleeding On Probing (BOP), Probing Pocket Depth (PPD), and Clinical Attachment Level (CAL) with serum lipid profile and Lp-PLA2 level in Chronic Periodontitis (CP) subjects. A total of 75 subjects were selected and divided into three groups; based on the inclusion and exclusion criteria: Group I - 25 subjects with severe generalized CP with CAL ≥ 5 mm in more than 30% of sites. Group II -25 subjects with moderate generalized CD with clinical CAL 3 mm-4 mm in more than 30% of sites. Group III- 25 systemically and periodontally healthy volunteers who served as control. Clinical parameters such as Plaque Index (PI), BOP, Probing Depth (PD) and CAL, lipid profile such as Total Cholesterol (TC), Triglyceride (TG), High density Lipoprotein (HDL), Low Density Lipoprotein (LDL), Very Low Density Lipoprotein (VLDL) as well as Lp-PLA2 were assessed. One-way ANOVA and Tukey's Post-hoc test were used for data analysis. Age was higher in Group I and II when compared to Group III. Group II showed a significant correlation between PI and LDL. In Group III, lower PI was significantly correlated with high HDL and low LDL and BOP was positively correlated with TG and HDL. A significant association of Lp-PLA2 was found to be higher with increase in the TG and VLDL level in Group I and Group II when compared to Group III. The study concluded that Lp-PLA2, TG and VLDL already being a predictor biomarker for atherosclerotic disease can be an inflammatory marker for periodontitis.


Local and systemic immunological parameters associated with remission of asthma symptoms in children.
The immunological and clinical parameters that are associated with asthma remission are poorly understood. The cytokine and local mediator changes associated with the resolution of asthma symptoms were examined in three groups of subjects 12-18 years of age (n = 15 in each group): (a) continuing asthma group (CA) who had persistent symptoms since early childhood, (b) an age, sex and atopic status-matched group who had persistent symptoms in early childhood but in whom these had resolved (RA), and (c) a non-atopic, non-asthmatic control group. Clinical parameters, sputum cell counts, peripheral blood mononuclear cell (PBMC) cytokine production and activation marker expression were determined. All of the CA had methacholine airway hyperresponsiveness compared with only half of the RA subjects. The CA showed elevated numbers of eosinophils and increased ECP and IL-5 in sputum, which were not observed in the RA. PBMC cytokine studies revealed increased production of the type 1 cytokines IL-12, IFN-γ and TNF-α in the CA group compared with the RA group, under a range of activation conditions, however, the production of IL-4 and IL-5 were unchanged. These findings suggest that decreased type 1 cytokine expression as well as decreased eosinophilic inflammation is associated with the resolution of asthma symptoms.


A randomized trial of basing treatment on human papillomavirus and/or cytology results in low-grade cervical lesion triage.
This study was undertaken to compare management algorithms that base treatment with loop electrosurgical excision procedure on human papillomavirus and/or repeat Papanicolaou test smear results. A randomized trial that referred 674 women with either atypical squamous cells of undetermined significance or low-grade squamous intraepithelial lesions cytology results, detected in organized screening to treatment either (1) if they were positive in a repeat Papanicolaou test smear and/or a human papillomavirus test or (2) if they were positive in the repeat Papanicolaou test smear test only. Women who tested positive were treated, regardless of colposcopic findings. There were 208 of 337 (62%) women who were treated in the human papillomavirus /Papanicolaou test smear group (187/337 because of HPV positivity) and 138 of 337 (41%) in the Papanicolaou test smear only group. Histopathologically diagnosed cervical intraepithelial neoplasia grade 2 or worse was found among 112 of 337 (33.2%) women in the human papillomavirus/Papanicolaou test smear group compared with 85 of 337 (25.2%) women in the Papanicolaou test smear only group (P < .05). Twenty-one women with cervical intraepithelial neoplasia 2+ had normal colposcopy. For adequate cervical intraepithelial neoplasia 2+ sensitivity, the decision to use loop electrosurgical excision procedure needs to be based on human papillomavirus testing results and should not exclude women with normal colposcopy.


Effect of body size on operative risk of carotid endarterectomy.
Many studies have found that women have a higher risk of perioperative stroke or death from carotid endarterectomy. Other vascular surgical procedures have demonstrated that body size and morphology impact on operative risk. We correlated the 30 day operative risk of stroke and death in the European Carotid Surgery Trial (ECST) with height, weight, body surface area (BSA), and body mass index using single variable analyses and multivariable logistic regression. Women were at significantly higher risk of perioperative stroke and death in the ECST. Both height and BSA confounded the effect of sex, implying that the generally smaller size of women may contribute to their increased risk. This finding should be validated in other large datasets.


Differential role of NADP+ and NADPH in the activity and structure of GDP-D-mannose 4,6-dehydratase from two chlorella viruses.
GDP-D-mannose 4,6-dehydratase (GMD) is a key enzyme involved in the synthesis of 6-deoxyhexoses in prokaryotes and eukaryotes. Paramecium bursaria chlorella virus-1 (PBCV-1) encodes a functional GMD, which is unique among characterized GMDs because it also has a strong stereospecific NADPH-dependent reductase activity leading to GDP-D-rhamnose formation (Tonetti, M., Zanardi, D., Gurnon, J., Fruscione, F., Armirotti, A., Damonte, G., Sturla, L., De Flora, A., and Van Etten, J.L. (2003) J. Biol. Chem. 278, 21559-21565). In the present study we characterized a recombinant GMD encoded by another chlorella virus, Acanthocystis turfacea chlorella virus 1 (ATCV-1), demonstrating that it has the expected dehydratase activity. However, it also displayed significant differences when compared with PBCV-1 GMD. In particular, ATCV-1 GMD lacks the reductase activity present in the PBCV-1 enzyme. Using recombinant PBCV-1 and ATCV-1 GMDs, we determined that the enzymatically active proteins contain tightly bound NADPH and that NADPH is essential for maintaining the oligomerization status as well as for the stabilization and function of both enzymes. Phylogenetic analysis indicates that PBCV-1 GMD is the most evolutionary diverged of the GMDs. We conclude that this high degree of divergence was the result of the selection pressures that led to the acquisition of new reductase activity to synthesize GDP-D-rhamnose while maintaining the dehydratase activity in order to continue to synthesize GDP-L-fucose.


Rates of local procedural-related structural injury following clipping or coiling of anterior communicating artery aneurysms.
Surgical clipping and endovascular coiling yield similar functional outcomes for the treatment of saccular aneurysms of the anterior communicating (ACOM) artery. However, surgical treatment may be associated with greater rates of cognitive impairment due to injury of adjacent structures. We aimed to quantify the rates of injury (infarction/hemorrhage) for both clipping and coiling of ACOM aneurysms. This was a retrospective dual-center radiological investigation of a consecutive series of patients with ruptured and unruptured ACOM aneurysms treated between January 2011 and October 2014. Post-treatment CT or MRI was assessed for new ischemic or hemorrhagic injury. Injury relating to the primary hemorrhage or vasospasm was differentiated. Univariate analysis using χ(2) tests and multivariate analysis using binary logistic regression was used. 66 patients treated with clipping were compared with 93 patients treated with coiling. 32/66 (48.5%) patients in the clipping group suffered treatment-related injury (31 ischemic, 1 hemorrhagic) compared with 4/93 (4.4%) patients in the coiling group (3 ischemic, 1 hemorrhagic) (p<0.0001). For patients with subarachnoid hemorrhage, the multivariate OR for infarction for clipping over coiling was 24.42 (95% CI 5.84 to 102.14), p<0.0001. The most common site of infarction was the basal forebrain (28/66 patients, 42.4%), with bilateral infarction in 4. There was injury of the septal/subcallosal region in 12/66 patients (18%). Clipping of ACOM aneurysms is associated with significantly higher rates of structural injury than coiling, and this may be a reason for superior cognitive outcomes in patients treated with coiling in previously published studies.


Discrete cosine transform-based shift estimation for fringe pattern profilometry using a generalized analysis model.
What is believed to be a new analysis algorithm to carry out profile measurement with low computational complexity and less noise sensitivity is presented. First, a discrete cosine transform (DCT)-based representation method is introduced to express the height distribution of a 3D surface. Then a novel shift estimation algorithm, called the DCT-based shift estimation (DCT-SE), is presented to reconstruct 3D object surfaces by using the proposed expression and the generalized analysis model. The advantage of DCT-SE is that without loss of measurement precision it provides lower computational complexity to implement 3D reconstruction from nonlinearly distorted fringe patterns and, at the same time, survives the random noise. Simulations and experiments show that the proposed DCT-SE is a fast, accurate, and efficient reconstruction algorithm for digital projection- based fringe pattern profilometry techniques.


Heterogeneity of response to exercise of rat muscle pyruvate dehydrogenase complex.
Muscle glucose uptake is greatly stimulated by moderate exercise, but full oxidation of the glucose to CO2 depends on the activity of the pyruvate dehydrogenase (PDH) complex. Our aim was to determine how PDH complex in different muscle groups responds to varying periods of moderate exercise. Rats were run on a motor-driven treadmill for 5-30 min and muscle PDH complex activity was determined in heart, diaphragm and red quadriceps muscles after isolation of mitochondria in the presence of inhibitors of PDH complex interconversion. In heart and diaphragm muscle, exercise caused an increase in PDH complex activity after 5 min, but this was followed by a significant decrease in activity as exercise progressed. In red quadriceps muscle, PDH complex activity was reduced after 5 min of exercise and was decreased further as exercise continued. We conclude that increased duration of exercise can lead to reduced PDH complex activity in rat muscles. We propose that this is a consequence of elevated fatty acid oxidation, the products of which stimulate PDH kinase. This implies that increased glycolysis to lactate and increased fatty acid oxidation can simultaneously provide energy for contracting muscle.


Long-term risk of carotid restenosis in patients randomly assigned to endovascular treatment or endarterectomy in the Carotid and Vertebral Artery Transluminal Angioplasty Study (CAVATAS): long-term follow-up of a randomised trial.
In the Carotid and Vertebral Artery Transluminal Angioplasty Study (CAVATAS), early recurrent carotid stenosis was more common in patients assigned to endovascular treatment than it was in patients assigned to endarterectomy (CEA), raising concerns about the long-term effectiveness of endovascular treatment. We aimed to investigate the long-term risks of restenosis in patients included in CAVATAS. 413 patients who were randomly assigned in CAVATAS and completed treatment for carotid stenosis (200 patients had endovascular treatment and 213 patients had endarterectomy) had prospective clinical follow-up at a median of 5 years and carotid duplex ultrasound at a median of 4 years. We investigated the cumulative long-term incidence of carotid restenosis after endovascular treatment and endarterectomy, the effect of the use of stents on restenosis after endovascular treatment, risk factors for the development of restenosis, and the effect of carotid restenosis on the risk of recurrent cerebrovascular events. Analysis was by intention to treat. This study is registered, number ISRCTN01425573. Severe carotid restenosis (>or=70%) or occlusion occurred significantly more often in patients in the endovascular arm than in patients in the endarterectomy arm (adjusted hazard ratio [HR] 3.17, 95% CI 1.89-5.32; p<0.0001). The estimated 5-year incidence of restenosis was 30.7% in the endovascular arm and 10.5% in the endarterectomy arm. Patients in the endovascular arm who were treated with a stent (n=50) had a significantly lower risk of developing restenosis of 70% or greater compared with those treated with balloon angioplasty alone (n=145; HR 0.43, 0.19-0.97; p=0.04). Current smoking or a history of smoking was a predictor of restenosis of 70% or more (2.32, 1.19-4.54; p=0.01) and the early finding of moderate stenosis (50-69%) up to 60 days after treatment was associated with the risk of progression to restenosis of 70% or more (3.76, 1.88-7.52; p=0.0002). The composite endpoint of ipsilateral non-perioperative stroke or transient ischaemic attack occurred more often in patients in whom restenosis of 70% or more was diagnosed in the first year after treatment compared with patients without restenosis of 70% or more (5-year incidence 23%vs 11%; HR 2.18, 1.04-4.54; p=0.04), but the increase in ipsilateral stroke alone was not significant (10%vs 5%; 1.67, 0.54-5.11). Restenosis is about three times more common after endovascular treatment than after endarterectomy and is associated with recurrent ipsilateral cerebrovascular symptoms; however, the risk of recurrent ipsilateral stroke is low. Further data are required from on-going trials of stenting versus endarterectomy to ascertain whether long-term ultrasound follow-up is necessary after carotid revascularisation. British Heart Foundation; UK National Health Service Management Executive; UK Stroke Association.


Canine-assisted Therapy as an Adjunct Tool in the Care of the Surgical Patient: A Literature Review and Opportunity for Research.
The use of canine-assisted therapy (CAT) in healthcare is expanding and the purpose of this review is to highlight its potential use in the surgical patient. While CAT literature to date has detailed widespread benefits in blood pressure control and improving pain, anxiety, and stress, little research has been performed specifically in surgical patients who may benefit significantly from CAT interventions. Critical points highlighted herein are as follows: (1) Hypertension is common and significantly increases morbidity and mortality associated with elective surgery. Pet ownership and brief CAT interventions (5-20 min) have demonstrated significant reductions in blood pressure and blood pressure variability in both adult and pediatric populations. (2) Pain management is of utmost importance in hospitalized, surgical patients and unfortunately the growing opioid addiction epidemic has complicated our ability to treat postoperative pain. CAT interventions have been shown to reduce self-reported pain. Therefore, CAT represents a cost-effective, safe, and noninvasive approach to pain management. (3) Patient satisfaction is of growing concern as reimbursement by Medicare, Medicaid, and other insurers is now linked to patient reported satisfaction with their hospital stay. While very limited data is available on this subject, some studies have showed that CAT intervention, specifically, improved patient reported satisfaction in multiple categories of the HCAHPS survey compared with patients who did not receive CAT. Overall, this is a novel narrative review detailing the therapeutic efficacy of CAT, highlighting the specific indications of CAT in the surgical patient, and urging further research of CAT in the surgical patient.


Meta-coexpression conservation analysis of microarray data: a "subset" approach provides insight into brain-derived neurotrophic factor regulation.
Alterations in brain-derived neurotrophic factor (BDNF) gene expression contribute to serious pathologies such as depression, epilepsy, cancer, Alzheimer's, Huntington and Parkinson's disease. Therefore, exploring the mechanisms of BDNF regulation represents a great clinical importance. Studying BDNF expression remains difficult due to its multiple neural activity-dependent and tissue-specific promoters. Thus, microarray data could provide insight into the regulation of this complex gene. Conventional microarray co-expression analysis is usually carried out by merging the datasets or by confirming the re-occurrence of significant correlations across datasets. However, co-expression patterns can be different under various conditions that are represented by subsets in a dataset. Therefore, assessing co-expression by measuring correlation coefficient across merged samples of a dataset or by merging datasets might not capture all correlation patterns. In our study, we performed meta-coexpression analysis of publicly available microarray data using BDNF as a "guide-gene" introducing a "subset" approach. The key steps of the analysis included: dividing datasets into subsets with biologically meaningful sample content (e.g. tissue, gender or disease state subsets); analyzing co-expression with the BDNF gene in each subset separately; and confirming co- expression links across subsets. Finally, we analyzed conservation in co-expression with BDNF between human, mouse and rat, and sought for conserved over-represented TFBSs in BDNF and BDNF-correlated genes. Correlated genes discovered in this study regulate nervous system development, and are associated with various types of cancer and neurological disorders. Also, several transcription factor identified here have been reported to regulate BDNF expression in vitro and in vivo. The study demonstrates the potential of the "subset" approach in co-expression conservation analysis for studying the regulation of single genes and proposes novel regulators of BDNF gene expression.


Transitions in living arrangements are associated with changes in dietary patterns in young women.
Household composition influences people's diet, so typical transitions in young women's lives, including cohabitation, marriage, and motherhood, might be expected to influence their subsequent dietary behavior. The objective was to examine associations between transitions in living arrangements and changes in energy intake and dietary patterns for women in their 20s and 30s using longitudinal data collected in 2003 and 2009. FFQ were collected twice from 6534 women born in 1973-1978 participating in the Australian Longitudinal Study on Women's Health. Transition groups were defined from changes in their living arrangements. Factor analysis was used to identify dietary patterns. Associations between transitions in living arrangements and changes in energy intake and dietary pattern scores were analyzed using multiple linear regression. Women living with children had greater energy intake than other women initially and those who started a family had the greatest increases over time. Five similar dietary patterns were derived from both surveys. Women living in a family at both times had higher scores on the high-fat and sugar, meat, and cooked vegetables patterns and lower scores on the Mediterranean-style and fruit patterns than other women. Women starting a family increased their consumption of the high-fat and sugar, fruit, and cooked vegetables patterns. Women not living with children at both times had increased scores on the Mediterranean-style pattern and decreased scores on the high-fat and sugar and cooked vegetables patterns compared with other women. In conclusion, starting a family is associated with changes in women's diet that are mainly unhealthy.


Overwhelming infections in trauma.
Following trauma, wound contamination with aerobic and/or anaerobic bacteria should always be suspected. Treatment with antibacterial antibiotics should begin immediately in the emergency room, particularly for those patients with fractures. Patients with serious trauma are best treated by a team of specialists including general surgeons, orthopedists, infectious disease specialists, and intensive care specialists. The author recommends transport of seriously injured patients to major hospitals specializing in the care of trauma. Particularly when gas gangrene secondary to clostridial infection is suspected, the patient should be moved to a major trauma center with the capability for hyperbaric oxygen therapy.


Cyclin D1 overexpression permits the reproducible detection of senescent human vascular smooth muscle cells.
The senescence of mitotic cells is hypothesized to play a causal role in organismal aging. Cultures of normal human cells become senescent in vitro as a result of a continuous decline in the mitotic fraction from cell turnover. However, one potential barrier to the evaluation of the frequency and distribution of senescent cells in tissues is the absence of a panel of robust markers for the senescent state. In parallel with an analysis of the growth kinetics of human vascular smooth muscle cells, we have undertaken transcriptomic comparisons of early- and late-passage cultures of human vascular smooth muscle cells to identify potential markers that can distinguish between senescent and growth-competent cells. A wide range of genes are upregulated at senescence in human vascular smooth muscle cells. In particular, we have identified a 12-fold upregulation of expression in the cyclin D1 message, which is reflected in a concomitant upregulation at the protein level. Quantitative cytochemical analysis of senescent and growing vascular smooth muscle cells indicates that cyclin D1 reactivity is a considerably better marker of replicative senescence than senescence-associated beta-galactosidase activity. We have applied this new marker (in combination with Ki67, COMET, and TUNEL staining) to the study of human vascular smooth muscle cells treated with resveratrol, a putative anti-aging molecule known to have significant effects on cell growth.


Incorporating social anxiety into a model of college student problematic drinking.
College problem drinking and social anxiety are significant public health concerns with highly negative consequences. College students are faced with a variety of novel social situations and situations encouraging alcohol consumption. The current study involved developing a path model of college problem drinking, including social anxiety, in 316 college students referred to an alcohol intervention due to a campus alcohol violation. Contrary to hypotheses, social anxiety generally had an inverse relationship with problem drinking. As expected, perceived drinking norms had important positive, direct effects on drinking variables. However, the results generally did not support the hypotheses regarding the mediating or moderating function of the valuations of expected effects and provided little support for the mediating function of alcohol expectancies in the relations among social anxiety and alcohol variables. Therefore, it seems that the influence of peers may be more important for college students than alcohol expectancies and valuations of alcohol's effects are. College students appear to be a unique population in respect to social anxiety and problem drinking. The implications of these results for college prevention and intervention programs were discussed.


Regulation of PIDD auto-proteolysis and activity by the molecular chaperone Hsp90.
In response to DNA damage, p53-induced protein with a death domain (PIDD) forms a complex called the PIDDosome, which either consists of PIDD, RIP-associated protein with a death domain and caspase-2, forming a platform for the activation of caspase-2, or contains PIDD, RIP1 and NEMO, important for NF-κB activation. PIDDosome activation is dependent on auto-processing of PIDD at two different sites, generating the fragments PIDD-C and PIDD-CC. Despite constitutive cleavage, endogenous PIDD remains inactive. In this study, we screened for novel PIDD regulators and identified heat shock protein 90 (Hsp90) as a major effector in both PIDD protein maturation and activation. Hsp90, together with p23, binds PIDD and inhibition of Hsp90 activity with geldanamycin efficiently disrupts this association and impairs PIDD auto-processing. Consequently, both PIDD-mediated NF-κB and caspase-2 activation are abrogated. Interestingly, PIDDosome formation itself is associated with Hsp90 release. Characterisation of cytoplasmic and nuclear pools of PIDD showed that active PIDD accumulates in the nucleus and that only cytoplasmic PIDD is bound to Hsp90. Finally, heat shock induces Hsp90 release from PIDD and PIDD nuclear translocation. Thus, Hsp90 has a major role in controlling PIDD functional activity.


A prospective study of toddlers with ASD: short-term diagnostic and cognitive outcomes.
Despite recent increases in the number of toddlers referred for a differential diagnosis of autism spectrum disorders (ASD), knowledge of short-term stability of the early diagnosis as well as cognitive outcomes in this cohort is still limited. Cognitive, social, and communication skills of 89 clinic-referred toddlers were assessed at the average age of 21.5 (SD = 4.9) months, and reassessed at 46.9 (SD = 7.7) months. Groups with stable and unstable diagnostic presentation were identified and compared on their profile of cognitive and social-communicative skills obtained at the time of initial diagnosis. Stability of the ASD diagnosis was 100%; diagnosis of autism was stable in 74% of cases as compared to 83% and 81% in PDD-NOS and Non-ASD groups, respectively. Worsening of social disability symptoms resulting in autism diagnosis was noted in 17% of toddlers initially diagnosed with PDD-NOS and in 19% of toddlers with initial diagnosis of non-ASD disorder. However, marked improvement was noted in approximately 1/4 of children initially presenting with autism, warranting diagnostic reassignment to PDD-NOS at follow-up. An analysis of developmental skills profiles suggests particular relevance of the assessment of verbal and nonverbal communication skills to diagnostic differentiation between subtypes within ASD in the second year of life. Stability of ASD diagnosis in toddlers is high, though marked changes in severity of symptoms is to be expected in a minority of cases. Simultaneous consideration of cognitive, social, and communication skills profiles enhances accuracy of diagnostic classification and prediction of outcome.


The good and bad of targeting cancer-associated extracellular matrix.
The maintenance of tissue homeostasis requires extracellular matrix (ECM) remodeling. Immune cells actively participate in regenerating damaged tissues contributing to ECM deposition and shaping. Dysregulated ECM deposition characterizes fibrotic diseases and cancer stromatogenesis, where a chronic inflammatory state sustains the ECM increase. In cancer, the ECM fosters several steps of tumor progression, providing pro-survival and proliferative signals, promoting tumor cell dissemination via collagen fibers or acting as a barrier to impede drug diffusion. Interfering with processes leading to chronic ECM deposition, as occurring in cancer, might allow the simultaneous targeting of both primary tumors and metastatic lesions. However, a note of caution comes from data showing that defective ECM deposition is associated with an exacerbated inflammatory and autoimmune phenotype and to lymphomagenesis. Immune cells display ITIM-inhibitory receptors recognizing collagens as counter ligands, which negatively regulate the immune response. This is in line with the idea that ECM components can provide homeostatic signals to immune cells to regulate and prevent unwanted activation, a concept particularly relevant in cancer where these mechanisms could be in place to keep infiltrating immune cells in a suppressive pro-tumoral state. In this context, the pharmacological targeting of myeloid cells, for which both direct and indirect roles in ECM deposition have been shown, can be a relevant option to this purpose.


Biogeographical patterns and speciation of the genus Pinguicula (Lentibulariaceae) inferred by phylogenetic analyses.
Earlier phylogenetic studies in the genus Pinguicua (Lentibulariaceae) suggested that the species within a geographical region was rather monophyletic, although the sampling was limited or was restricted to specific regions. Those results conflicted with the floral morphology-based classification, which has been widely accepted to date. In the current study, one nuclear ribosomal DNA (internal transcribed spacer; ITS) and two regions of chloroplast DNA (matK and rpl32-trnL), from up to ca. 80% of the taxa in the genus Pinguicula, covering all three subgenera, were sequenced to demonstrate the inconsistency and explore a possible evolutionary history of the genus. Some incongruence was observed between nuclear and chloroplast topologies and the results from each of the three DNA analyses conflicted with the morphology-based subgeneric divisions. Both the ITS tree and network, however, corresponded with the biogeographical patterns of the genus supported by life-forms (winter rosette or hibernaculum formation) and basic chromosome numbers (haploidy). The dormant strategy evolved in a specific geographical region is a phylogenetic constraint and a synapomorphic characteristic within a lineage. Therefore, the results denied the idea that the Mexican group, morphologically divided into the three subgenera, independently acquired winter rosette formations. Topological incongruence among the trees or reticulations, indicated by parallel edges in phylogenetic networks, implied that some taxa originated by introgressive hybridisation. Although there are exceptions, species within the same geographical region arose from a common ancestor. Therefore, the classification by the floral characteristics is rather unreliable. The results obtained from this study suggest that evolution within the genus Pinguicula has involved; 1) ancient expansions to geographical regions with gene flow and subsequent vicariance with genetic drift, 2) acquirement of a common dormant strategy within a specific lineage to adapt a local climate (i.e., synapomorphic characteristic), 3) recent speciation in a short time span linked to introgressive hybridisation or multiplying the ploidy level (i.e., divergence), and 4) parallel evolution in floral traits among lineages found in different geographical regions (i.e., convergence). As such, the floral morphology masks and obscures the phylogenetic relationships among species in the genus.


Valley Polarization by Spin Injection in a Light-Emitting van der Waals Heterojunction.
The band structure of transition metal dichalcogenides (TMDCs) with valence band edges at different locations in the momentum space could be harnessed to build devices that operate relying on the valley degree of freedom. To realize such valleytronic devices, it is necessary to control and manipulate the charge density in these valleys, resulting in valley polarization. While this has been demonstrated using optical excitation, generation of valley polarization in electronic devices without optical excitation remains difficult. Here, we demonstrate spin injection from a ferromagnetic electrode into a heterojunction based on monolayers of WSe2 and MoS2 and lateral transport of spin-polarized holes within the WSe2 layer. The resulting valley polarization leads to circularly polarized light emission that can be tuned using an external magnetic field. This demonstration of spin injection and magnetoelectronic control over valley polarization provides a new opportunity for realizing combined spin and valleytronic devices based on spin-valley locking in semiconducting TMDCs.