Loss of nucleosomal DNA condensation coincides with appearance of a novel nuclear protein in dinoflagellates.
The packaging, expression, and maintenance of nuclear genomes using histone proteins is a ubiquitous and fundamental feature of eukaryotic cells, yet the phylum Dinoflagellata has apparently abandoned this model of nuclear organization. Their nuclei contain permanently condensed, liquid crystalline chromosomes that seemingly lack histone proteins, and contain remarkably large genomes. The molecular basis for this reorganization is poorly understood, as is the sequence of evolutionary events that led to such radical change. We have investigated nuclear organization in the closest relative to dinoflagellates, Perkinsus marinus, and an early-branching dinoflagellate, Hematodinium sp., to identify early changes that occurred during dinoflagellate nuclear evolution. We show that P. marinus has a typical nuclear organization that is based on the four core histones. By the early divergence of Hematodinium sp., however, dinoflagellate genome size is dramatically enlarged, chromosomes are permanently condensed, and histones are scarcely detectable. In place of histones, we identify a novel, dominant family of nuclear proteins that is only found in dinoflagellates and, surprisingly, in a family of large algal viruses, the Phycodnaviridae. These new proteins, which we call DVNPs (dinoflagellate/viral nucleoproteins), are highly basic, bind DNA with similar affinity to histones, and occur in multiple posttranslationally modified forms. We find these proteins throughout all dinoflagellates, including early- and late-branching taxa, but not in P. marinus. Gain of a major novel family of nucleoproteins, apparently from an algal virus, occurred early in dinoflagellate evolution and coincides with rapid and dramatic reorganization of the dinoflagellate nucleus.


Genetic variance components estimation for binary traits using multiple related individuals.
Understanding and modeling genetic or nongenetic factors that influence susceptibility to complex traits has been the focus of many genetic studies. Large pedigrees with known complex structure may be advantageous in epidemiological studies since they can significantly increase the number of factors whose influence on the trait can be estimated. We propose a likelihood approach, developed in the context of generalized linear mixed models, for modeling dichotomous traits based on data from hundreds of individuals all of whom are potentially correlated through either a known pedigree or an estimated covariance matrix. Our approach is based on a hierarchical model where we first assess the probability of each individual having the trait and then formulate a likelihood assuming conditional independence of individuals. The advantage of our formulation is that it easily incorporates information from pertinent covariates as fixed effects and at the same time takes into account the correlation between individuals that share genetic background or other random effects. The high dimensionality of the integration involved in the likelihood prohibits exact computations. Instead, an automated Monte Carlo expectation maximization algorithm is employed for obtaining the maximum likelihood estimates of the model parameters. Through a simulation study we demonstrate that our method can provide reliable estimates of the model parameters when the sample size is close to 500. Implementation of our method to data from a pedigree of 491 Hutterites evaluated for Type 2 diabetes (T2D) reveal evidence of a strong genetic component to T2D risk, particularly for younger and leaner cases.


Early Assessment of Tumor Response to Radiation Therapy using High-Resolution Quantitative Microvascular Ultrasound Imaging.
Measuring changes in tumor volume using anatomical imaging weeks to months post radiation therapy (RT) is currently the clinical standard for indicating treatment response to RT. For patients whose tumors do not respond successfully to treatment, this approach is suboptimal as timely modification of the treatment approach may lead to better clinical outcomes. We propose to use tumor microvasculature as a biomarker for early assessment of tumor response to RT. Acoustic angiography is a novel contrast ultrasound imaging technique that enables high-resolution microvascular imaging and has been shown to detect changes in microvascular structure due to cancer growth. Data suggest that acoustic angiography can detect longitudinal changes in the tumor microvascular environment that correlate with RT response. Methods: Three cohorts of Fisher 344 rats were implanted with rat fibrosarcoma tumors and were treated with a single fraction of RT at three dose levels (15 Gy, 20 Gy, and 25 Gy) at a dose rate of 300 MU/min. A simple treatment condition was chosen for testing the feasibility of our imaging technique. All tumors were longitudinally imaged immediately prior to and after treatment and then every 3 days after treatment for a total of 30 days. Both acoustic angiography (using in-house produced microbubble contrast agents) and standard b-mode imaging was performed at each imaging time point using a pre-clinical Vevo770 scanner and a custom modified dual-frequency transducer. Results: Results show that all treated tumors in each dose group initially responded to treatment between days 3-15 as indicated by decreased tumor growth accompanied with decreased vascular density. Untreated tumors continued to increase in both volume and vascular density until they reached the maximum allowable size of 2 cm in diameter. Tumors that displayed complete control (no tumor recurrence) continued to decrease in size and vascular density, while tumors that progressed after the initial response presented an increase in tumor volume and volumetric vascular density. The increase in tumor volumetric vascular density in recurring tumors can be detected 10.25 ± 1.5 days, 6 ± 0 days, and 4 ± 1.4 days earlier than the measurable increase in tumor volume in the 15, 20, and 25 Gy dose groups, respectively. A dose-dependent growth rate for tumor recurrence was also observed. Conclusions: In this feasibility study we have demonstrated the ability of acoustic angiography to detect longitudinal changes in vascular density, which was shown to be a potential biomarker for tumor response to RT.


Peptidergic innervation of human epicardial coronary arteries.
The peptidergic innervation of proximal (internal diameter, > 0.8 mm) and distal (internal diameter, < 0.8 mm) regions of human epicardial coronary arteries was investigated by means of immunohistochemical, chromatographic, radioimmunological, and in vitro pharmacological techniques. The use of an antiserum to the general neuronal marker protein gene product 9.5 revealed that the proximal part of epicardial arteries possessed a relatively sparse supply of nerve fibers forming a loose network in the adventitia. The perivascular network increased in density as the vessels were followed distally. In both proximal and distal regions, the majority of nerve fibers possessed neuropeptide Y and tyrosine hydroxylase immunoreactivity. Calcitonin gene-related peptide (CGRP)- and substance P-immunoreactive nerve fibers were very sparse in the proximal region of the arteries and increased in number distally. Only a few scattered vasoactive intestinal peptide (VIP)-immunoreactive nerve fibers were detected in both arterial regions. The use of high-performance liquid chromatography and radioimmunoassay revealed that the immunoreactive material present in coronary artery extracts closely resembled synthetic peptides. An in vitro pharmacological method demonstrated that neuropeptide Y elicited no detectable response in either proximal or distal arterial segments. In contrast, CGRP, substance P, and VIP all produced a concentration-dependent relaxation of both arterial regions. CGRP and substance P were stronger and more potent than VIP. CGRP and substance P induced a more potent response in distal compared with proximal regions of the arteries. These results suggest that the peptidergic nerves supplying human large epicardial coronary arteries may be predominantly involved in mediating vasodilation.


Changes in Attitudes toward Mental Illness in Healthcare Professionals and Students.
Mental-illness-related stigma not only exists in the public but also in healthcare systems. Healthcare providers (HCPs) who have stigmatizing attitudes or behaviors might be thought of as a key barrier to mental health service use, and influence the quality of healthcare. Although cumulative projects have been conducted to reduce stigma related to mental illness among HCPs around the world, little is known about whether the attitudes of HCPs toward mental illness have changed over time. Research on this topic is mixed with respect to whether attitudes of HCPs toward mental illness have become more or less positive. The aim of the current study was to help clarify this issue using a cross-temporal meta-analysis of scores on the Social Distance Scale (SDS), Opinions about Mental Illness (OMI), and Community Attitudes towards Mental Illness (CAMI) measures among health care professionals and students (N = 15,653) from 1966 to 2016. Our results indicated that both social distance (β = -0.32, p < 0.001) and attitudes (β = 0.43, p = 0.007) of HCPs toward mental illness have become increasingly positive over time. These findings provide empirical evidence to support that the anti-stigma programs and courses have positive effects on HCPs and can inform future anti-stigma programs focusing on improving the attitudes of HCPs toward mental illness, thereby improving the quality of healthcare provided.


Isolation and characterization of Neisseria musculi sp. nov., from the wild house mouse.
Members of the genus Neisseria have been isolated from or detected in a wide range of animals, from non-human primates and felids to a rodent, the guinea pig. By means of selective culture, biochemical testing, Gram staining and PCR screening for the Neisseria-specific internal transcribed spacer region of the rRNA operon, we isolated four strains of the genus Neisseria from the oral cavity of the wild house mouse, Mus musculus subsp. domesticus. The isolates are highly related and form a separate clade in the genus, as judged by tree analyses using either multi-locus sequence typing of ribosomal genes or core genes. One isolate, provisionally named Neisseria musculi sp. nov. (type strain AP2031T=DSM 101846T=CCUG 68283T=LMG 29261T), was studied further. Strain AP2031T/N. musculi grew well in vitro. It was naturally competent, taking up DNA in a DNA uptake sequence and pilT-dependent manner, and was amenable to genetic manipulation. These and other genomic attributes of N. musculi sp. nov. make it an ideal candidate for use in developing a mouse model for studying Neisseria-host interactions.


A case of delayed-onset pulmonary barotrauma in a scuba diver.
A 23-yr-old male scuba diver was admitted to the hyperbaric chamber of the Polish Army Training Centre of Divers and SCUBA Divers for treatment of pneumomediastinum and possible decompression sickness. Medical history suggested a case of pulmonary barotrauma with delayed and exacerbated symptoms. About 36 h after the onset of symptoms the patient was treated successfully according to U.S. Navy Treatment Table 6-A. A recurrence of symptoms was observed more than 3 d after the initial hyperbaric therapy. Signs and symptoms suggested the presence of air in the vessels of the nervous system, and possibly in the heart muscle and bones of the upper limbs. A second recompression therapy began according to method III of the Polish Navy Treatment Tables. Adjunctive therapy included intravenous aspirin, steroids, isosorbide dinitrate, and fluids. Symptoms resolved and did not recur after this second hyperbaric therapy.


DISC1 and Huntington's disease--overlapping pathways of vulnerability to neurological disorder?
We re-annotated the interacting partners of the neuronal scaffold protein DISC1 using a knowledge-based approach that incorporated recent protein interaction data and published literature to. This revealed two highly connected networks. These networks feature cellular function and maintenance, and cell signaling. Of potentially greatest interest was the novel finding of a high degree of connectivity between the DISC1 scaffold protein, linked to psychiatric illness, and huntingtin, the protein which is mutated in Huntington's disease. The potential link between DISC1, huntingtin and their interacting partners may open new areas of research into the effects of pathway dysregulation in severe neurological disorders.


A deep analysis of the small non-coding RNA population in Schistosoma japonicum eggs.
Schistosoma japonicum is a parasitic flatworm that causes zoonotic schistosomiasis. The typical outcome of schistosomiasis is hepatic granuloma and fibrosis, which is primarily induced by soluble egg-derived antigens. Although schistosomal eggs represent an important pathogenic stage to the host, the biology of this critical stage is largely unknown. We previously investigated the expression profiles of sncRNAs during different developmental stages of this parasite. However, using small RNA extracted from egg-deposited liver tissues generated limited information about sncRNAs in eggs. Here, we characterized the complete small RNAome in this stage of the parasite after optimization of RNA purification. A library, SjE, was constructed with the small RNA extracted from S. japonicum eggs and subjected to high-throughput sequencing. The data were depicted by comprehensive bioinformatic analysis to explore the expression features of sncRNAs in the egg stage. MicroRNAs accounted for about one quarter of the total small RNA population in this stage, with a strongly biased expression pattern of certain miRNA family members. Sja-miR-71, sja-miR-71-5p, and sja-miR-36-3p were suggested to play important roles in embryo development. A panel of transfer RNA fragments (tRFs) precisely processed from the 5' end of mature tRNAs was identified for the first time, which represented a strong egg stage-biased expression. The tRNA-Ala derived small RNAs were the most highly expressed Sj-tRFs in eggs. Further, the expression of siRNAs from 29 types of well-defined transposable elements (TEs) was observed to be relatively stable among different developmental stages. In this study, we characterized the sncRNA profile in the egg stage of S. japonicum. Featured expression of sncRNAs, especially the tRNA-derived small RNAs, was identified, which was further compared with that of other developmental stages. These novel findings would facilitate a deeper understanding of the biology of schistosomal parasites.


Role of the N-terminal domain of the calcitonin receptor-like receptor in ligand binding.
Calcitonin receptor-like receptor (CRLR) is a seven-transmembrane (7-TM) domain class B G protein-coupled receptor (GPCR) which requires coexpression of different receptor activity modifying proteins (RAMP) to become a functional calcitonin gene-related peptide (CGRP) receptor or an adrenomedullin (AM) receptor. The N-terminal (Nt) extracellular region of class B GPCRs in ligand binding has been reported for receptors such as glucagon and parathyroid hormone. We hypothesize that the Nt-domain of CRLR (Nt-CRLR) is an autonomously folded unit possessing a well-defined structure and is involved in ligand binding and specificity. To obtain structural and functional information on the Nt-CRLR, we cloned and expressed the Nt-CRLR as a fusion protein in Escherichia coli. Overexpressed protein formed an inclusion body, which was refolded and purified, resulting in a soluble monomeric protein. Far-UV CD and fluorescence spectra of Nt-CRLR showed characteristics of a folded protein. The ability of Nt-CRLR to bind CGRP and AM independent of RAMPs was determined by studying inhibition of (125)I-CGRP and (125)I-AM binding to pregnant rat uterine membrane in the presence of Nt-CRLR protein. We observe that Nt-CRLR inhibits (125)I-CGRP and (125)I-AM binding to rat uterus in a dose-dependent fashion (IC(50) = 0.25 and 0.29 muM, respectively). Taken together, our data provide evidence that Nt-CRLR is structured and further that a significant part of the binding affinity comes from binding to the Nt-domain.


X-ray holographic microscopy with zone plates applied to biological samples in the water window using 3rd harmonic radiation from the free-electron laser FLASH.
The imaging of hydrated biological samples - especially in the energy window of 284-540 eV, where water does not obscure the signal of soft organic matter and biologically relevant elements - is of tremendous interest for life sciences. Free-electron lasers can provide highly intense and coherent pulses, which allow single pulse imaging to overcome resolution limits set by radiation damage. One current challenge is to match both the desired energy and the intensity of the light source. We present the first images of dehydrated biological material acquired with 3rd harmonic radiation from FLASH by digital in-line zone plate holography as one step towards the vision of imaging hydrated biological material with photons in the water window. We also demonstrate the first application of ultrathin molecular sheets as suitable substrates for future free-electron laser experiments with biological samples in the form of a rat fibroblast cell and marine biofouling bacteria Cobetia marina.


The In Vivo Potential-Regulated Protective Protein of Nitrogenase in Azotobacter vinelandii Supports Aerobic Bioelectrochemical Dinitrogen Reduction In Vitro.
Nitrogenase, the only enzyme known to be able to reduce dinitrogen (N2) to ammonia (NH3), is irreversibly damaged upon exposure to molecular oxygen (O2). Several microbes, however, are able to grow aerobically and diazotrophically (fixing N2 to grow) while containing functional nitrogenase. The obligate aerobic diazotroph, Azotobacter vinelandii, employs a multitude of protective mechanisms to preserve nitrogenase activity, including a "conformational switch" protein (FeSII, or "Shethna") that reversibly locks nitrogenase into a multicomponent protective complex upon exposure to low concentrations of O2. We demonstrate in vitro that nitrogenase can be oxidatively damaged under anoxic conditions and that the aforementioned conformational switch can protect nitrogenase from such damage, confirming that the conformational change in the protecting protein can be achieved solely by regulating the potential of its [2Fe-2S] cluster. We further demonstrate that this protective complex preserves nitrogenase activity upon exposure to air. Finally, this protective FeSII protein was incorporated into an O2-tolerant bioelectrosynthetic cell whereby NH3 was produced using air as a substrate, marking a significant step forward in overcoming the crippling limitation of nitrogenase's sensitivity toward O2.


Regulation of follicular B cell differentiation by the related E26 transformation-specific transcription factors PU.1, Spi-B, and Spi-C.
Splenic B-2 cells can be divided into two major subsets: follicular (FO) and marginal zone (MZ) B cells. FO and MZ B cells are generated from immature transitional B cells. Few transcription factors have been identified that regulate FO B cell differentiation. The highly related proteins PU.1, Spi-B, and Spi-C are transcription factors of the E26-transformation-specific family and are important for B cell differentiation and function. To determine whether these proteins play a role in the differentiation of FO B cells, we performed a detailed analysis of splenic B cells in mice with inactivating mutations in the genes encoding PU.1 (Sfpi1) or Spi-B (Spib). Sfpi1(+/-) Spib(-/-) (PUB) mice had a 9-fold reduction in the frequency of CD23(+) FO B cells compared with that of wild-type mice. In contrast, PUB mice had a 2-fold increase in the frequency of MZ B cells that was confirmed by immunofluorescence staining. Expression of Spi-C in Eμ-Spi-C transgenic PUB mice partially rescued frequencies of CD23(+) B cells. Gene expression analysis, in vitro reporter assays, and chromatin immunoprecipitation experiments showed that transcription of the Fcer2a gene encoding CD23 is activated by PU.1, Spi-B, and Spi-C. These results demonstrate that FO B cell differentiation is regulated by the E26-transformation-specific transcription factors PU.1, Spi-B, and Spi-C.


The response of a co-culture lung model to fine and ultrafine particles of incinerator fly ash at the air-liquid interface.
Elevated concentrations of particulate matter in the environmental atmosphere constitute a potential risk to human health. In vitro cell-based assays are therefore necessary to evaluate the toxicological potential of inhaled particulate emissions. In this study, the exposure of a co-culture cell model at the air-liquid interface was used to evaluate the dose-dependent biological effects of a test aerosol. The CULTEX system was used to expose human cells to an environmentally-relevant aerosol, generated from fly ash collected in a commercial municipal waste incinerator and resuspended in filtered air. Human bronchial epithelial cells, BEAS-2B, co-cultured with differentiated THP-1 macrophages growing on Transwell inserts, were employed in the bioassay. Analyses of cell viability, interleukin-8 (IL-8) release, intracellular glutathione, and haeme oxygenase-1 enzyme expression were performed. Transportation of the cells and exposure to humidified filtered air or the test aerosol, at 100 ml/min for 1 to 6 hours, were well tolerated by the cells and had no effect on their viability. Levels of IL-8 release and haeme oxygenase-1 expression were elevated by exposure to fly ash aerosol as a function of time, but not by exposure to clean air. For IL-8 release, a dose-dependent effect was demonstrated with the assumption that the deposited mass of the particles correlated with exposure time. Exposure to the test aerosol did not affect the intracellular glutathione concentration. This in vitro approach simulates particle deposition in the human lung more realistically than does submerged exposure, and it preserves the inherent properties of the particles. It shows promise for use to detect particulate emissions which are potentially detrimental to human health.


Prenatal nicotine exposure and child behavioural problems.
In utero exposure to tobacco smoke has been related to numerous adverse health effects in new-borns, infants, children, adolescents and adults. The aim of this review was to summarise findings on prenatal nicotine exposure and its relationship with behavioural problems in the offspring. The majority of studies, and especially several recent epidemiological studies, observed a higher likelihood for attention-deficit/hyperactivity disorder (ADHD) or ADHD symptoms in exposed subjects. However, both human and animal studies have failed to provide clear evidence on causality. Existing literature on studies investigating the association between prenatal nicotine exposure and conduct or externalising problems in the offspring suggests a causal effect. The establishment of a final conclusion concerning the relationship between prenatal nicotine exposure and internalising problems in the offspring is complicated by insufficient data and mixed results in epidemiological studies. Prenatal nicotine exposure has been associated with altered brain structure and function in human offspring, and a proposed biological mechanism is related to nicotine's adverse influence on neurotransmitter systems during brain development. In conclusion, establishing a statement on the causality of the relationship between prenatal nicotine exposure and behavioural problems in children remains a challenging task. Nevertheless, considering the results of an increasing number of studies which link prenatal exposure to nicotine to externalising problems applying different methodologies to account for confounding and in view of other adverse health effects known to be caused by this exposure, parents should consider smoking cessation.


National Radio Astronomy Observatory: The early history and development of the observatory at Green Bank, West Virginia, are reviewed.
The existence of the National Radio Astronomy Observatory and the researches already accomplished there are the result of the foresight and wisdom of United States scientists, the National Science Board, and the Congress, who joined forces to make possible this new national asset. Continued effort will be needed td insure that the observatory will always have the finest possible research instruments and that the site will be a haven of radio quiet. Visiting scientists in some instances may wish to bring equipment with them for studying special problems. Within its means, the observatory will provide supporting facilities, including receivers and other electronic devices, computers, laboratories and shops, and housing. Scientists interested in more details concerning arrangements for visitors should direct their inquiries to the National Radio Astronomy Observatory, P. O. Box 2, Green Bank, West Virginia.


Modeling and evaluating the performance of Brillouin distributed optical fiber sensors.
A thorough analysis of the key factors impacting on the performance of Brillouin distributed optical fiber sensors is presented. An analytical expression is derived to estimate the error on the determination of the Brillouin peak gain frequency, based for the first time on real experimental conditions. This expression is experimentally validated, and describes how this frequency uncertainty depends on measurement parameters, such as Brillouin gain linewidth, frequency scanning step and signal-to-noise ratio. Based on the model leading to this expression and considering the limitations imposed by nonlinear effects and pump depletion, a figure-of-merit is proposed to fairly compare the performance of Brillouin distributed sensing systems. This figure-of-merit offers to the research community and to potential users the possibility to evaluate with an objective metric the real performance gain resulting from any proposed configuration.


Comparison of a multiplexed MassARRAY system with real-time allele-specific PCR technology for genotyping of methicillin-resistant Staphylococcus aureus.
The Sequenom MassARRAY iPLEX single-nucleotide polymorphism (SNP) typing platform uses matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) coupled with single-base extension PCR for high-throughput multiplex SNP detection. In this study, we investigated the use of iPLEX MassARRAY technology for methicillin-resistant Staphylococcus aureus (MRSA) genotyping. A 16-plex MassARRAY iPLEX GOLD assay (MRSA-iPLEX) was developed that targets a set of informative SNPs and binary genes for MRSA characterization. The method was evaluated with 147 MRSA isolates, and the results were compared with those of an established SYBR Green-based real-time PCR system utilizing the same SNP-binary markers. A total of 2352 markers belonging to 44 SNP-binary profiles were analysed by both real-time PCR and MRSA-iPLEX. With real-time PCR as the reference standard, MRSA-iPLEX correctly assigned 2298 of the 2352 (97.7%) markers. Sequence variation in the MRSA-iPLEX primer targets accounted for the majority of MRSA-iPLEX erroneous results, highlighting the importance of primer target selection. MRSA-iPLEX provided optimal throughput for MRSA genotyping, and was, on a reagent basis, more cost-effective than the real-time PCR methods. The 16-plex MRSA-iPLEX is a suitable alternative to SYBR Green-based real-time PCR typing of major sequence types and clonal complexes of MRSA.


Immune checkpoint-related serum proteins and genetic variants predict outcomes of localized prostate cancer, a cohort study.
The clinical predictors and biological mechanisms for localized prostate cancer (PCa) outcomes remain mostly unknown. We aim to evaluate the role of serum immune-checkpoint-related (ICK) proteins and genetic variations in predicting outcomes of localized PCa. We profiled the serum levels of 14 ICK-related proteins (BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, PD-L2, Tim-3, CD28, CD80, 4-1BB, CD27, and CTLA-4) in 190 patients with localized PCa. The genotypes of 97 single nucleotide polymorphisms (SNPs) from 19 ICK-related genes were analyzed in an extended population (N = 1762). Meta-data from ArrayExpress and TCGA was employed to validate and to probe functional data. Patients were enrolled and tumor aggressiveness, biochemical recurrence (BCR), and progression information were obtained. Statistical analyses were performed analyzing associations between serum biomarkers, genotypes, mRNA and outcomes. We showed that serum (s)BTLA and sTIM3 levels were associated with PCa aggressiveness (P < 0.05). sCD28, sCD80, sCTLA4, sGITR, sHVEM and sIDO correlated with both BCR and progression risks (all P < 0.05). We further identified ICK variants were significantly associated with aggressiveness, BCR and progression. Among them, 4 SNPs located in CD80 (rs7628626, rs12695388, rs491407, rs6804441) were not only associated with BCR and progression risk, but also correlated with sCD80 level (P < 0.01). rs491407 was further validated in an independent cohort. The CD80 mRNA expression was associated with BCR (HR, 1.85, 95% CI 1.06-3.22, P = 0.03) in meta-analysis of validation cohorts. We highlight the prognostic value of serum ICK-related proteins for predicting aggressiveness, BCR and progression of PCa. The genetic variations and mRNA expression in CD80 could be predictors and potential targets of localized PCa.


Atypical brain response to novelty in rural African children with a history of severe falciparum malaria.
Plasmodium falciparum is the most common parasitic infection of the central nervous system causing neuro-cognitive deficits in 5-26% of paediatric cases. The burden cannot be reliably estimated because of lack of sensitive, culture-fair and robust assessments in rural settings. Auditory and visual brain event related potentials (ERPs) are used to compare novelty processing in children exposed to severe malaria with community controls. Fifty children previously admitted and discharged from Kilifi District Hospital with severe falciparum malaria were selected and compared with 77 unexposed age matched children. The results showed that up to 14% of children exposed to severe malaria had significantly different responses to novelty compared to unexposed children. Children exposed to severe malaria had smaller P3a amplitudes to novelty in both auditory [F (3, 119)=4.545, p=0.005] and visual [F (3, 119)=6.708, p<0.001] paradigms compared to unexposed children. In the auditory domain the differences in processing of novelty were not related to early component processing. The percentage of children with severe malaria showing impaired performance using ERPs is within the range previously reported using neuropsychological tests. The overall pattern suggests that severe malaria affects prefrontal and temporal cortices normally activated by stimulus novelty.


Energy scaling of mode-locked fiber lasers with chirally-coupled core fiber.
We report a mode-locked dissipative soliton laser based on large-mode-area chirally-coupled-core Yb-doped fiber. This demonstrates scaling of a fiber oscillator to large mode area in a format that directly holds the lowest-order mode and that is also compatible with standard fiber integration. With an all-normal-dispersion cavity design, chirped pulse energies above 40 nJ are obtained with dechirped durations below 200 fs. Using a shorter fiber, dechirped durations close to 100 fs are achieved at pump-limited energies. The achievement of correct energy scaling is evidence of single-transverse-mode operation, which is confirmed by beam-quality and spectral-interference measurements.


Variable structure motifs for transcription factor binding sites.
Classically, models of DNA-transcription factor binding sites (TFBSs) have been based on relatively few known instances and have treated them as sites of fixed length using position weight matrices (PWMs). Various extensions to this model have been proposed, most of which take account of dependencies between the bases in the binding sites. However, some transcription factors are known to exhibit some flexibility and bind to DNA in more than one possible physical configuration. In some cases this variation is known to affect the function of binding sites. With the increasing volume of ChIP-seq data available it is now possible to investigate models that incorporate this flexibility. Previous work on variable length models has been constrained by: a focus on specific zinc finger proteins in yeast using restrictive models; a reliance on hand-crafted models for just one transcription factor at a time; and a lack of evaluation on realistically sized data sets. We re-analysed binding sites from the TRANSFAC database and found motivating examples where our new variable length model provides a better fit. We analysed several ChIP-seq data sets with a novel motif search algorithm and compared the results to one of the best standard PWM finders and a recently developed alternative method for finding motifs of variable structure. All the methods performed comparably in held-out cross validation tests. Known motifs of variable structure were recovered for p53, Stat5a and Stat5b. In addition our method recovered a novel generalised version of an existing PWM for Sp1 that allows for variable length binding. This motif improved classification performance. We have presented a new gapped PWM model for variable length DNA binding sites that is not too restrictive nor over-parameterised. Our comparison with existing tools shows that on average it does not have better predictive accuracy than existing methods. However, it does provide more interpretable models of motifs of variable structure that are suitable for follow-up structural studies. To our knowledge, we are the first to apply variable length motif models to eukaryotic ChIP-seq data sets and consequently the first to show their value in this domain. The results include a novel motif for the ubiquitous transcription factor Sp1.


Anticipatory and Compensatory Postural Adjustments in Response to External Lateral Shoulder Perturbations in Subjects with Parkinson's Disease.
The purpose of this study was to investigate the anticipatory (APA) and compensatory (CPA) postural adjustments in individuals with Parkinson's disease (PD) during lateral instability of posture. Twenty-six subjects (13 individuals with PD and 13 healthy matched controls) were exposed to predictable lateral postural perturbations. The electromyographic (EMG) activity of the lateral muscles and the displacement of the center of pressure (COP) were recorded during four time intervals that are typical for postural adjustments, i.e., immediately before (APA1, APA2) and after (CPA1 and CPA2) the postural disturbances. The magnitude of the activity of the lateral muscles in the group with PD was lower only during the CPA time intervals and not during the anticipatory adjustments (APAs). Despite this finding, subjects with PD exhibit smaller COP excursions before and after the disturbance, probably due to lack of flexibility and proprioceptive impairments. The results of this study suggest that postural instability in subjects with PD can be partially explained by decreased postural sway, before and after perturbations, and reduced muscular activity after body disturbances. Our findings can motivate new studies to investigate therapeutic interventions that optimize the use of postural adjustment strategies in subjects with PD.


Attachment and fusion inhibitors potently prevent dendritic cell-driven HIV infection.
Dendritic cells (DCs) efficiently transfer captured (trans) or de novo-produced (cis) virus to CD4 T cells. Using monocyte-derived DCs, we evaluated entry inhibitors targeting HIV envelope (BMS-C, T-1249) or CCR5 (CMPD167) for their potency to prevent DC infection, DC-driven infection in T cells in trans and cis, and direct infection of DC-T-cell mixtures. Immature DC-T-cell cultures with distinct mechanisms of viral transfer yielded similar levels of infection and produced more proviral DNA compared with matched mature DC-T-cell cultures or infected immature DCs. Although all compounds completely blocked HIV replication, 16 times more of each inhibitor (250 vs 15.6 nM) was required to prevent low-level infection of DCs compared with the productive DC-T-cell cocultures. Across all cell systems tested, BMS-C blocked infection most potently. BMS-C was significantly more effective than CMPD167 at preventing DC infection. In fact, low doses of CMPD167 significantly enhanced DC infection. Elevated levels of CCL4 were observed when immature DCs were cultured with CMPD167. Viral entry inhibitors did not interfere with Candida albicans-specific DC cytokine/chemokine responses. These findings indicate that an envelope-binding small molecule is a promising tool for topical microbicide design to prevent the infection of early targets needed to establish and disseminate HIV infection.


Does the pretherapeutic tumor SUV in 68Ga DOTATOC PET predict the absorbed dose of 177Lu octreotate?
Selection of candidates for peptide receptor radionuclide therapy (PRRT) is increasingly based on receptor positron emission tomography (PET) imaging, including the common tracer 68Ga DOTATOC. However, no studies have yet compared standardized uptake values (SUVs) and absorbed doses in this field. We retrospectively analyzed a consecutive cohort of 21 patients with 61 evaluable tumor lesions undergoing both pretherapeutic 68Ga DOTATOC-PET/CT (Biograph Duo [Siemens Medical Solutions, Erlangen, Germany]; PET acquisition, 75.3 ± 15.4 minutes postinjection; 117.3 ± 33.9 MBq 68Ga DOTATOC) and PRRT with Lu octreotate (7.47 ± 1.39 GBq; intratherapeutic tumor dosimetry with serial whole-body scans; 1, 2, and 4 days postinjection) at our institution. SUVs were compared with the tumor-absorbed doses per injected activity (D/A0) of the subsequent first treatment cycle. The correlation of SUV and D/A0 was r = 0.72 (SUVmean) and r = 0.71 (SUVmax), both P < 0.001. Pancreatic origin and hepatic localization were associated with higher D/A0, and chromogranin A level and Ki-67 index had no influence on SUV or D/A0. High-SUV lesions (SUVmean >15; SUVmax >25) resulted in high D/A0 (>10 Gy/GBq) in 66.7% to 70.8% and low D/A0 (<5 Gy/GBq) in only 8.3% to 12.5% on subsequent PRRT. The mentioned low D/A0 range, on the other hand, was achieved by all lesions with SUVmean <7 or SUVmax <9. Somatostatin receptor PET imaging may predict tumor-absorbed doses. The ability to indicate insufficient target irradiation by a low SUV could aid in selection of appropriate candidates for PRRT. However, larger series are needed to confirm and validate these initial findings.


Basic life support refresher training of nurses: individual training and group training are equally effective.
Basic life support (BLS) skills of hospital nurses are often poor. We compared individual BLS refresher training (IT; one instructor to one trainee) with group refresher training (GT; one instructor to six trainees). We hypothesised that IT would result in better skill acquisition and retention. Nurses from non-critical care wards (n=120) were randomised to IT or GT. Skills were assessed by a 3 min BLS test on a computerised manikin (Laerdal, Norway) immediately before training (T0), immediately after training (T1), and 10 months after training (T2). Results are expressed as median and [interquartile range]. The study was completed by 103 nurses (IT 56, GT 47). For GT the median group size was 5 [4-5]. The median duration of IT was 20 [17-21] min. The median duration of GT was 90 [84-95] or 19 min per trainee. Baseline skills did not differ between GT and IT, except for less compressions with correct depth for IT. At T1 and T2 there were no clinically significant differences between GT and IT for number of ventilations, ventilation volume, number of compressions, compression depth, compression rate and hands off time. Total instructor time was similar for IT and GT training strategies. There was no difference in IT and GT immediately and 10 months after training. However, training time per nurse for IT was only one fifth, whereas total instructor time did not increase. Although not superior in outcome, IT may be a cost-effective alternative for GT.


Identification of LIFR, PIK3R1, and MMP12 as Novel Prognostic Signatures in Gallbladder Cancer Using Network-Based Module Analysis.
Background: Gallbladder cancer (GBC) is a rare and aggressive malignancy of the biliary tract with a dismal survival rate. Effective biomarkers and therapeutic targets are urgently needed. Methods: We analyzed gene expression profiles of GBC to identify differentially expressed genes (DEGs) and then used these DEGs to identify functional module biomarkers based on protein functional interaction (FI) networks. We further evaluated the module-gene protein expression and clinical significance with immunohistochemistry staining (IHC) in a tissue microarray (TMA) from 80 GBC samples. Results: Five functional modules were identified. Module 0 included classical cancer signaling pathways, such as Ras and PI3K-Akt; and modules 1-4 included genes associated with muscle cells, fibrinogen, extracellular matrix, and integrins, respectively. We validated the expression of LIFR, PIK3R1, and MMP12, which were hubs or functional nodes in modules. Compared with paired peritumoural tissues, we found that the expression of LIFR (P = 0.002) and PIK3R1 (P = 0.046) proteins were significantly downregulated, and MMP12 (P = 0.006) was significantly upregulated. Further prognostic analysis showed that patients with low expression of LIFR had shorter overall survival than those with high expression (log-rank test P = 0.028), the same trend as for PIK3R1 (P = 0.053) and MMP12 (P = 0.006). Multivariate analysis indicated that expression of MMP12 protein (hazard ratio [HR] = 0.429; 95% confidence interval [CI] 0.198, 0.930; P = 0.032) was one of the significant independent prognostic factors for overall survival. Conclusions: We found a highly reliable FI network, which revealed LIFR, PIK3R1, and MMP12 as novel prognostic biomarker candidates for GBC. These findings could accelerate biomarker discovery and therapeutic development in this cancer.


Astragalus membranaceus and Punica granatum alleviate infertility and kidney dysfunction induced by aging in male rats.
By aging, male fertility and kidney function decline. Therefore, the investigation of health span-extending agents becomes more urgent to overcome aging-induced infertility and kidney dysfunction. The current research was undertaken to investigate the antiaging efficacy of Astragalus membranaceus telomerase activator-65 (Ta-65) and pomegranate supplements. Forty male Wistar rats were divided into young rats, aged rats, aged rats treated with Ta-65 (500mg/kg/day), and aged rats treated with pomegranate (250mg/kg/day). Testosterone, FSH, LH, and kidney functions were measured in serum. Sperm analysis as well as testicular histological examination was performed. Aging caused an imbalance in male sex hormones resulting in sperm abnormality and reductions in the sperm count and motility. Elevations in serum creatinine, uric acid, sodium, and potassium were reported in aged rats. Treatment with Ta-65 or pomegranate effectively ameliorated all the deteriorations induced by normal aging in male fertility and renal function. Ta-65 and pomegranate possessed strong antiaging activity by alleviating aging-induced male infertility through reestablishing the hormonal balance and testis architecture. They also alleviated the kidney dysfunction. On comparing Ta-65 with pomegranate, the improvement in FSH, LH, and sperm abnormalities caused by Ta-65 was much better than that caused by pomegranate.


R-CHIE: a web server and R package for visualizing RNA secondary structures.
Visually examining RNA structures can greatly aid in understanding their potential functional roles and in evaluating the performance of structure prediction algorithms. As many functional roles of RNA structures can already be studied given the secondary structure of the RNA, various methods have been devised for visualizing RNA secondary structures. Most of these methods depict a given RNA secondary structure as a planar graph consisting of base-paired stems interconnected by roundish loops. In this article, we present an alternative method of depicting RNA secondary structure as arc diagrams. This is well suited for structures that are difficult or impossible to represent as planar stem-loop diagrams. Arc diagrams can intuitively display pseudo-knotted structures, as well as transient and alternative structural features. In addition, they facilitate the comparison of known and predicted RNA secondary structures. An added benefit is that structure information can be displayed in conjunction with a corresponding multiple sequence alignments, thereby highlighting structure and primary sequence conservation and variation. We have implemented the visualization algorithm as a web server R-chie as well as a corresponding R package called R4RNA, which allows users to run the software locally and across a range of common operating systems.


Using electronic health record data for environmental and place based population health research: a systematic review.
We conducted a systematic review of literature published on January 2000-May 2017 that spatially linked electronic health record (EHR) data with environmental information for population health research. We abstracted information on the environmental and health outcome variables and the methods and data sources used. The automated search yielded 669 articles; 128 articles are included in the full review. The number of articles increased by publication year; the majority (80%) were from the United States, and the mean sample size was approximately 160,000. Most articles used cross-sectional (44%) or longitudinal (40%) designs. Common outcomes were health care utilization (32%), cardiometabolic conditions/obesity (23%), and asthma/respiratory conditions (10%). Common environmental variables were sociodemographic measures (42%), proximity to medical facilities (15%), and built environment and land use (13%). The most common spatial identifiers were administrative units (59%), such as census tracts. Residential addresses were also commonly used to assign point locations, or to calculate distances or buffer areas. Future research should include more detailed descriptions of methods used to geocode addresses, focus on a broader array of health outcomes, and describe linkage methods. Studies should also explore using longitudinal residential address histories to evaluate associations between time-varying environmental variables and health outcomes.


Sublimation not an innocent technique: a case of bis-cyclometalated iridium emitter for OLED.
Isomerization of a neutral bis-cyclometalated iridium(III) complex has been observed for the first time during the preparation of vacuum-processed organic light-emitting devices (OLEDs) and reproduced in solution. Isolation of the isomer revealed a cis organization of the two pyridine rings of the cyclometalating ligands. Photophysical studies show very similar emission properties of the two isomers. However, due to in situ isomerization, it is only possible to prepare vacuum-processed OLED devices having a mixture of isomers.


Alterations in circadian rhythms are associated with increased lipid peroxidation in females with bipolar disorder.
Disturbances in both circadian rhythms and oxidative stress systems have been implicated in the pathophysiology of bipolar disorder (BD), yet no studies have investigated the relationship between these systems in BD. We studied the impact of circadian rhythm disruption on lipid damage in 52 depressed or euthymic BD females, while controlling for age, severity of depressive symptoms and number of psychotropic medications, compared to 30 healthy controls. Circadian rhythm disruption was determined by a self-report measure (Biological Rhythm Interview of Assessment in Neuropsychiatry; BRIAN), which measures behaviours such as sleep, eating patterns, social rhythms and general activity. Malondialdehyde (MDA) levels were measured as a proxy of lipid peroxidation. We also measured the activity of total and extracellular superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST). Multiple linear regressions showed that circadian rhythm disturbance was independently associated with increased lipid peroxidation in females with BD (p < 0.05). We found decreased extracellular SOD (p < 0.05), but no differences in total SOD, CAT or GST activity between bipolar females and controls. Circadian rhythms were not associated with lipid peroxidation in healthy controls, where aging was the only significant predictor. These results suggest an interaction between the circadian system and redox metabolism, in that greater disruption in daily rhythms was associated with increased lipid peroxidation in BD only. Antioxidant enzymes have been shown to follow a circadian pattern of expression, and it is possible that disturbance of sleep and daily rhythms experienced in BD may result in decreased antioxidant defence and therefore increased lipid peroxidation. This study provides a basis for further investigation of the links between oxidative stress and circadian rhythms in the neurobiology of BD.


A pathophysiological view of the long non-coding RNA world.
Because cells are constantly exposed to micro-environmental changes, they require the ability to adapt to maintain a dynamic equilibrium. Proteins are considered critical for the regulation of gene expression, which is a fundamental process in determining the cellular responses to stimuli. Recently, revolutionary findings in RNA research and the advent of high-throughput genomic technologies have revealed a pervasive transcription of the human genome, which generates many long non-coding RNAs (lncRNAs) whose roles are largely undefined. However, there is evidence that lncRNAs are involved in several cellular physiological processes such as adaptation to stresses, cell differentiation, maintenance of pluripotency and apoptosis. The correct balance of lncRNA levels is crucial for the maintenance of cellular equilibrium, and the dysregulation of lncRNA expression is linked to many disorders; certain transcripts are useful prognostic markers for some of these pathologies. This review revisits the classic concept of cellular homeostasis from the perspective of lncRNAs specifically to understand how this novel class of molecules contributes to cellular balance and how its dysregulated expression can lead to the onset of pathologies such as cancer.


Sexual behavior of single adult American women.
Public policies promoting abstinence until marriage attempt to influence the sexual behavior of the more than 18 million American women who are currently single. An analysis of these women's behavior is needed to inform policies that are responsive to their sexual and reproductive health needs. Sexual behaviors, risk factors and reproductive health needs were examined among a nationally representative sample of 6,493 women aged 20-44 from the 2002 National Survey of Family Growth. Paired t tests were used to assess differences among single, married and cohabiting women by selected demographic, behavioral and risk measures. Thirty-six percent of women aged 20-44 are single, and nine in 10 single women are sexually experienced. Seventy percent of the latter women are currently sexually active; on average, they had intercourse in seven of the last 12 months. A higher proportion of single women (22%) than of cohabiting (9%) or married women (2%) have had two or more partners in the past year, and half of single women are at risk of unintended pregnancy. Furthermore, single women and cohabiting women are more likely to lack health insurance than are married women (21-25% vs. 12%). Because of the high level of sexual activity among single adult women, providers must address their reproductive health care needs and offer appropriate counseling and services. Government policies aimed at encouraging adult women to have sex only within marriage appear out of touch with the reality of the sexual behavior of single women.


Medical Eschatologies: The Christian Spirit of Hospital Protocol.
If much has been written of the forms of bodiliness reinforced by hospitals, less attention has been paid to the medicalization of the soul. The medical management of death institutionalizes divisions between body and soul, and matter and spirit, infusing end-of-life care with latent Christian theological presumptions. The invisibility of these presumptions is partly sustained by projecting religiosity on those who endorse other cosmologies, while retaining for medicine a mask of secular science. Stories of conflict with non-Christian patients force these presumptions into visibility, suggesting alternative ethics of care and mourning rooted in other understandings. In this article, I explore one such story. Considering the story as an allegory for how matter and spirit figure in contemporary postmortem disciplines, I suggest that it exposes both the operation of a taboo against mixing material and spiritual agendas, and an assumption that appropriate mourning is oriented toward symbolic homage, rather than concern for the material welfare of the dead.


Phase II trial of CoQ10 for ALS finds insufficient evidence to justify phase III.
Amyotrophic lateral sclerosis (ALS) is a devastating, and currently incurable, neuromuscular disease in which oxidative stress and mitochondrial impairment are contributing to neuronal loss. Coenzyme Q10 (CoQ10), an antioxidant and mitochondrial cofactor, has shown promise in ALS transgenic mice, and in clinical trials for neurodegenerative diseases other than ALS. Our aims were to choose between two high doses of CoQ10 for ALS, and to determine if it merits testing in a Phase III clinical trial. We designed and implemented a multicenter trial with an adaptive, two-stage, bias-adjusted, randomized, placebo-controlled, double-blind, Phase II design (n = 185). The primary outcome in both stages was a decline in the ALS Functional Rating Scale-revised (ALSFRSr) score over 9 months. Stage 1 (dose selection, 35 participants per group) compared CoQ10 doses of 1,800 and 2,700 mg/day. Stage 2 (futility test, 75 patients per group) compared the dose selected in Stage 1 against placebo. Stage 1 selected the 2,700 mg dose. In Stage 2, the pre-specified primary null hypothesis that this dose is superior to placebo was not rejected. It was rejected, however, in an accompanying prespecified sensitivity test, and further supplementary analyses. Prespecified secondary analyses showed no significant differences between CoQ10 at 2,700 mg/day and placebo. There were no safety concerns. CoQ10 at 2,700 mg daily for 9 months shows insufficient promise to warrant Phase III testing. Given this outcome, the adaptive Phase II design incorporating a dose selection and a futility test avoided the need for a much larger conventional Phase III trial.


Penetration Depth Measurement of Near-Infrared Hyperspectral Imaging Light for Milk Powder.
The increasingly common application of the near-infrared (NIR) hyperspectral imaging technique to the analysis of food powders has led to the need for optical characterization of samples. This study was aimed at exploring the feasibility of quantifying penetration depth of NIR hyperspectral imaging light for milk powder. Hyperspectral NIR reflectance images were collected for eight different milk powder products that included five brands of non-fat milk powder and three brands of whole milk powder. For each milk powder, five different powder depths ranging from 1 mm-5 mm were prepared on the top of a base layer of melamine, to test spectral-based detection of the melamine through the milk. A relationship was established between the NIR reflectance spectra (937.5-1653.7 nm) and the penetration depth was investigated by means of the partial least squares-discriminant analysis (PLS-DA) technique to classify pixels as being milk-only or a mixture of milk and melamine. With increasing milk depth, classification model accuracy was gradually decreased. The results from the 1-mm, 2-mm and 3-mm models showed that the average classification accuracy of the validation set for milk-melamine samples was reduced from 99.86% down to 94.93% as the milk depth increased from 1 mm-3 mm. As the milk depth increased to 4 mm and 5 mm, model performance deteriorated further to accuracies as low as 81.83% and 58.26%, respectively. The results suggest that a 2-mm sample depth is recommended for the screening/evaluation of milk powders using an online NIR hyperspectral imaging system similar to that used in this study.


Trehalose click polymers inhibit nanoparticle aggregation and promote pDNA delivery in serum.
Herein, three new glycopolymers have been synthesized via "click polymerization" to promote nucleic acid delivery in the presence of biological media containing serum. These structures were designed to contain a trehalose moiety to promote biocompatibility, water solubility, and stability against aggregation, amide-triazole groups to enhance DNA binding affinity, and an oligoamine unit to facilitate DNA encapsulation, phosphate neutralization, and interactions with cell surfaces. A 2,3,4,2',3',4'-hexa-O-acetyl-6,6'-diazido-6,6'-dideoxy-D-trehalose (4) monomer was polymerized via copper(I)-catalyzed azide-alkyne cycloaddition with a series of dialkyne-amide comonomers that contain either one, two, or three Boc-protected secondary amines (7a, 7b, or 7c, respectively). After deprotection, three water-soluble polycations (9a, 9b, or 9c) were obtained with similar degrees of polymerization (n = 56-61) to elucidate the role of amine number on nucleic acid binding, complex formation, stability, and cellular delivery. Gel electrophoresis and ethidium bromide experiments showed that 9a-9c associated with plasmid DNA (pDNA) and formed complexes (polyplexes) at N/P ratios dependent on the amine number. TEM experiments revealed that 9a-9c polyplexes were small (50-120 nm) and had morphologies (spherical and rodlike) associated with the polymer chain stiffness. Dynamic light scattering studies in the presence of media containing serum demonstrated that 9c polyplexes had a low degree of flocculation, whereas 9a and 9b polyplexesd aggregate rapidly. Further biological studies revealed that these structures were biocompatible and deliver pDNA into HeLa cells. Particularly, 9c polyplexes promoted high delivery efficacy and gene expression profiles in the presence of serum.


Causes, prevention and management of surgical site infection.
Superficial surgical site infection (SSI) usually resolves quickly causing only short-term discomfort. However, SSI involving deeper tissues is a leading cause of morbidity and mortality and increases the overall cost of care. Although SSI commonly originates in the operating theatre, signs and symptoms of infection often do not appear until patients have been discharged from hospital. Nurses in a range of clinical settings are involved in the care of patients who either develop or are at risk of developing SSI and therefore need to know how to advise and manage these patients.


Effects of atrial natriuretic factor on hormone-induced renin release.
The purpose of this study was to determine whether or not atrial natriuretic factor can act directly on the juxtaglomerular cell in vivo to inhibit hormone-induced renin release. To achieve this objective the interaction between synthetic atrial natriuretic factor and two different renin secretagogues was examined. To exclude any indirect effect of atrial natriuretic factor on renin release due to changes in sodium delivery to the macula densa, all studies were conducted in nonfiltering, canine kidneys. In one series of studies renin release was stimulated by intrarenal infusions of norepinephrine (3 micrograms/kg/min), and in a second series of studies renin release was induced by intrarenal infusions of prostacyclin (0.1 micrograms/kg/min). In both studies intrarenal infusions of atrial natriuretic factor (0.3 micrograms/kg/min), which provided supraphysiological levels of atrial natriuretic factor in the renal arterial plasma, failed to attenuate hormone-induced renin release. In contrast, adenosine, a well-known inhibitor of renin release, abolished the renin release response to both hormones. These data indicate that, at the dose used in this study, synthetic atrial natriuretic factor does not act directly on the juxtaglomerular cell to attenuate hormone-induced renin release. Further, these results imply that circulating endogenous atrial natriuretic factor cannot directly attenuate juxtaglomerular cell responsiveness.


[Malocclusion prevalence and orthodontic treatment need in 10-14-year-old schoolchildren in Belo Horizonte, Minas Gerais State, Brazil: a psychosocial focus].
The objectives of this cross-sectional study were to assess the prevalence of malocclusion and to verify the association between normative orthodontic treatment need and certain psychosocial aspects. The sample (n = 333) was representative of the population of adolescents (10-14 years) in Belo Horizonte, Minas Gerais, Brazil. The dependent variable "normative orthodontic treatment need" was evaluated through the Dental Aesthetic Index (DAI) and the independent variables (gender, age, mother's schooling, economic status, parent's perception of the child's oral aesthetics, parent's perception of the child's orthodontic treatment need, and the adolescent's desire for orthodontic treatment) were evaluated through questionnaires. Descriptive, bivariate, and logistic regression (stepwise forward procedure, p <or= 0.05) analyses were performed. Malocclusion prevalence was 62.0% and normative orthodontic treatment need was 52.2%. The variables adolescent's orthodontic treatment desire and parents' perception of the child's oral aesthetics were significantly associated with normative orthodontic treatment need. These results suggest that psychosocial factors should be considered in making the decision to undergo orthodontic treatment.


Spatio-temporal patterns of genetic variation in Arbacia lixula, a thermophilous sea urchin in expansion in the Mediterranean.
The genetic structure of 13 populations of the amphiatlantic sea urchin Arbacia lixula, as well as temporal genetic changes in three of these localities, were assessed using ten hypervariable microsatellite loci. This thermophilous sea urchin is an important engineer species triggering the formation of barren grounds through its grazing activity. Its abundance seems to be increasing in most parts of the Mediterranean, probably favoured by warming conditions. Significant genetic differentiation was found both spatially and temporally. The main break corresponded to the separation of western Atlantic populations from those in eastern Atlantic and the Mediterranean Sea. A less marked, but significant differentiation was also found between Macaronesia (eastern Atlantic) and the Mediterranean. In the latter area, a signal of differentiation between the transitional area (Alboran Sea) and the rest of the Mediterranean was detected. However, no genetic structure is found within the Mediterranean (excluding Alboran) across the Siculo-Tunisian Strait, resulting from either enough gene flow to homogenize distance areas or/and a recent evolutionary history marked by demographic expansion in this basin. Genetic temporal variation at the Alboran Sea is as important as spatial variation, suggesting that temporal changes in hydrological features can affect the genetic composition of the populations. A picture of genetic homogeneity in the Mediterranean emerges, implying that the potential expansion of this keystone species will not be limited by intraspecific genetic features and/or potential impact of postulated barriers to gene flow in the region.


Ultrathin palladium nanosheets with selectively controlled surface facets.
We report a facile bottom-up synthetic approach to preparing ultrathin two-dimensional (2D) palladium nanosheets (PdNSs) with selectively exposed surface facets. Our synthetic strategy is based on the utilization of the nanoconfined lamellar mesophases of amphiphilic functional surfactants to template the growth of PdNSs in aqueous solution. Preferential adsorption of functional groups (e.g., COOH, pyridyl and quaternary ammonium) and halide counter ions (e.g., Br- and Cl-) in the long-chain surfactants onto different Pd planes results in the epitaxial growth of {100}, {110} and {111}-exposed surface facets of ultrathin PdNSs. Our synthetic approach is a general, powerful and scalable method to precisely control the surface facets of ultrathin 2D PdNSs, thus providing an opportunity to evaluate facet-dependent catalytic performance of Pd nanocrystals. Ultrathin PdNSs have been examined as the electrocatalysts for hydrogen evolution reactions (HERs). We show that {100}-exposed PdNSs display superior catalytic activity and stability for HERs, compared to that of {110} and {111}-exposed ones as well as their bulk counterparts. Conceivably, our findings will offer a general guideline in rational design of surfactant templates for other 2D metal nanosheets.


Craniofacial landmarks in young children: how reliable are measurements based on 3-dimensional imaging?
Different approaches for 3-dimensional (3D) data acquisition of the facial surface are common nowadays. Meticulous evaluation has proven their level of precision and accuracy. However, the question remains as to which level of craniofacial landmarks, especially in young children, are reliable if identified in 3D images. Potential sources of error, aside from the systems technology itself, need to be identified and addressed. Reliable and unreliable landmarks have to be identified. The 3dMDface System was used in a clinical setting to evaluate the intraobserver repeatability of 27 craniofacial landmarks in 7 young children between 6 and 18 months of age with a total of 1134 measurements. The handling of the system was mostly unproblematic. The mean 3D repeatability error was 0.82 mm, with a range of 0.26 mm to 2.40 mm, depending on the landmark. Single landmarks that have been shown to be relatively imprecise in 3D analysis could still provide highly accurate data if only 1 of the 3 spatial planes was relevant. There were no statistical differences from 1 patient to another. Reliability in craniofacial measurements can be achieved by such 3D soft-tissue imaging techniques as the 3dMDface System, but one must always be aware that the degree of precision is strictly dependent on the landmark and axis in question.For further clinical investigations, the degree of reliability for each landmark evaluated must be addressed and taken into account.


The relationship between sleep disturbance, symptoms and daytime functioning in psoriasis: a prospective study integrating actigraphy and experience sampling methodology.
Sleep disturbance is common in individuals with psoriasis and appears to be related to both physical and psychological factors. We sought to examine whether psoriasis symptoms, night-time arousal and low mood predicted subsequent objective and self-reported sleep; and whether objective and self-reported sleep predicted next-day psoriasis symptoms and day-time functioning. A total of 19 individuals (Female: 11 [57.9%], median age: 39 years) with chronic plaque psoriasis and poor sleep quality (mean Pittsburgh Sleep Quality Index, PSQI = 9.11) participated. Momentary assessments of psoriasis symptoms, mood and daytime functioning were completed at five pseudo-random intervals each day for 15 days using time-stamped digital diary entry. Objective sleep was estimated using wrist-worn actigraphy. Self-reported sleep and night-time arousal were assessed each morning using validated measures. Two-level random intercept models showed that increased night-time arousal was associated with poorer diary-reported sleep. Neither self-reported nor objective sleep parameters were associated with daytime psoriasis symptoms in bi-directional analyses. Diary-reported sleep predicted next-day functioning, specifically sleepiness, concentration, and fatigue. Actigraphy-defined total sleep time predicted next-day fatigue. Night-time arousal is associated with poorer self-reported sleep in people with psoriasis, and sleep predicts next-day functioning. Contrary to our hypothesis, sleep disturbance does not appear to be associated with momentary assessments of psoriasis symptoms.


Image-Based Morphological Profiling Identifies a Lysosomotropic, Iron-Sequestering Autophagy Inhibitor.
Chemical proteomics is widely applied in small-molecule target identification. However, in general it does not identify non-protein small-molecule targets, and thus, alternative methods for target identification are in high demand. We report the discovery of the autophagy inhibitor autoquin and the identification of its molecular mode of action using image-based morphological profiling in the cell painting assay. A compound-induced fingerprint representing changes in 579 cellular parameters revealed that autoquin accumulates in lysosomes and inhibits their fusion with autophagosomes. In addition, autoquin sequesters Fe2+ in lysosomes, resulting in an increase of lysosomal reactive oxygen species and ultimately cell death. Such a mechanism of action would have been challenging to unravel by current methods. This work demonstrates the potential of the cell painting assay to deconvolute modes of action of small molecules, warranting wider application in chemical biology.


Online Support and Intervention for Child Anxiety (OSI): Development and Usability Testing.
Internet-based treatments for child anxiety may help to increase access to evidence-based therapies; however, user engagement, uptake, and adherence within routine clinical practice remain as challenges. Involving the intended end users in the development process through user-centered design and usability testing is crucial for maximizing user engagement and adoption of internet-based treatments, but so far this has been lacking for internet-based treatments for child anxiety. The aim of this study is to develop an internet-based treatment for child anxiety through a process of user-centered design (phase 1) and usability testing (phase 2), based on an existing evidence-based, face-to-face, therapist-supported, parent-led cognitive behavioral therapy intervention. It is intended that the internet-based version of this treatment would consist of a parent website, case management system for clinicians, and mobile game app for children. Parents, children, and clinicians who were familiar with the face-to-face version of the treatment were recruited from 2 National Health Service clinics. In phase 1, participants participated in 3 workshops to gain feedback on the overall concept, explore their wants and needs for the websites and game, generate ideas on how the treatment may look, and gain feedback on initial mock-ups of the websites and game. In phase 2, participants attended 3 individual usability testing sessions where they were presented with working prototypes of the website or game and asked to perform a series of tasks on the website (parents and clinicians) or play the game (children). The frequency and details on usability errors were recorded. Participants were asked for their feedback on the website and game using a standardized usability questionnaire and semistructured interviews. The websites and game were iterated after each round of usability testing in response to this feedback. In phase 1, participants approved the general concept and rated the initial mock-ups of the website and game positively. In phase 2, working prototypes were rated positively and usability errors declined across the iterations and were mainly cosmetic or minor issues relating to esthetic preference, with few issues regarding ability to navigate the website or technical issues affecting functionality. Feedback from the semistructured interviews further supported the positive response of participants to the website and game, and helped identify areas for improvement during the iteration process. The final iteration of the website and game are presented. Taking an iterative approach to development through user-centered design and usability testing has resulted in an internet-based treatment for child anxiety (Online Support and Intervention for child anxiety) that appears to meet the needs and expectations of the intended users (parents, children, and clinicians) and is easy and enjoyable to use.


Comparing Next-Generation Robotic Technology with 3-Dimensional Computed Tomography Navigation Technology for the Insertion of Posterior Pedicle Screws.
To study the differences between robot-guided (Mazor X, Mazor Robotics Ltd., Caesarea, Israel) and 3-dimensional (3D) computed tomography (CT) navigation (O-arm Surgical Imaging System, Medtronic, Minneapolis, Minnesota, USA) for the insertion of pedicle screws. We reviewed the charts of 50 patients who underwent robot-guided pedicle screw insertion (between May 2017-October 2017), and 49 patients who underwent 3D-CT navigation pedicle screw insertion (between September 2015-August 2016). Variables included were age, sex, body mass index, blood loss, length of stay, lumbar level(s), operation time, fluoroscopy time, radiation dose, accuracy, and time-per-screw placement. Mean ages were 59.3 years in the robotic group and 58.2 years in the 3D-CT navigation group. Mean was 30.7 kg/m2 in the robotic group and 32.1 kg/m2 in the 3D-CT navigation group. Mean time-per-screw placement was 3.7 minutes for the robotic group and 6.8 minutes for the 3D-CT navigation group, P < 0.001. In the robotic group, 189 of 190 screws were placed with Ravi grade I accuracy, and 1 was grade II. In the 3D-CT navigation group, 157 of 165 screws were Ravi grade I, and 8 were grade II (P = 0.11). Fluoroscopy time (P < 0.001), time-per-screw placement (P < 0.001), and length of stay (P < 0.001) were significantly lower in the robotic group. Both technologies are safe and accurate. Robotic technology exposed patients to less fluoroscopy time, decreased time-per-screw placement and shorter hospital stay than 3D-CT navigation. Further studies are warranted to verify our results.


Profiling of UGT1A1*6, UGT1A1*60, UGT1A1*93, and UGT1A1*28 Polymorphisms in Indonesian Neonates With Hyperbilirubinemia Using Multiplex PCR Sequencing.
Background: Single nucleotide polymorphism (SNP) variants of the uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) gene have been studied as an important factor in neonatal hyperbilirubinemia (jaundice) severity. Specific ethnicities, including Asians, have certain SNPs that appear more frequently than others. Aim: To identify the most common SNPs in Indonesian neonates and their association with the severity of neonatal hyperbilirubinemia. Methods: Eighty-eight inborn and outborn jaundiced infants from three different hospitals (Bengkulu, Jakarta, Biak Papua) across Indonesia were enrolled in this cross-sectional study and their peak total serum bilirubin (TSB) levels assessed. SNP variant analyses of the TATAA box, promoter, and exon 1 regions of UGT1A1 gene from 78 of the 88 infants were carried out using the SNaPshotR Multiplex Polymerase Chain Reaction (PCR) System followed by DNA sequencing. Results: We detected SNP variants UGT1A1 * 28, UGT1A1 * 60, UGT1A1 * 93, and UGT1A1 * 6 in our population. Mean total serum bilirubin (TSB) was 14.59 ± 5.57 mg/dL. Bivariate analyses using delivery location, gestational age, birth weight, mother's age, and ethnicity were shown to be associated with moderate-to-severe hyperbilirubinemia (p < 0.05). None of the four SNPs appeared to be associated with moderate-to-severe hyperbilirubinemia. In multivariate analysis, however, only the "other ethnic group" (e.g., Chinese, Bengkulu, Papua, Bima) category showed an association with moderate-to-severe hyperbilirubinemia, with an odds ratio of 6.49 (95% CI 1.01-41.67; p < 0.05). Conclusions: We found that the UGT1A1 * 60 is the most common SNP detected in neonates with hyperbilirubinemia in the Indonesian population. Interestingly, in Indonesia, UGT1A1 polymorphisms do not appear to be associated with differences in the severity of hyperbilirubinemia.


Evaluation of left ventricular ejection fraction in non-ST-segment elevation acute coronary syndromes and its relationship to treatment.
In-hospital assessment of left ventricular ejection fraction (LVEF) in non-ST-segment elevation acute coronary syndrome (NSTE-ACS) is emphasized in current practice guidelines. There are limited data regarding the evaluation of LVEF and clinical characteristics and in-hospital management in the "real world." Registries including the Canadian Acute Coronary Syndrome (ACS) I and II, Global Registry of Acute Coronary Events (main GRACE/expanded GRACE(2)), and Canadian Registry of Acute Coronary Events (CANRACE) enrolled 13,703 NSTE-ACS patients across Canada between 1999 and 2008. Patients were stratified by in-hospital LVEF measurement, and LVEF was categorized as normal, mildly, or moderately to severely impaired. We compared clinical characteristics, cardiac procedures, and clinical outcomes across these groups. Multivariable logistic regression identified factors independently associated with the assessment of LVEF. Overall, 8,116 patients (59.2%) had LVEF measurement, and of the 7,667 patients with available LVEF data, 4,470 (58.3%) had normal, 1,916 (25%) mildly impaired, and 1,281 (16.7%) moderately to severely impaired LVEF. Patients with LVEF assessment more frequently (all P < .001) underwent cardiac catheterization, percutaneous coronary intervention or coronary bypass surgery, and had higher (both P < .001) rates of myocardial (re) infarction and heart failure. In-hospital reinfarction, higher Killip class, abnormal biomarker, hospital stay >10 days, and on-site cardiac catheterization facility were independently associated with LVEF assessment. Despite increasing LVEF assessment over time (P for trend < .001), 31.2% of patients in the most recent registry (2008) had no in-hospital LVEF assessment. In-hospital LVEF assessment is not performed in many NSTE-ACS patients. The LVEF assessment, associated with increased use of evidence-based therapies and invasive cardiac procedures, was obtained more frequently in patients with myocardial (re) infarction, heart failure on presentation, and prolonged hospital stay.


α-Synuclein occurs physiologically as a helically folded tetramer that resists aggregation.
Parkinson's disease is the second most common neurodegenerative disorder. Growing evidence indicates a causative role of misfolded forms of the protein α-synuclein in the pathogenesis of Parkinson's disease. Intraneuronal aggregates of α-synuclein occur in Lewy bodies and Lewy neurites, the cytopathological hallmarks of Parkinson's disease and related disorders called synucleinopathies. α-Synuclein has long been defined as a 'natively unfolded' monomer of about 14 kDa (ref. 6) that is believed to acquire α-helical secondary structure only upon binding to lipid vesicles. This concept derives from the widespread use of recombinant bacterial expression protocols for in vitro studies, and of overexpression, sample heating and/or denaturing gels for cell culture and tissue studies. In contrast, we report that endogenous α-synuclein isolated and analysed under non-denaturing conditions from neuronal and non-neuronal cell lines, brain tissue and living human cells occurs in large part as a folded tetramer of about 58 kDa. Several methods, including analytical ultracentrifugation, scanning transmission electron microscopy and in vitro cell crosslinking confirmed the occurrence of the tetramer. Native, cell-derived α-synuclein showed α-helical structure without lipid addition and had much greater lipid-binding capacity than the recombinant α-synuclein studied heretofore. Whereas recombinantly expressed monomers readily aggregated into amyloid-like fibrils in vitro, native human tetramers underwent little or no amyloid-like aggregation. On the basis of these findings, we propose that destabilization of the helically folded tetramer precedes α-synuclein misfolding and aggregation in Parkinson's disease and other human synucleinopathies, and that small molecules that stabilize the physiological tetramer could reduce α-synuclein pathogenicity.


Do patients with OCD and pathological gambling have similar dysfunctional cognitions?
The obsessive-compulsive spectrum disorder (OCSD) theory postulates that a wide range of disorders is closely related to OCD. Current cognitive models ascertain that certain beliefs leading to misinterpretation of the significance of intrusions are important in the etiology and maintenance of OCD. This study examined whether pathological gambling, a disorder belonging to the OC spectrum, is characterized by similar dysfunctional cognitions as OCD. Dysfunctional beliefs of OCD patients were compared to those of patients with pathological gambling, panic disorder and normal controls. These beliefs were measured by the Obsessive-compulsive Beliefs Questionnaire-87 (OBQ-87), which was developed by a group of leading OCD researchers [Behav. Res. Ther. 35 (1997) 667]. It was hypothesized that according to the OCSD theory, pathological gamblers would exhibit similar cognitions to OCD patients, as well as increased levels of OCD symptoms. Analysis showed that OCD patients exhibited higher OBQ-87 scores than both panic patients and normal controls, but equal to pathological gambling patients. Pathological gamblers exhibited, however, no increase in OCD symptoms. These mixed results do not seem to support the OC spectrum theory for pathological gambling, moreover being contradictory to contemporary cognitive OCD models.


Differential glycolytic profile and Warburg effect in papillary thyroid carcinoma cell lines.
Acceleration of glycolysis is a characteristic of neoplasia. Previous studies have shown that a metabolic shift occurs in many tumors and correlates with a negative prognosis. The present study aimed to investigate the glycolytic profile of thyroid carcinoma cell lines. We investigated glycolytic and oxidative parameters of two thyroid carcinoma papillary cell lines (BCPAP and TPC1) and the non-tumor cell line NTHY-ori. All carcinoma cell lines showed higher rates of glycolysis efficiency, when compared to NTHY-ori, as assessed by a higher rate of glucose consumption and lactate production. The BCPAP cell line presented higher rates of growth, as well as elevated intracellular ATP levels, compared to the TPC1 and NTHY-ori cells. We found that glycolysis and activities of pentose phosphate pathway (PPP) regulatory enzymes were significantly different among the carcinoma cell lines, particularly in the mitochondrial hexokinase (HK) activity which was higher in the BCPAP cells than that in the TPC1 cell line which showed a balanced distribution of HK activity between cytoplasmic and mitochondrial subcellular localizations. However, TPC1 had higher levels of glucose‑6-phosphate dehydrogenase activity, suggesting that the PPP is elevated in this cell type. Using high resolution respirometry, we observed that the Warburg effect was present in the BCPAP and TPC1 cells, characterized by low oxygen consumption and high reactive oxygen species production. Overall, these results indicate that both thyroid papillary carcinoma cell lines showed a glycolytic profile. Of note, BCPAP cells presented some relevant differences in cell metabolism compared to TPC1 cells, mainly related to higher mitochondrial-associated HK activity.


Nonlinear optical detection of terahertz-wave radiation from resonant tunneling diodes.
The sensitive detection of terahertz (THz)-wave radiation from compact sources at room temperature is crucial for real-world THz-wave applications. Here, we demonstrate the nonlinear optical detection of THz-wave radiation from continuous-wave (CW) resonant tunneling diodes (RTDs) at 0.58, 0.78, and 1.14 THz. The up-conversion process in a MgO:LiNbO3 crystal under the noncollinear phase-matching condition offers efficient wavelength conversion from a THz wave to a near-infrared (NIR) wave that is detected using a commercial NIR photodetector. The minimum detection limit of CW THz-wave power is as low as 5 nW at 1.14 THz, corresponding to 2-aJ energy and 2.7 × 103 photons within the time window of a 0.31-ns pump pulse. Our results show that the input frequency and power of RTD devices can be calibrated by measuring the output wavelength and energy of up-converted waves, respectively. This optical detection technique for compact electronic THz-wave sources will open up a new opportunity for the realization of real-world THz-wave applications.


Microbe-microbe and host-microbe interactions drive microbiome dysbiosis and inflammatory processes.
An extensive microbiome comprised of bacteria, viruses, bacteriophages, and fungi is now understood to persist in nearly every human body site, including tissue and blood. The genomes of these microbes continually interact with the human genome in order to regulate host metabolism. Many components of this microbiome are capable of both commensal and pathogenic activity. They are additionally able to persist in both 'acute' and chronic forms. Inflammatory conditions historically studied separately (autoimmune, neurological and malignant) are now repeatedly tied to a common trend: imbalance or dysbiosis of these microbial ecosystems. Population-based studies of the microbiome can shed light on this dysbiosis. However, it is the collective activity of the microbiome that drives inflammatory processes via complex microbe-microbe and host-microbe interactions. Many microbes survive as polymicrobial entities in order to evade the immune response. Pathogens in these communities alter their gene expression in ways that promote community-wide virulence. Other microbes persist inside the cells of the immune system, where they directly interfere with host transcription, translation, and DNA repair mechanisms. The numerous proteins and metabolites expressed by these pathogens further dysregulate human gene expression in a manner that promotes imbalance and immunosuppression. Molecular mimicry, or homology between host and microbial proteins, complicates the nature of this interference. When taken together, these microbe-microbe and host-microbe interactions are capable of driving the large-scale failure of human metabolism characteristic of many different inflammatory conditions.


The haggis tolerance test in Scots and Sassenachs.
To find out if the Scottish national dish, haggis, contributes to the high incidence of coronary heart disease in Scotland the lipaemic effect of a meal of 200 g of haggis was measured in six Scottish and 10 Sassenach men. The Scots had higher fasting cholesterol and triglyceride concentrations and a lower proportion of high density lipoprotein cholesterol than the Sassenachs. Four subjects were found to have hyperlipoproteinaemia, which had been unrecognised previously. Serum cholesterol concentrations did not change after haggis was eaten (mean dose 2.6 g/kg body weight). Serum concentrations of triglycerides increased by 51% at 90 minutes in the Sassenachs but were unaltered in the Scots. There were no serious adverse effects. This study shows that Scots have higher lipid concentrations than Sassenachs but seem to be resistant to the lipaemic effect of haggis. The haggis tolerance test may be useful in Sassenachs.


Evidence for a dominant-negative effect in ACTA1 nemaline myopathy caused by abnormal folding, aggregation and altered polymerization of mutant actin isoforms.
We have studied a cohort of nemaline myopathy (NM) patients with mutations in the muscle alpha-skeletal actin gene (ACTA1). Immunoblot analysis of patient muscle demonstrates increased gamma-filamin, myotilin, desmin and alpha-actinin in many NM patients, consistent with accumulation of Z line-derived nemaline bodies. We demonstrate that nebulin can appear abnormal secondary to a primary defect in actin, and show by isoelectric focusing that mutant actin isoforms are present within insoluble actin filaments isolated from muscle from two ACTA1 NM patients. Transfection of C2C12 myoblasts with mutant actin(EGFP) constructs resulted in abnormal cytoplasmic and intranuclear actin aggregates. Intranuclear aggregates were observed with V163L-, V163M- and R183G-actin(EGFP) constructs, and modeling shows these residues to be adjacent to the nuclear export signal of actin. V163L and V163M actin mutants are known to cause intranuclear rod myopathy, however, intranuclear bodies were not reported in patient R183G. Transfection studies in C2C12 myoblasts showed significant alterations in the ability of V136L and R183G actin mutants to polymerize and contribute to insoluble actin filaments. Thus, we provide direct evidence for a dominant-negative effect of mutant actin in NM. In vitro studies suggest that abnormal folding, altered polymerization and aggregation of mutant actin isoforms are common properties of NM ACTA1 mutants. Some of these effects are mutation-specific, and likely result in variations in the severity of muscle weakness seen in individual patients. A combination of these effects contributes to the common pathological hallmarks of NM, namely intranuclear and cytoplasmic rod formation, accumulation of thin filaments and myofibrillar disorganization.


Lissamphibian origins: possible protolissamphibian from the lower permian of oklahoma.
A new genus and family of rhachitomous labyrinthodont amphibian, related to the Upper Paleozoic Dissorophidae and Trematopsidae, may be ancestral to some or all of the modern Amphibia. Doleserpeton occurs in Lower Permian fissure fill deposits in southwestern Oklahoma. It is unique among nonlissamphibian tetrapods in that it possesses pedicellate, bicuspid teeth together with nearly monospondylous vertebrae in which the main central element is a pleurocentrum. Doleserpeton may have been utilizing the food resources of the upland, terrestrial environment in a novel fashion for rhachitomes.


Vaccination with NY-ESO-1 protein and CpG in Montanide induces integrated antibody/Th1 responses and CD8 T cells through cross-priming.
The use of recombinant tumor antigen proteins is a realistic approach for the development of generic cancer vaccines, but the potential of this type of vaccines to induce specific CD8(+) T cell responses, through in vivo cross-priming, has remained unclear. In this article, we report that repeated vaccination of cancer patients with recombinant NY-ESO-1 protein, Montanide ISA-51, and CpG ODN 7909, a potent stimulator of B cells and T helper type 1 (Th1)-type immunity, resulted in the early induction of specific integrated CD4(+) Th cells and antibody responses in most vaccinated patients, followed by the development of later CD8(+) T cell responses in a fraction of them. The correlation between antibody and T cell responses, together with the ability of vaccine-induced antibodies to promote in vitro cross-presentation of NY-ESO-1 by dendritic cells to vaccine-induced CD8(+) T cells, indicated that elicitation of NY-ESO-1-specific CD8(+) T cell responses by cross-priming in vivo was associated with the induction of adequate levels of specific antibodies. Together, our data provide clear evidence of in vivo cross-priming of specific cytotoxic T lymphocytes by a recombinant tumor antigen vaccine, underline the importance of specific antibody induction for the cross-priming to occur, and support the use of this type of formulation for the further development of efficient cancer vaccines.


[Structural analysis of the PKR-binding region of HCV 1b samples from patients with chronic hepatitis C and the correlation with IFN-sensitivity].
91.5% of all chronic HCV infections are of 1b-type in Hungary. Japanese researchers found a correlation between the outcome of interferon (IFN) therapy and the structure of the PKR-Binding Region (aa: 2209-2274) of the viral NS5A domain, especially a particular subsection of the PKR-BR, the Interferon Sensitivity Determining Region (ISDR: aa 2209-2248). Several international studies could not confirm these findings. The objectives of this study were 1. to determine the Hungarian prototype of HCV 1b based on the nucleotide sequence analysis of the PKR-BR of HCV 1b samples from patients with chronic hepatitis C; and 2. to investigate the relationship between the phenotypically expressed mutations of ISDR and the response to IFN therapy. Pre-treatment serum samples of 21 chronic hepatitis C patients (13 women, 8 men), infected with HCV 1b and treated with IFN-alpha (3 sustained responders, 18 non-responders), were analysed retrospectively. Nested reverse transcriptase-polymerase chain reaction (RT-PCR) and direct nucleotide sequencing were applied. 1. The dominant Hungarian quasispecies of HCV 1b differs from the Japan HCV 1b-J, the internationally accepted prototype. 2. The results showed significant correlation between the type of ISDR and the interferon response. Mutant type ISDRs (4 > or = amino acid substitutions) have predictive value for sustained response (p = 0.012). 3. The types and locations of substitutions are not characteristic, except that arginine in position 2218 has predictive value for therapy resistance. The authors' study results are in accordance with the earlier Japanese findings and confirm the predictive value of pre-treatment nucleotide sequencing in Hungary. Full adaptation of international research results to a Hungarian context should be treated with caution due to the between-countries virus prototype differences.


Working definitions, subjective and objective assessments and experimental paradigms in a study exploring social withdrawal in schizophrenia and Alzheimer's disease.
Social withdrawal is one of the first and common signs of early social dysfunction in a number of important neuropsychiatric disorders, likely because of the enormous amount and complexity of brain processes required to initiate and maintain social relationships (Adolphs, 2009). The Psychiatric Ratings using Intermediate Stratified Markers (PRISM) project focusses on the shared and unique neurobiological basis of social withdrawal in schizophrenia, Alzheimer and depression. In this paper, we discuss the working definition of social withdrawal for this study and the selection of objective and subjective rating scales to assess social withdrawal chosen or adapted for this project. We also discuss the MRI and EEG paradigms selected to study the systems and neural circuitry thought to underlie social functioning and more particularly to be involved in social withdrawal in humans, such as the social perception and the social affiliation networks. A number of behavioral paradigms were selected to assess complementary aspects of social cognition. Also, a digital phenotyping method (a smartphone application) was chosen to obtain real-life data.


Reading front to back: MEG evidence for early feedback effects during word recognition.
Magnetoencephalography studies in humans have shown word-selective activity in the left inferior frontal gyrus (IFG) approximately 130 ms after word presentation ( Pammer et al. 2004; Cornelissen et al. 2009; Wheat et al. 2010). The role of this early frontal response is currently not known. We tested the hypothesis that the IFG provides top-down constraints on word recognition using dynamic causal modeling of magnetoencephalography data collected, while subjects viewed written words and false font stimuli. Subject-specific dipoles in left and right occipital, ventral occipitotemporal and frontal cortices were identified using Variational Bayesian Equivalent Current Dipole source reconstruction. A connectivity analysis tested how words and false font stimuli differentially modulated activity between these regions within the first 300 ms after stimulus presentation. We found that left inferior frontal activity showed stronger sensitivity to words than false font and a stronger feedback connection onto the left ventral occipitotemporal cortex (vOT) in the first 200 ms. Subsequently, the effect of words relative to false font was observed on feedforward connections from left occipital to ventral occipitotemporal and frontal regions. These findings demonstrate that left inferior frontal activity modulates vOT in the early stages of word processing and provides a mechanistic account of top-down effects during word recognition.