Huntington's disease-like disorders in Latin America and the Caribbean.
Diseases with a choreic phenotype can be due to a variety of genetic etiologies. As testing for Huntington's disease (HD) becomes more available in previously resource-limited regions, it is becoming apparent that there are patients in these areas with other rare genetic conditions which cause an HD-like phenotype. Documentation of the presence of these conditions is important in order to provide appropriate diagnostic and clinical care for these populations. Information for this article was gathered in two ways; the literature was surveyed for publications reporting a variety of genetic choreic disorders, and movement disorders specialists from countries in Latin America and the Caribbean were contacted regarding their experiences with chorea of genetic etiology. Here we discuss the availability of molecular diagnostics for HD and for other choreic disorders, along with a summary of the published reports of affected subjects, and authors' personal experiences from the regions. While rare, patients affected by non-HD genetic choreas are evidently present in Latin America and the Caribbean. HD-like 2 is particularly prevalent in countries where the population has African ancestry. The incidence of other conditions is likely determined by other variations in ethnic background and settlement patterns. As genetic resources and awareness of these disorders improve, more patients are likely to be identified, and have the potential to benefit from education, support, and ultimately molecular therapies.


Next-generation hypomethylating agent SGI-110 primes acute myeloid leukemia cells to IAP antagonist by activating extrinsic and intrinsic apoptosis pathways.
Therapeutic efficacy of first-generation hypomethylating agents (HMAs) is limited in elderly acute myeloid leukemia (AML) patients. Therefore, combination strategies with targeted therapies are urgently needed. Here, we discover that priming with SGI-110 (guadecitabine), a next-generation HMA, sensitizes AML cells to ASTX660, a novel antagonist of cellular inhibitor of apoptosis protein 1 and 2 (cIAP1/2) and X-linked IAP (XIAP). Importantly, SGI-110 and ASTX660 synergistically induced cell death in a panel of AML cell lines as well as in primary AML samples while largely sparing normal CD34+ human progenitor cells, underlining the translational relevance of this combination. Unbiased transcriptome analysis revealed that SGI-110 alone or in combination with ASTX660 upregulated the expression of key regulators of both extrinsic and intrinsic apoptosis signaling pathways such as TNFRSF10B (DR5), FAS, and BAX. Individual knockdown of the death receptors TNFR1, DR5, and FAS significantly reduced SGI-110/ASTX660-mediated cell death, whereas blocking antibodies for tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) or FAS ligand (FASLG) failed to provide protection. Also, TNFα-blocking antibody Enbrel had little protective effect on SGI-110/ASTX660-induced cell death. Further, SGI-110 and ASTX660 acted in concert to promote cleavage of caspase-8 and BID, thereby providing a link between extrinsic and intrinsic apoptotic pathways. Consistently, sequential treatment with SGI-110 and ASTX660-triggered loss of mitochondrial membrane potential (MMP) and BAX activation which contributes to cell death, as BAX silencing significantly protected from SGI-110/ASTX660-mediated apoptosis. Together, these events culminated in the activation of caspases-3/-7, nuclear fragmentation, and cell death. In conclusion, SGI-110 and ASTX660 cooperatively induced apoptosis in AML cells by engaging extrinsic and intrinsic apoptosis pathways, highlighting the therapeutic potential of this combination for AML.


Association between SDF1-3'A or CXCR4 gene polymorphisms with predisposition to and clinicopathological characteristics of prostate cancer with or without metastases.
In the present study, we aimed to investigate the association between SDF1-3'A and CXCR4 gene polymorphisms and the susceptibility and clinicopathological development of prostate cancer. SDF1-3'A and CXCR4 gene polymorphisms were assessed by polymerase chain reaction restriction-fragment length polymorphism (PCR-RFLP) in 149 healthy subjects and 152 patients with prostate cancer. There were no significant differences in the distributions of SDF-1 and CXCR4 genotypes between controls and prostate cancer patients. However, the patients with AA genotype of SDF1-3'A gene presented a higher risk for developing an advanced disease status as compared to patients with GG homozygotes (aOR = 2.02; 95 % CI = 1.05-3.90; P = 0.035). In addition, the distribution of AA genotype of SDF1-3'A gene was found significantly increased in the patients with bone metastasis in comparison to those without bone metastasis (aOR = 2.94; 95 % CI = 1.26-6.82; P = 0.012). On the other hand, CXCR4 gene polymorphism was not associated with the clinicopathological characteristics of prostate cancer. Our results suggest that SDF1-3'A and CXCR4 gene polymorphisms may not be risk factors for the susceptibility to prostate cancer. However, SDF1-3'A gene polymorphism may be associated with the progression and bone metastasis of prostate cancer in a Turkish men population.


Genomic profiling defines subtypes of prostate cancer with the potential for therapeutic stratification.
The remarkable variation in prostate cancer clinical behavior represents an opportunity to identify and understand molecular features that can be used to stratify patients into clinical subgroups for more precise outcome prediction and treatment selection. Significant progress has been made in recent years in establishing the composition of genomic and epigenetic alterations in localized and advanced prostate cancers using array-based technologies and next-generation sequencing approaches. The results of these efforts shed new light on our understanding of this disease and point to subclasses of prostate cancer that exhibit distinct vulnerabilities to therapeutics. The goal of this review is to categorize the genomic data and, where available, corresponding expression, functional, or related therapeutic information, from recent large-scale and in-depth studies that show a new appreciation for the molecular complexity of this disease. We focus on how these results inform our growing understanding of the mechanisms that promote genetic instability, as well as routes by which specific genes and biologic pathways may serve as biomarkers or potential targets for new therapies. We summarize data that indicate the presence of genetic subgroups of prostate cancers and show the high level of intra- and intertumoral heterogeneity, as well as updated information on disseminated and circulating tumor cells. The integrated analysis of all types of genetic alterations that culminate in altering critical biologic pathways may serve as the impetus for developing new therapeutics, repurposing agents used currently for treating other malignancies, and stratifying early and advanced prostate cancers for appropriate interventions.


Influence of the Atlantic subpolar gyre on the thermohaline circulation.
During the past decade, record-high salinities have been observed in the Atlantic Inflow to the Nordic Seas and the Arctic Ocean, which feeds the North Atlantic thermohaline circulation (THC). This may counteract the observed long-term increase in freshwater supply to the area and tend to stabilize the North Atlantic THC. Here we show that the salinity of the Atlantic Inflow is tightly linked to the dynamics of the North Atlantic subpolar gyre circulation. Therefore, when assessing the future of the North Atlantic THC, it is essential that the dynamics of the subpolar gyre and its influence on the salinity are taken into account.


Skeletal muscle glucose uptake during contraction is regulated by nitric oxide and ROS independently of AMPK.
Reactive oxygen species (ROS) and nitric oxide (NO) have been implicated in the regulation of skeletal muscle glucose uptake during contraction, and there is evidence that they do so via interaction with AMP-activated protein kinase (AMPK). In this study, we tested the hypothesis that ROS and NO regulate skeletal muscle glucose uptake during contraction via an AMPK-independent mechanism. Isolated extensor digitorum longus (EDL) and soleus muscles from mice that expressed a muscle-specific kinase dead AMPKalpha2 isoform (AMPK-KD) and wild-type litter mates (WT) were stimulated to contract, and glucose uptake was measured in the presence or absence of the antioxidant N-acetyl-l-cysteine (NAC) or the nitric oxide synthase (NOS) inhibitor N(G)-monomethyl-l-arginine (l-NMMA). Contraction increased AMPKalpha2 activity in WT but not AMPK-KD EDL muscles. However, contraction increased glucose uptake in the EDL and soleus muscles of AMPK-KD and WT mice to a similar extent. In EDL muscles, NAC and l-NMMA prevented contraction-stimulated increases in oxidant levels (dichloroflourescein fluorescence) and NOS activity, respectively, and attenuated contraction-stimulated glucose uptake in both genotypes to a similar extent. In soleus muscles of AMPK-KD and WT mice, NAC prevented contraction-stimulated glucose uptake and l-NMMA had no effect. This is likely attributed to the relative lack of neuronal NOS in the soleus muscles compared with EDL muscles. Contraction increased AMPKalpha Thr(172) phosphorylation in EDL and soleus muscles of WT but not AMPK-KD mice, and this was not affected by NAC or l-NMMA treatment. In conclusion, ROS and NO are involved in regulating skeletal muscle glucose uptake during contraction via an AMPK-independent mechanism.


A red-fluorescent substrate microarray for lipase fingerprinting.
A lipase substrate microarray was obtained by printing aliphatic C2-C12 monoesters of (5R)- and (5S)-3-(5,6-dihydroxyhexyloxy)benzaldehyde by reductive alkylation on amine-functionalized glass slides coated with bovine serum albumin and a short PEG linker. The microarray features 12 substrates and their 66 possible binary mixtures spotted in a 9 x 36 spot array. Lipase reactions are detected by chemoselective NaIO(4)-oxidation of the 1,2-diol hydrolysis product to form an aldehyde, which is then tagged with the red-fluorescent dye rhodamine B sulfohydrazide . Specific fingerprints are produced by active enzymes. These experiments provide the first example of lipase fingerprinting using microarrays.


Household survey of pesticide practice, deliberate self-harm, and suicide in the Sundarban region of West Bengal, India.
The toxicological impact and intentional ingestion of pesticides are major public health concerns globally. This study aimed to estimate the extent of deliberate self-harm (DSH) and suicides (suicidal behaviour) and document pesticide practices in Namkhana block of the Sundarban region, India. A cross-sectional study was conducted in 1680 households (21 villages) following a mixed random and cluster design sampling. The survey questionnaire (Household Information on Pesticide Use and DSH) was developed by the research team to elicit qualitative and quantitative information. The Kappa statistic and McNemar's test were used to assess the level of agreement and association between respondents' and investigators' opinions about safe storage of pesticides. Over five years, 1680 households reported 181 incidents of suicidal behaviour. Conflict with family members was the most frequently reported reason for suicidal behaviour (53.6%). The Kappa statistic indicated poor agreement between respondents and investigators about safe storage of pesticides. The pesticide-related annual DSH rate was 158.1 (95% CI 126.2-195.5), and for suicide it was 73.4 (95% CI 52.2-100.3) per 100,000. Unsafe pesticide practice and psychosocial stressors are related to the high rates of suicidal behaviour. An intersectoral approach involving the local governments, agricultural department and the health sector would help to reduce the magnitude of this public health problem.


Attention periodically samples competing stimuli during binocular rivalry.
The attentional sampling hypothesis suggests that attention rhythmically enhances sensory processing when attending to a single (~8 Hz), or multiple (~4 Hz) objects. Here, we investigated whether attention samples sensory representations that are not part of the conscious percept during binocular rivalry. When crossmodally cued toward a conscious image, subsequent changes in consciousness occurred at ~8 Hz, consistent with the rates of undivided attentional sampling. However, when attention was cued toward the suppressed image, changes in consciousness slowed to ~3.5 Hz, indicating the division of attention away from the conscious visual image. In the electroencephalogram, we found that at attentional sampling frequencies, the strength of inter-trial phase-coherence over fronto-temporal and parieto-occipital regions correlated with changes in perception. When cues were not task-relevant, these effects disappeared, confirming that perceptual changes were dependent upon the allocation of attention, and that attention can flexibly sample away from a conscious image in a task-dependent manner.


Systemic mastocytosis in adults: 2013 update on diagnosis, risk stratification, and management.
Systemic mastocytosis (SM) results from a clonal proliferation of abnormal mast cells (MC) in one or more extracutaneous organs. The major criterion is presence of multifocal clusters of morphologically abnormal MC in the bone marrow. Minor diagnostic criteria include elevated serum tryptase level, abnormal MC expression of CD25 and/or CD2, and presence of KITD816V. The 2008 World Health Organization (WHO) classification of SM has been shown to be prognostically relevant. Classification of SM patients into indolent (SM), aggressive SM (ASM), SM associated with a clonal non-MC lineage disease (SM-AHNMD) and mast cell leukemia (MCL) subgroups is a useful first step in establishing prognosis. SM treatment is generally palliative. ISM patients have a normal life expectancy and receive symptom-directed therapy; infrequently, cytoreductive therapy may be indicated for refractory symptoms. ASM patients have disease-related organ dysfunction; interferon-α (±corticosteroids) can control dermatological, hematological, gastrointestinal, skeletal, and mediator-release symptoms, but is hampered by poor tolerability. Similarly, cladribine has broad therapeutic activity, with particular utility when rapid MC debulking is indicated; the main toxicity is myelosuppression. Imatinib has a therapeutic role in the presence of an imatinib-sensitive KIT mutation or in KITD816-unmutated patients. Treatment of SM-AHNMD is governed primarily by the non-MC neoplasm; hydroxyurea has modest utility in this setting. Dasatinib's in vitro anti- KITD816V activity has not translated into significant therapeutic activity in most SM patients. In contrast, recently updated data confirms Midostaurin's significant anti-MC activity in patients with advanced SM.


Diagnosis and treatment of cutaneous vascular lesions.
Cutaneous vascular lesions are the most common pediatric birthmarks. Flat vascular malformations tend to persist, but raised vascular lesions, known as hemangiomas, generally involute. Although not always necessary, treatment of flat lesions, if desired, is best accomplished with flash-lamp pumped pulsed dye laser. Therapy of hemangiomas varies depending on the presence of associated symptoms or syndromes. Specifically, hemangiomas that are likely to lead to loss of function or life ( e.g. lesions of internal organs, lesions associated with coagulopathy) should be treated promptly. Treatment may also be required for hemangiomas that are likely to lead to scarring when the lesion involutes, such as hemangiomas of the nose and lip. The natural history of hemangiomas includes proliferative, stationary and involutional phases. Many superficial hemangiomas resolve with minimal sequelae.


Identification of a PA-binding peptide with inhibitory activity against influenza A and B virus replication.
There is an urgent need for new drugs against influenza type A and B viruses due to incomplete protection by vaccines and the emergence of resistance to current antivirals. The influenza virus polymerase complex, consisting of the PB1, PB2 and PA subunits, represents a promising target for the development of new drugs. We have previously demonstrated the feasibility of targeting the protein-protein interaction domain between the PB1 and PA subunits of the polymerase complex of influenza A virus using a small peptide derived from the PA-binding domain of PB1. However, this influenza A virus-derived peptide did not affect influenza B virus polymerase activity. Here we report that the PA-binding domain of the polymerase subunit PB1 of influenza A and B viruses is highly conserved and that mutual amino acid exchange shows that they cannot be functionally exchanged with each other. Based on phylogenetic analysis and a novel biochemical ELISA-based screening approach, we were able to identify an influenza A-derived peptide with a single influenza B-specific amino acid substitution which efficiently binds to PA of both virus types. This dual-binding peptide blocked the viral polymerase activity and growth of both virus types. Our findings provide proof of principle that protein-protein interaction inhibitors can be generated against influenza A and B viruses. Furthermore, this dual-binding peptide, combined with our novel screening method, is a promising platform to identify new antiviral lead compounds.


The global campaign to eliminate HBV and HCV infection: International Viral Hepatitis Elimination Meeting and core indicators for development towards the 2030 elimination goals.
Hepatitis B virus (HBV) and hepatitis C virus (HCV) affect more than 320 million people worldwide, which is more than HIV, tuberculosis (TB) and malaria combined. Elimination of HBV and HCV will, therefore, produce substantial public health and economic benefits and, most importantly, the prevention of 1.2 million deaths per year. In 2016, member states of the World Health Assembly unanimously adopted a resolution declaring that viral hepatitis should be eliminated by 2030. Currently, few countries have elimination programmes in place and even though the tools to achieve elimination are available, the right resources, commitments and allocations are lacking. During the fifth International Viral Hepatitis Elimination Meeting (IVHEM), 7-8 December 2018, Amsterdam, the Netherlands, an expert panel of clinicians, virologists and public health specialists discussed the current status of viral hepatitis elimination programmes across multiple countries, challenges in achieving elimination and the core indicators for monitoring progress, approaches that have failed and successful elimination plans.


Removal of the X-ray contrast media diatrizoate by electrochemical reduction and oxidation.
Due to their resistance to biological wastewater treatment, iodinated X-ray contrast media (ICM) have been detected in municipal wastewater effluents at relatively high concentrations (i.e., up to 100 μg L(-1)), with hospitals serving as their main source. To provide a new approach for reducing the concentrations of ICMs in wastewater, electrochemical reduction at three-dimensional graphite felt and graphite felt doped with palladium nanoparticles was examined as a means for deiodination of the common ICM diatrizoate. The presence of palladium nanoparticles significantly enhanced the removal of diatrizoate and enabled its complete deiodination to 3,5-diacetamidobenzoic acid. When the system was employed in the treatment of hospital wastewater, diatrizoate was reduced, but the extent of electrochemical reduction decreased as a result of competing reactions with solutes in the matrix. Following electrochemical reduction of diatrizoate to 3,5-diacetamidobenzoic acid, electrochemical oxidation with boron-doped diamond (BDD) anodes was employed. 3,5-Diacetamidobenzoic acid disappeared from solution at a rate that was similar to that of diatrizoate, but it was more readily mineralized than the parent compound. When electrochemical reduction and oxidation were coupled in a three-compartment reactor operated in a continuous mode, complete deiodination of diatrizoate was achieved at an applied cathode potential of -1.7 V vs SHE, with the released iodide ions electrodialyzed in a central compartment with 80% efficiency. The resulting BDD anode potential (i.e., +3.4-3.5 V vs SHE) enabled efficient oxidation of the products of the reductive step. The presence of other anions (e.g., chloride) was likely responsible for a decrease in I(-) separation efficiency when hospital wastewater was treated. Reductive deiodination combined with oxidative degradation provides benefits over oxidative treatment methods because it does not produce stable iodinated intermediates. Nevertheless, the process must be further optimized for the conditions encountered in hospital wastewater to improve the separation efficiency of halide ions prior to the electrooxidation step.


Theory and applications of guided-mode resonance filters.
The guided-mode resonance properties of planar dielectric waveguide gratings are presented and explained. It is shown that these structures function as filters that produce complete exchange of energy between forward- and backward-propagating diffracted waves with smooth line shapes and arbitrarily narrow filter linewidths. Simple expressions based on rigorous coupled-wave theory and on classical slab waveguide theory give a clear view and quantification of the inherent TE/TM polarization separation and the free spectral ranges of the filters. Furthermore, the resonance regimes, defining the parametric regions of the guided-mode resonances, can be directly visualized. It is shown that the linewidths of the resonances can be controlled by the grating modulation amplitude and by the degree of mode confinement (refractive-index difference at the boundaries). Examples presented of potential uses for these elements include a narrow-line polarized laser, a tunable polarized laser, a photorefractive tunable filter, and an electro-optic switch. The guided-mode resonance filter represents a basic new optical element with significant potential for practical applications.


All lactase preparations are not the same: results of a prospective, randomized, placebo-controlled trial.
To compare the efficacy of three commercially available oral lactase preparations in adults with lactose intolerance. Design--Prospective, randomized, placebo-controlled trial. Setting--Outpatient study in a General Clinical Research Center. Subjects--Ten lactose-intolerant healthy volunteers were challenged with ice cream containing 18 g of lactose. Lactase or placebo was given immediately prior to challenge. Measurements--Symptoms and breath hydrogen excretion were recorded for 3 h following lactose challenge. The three products differed in their abilities to influence symptoms and breath hydrogen excretion. Only Lactaid reduced the breath hydrogen excretion with lactose (mean peak, area under the curve and cumulative breath hydrogen excretion) (p < 0.05). Lactrase and Dairy Ease influenced symptoms: Lactrase reduced pain, bloating and total symptomatic scores (p < 0.05), whereas Dairy Ease only reduced pain (p < 0.05). Lactaid administration did not reduce symptoms. In lactose-intolerant subjects, the available lactase preparations differ in their ability to improve both breath hydrogen excretion and symptoms. Lactrase may be the product of choice for achieving symptomatic improvement.


Feeling we're biased: autonomic arousal and reasoning conflict.
Human reasoning is often biased by intuitive beliefs. A key question is whether the bias results from a failure to detect that the intuitions conflict with logical considerations or from a failure to discard these tempting intuitions. The present study addressed this unresolved debate by focusing on conflict-related autonomic nervous system modulation during biased reasoning. Participants' skin conductance responses (SCRs) were monitored while they solved classic syllogisms in which a cued intuitive response could be inconsistent or consistent with the logical correct response. Results indicated that all reasoners showed increased SCRs when solving the inconsistent conflict problems. Experiment 2 validated that this autonomic arousal boost was absent when people were not engaged in an active reasoning task. The presence of a clear autonomic conflict response during reasoning lends credence to the idea that reasoners have a "gut" feeling that signals that their intuitive response is not logically warranted. Supplemental materials for this article may be downloaded from http://cabn.psychonomic-journals.org/content/supplemental.


Reference genome and comparative genome analysis for the WHO reference strain for Mycobacterium bovis BCG Danish, the present tuberculosis vaccine.
Mycobacterium bovis bacillus Calmette-Guérin (M. bovis BCG) is the only vaccine available against tuberculosis (TB). In an effort to standardize the vaccine production, three substrains, i.e. BCG Danish 1331, Tokyo 172-1 and Russia BCG-1 were established as the WHO reference strains. Both for BCG Tokyo 172-1 as Russia BCG-1, reference genomes exist, not for BCG Danish. In this study, we set out to determine the completely assembled genome sequence for BCG Danish and to establish a workflow for genome characterization of engineering-derived vaccine candidate strains. By combining second (Illumina) and third (PacBio) generation sequencing in an integrated genome analysis workflow for BCG, we could construct the completely assembled genome sequence of BCG Danish 1331 (07/270) (and an engineered derivative that is studied as an improved vaccine candidate, a SapM KO), including the resolution of the analytically challenging long duplication regions. We report the presence of a DU1-like duplication in BCG Danish 1331, while this tandem duplication was previously thought to be exclusively restricted to BCG Pasteur. Furthermore, comparative genome analyses of publicly available data for BCG substrains showed the absence of a DU1 in certain BCG Pasteur substrains and the presence of a DU1-like duplication in some BCG China substrains. By integrating publicly available data, we provide an update to the genome features of the commonly used BCG strains. We demonstrate how this analysis workflow enables the resolution of genome duplications and of the genome of engineered derivatives of the BCG Danish vaccine strain. The BCG Danish WHO reference genome will serve as a reference for future engineered strains and the established workflow can be used to enhance BCG vaccine standardization.


Xanthones as potential antioxidants.
Xanthones (dibenzo-γ-pyrones) constitutes an important class of oxygenated heterocycles and occur as secondary metabolites in plants and microorganisms. They are known for various biological activities such as antioxidant, monoamine oxidase inhibitor, antihypertensive, hepatoprotective, antithrombotic, antifungal and anticancer. The tricyclic scaffold as well as the nature and/or position of the substituents present on it play an important role in displaying various biological activities. The unique structural scaffold and medicinal importance of xanthones have therefore attracted many Scientists in the past, to isolate or synthesize xanthones or their analogs as potential drug candidates. It would not be wrong to call them as close cousins to the polyphenol family that are known to possess strong antioxidant effects on the nervous system. The main two sources of xanthones are: Isolation from natural resources or synthesis. Though few reviews have been published in the past, mainly focusing on the anticancer activities of xanthone derivatives, but there is not a single review which is based on their antioxidant activities. We therefore have made efforts to briefly summarize natural and synthetic xanthones possessing antioxidant activity in this review.


UBXD4, a UBX-containing protein, regulates the cell surface number and stability of alpha3-containing nicotinic acetylcholine receptors.
Adaptor proteins are likely to modulate spatially and temporally the trafficking of a number of membrane proteins, including neuronal nicotinic acetylcholine receptors (nAChRs). A yeast two-hybrid screen identified a novel UBX-containing protein, UBXD4, as one of the cytosolic proteins that interact directly with the alpha3 and alpha4 nAChR subunits. The function of UBX-containing proteins is largely unknown. Immunoprecipitation and confocal microscopy confirmed the interaction of UBXD4 with alpha3-containing nAChRs (alpha3* nAChRs) expressed in HEK293 cells, PC12 cells, and rat cortical neurons. Overexpression of UBXD4 in differentiated PC12 cells (dPC12) increased nAChR cell surface expression, especially that of the alpha3beta2 subtype. These findings were corroborated by electrophysiology, immunofluorescent staining, and biotinylation of surface receptors. Silencing of UBXD4 led to a significant reduction of alpha3* nAChRs in rat cortical neurons and dPC12 cells. Biochemical and immunofluorescence studies of endogenous UBXD4 showed that the protein is located in both the ER and cis-Golgi compartments. Our investigations also showed that the alpha3 subunit is ubiquitinated and that UBXD4 can interfere with its ubiquitination and consequent degradation by the proteasome. Our data suggest that UBXD4 modulates the distribution of alpha3* nAChRs between specialized intracellular compartments and the plasma membrane. This effect is achieved by controlling the stability of the alpha3 subunit and, consequently, the number of receptors at the cell surface.


Mutations in the alternatively spliced exons of USH1C cause non-syndromic recessive deafness.
We have recently shown that USH1C underlies Usher syndrome type 1c (USH1C), an USH1 subtype characterized by profound deafness, retinitis pigmentosa, and vestibular dysfunction. USH1C encodes a PDZ-domain-containing protein, harmonin. Eight different Ush1c transcripts were identified in the mouse inner ear. Moreover, transcripts containing seven alternatively spliced exons (A-F, G/G) were found to be expressed in the inner ear, but not in the eye. These findings suggested that mutations involving USH1C might also be the cause of DFNB18, a form of non-syndromic deafness, which maps to a chromosomal region that includes USH1C. We screened 32 Chinese multiplex families with non-syndromic recessive deafness for USH1C mutations. In one family, congenital profound deafness without RP was associated with a C to G transversion in the alternatively spliced exon D, predicting an arginine to proline substitution at codon 608 in the proline-, serine- and threonine-rich region of harmonin. We also screened 320 deaf probands from other ethnic background and found three who were heterozygous for changes in the alternately spliced exons including Gly431Val in exon B, Arg620Leu and Arg636Cys in exon D. None of these mutations were detected in DNA from 200 control subjects with normal hearing including 110 Chinese. We also screened 121 non-Acadian probands with type 1 Usher syndrome. None carried any mutations in these exons of USH1C. Our findings show that USH1C mutations can also cause non-syndromic deafness and that some harmonin isoforms are specifically required for inner ear function.


Gpi19, the Saccharomyces cerevisiae homologue of mammalian PIG-P, is a subunit of the initial enzyme for glycosylphosphatidylinositol anchor biosynthesis.
Glycosylphosphatidylinositols (GPIs) are attached to the C termini of some glycosylated secretory proteins, serving as membrane anchors for many of those on the cell surface. Biosynthesis of GPIs is initiated by the transfer of N-acetylglucosamine (GlcNAc) from UDP-GlcNAc to phosphatidylinositol. This reaction is carried out at the endoplasmic reticulum (ER) by an enzyme complex called GPI-N-acetylglucosaminyltransferase (GPI-GlcNAc transferase). The human enzyme has six known subunits, at least four of which, GPI1, PIG-A, PIG-C, and PIG-H, have functional homologs in the budding yeast Saccharomyces cerevisiae. The uncharacterized yeast gene YDR437w encodes a protein with some sequence similarity to human PIG-P, a fifth subunit of the GPI-GlcNAc transferase. Here we show that Ydr437w is a small but essential subunit of the yeast GPI-GlcNAc transferase, and we designate its gene GPI19. Similar to other mutants in the yeast enzyme, temperature-sensitive gpi19 mutants display cell wall defects and hyperactive Ras phenotypes. The Gpi19 protein associates with the yeast GPI-GlcNAc transferase in vivo, as judged by coimmuneprecipitation with the Gpi2 subunit. Moreover, conditional gpi19 mutants are defective for GPI-GlcNAc transferase activity in vitro. Finally, we present evidence for the topology of Gpi19 within the ER membrane.


On summary measure analysis of linear trend repeated measures data: performance comparison with two competing methods.
The summary measure approach (SMA) is sometimes the only applicable tool for the analysis of repeated measurements in medical research, especially when the number of measurements is relatively large. This study aimed to describe techniques based on summary measures for the analysis of linear trend repeated measures data and then to compare performances of SMA, linear mixed model (LMM), and unstructured multivariate approach (UMA). Practical guidelines based on the least squares regression slope and mean of response over time for each subject were provided to test time, group, and interaction effects. Through Monte Carlo simulation studies, the efficacy of SMA vs. LMM and traditional UMA, under different types of covariance structures, was illustrated. All the methods were also employed to analyze two real data examples. Based on the simulation and example results, it was found that the SMA completely dominated the traditional UMA and performed convincingly close to the best-fitting LMM in testing all the effects. However, the LMM was not often robust and led to non-sensible results when the covariance structure for errors was misspecified. The results emphasized discarding the UMA which often yielded extremely conservative inferences as to such data. It was shown that summary measure is a simple, safe and powerful approach in which the loss of efficiency compared to the best-fitting LMM was generally negligible. The SMA is recommended as the first choice to reliably analyze the linear trend data with a moderate to large number of measurements and/or small to moderate sample sizes.


Novel epitope begets a novel pathway in type 1 diabetes progression.
While CD8+ T cells are critical to diabetogenesis in NOD mice, evidence of their involvement in human type 1 diabetes (T1D) has been circumstantial. The existence of CD8+ T cells specific for beta cell peptides has been demonstrated, but functional data regarding the role of these cells in T1D have been lacking. In this issue of the JCI, Skowera et al. describe an unusual self-peptide epitope derived from the leader sequence of preproinsulin (PPI) and show that 50% of HLA-A2+ patients with new-onset T1D possessed circulating CD8+ T cells specific for this epitope, suggesting that PPI plays a critical role in the development of T1D (see the related article beginning on page 3390). They also report that beta cells upregulate PPI expression in the presence of high glucose levels, rendering these cells more susceptible to lysis and potentially accelerating disease. This suggests that interventions aimed at decreasing the PPI-specific CD8+ T cell response early after T1D diagnosis may be efficacious in ameliorating the disease process.


Bilateral globus pallidus internus deep brain stimulation in Parkinson's disease with periodic limb movement disorder.
Deep brain stimulation for Parkinson's disease is a well-established therapeutic intervention for refractory disease. The main nuclear targets are the subthalamic nucleus and the globus pallidus internus. Periodic limb movements are often an associated condition in patients with Parkinson's disease. Both conditions can respond to dopaminergic agents. The case of a 57-year-old man with severe Parkinson's disease and periodic limb movement who underwent bilateral globus pallidus deep brain stimulation surgery for his Parkinson's disease is presented and discussed. Intraoperatively the patient's periodic limb movements responded to voltage stimulation higher than those required for improvement of his parkinsonian symptoms. The immediate and reversible improvement in these symptoms contralateral to the stimulation site suggests pallidal stimulation may directly ameliorate periodic limb movement symptoms.


Isomerization of Internal Alkynes to Iridium(III) Allene Complexes via C-H Bond Activation: Expanded Substrate Scope, and Progress towards a Catalytic Methodology.
The synthesis of a series of allene complexes (POCOP)Ir(η²-RC=(.)=CR') 1b-4b (POCOP = 2,6-bis(di-tert-butylphosphonito)benzene) via isomerization of internal alkynes is reported. We have demonstrated that the application of this methodology is viable for the isomerization of a wide variety of alkyne substrates. Deuterium labeling experiments support our proposed mechanism. The structures of the allene complexes 1b-4b were determined using spectroscopic data analysis. Additionally, the solid-state molecular structure of complex 2b was determined using single crystal X-ray diffraction studies and it confirmed the assignment of an iridium-bound allene isomerization product. The rates of isomerization were measured using NMR techniques over a range of temperatures to allow determination of thermodynamic parameters. Finally, we report a preliminary step towards developing a catalytic methodology; the allene may be liberated from the metal center by exposure of the complex to an atmosphere of carbon monoxide.


Possible association between common variants of the phenylalanine hydroxylase (PAH) gene and memory performance in healthy adults.
Phenylalanine hydroxylase (PAH) is the enzyme that metabolizes phenylalanine, an essential amino acid required for catecholamine synthesis. Rare mutations in PAH are causal to phenylketonuria (PKU), an autosomal recessive disease characterized by neuropsychiatric symptoms including intellectual disability. We examined whether there is an association between common single nucleotide polymorphisms (SNPs) of PAH and memory performance in the Japanese population. Subjects were 599 healthy adults (166 males and 433 females; mean age 43.8 ± 15.5 years). The Wechsler Memory Scale-Revised (WMS-R) was administered to all participants to assess memory performance. Genotyping was performed for 6 selected tagging SNPs of PAH (rs1722387, rs3817446, rs1718301, rs2037639, rs10860936 and rs11111419). Analyses of covariance controlling for sex and education years, indicated a significant association between a SNP (rs2037639) and age-corrected verbal memory index of WMS-R (nominal p = 0.0013) which remained significant after correction for multiple testing ( p = 0.0013 < 0.0017 = 0.05/30tests). Individuals with the GG genotype showed a significantly lower mean verbal memory score, compared with those individuals carrying the AA/AG genotype (106.0 ± 16.0 vs. 111.7 ± 13.4; p = 0.00099). A haplotype block containing two markers of rs2037639 and rs10860936 was associated with verbal memory index (permutation global p = 0.0091). Our findings suggest that common genetic variations in PAH are associated with verbal memory in healthy adults. Unknown functional polymorphisms in PAH or those in other genes nearby might affect memory performance.


Nutritional management of (some) autism: a case for gluten- and casein-free diets?
Autism spectrum disorders represent a diverse and heterogeneous array of conditions unified by the variable presence of specific behaviours impacting social and communicative functions (social affect) alongside other presentation. Common overt characteristics may come about as a consequence of several different genetic and biological processes differentially manifesting across different people or groups. The concept of plural 'autisms' is evolving, strengthened by an increasingly important evidence base detailing different developmental trajectories across the autism spectrum and the appearance of comorbidity variably interacting with core symptoms and onwards influencing quality of life. Reports that dietary intervention, specifically the removal of foods containing gluten and/or casein from the diet, may impact on the presentation of autism for some, complement this plural view of autism. Evidence suggestive of differing responses to the use of a gluten- and casein-free diet, defined as best- and non-response, has combined with some progress on determining the underlying genetic and biological correlates potentially related to such dietary elements. The preliminary suggestion of a possible diet-related autism phenotype is the result. This review will highlight several pertinent aspects onwards to an effect of food in some cases of autism including research on the pharmacological activity of food metabolites, immune response, issues with gut barrier function and some contribution from the gut microbiota. These represent promising areas in need of far greater research inspection in order to potentially define such a diet-related subgroup on the autism spectrum.


On the role of benchmarking data sets and simulations in method comparison studies.
Method comparisons are essential to provide recommendations and guidance for applied researchers, who often have to choose from a plethora of available approaches. While many comparisons exist in the literature, these are often not neutral but favor a novel method. Apart from the choice of design and a proper reporting of the findings, there are different approaches concerning the underlying data for such method comparison studies. Most manuscripts on statistical methodology rely on simulation studies and provide a single real-world data set as an example to motivate and illustrate the methodology investigated. In the context of supervised learning, in contrast, methods are often evaluated using so-called benchmarking data sets, that is, real-world data that serve as gold standard in the community. Simulation studies, on the other hand, are much less common in this context. The aim of this paper is to investigate differences and similarities between these approaches, to discuss their advantages and disadvantages, and ultimately to develop new approaches to the evaluation of methods picking the best of both worlds. To this aim, we borrow ideas from different contexts such as mixed methods research and Clinical Scenario Evaluation.


Robust parameter estimation techniques for stochastic within-host macroparasite models.
We present a stochastic model of the within-host population dynamics of lymphatic filariasis, and use a simulated goodness-of-fit (GOF) method to estimate immunological parameters and their confidence intervals from experimental data. A variety of deterministic moment closure approximations to the stochastic system are explored and compared with simulation results. For the maximum GOF parameter estimates, none of the methods of closure accurately reproduce the behaviour of the stochastic model. However, direct analysis of the stochastic model demonstrates that the high levels of variation observed in the data can be reproduced without requiring parameters to vary between hosts. This indicates that the observed aggregation of parasite load may be dynamically generated by random variation in the development of an effective immune response against parasite larvae.


Differentiation of the Infarct Core from Ischemic Penumbra within the First 4.5 Hours, Using Diffusion Tensor Imaging-Derived Metrics: A Rat Model.
To investigate whether the diffusion tensor imaging-derived metrics are capable of differentiating the ischemic penumbra (IP) from the infarct core (IC), and determining stroke onset within the first 4.5 hours. All procedures were approved by the local animal care committee. Eight of the eleven rats having permanent middle cerebral artery occlusion were included for analyses. Using a 7 tesla magnetic resonance system, the relative cerebral blood flow and apparent diffusion coefficient maps were generated to define IP and IC, half hour after surgery and then every hour, up to 6.5 hours. Relative fractional anisotropy, pure anisotropy (rq) and diffusion magnitude (rL) maps were obtained. One-way analysis of variance, receiver operating characteristic curve and nonlinear regression analyses were performed. The evolutions of tensor metrics were different in ischemic regions (IC and IP) and topographic subtypes (cortical, subcortical gray matter, and white matter). The rL had a significant drop of 40% at 0.5 hour, and remained stagnant up to 6.5 hours. Significant differences (p < 0.05) in rL values were found between IP, IC, and normal tissue for all topographic subtypes. Optimal rL threshold in discriminating IP from IC was about -29%. The evolution of rq showed an exponential decrease in cortical IC, from -26.9% to -47.6%; an rq reduction smaller than 44.6% can be used to predict an acute stroke onset in less than 4.5 hours. Diffusion tensor metrics may potentially help discriminate IP from IC and determine the acute stroke age within the therapeutic time window.


Safety and therapeutic potential of human bone marrow-derived mesenchymal stromal cells in regenerative medicine.
Regenerative medicine is considered as an alternative approach to healthcare. Owing to their pluripotent abilities and their relative lack of ethical and legal issues, adult stem cells are considered as optimal candidates for use in the treatment of various diseases. Bone marrow-derived mesenchymal stem cells are among the most promising candidates for clinical applications as they have expressed a higher degree of plasticity in vitro. Many investigators have begun to examine how bone marrow stem cells might be used to rebuild damaged tissues. The systemic administration of cells for therapeutic applications requires efficient migration and homing of cells to the target site. Cell adhesion molecules and their ligands, chemokines, extracellular matrix components and specialized bone marrow niches all participate in the proper regulation of this process. MSCs suppress the pathophysiological process that is mediated by chronic inflammation and contributes to a modification of the microenvironment and tissue regeneration. Due to the intricacy of the mesenchymal stem cell, there is ever-increasing amount of data emerging about their migration and regenerative mechanisms. Many factors influence MSC mobilization and their homing to injured tissues. This review summarizes the current clinical and pre-clinical data available in literature regarding the use of MSC in tissue repair and their prospective therapeutic role in various diseases and the underlying repair mechanisms will be discussed.


Urinary and dietary analysis of 18,470 bangladeshis reveal a correlation of rice consumption with arsenic exposure and toxicity.
We utilized data from the Health Effects of Arsenic Longitudinal Study (HEALS) in Araihazar, Bangladesh, to evaluate the association of steamed rice consumption with urinary total arsenic concentration and arsenical skin lesions in the overall study cohort (N=18,470) and in a subset with available urinary arsenic metabolite data (N=4,517). General linear models with standardized beta coefficients were used to estimate associations between steamed rice consumption and urinary total arsenic concentration and urinary arsenic metabolites. Logistic regression models were used to estimate prevalence odds ratios (ORs) and their 95% confidence intervals (CIs) for the associations between rice intake and prevalent skin lesions at baseline. Discrete time hazard models were used to estimate discrete time (HRs) ratios and their 95% CIs for the associations between rice intake and incident skin lesions. Steamed rice consumption was positively associated with creatinine-adjusted urinary total arsenic (β=0.041, 95% CI: 0.032-0.051) and urinary total arsenic with statistical adjustment for creatinine in the model (β=0.043, 95% CI: 0.032-0.053). Additionally, we observed a significant trend in skin lesion prevalence (P-trend=0.007) and a moderate trend in skin lesion incidence (P-trend=0.07) associated with increased intake of steamed rice. This study suggests that rice intake may be a source of arsenic exposure beyond drinking water.


Evidence that the UL84 gene product of human cytomegalovirus is essential for promoting oriLyt-dependent DNA replication and formation of replication compartments in cotransfection assays.
The protein products of 11 viral genomic loci cooperate in a transient cotransfection assay to mediate lytic-phase DNA replication of oriLyt, the human cytomegalovirus (HCMV) origin of replication. Six of these genes have homology with the well-characterized herpes simplex virus replication genes and encode core replication machinery proteins that are typically essential for DNA synthesis. The remaining five HCMV gene loci, initially referred to as auxiliary components, include several known immediate-early (IE) transcriptional regulatory proteins as well as genes encoding functionally uncharacterized polypeptides. Some or all of the auxiliary components may be necessary in trans to replicate the HCMV oriLyt only because they are required for efficient expression or transactivation of the native early promoters and 3' processing elements included in the genomic clones. Therefore, we reassessed the requirements for the auxiliary components by adding constitutive heterologous promoters and control signals to the coding regions and carrying out transient DpnI replication assays in cotransfected Vero cells. The results revealed that in the presence of the UL69 posttranscriptional activator and the remaining auxiliary polypeptides, UL84 was the only auxiliary component that could not be omitted to obtain oriLyt-dependent DNA replication. Nevertheless, in human diploid fibroblasts, some additional auxiliary loci as well as UL84 were critical. There was also an obligatory requirement for UL84, in cooperation with two other auxiliary factors, UL112-113 and IE2, and the core machinery, to constitute the minimal HCMV proteins necessary to direct oriLyt-dependent DNA amplification. However, the Epstein-Barr virus core replication genes could substitute for the HCMV core genes, and in these circumstances, UL84 alone directed amplification of HCMV oriLyt. Moreover, there was also an absolute requirement for UL84 along with the core and other auxiliary factors for the formation of intranuclear replication compartments as assayed by immunofluorescence in transient DNA cotransfection assays. These compartments were typical of those associated with active viral DNA replication in HCMV-infected cells, they incorporated pulse-labeled bromodeoxyuridine, and their formation was both phosphonoacetic acid sensitive and oriLyt dependent. These results demonstrate that UL84 is obligatory for both intranuclear replication compartment formation and origin-dependent DNA amplification and suggest that it is a key viral component in promoting the initiation of HCMV oriLyt-directed DNA replication.


Activity of interpositus neurons during a visually guided reach.
Neurons in the cerebellar interpositus nucleus greatly increase their discharge rates when a monkey reaches out to grasp an object. However, when the monkey is required to track a target on a screen by moving a manipulandum, the increase in discharge rate is relatively small or nonexistent. Moving the hand directly to a target is a visuomotor task that may be fundamentally different from a remote tracking task. We hypothesize that the interpositus nucleus is specialized for direct visual guidance of the limb or, alternatively, interpositus is specialized for controlling hand movements required to grasp an object. A monkey was trained to hold a sensor and move it directly over a visual target to obtain water reward. Small drawers were mounted next to two of the targets; on some trials a drawer would open so that the monkey would reach out and retrieve a raisin that had been placed in it. Interpositus neurons discharged strongly during reach to grasp the raisin but not when the monkey was positioning the sensor over the target. For individual cells, discharge pattern and amplitude were largely independent of the size and direction of the reach to grasp, suggesting that interpositus does not control direction or amplitude of the reach. The results are consistent with the hypothesis that neurons in forelimb regions of interpositus participate in the control of hand movements used in grasping, but they are not consistent with the hypothesis that interpositus neurons participate in direct visual guidance of the limb.


Methotrexate/6-mercaptopurine maintenance therapy influences the risk of a second malignant neoplasm after childhood acute lymphoblastic leukemia: results from the NOPHO ALL-92 study.
Among 1614 children with acute lymphoblastic leukemia (ALL) treated with the Nordic Society for Paediatric Haematology and Oncology (NOPHO) ALL-92 protocol, 20 patients developed a second malignant neoplasm (SMN) with a cumulative risk of 1.6% at 12 years from the diagnosis of ALL. Nine of the 16 acute myeloid leukemias or myelodysplastic syndromes had monosomy 7 (n = 7) or 7q deletions (n = 2). In Cox multivariate analysis, longer duration of oral 6-mercaptopurine (6MP)/methotrexate (MTX) maintenance therapy (P = .02; longest for standard-risk patients) and presence of high hyperdiploidy (P = .07) were related to increased risk of SMN. Thiopurine methyltransferase (TPMT) methylates 6MP and its metabolites, and thus reduces cellular levels of cytotoxic 6-thioguanine nucleotides. Of 524 patients who had erythrocyte TPMT activity measured, the median TPMT activity in 9 patients developing an SMN was significantly lower than in the 515 that did not develop an SMN (median, 12.1 vs 18.1 IU/mL; P = .02). Among 427 TPMT wild-type patients for whom the 6MP dose was registered, those who developed SMN received higher average 6MP doses than the remaining patients (69.7 vs 60.4 mg/m2; P = .03). This study indicates that the duration and intensity of 6MP/MTX maintenance therapy of childhood ALL may influence the risk of SMNs in childhood ALL.


Key symptoms in the detection of affective disorders in medical patients.
Psychiatric disorder in medical patients often goes undetected. In this study we have attempted to identify a minimum set of key symptoms from an extensive research interview [Present State Examination (PSE)] that might assist in the identification of psychiatric disorders among general medical patients. Stepwise logistic regression analysis was used to identify PSE-items that contributed most to the PSE-CATEGO classification of psychiatric disorders in 192 Dutch medical out-patients. A risk score based on two core symptoms (panic and depressed mood) and five supplementary symptoms classified patients with a sensitivity of 0.94 and a specificity of 0.91. In a separate U.K. sample of medical in-patients, consisting of 37 PSE cases and age and sex matched controls, the risk score yielded a sensitivity and specificity of 0.89 and 0.97 respectively. The results suggest that a few questions concerning psychiatric symptoms may facilitate the identification of the majority of patients with anxiety and depressive disorders in medical populations.


Structural insights and biomedical potential of IgNAR scaffolds from sharks.
In addition to antibodies with the classical composition of heavy and light chains, the adaptive immune repertoire of sharks also includes a heavy-chain only isotype, where antigen binding is mediated exclusively by a small and highly stable domain, referred to as vNAR. In recent years, due to their high affinity and specificity combined with their small size, high physicochemical stability and low-cost of production, vNAR fragments have evolved as promising target-binding scaffolds that can be tailor-made for applications in medicine and biotechnology. This review highlights the structural features of vNAR molecules, addresses aspects of their generation using immunization or in vitro high throughput screening methods and provides examples of therapeutic, diagnostic and other biotechnological applications.


SHIP1 negatively regulates proliferation of osteoclast precursors via Akt-dependent alterations in D-type cyclins and p27.
Osteoclasts arise from macrophage progenitors in bone marrow (BMMs) as a consequence of signaling events elicited by M-CSF and receptor activator of NF-kappaB ligand, acting on their unique receptors, via c-Fms and receptor activator of NF-kappaB. Both receptors activate the PI3K and MAPK pathways, which promote cell proliferation and survival. SHIP1 is essential for normal bone homeostasis, as mice lacking the protein exhibit osteoporosis resulting from increased numbers of hyper-resorptive osteoclasts. In this study, we show that BMMs from SHIP1 null mice respond to M-CSF, but not receptor activator of NF-kappaB ligand, by increasing Akt activation. In consequence, there are up-regulation of D-type cyclins, down-regulation of the cyclin-dependent kinase inhibitor p27, and, therefore, increased phosphorylation of the retinoblastoma protein and cell proliferation. Surprisingly, cell survival of wild-type and knockout BMMs is unaltered. Finally, osteoclastogenesis and periarticular bone erosions are markedly increased in SHIP1(-/-) mice with inflammatory arthritis, a condition characterized by increased M-CSF expression. The SHIP1/Akt pathway therefore suppresses bone loss in pathological states associated with an excess of the cytokine.


Simple model of corner reflector phenomena.
The principle of cube corner reflector operation is simple, and a careful examination of operational details is valuable in promoting an understanding of the potentials and limitations in reflector use. This paper describes an easily visualized model by which these principles may be understood. From this model, simple procedures may be derived for determining the effective aperture area variation of conventional corner reflector configurations. The variation of the effective aperture of some conventional configurations is then computed. The effects of angular errors in the alignment of the reflecting surfaces are also modeled.


Modulation of human sympathetic periodicity by mild, brief hypoxia and hypercapnia.
We determined the influence of brief mild normocapnic hyperoxia, hypoxia, and hyperoxic hypercapnia on human muscle sympathetic nerve activity and R-R intervals, as quantified by both time- and frequency-domain analyses. We obtained measurements in nine healthy young adult men and women during uncontrolled and frequency (but not tidal volume) controlled breathing. Responses were evaluated with forward selection and backward elimination statistical models, with muscle sympathetic nerve activity as the dependent variable, and power spectral techniques. Hyperoxia and hypoxia did not alter arterial pressure; hypercapnia increased diastolic pressure modestly. Average R-R intervals tended to increase during hyperoxia, and decrease during hypoxia and hypercapnia. During uncontrolled breathing, changes of inspiratory gases exerted only minor effects on muscle sympathetic nerve activity; during controlled breathing, both hypoxia and hypercapnia tended to increase muscle sympathetic nerve activity. Statistical modeling suggested that chemoreceptor stimulation increased muscle sympathetic neural outflows, but that increases of sympathetic traffic were opposed by secondary increases of ventilation. Inspiratory gases modulated the frequency distribution of muscle sympathetic nerve activity strikingly: hypoxia increased sympathetic power at respiratory frequencies and hypercapnia increased sympathetic power at both respiratory and (primarily in one subject) cardiac frequencies. Our data suggest that mild brief hypoxia and hypercapnia increase human muscle sympathetic nerve activity, but that this tendency is opposed by chemoreflex-induced increases of ventilation. Our results suggest also that chemoreceptor activity exerts important influences on the frequency content, as well as the quantity of sympathetic neural outflow.


Evaluation of the management of heart failure in primary care.
The extent to which guidelines for the treatment of heart failure are currently followed in primary care in the UK is unclear. To evaluate the prevalence of heart failure and the pharmacological management of heart failure in relation to European Society of Cardiology (ESC) and National Institute for Health and Clinical Excellence guidelines. Retrospective cohort study using routinely collected data from 163 general practices across Great Britain contributing data to the Doctors Independent Network (DIN-LINK) database over a 5-year period until December 31, 2006. From a patient population of nearly 1.43 million, 9311 patients with heart failure were identified [mean age 78 years (SD 12)], giving an estimated prevalence of 0.7%. Of these, 7410 (79.6%) were prescribed a loop diuretic, 6620 (71.1%) were prescribed an angiotensin-converting enzyme (ACE) inhibitor or ARB, 3403 (36.6%) were prescribed beta-blockers but only 2732 (29.3%) were prescribed an ACE inhibitor or ARB and a beta-blocker in combination. Thirty-five per cent of patients prescribed ACE inhibitor and 11.5% of those prescribed beta-blockers met ESC guideline target doses. Age, gender and comorbidity predicted whether patients received beta-blocker or ACE inhibitor with younger males being more likely to receive maximal therapy. These data suggest that while most patients with heart failure receive an ACE inhibitor/ARB in primary care, few are titrated to target dose and many do not receive a beta-blocker. Optimum treatment appears to be most likely for young men. New strategies are required to ensure equitable and optimal treatment for all.


Psychometric Evaluation of Disordered Eating Measures in Bariatric Surgery Candidates.
Assessment of disordered eating is common in bariatric surgery candidates, yet psychometric properties of disordered eating measures in this population are largely unknown. Measures were completed by 405 adult bariatric surgery candidates at pre-surgical consultation. Fit of the original scale structures was tested using confirmatory factor analysis (CFA) and alternative factor solutions were generated using exploratory factor analysis (EFA). Reliability (internal consistency), construct validity (convergent and divergent) and criterion validity (with the EDE as criterion) were assessed. The measures prioritised for evaluation are the following: Eating Disorder Examination Questionnaire (EDE-Q; n = 405), Three-Factor Eating Questionnaire (TFEQ; n = 405), Questionnaire of Eating and Weight Patterns Revised (QEWP-R; n = 204), Clinical Impairment Assessment (CIA; n = 204) and the Eating Disorder Examination clinical interview (EDE; n = 131). CFA revealed adequate fit for only the CIA in its current form (CFI = 0.925, RMSEA = 0.096). EFA produced revised scales with improved reliability for the EDE, EDE-Q and TFEQ. Reliability of revised subscales was improved (original scales α = 0.43-0.82; revised scales α = 0.67-0.93). Correlational analyses of the CIA and revised versions of remaining scales with measures of psychological wellbeing and impairment revealed adequate convergent validity. All measures differentiated an EDE-classified disordered eating group from a non-disordered eating group (criterion validity). Diagnostic concordance between the EDE, EDE-Q and QEWP-R was low, and identification of disordered eating behaviours was inconsistent across measures. Findings highlight the limitations of existing disordered eating questionnaires in bariatric surgery candidates. Results suggest revised assessments are required to overcome these limitations and ensure that measures informing clinical recommendations regarding patient care are reliable and valid.


Recovery of the soliton self-frequency shift by optical phase conjugation.
The recovery of the self-frequency shift effect on a soliton by optical phase conjugation is investigated. It is proved that, without third-order dispersion and fiber loss, the soliton self-frequency shift can be completely recovered. The recoveries for a fundamental soliton and a second-order soliton in the presence of third-order dispersion and of fiber loss, which is compensated by the distributed erbium-doped fiber amplifiers, are numerically studied.


Congenital anomalies of the aortic arch.
Congenital anomalies of the aortic arch include diverse subgroups of malformations that may be clinically silent or may present with severe respiratory or esophageal symptoms especially when associated with complete vascular rings. These anomalies may be isolated or may be associated with other congenital heart diseases. Volume rendered computed tomography (CT) and magnetic resonance angiography (MRA) help in preoperative surgical planning by providing information about the complex relationship of aortic arch and its branches to the trachea and esophagus. Three dimensional capabilities of both computed tomography angiography (CTA) and MRA are helpful in determining evidence of tracheal or esophageal compression or other high-risk features in patients with a complete vascular ring.


The role of chaperone-mediated autophagy in huntingtin degradation.
Huntington Disease (HD) is caused by an abnormal expansion of polyQ tract in the protein named huntingtin (Htt). HD pathology is featured by accumulation and aggregation of mutant Htt in striatal and cortical neurons. Aberrant Htt degradation is implicated in HD pathogenesis. The aim of this study was to investigate the regulatory role of chaperone-mediated autophagy (CMA) components, heat shock protein cognate 70 (Hsc70) and lysosome-associated protein 2A (LAMP-2A) in degradation of Htt fragment 1-552aa (Htt-552). A cell model of HD was produced by overexpression of Htt-552 with adenovirus. The involvement of CMA components in degradation of Htt-552 was determined with over-expression or silencing of Hsc70 and LAMP-2A. The results confirmed previous reports that both macroautophagy and CMA were involved in degradation of Htt-552. Changing the levels of CMA-related proteins affected the accumulation of Htt-552. The lysosomal binding and luminal transport of Htt-552 was demonstrated by incubation of Htt-552 with isolated lysosomes. Expansion of the polyQ tract in Htt-552 impaired its uptake and degradation by lysosomes. Mutation of putative KFERQ motif in wild-type Htt-552 interfered with interactions between Htt-552 and Hsc70. Endogenous Hsc70 and LAMP-2A interacted with exogenously expressed Htt-552. Modulating the levels of CMA related proteins degraded endogenous full-length Htt. These studies suggest that Hsc70 and LAMP-2A through CMA play a role in the clearance of Htt and suggest a novel strategy to target the degradation of mutant Htt.


The apoptotic actions of platelets in acute ischemic stroke.
Stroke is a disease that affects the blood vessels that supply blood to the brain. Although platelets are implicated in the pathophysiology of stroke the mechanism is still not clear and there antiplatelet agents available for the prevention and treatment of stroke. We herein examined the relationship between the potential cytokine, TNF-α platelet activation and apoptosis in acute ischemic stroke patients. We selected 60 patients (mean age 57.9 ± 10.2 years) who had not taken any antiplatelet drugs for 14 days. A group of 45 participants (mean age 51.05 ± 9.07 years) were selected as the control group. For both the patients and for the control group, P-selectin (CD62p) and Annexin-V binding, cytochrome-c levels, caspase-3 gene expression and caspase-3 releasing and plasma TNF-α levels were measured in platelets. The results showed significant increase in plasma TNF-α and platelet Annexin-V, CD62p, cytochrome-c and caspase-3 gene expression in stroke patients compared to the control group. The data of this work suggests that inflammation may have a role in platelet apoptosis in stroke which may suggest a new aspect of the role of inflammation in the development of acute ischemic stroke.


Selectivity for complex shapes in primate visual area V2.
To explore the role of visual area V2 in shape analysis, we studied the responses of neurons in area V2 of the alert macaque using a set of 128 grating and geometric line stimuli that varied in their shape characteristics and geometric complexity. Simple stimuli included oriented bars and sinusoidal gratings; complex stimuli included angles, arcs, circles, and intersecting lines, plus hyperbolic and polar gratings. We found that most V2 cells responded well to at least some of the complex stimuli, and in many V2 cells the most effective complex stimulus elicited a significantly larger response than the most effective bar or sinusoid. Approximately one-third of the V2 cells showed significant differential responsiveness to various complex shape characteristics, and many were also selective for the orientation, size, and/or spatial frequency of the preferred shape. These results indicate that V2 cells explicitly represent complex shape information and suggest specific types of higher order visual information that V2 cells extract from visual scenes.


Role of a polyphenol-enriched preparation on chemoprevention of mammary carcinoma through cancer stem cells and inflammatory pathways modulation.
Naturally occurring polyphenolic compounds from fruits, particularly from blueberries, have been reported to be significantly involved in cancer chemoprevention and chemotherapy. Biotransformation of blueberry juice by Serratia vaccinii increases its polyphenolic content and endows it with anti-inflammatory properties. This study evaluated the effect of a polyphenol-enriched blueberry preparation (PEBP) and its non-fermented counterpart (NBJ), on mammary cancer stem cell (CSC) development in in vitro, in vivo and ex vivo settings. Effects of PEBP on cell proliferation, mobility, invasion, and mammosphere formation were measured in vitro in three cell lines: murine 4T1 and human MCF7 and MDA-MB-231. Ex vivo mammosphere formation, tumor growth and metastasis observations were carried out in a BALB/c mouse model. Our research revealed that PEBP influence cellular signaling cascades of breast CSCs, regulating the activity of transcription factors and, consequently, inhibiting tumor growth in vivo by decreasing metastasis and controlling PI3K/AKT, MAPK/ERK, and STAT3 pathways, central nodes in CSC inflammatory signaling. PEBP significantly inhibited cell proliferation of 4T1, MCF-7 and MDA-MB-231. In all cell lines, PEBP reduced mammosphere formation, cell mobility and cell migration. In vivo, PEBP significantly reduced tumor development, inhibited the formation of ex vivo mammospheres, and significantly reduced lung metastasis. This study showed that polyphenol enrichment of a blueberry preparation by fermentation increases its chemopreventive potential by protecting mice against tumor development, inhibiting the formation of cancer stem cells and reducing lung metastasis. Thus, PEBP may represent a novel complementary alternative medicine therapy and a source for novel therapeutic agents against breast cancer.


Economic healthcare costs of Clostridium difficile infection: a systematic review.
Clostridium difficile infection (CDI) is the leading cause of infectious diarrhoea in hospitalised patients. CDI increases patient healthcare costs due to extended hospitalisation, re-hospitalisation, laboratory tests and medications. However, the economic costs of CDI on healthcare systems remain uncertain. The purpose of this study was to perform a systematic review to summarise available studies aimed at defining the economic healthcare costs of CDI. We conducted a literature search for peer-reviewed studies that investigated costs associated with CDI (1980 to present). Thirteen studies met inclusion and exclusion criteria. CDI costs in 2008 US dollars were calculated using the consumer price index. The total and incremental costs for primary and recurrent CDI were estimated. Of the 13, 10 were from the USA and one each from Canada, UK, and Ireland. In US-based studies incremental cost estimates ranged from $2,871 to $4,846 per case for primary CDI and from $13,655 to $18,067 per case for recurrent CDI. US-based studies in special populations (subjects with irritable bowel disease, surgical inpatients, and patients treated in the intensive care unit) showed an incremental cost range from $6,242 to $90,664. Non-US-based studies showed an estimated incremental cost of $5,243 to $8,570 per case for primary CDI and $13,655 per case for recurrent CDI. Economic healthcare costs of CDI were high for primary and recurrent cases. The high cost associated with CDI justifies the use of additional resources for CDI prevention and control.


Unmasking tandem site interaction in human acetylcholinesterase. Substrate activation with a cationic acetanilide substrate.
Acetylcholinesterase (AChE) contains a narrow and deep active site gorge with two sites of ligand binding, an acylation site (or A-site) at the base of the gorge, and a peripheral site (or P-site) near the gorge entrance. The P-site contributes to catalytic efficiency by transiently binding substrates on their way to the acylation site, where a short-lived acyl enzyme intermediate is produced. A conformational interaction between the A- and P-sites has recently been found to modulate ligand affinities. We now demonstrate that this interaction is of functional importance by showing that the acetylation rate constant of a substrate bound to the A-site is increased by a factor a when a second molecule of substrate binds to the P-site. This demonstration became feasible through the introduction of a new acetanilide substrate analogue of acetylcholine, 3-(acetamido)-N,N,N-trimethylanilinium (ATMA), for which a = 4. This substrate has a low acetylation rate constant and equilibrates with the catalytic site, allowing a tractable algebraic solution to the rate equation for substrate hydrolysis. ATMA affinities for the A- and P-sites deduced from the kinetic analysis were confirmed by fluorescence titration with thioflavin T as a reporter ligand. Values of a >1 give rise to a hydrolysis profile called substrate activation, and the AChE site-specific mutant W86F, and to a lesser extent wild-type human AChE itself, showed substrate activation with acetylthiocholine as the substrate. Substrate activation was incorporated into a previous catalytic scheme for AChE in which a bound P-site ligand can also block product dissociation from the A-site, and two additional features of the AChE catalytic pathway were revealed. First, the ability of a bound P-site ligand to increase the substrate acetylation rate constant varied with the structure of the ligand: thioflavin T accelerated ATMA acetylation by a factor a(2) of 1.3, while propidium failed to accelerate. Second, catalytic rate constants in the initial intermediate formed during acylation (EAP, where EA is the acyl enzyme and P is the alcohol leaving group cleaved from the ester substrate) may be constrained such that the leaving group P must dissociate before hydrolytic deacylation can occur.


Hollow metallic waveguides integrated with terahertz quantum cascade lasers.
We present the realization of a compact, monolithically integrated arrangement of terahertz quantum cascade lasers with hollow metallic cylindrical waveguides. By directly mounting a copper pipe to the end facet of a double metal waveguide, it was possible to significantly improve the far field emission from such a sub-wavelength plasmonic mode, while preserving the characteristic performance of the laser. Careful alignment of the quantum cascade laser and the hollow waveguide is required in order to prevent the excitation of higher order/mixed modes as predicted with a high degree of accuracy by a theoretical model. Finally, this approach proved to be a superior method of beam shaping when compared to other in situ arrangements, such as a silicon hyper-hemispherical lens glued to the facet, which are presented.


Facile production of nanocomposites of carbon nanotubes and polycaprolactone with high aspect ratios with potential applications in drug delivery.
The geometries and surface properties of nanocarriers greatly influence the interaction between nanomaterials and living cells. In this work we combine multiwalled carbon nanotubes (CNTs) with poly-ε-caprolactone (PCL) to produce non-spherical nanocomposites with high aspect ratios by using a facile emulsion solvent evaporation method. Particles were characterised by dynamic light scattering (DLS), scanning electron microscopy (SEM), atomic force microscopy (AFM) and asymmetric flow field flow fractionation (AF4). Different sizes and morphologies of nanoparticles were produced depending on the concentration of the sodium dodecyl sulphate (SDS), CNTs and PCL. Rod-like PCL-CNT nanostructures with low polydispersity were obtained with 1.5 mg mL-1 of SDS, 0.9 mg mL-1 of CNTs and 10 mg mL-1 PCL. AFM analysis revealed that the PCL and PCL-CNT nanocomposite had comparatively similar moduli of 770 and 560 MPa respectively, indicating that all the CNTs have been coated with at least 2 nm of PCL. Thermogravimetric analysis of the PCL-CNT nanocomposite indicated that they contained 9.6% CNTs by mass. The asymmetric flow field flow fractionation of the samples revealed that the PCL-CNT had larger hydrodynamic diameters than PCL alone. Finally, the drug loading properties of the nanocomposites were assessed using docetaxel as the active substance. The nanocomposites showed comparable entrapment efficiencies of docetaxel (89%) to the CNTs alone (95%) and the PCL nanoparticles alone (81%). This is a facile method for obtaining non-spherical nanocomposites that combines the properties of PCL and CNTs such as the high aspect ratio, modulus. The high drug entrapment efficiency of these nanocomposites may have promising applications in drug delivery.


Water soluble compounds of Rosmarinus officinalis L. improve the oxidative and inflammatory states of rats with adjuvant-induced arthritis.
Anti-inflammatory and antioxidant properties have been attributed to constituents of Rosmarinus officinalis. Considering the inflammatory nature of arthritis and the oxidative stress induced by the disease, this work aimed to investigate if R. officinalis is able to attenuate oxidative and inflammatory injuries caused by the disease. Rats with complete Freund's adjuvant induced arthritis were used. An aqueous extract of the rosemary leaves was administered orally. The parameters related to oxidative stress were measured in the liver, brain and plasma. The administration of the rosemary extract (RE) to arthritic rats diminished oxidative damage (e.g., less carbonylated proteins), improved the oxidative state (e.g., less reactive oxygen species) and also increased the antioxidant capacity by increasing the GSH level and the GSH/GSSG ratio and by almost normalizing the activity of several antioxidant enzymes. The treatment of arthritic rats with the extract also diminished paw edema, the number of leukocytes recruited in the femoro-tibial joint cavities and the weight of the lymph nodes and delayed the appearance of secondary lesions. Twelve phenolic compounds were identified and quantified in the aqueous RE, rosmarinic acid being the most abundant one. In addition, the extract also contains polysaccharides and proteins. The anti-inflammatory and antioxidant activities of the RE can be attributed, partly at least, to its content of polyphenolics with the strong possibility of synergistic interactions as well as metabolic activations. The results corroborate and amplify the general notion that rosemary aqueous extracts possess efficient anti-inflammatory agents and suggest that they are able to attenuate the oxidative stress inherent to arthritis.


Morphology and electrophysiological properties of immunocytochemically identified rat dopamine neurons recorded in vitro.
In vitro intracellular recordings were made from neurons in the rat midbrain slice. Two neuronal types could be distinguished in dopamine-containing (DA) midbrain regions based on electrophysiological criteria. One neuron type exhibited short duration action potentials (less than 1.5 msec), could fire at high frequencies (greater than 10 Hz), and exhibited either phasic or burst firing patterns. This neuron did not exhibit tyrosine hydroxylase immunoreactivity. A second neuronal type exhibited a unique set of electrophysiological properties, which included (1) a spontaneous pacemaker-like depolarizing potential, (2) a highly regular firing pattern, (3) long duration (greater than 2 msec) action potentials, and (4) a high (i.e., depolarized) spike threshold. This neuron was consistently double labeled using intracellular staining and immunocytochemical localization of the catecholamine-specific enzyme tyrosine hydroxylase, and thus represented the DA neuronal type. Midbrain DA neurons stained with Lucifer yellow could be separated into 3 classes based on their location and morphology: (1) fusiform neurons with laterally projecting dendrites in the dorsal substantia nigra zona compacta region, (2) multipolar cells with laterally and ventrally projecting dendrites in the ventral substantia nigra zona compacta, and (3) neurons with fusiform and multipolar somata and radially projecting dendrites in the ventral tegmental area. The dendrites also exhibited spine-like protrusions and ended with specialized forked processes. Spontaneously firing DA cells recorded in vitro had a number of distinguishing electrophysiological characteristics in common with those of DA neurons recorded in vivo, such as the presence of a slow depolarizing potential driving spike activity and a characteristic depolarized spike threshold (approximately-36 mV). However, in contrast to that found in vivo, the DA cells characterized here exhibited substantially higher input resistances and fired spontaneously in a very regular pacemaker pattern. Burst firing was not observed. Spike activity was apparently dependent on 4 depolarizing events: (1) a voltage-dependent TTX-sensitive slow depolarization, (2) a cobalt-sensitive low threshold depolarization that was activated during the rebound from brief membrane hyperpolarizations, (3) high threshold dendritic calcium spikes which gave rise to the spike afterhyperpolarization, and (4) a high threshold initial segment sodium spike. These depolarizations were modulated by several processes, including a 4-aminopyridine-insensitive delayed repolarization, an instantaneous and time-dependent anomalous rectifier, and an afterhyperpolarization. Although low threshold depolarizations and rebound action potentials could be triggered by the membrane repolarization following small membrane hyperpolarizations, comparatively larger hyperpolarizations attenuated this rebound activation, thereby suppressing anodal break excitation.(ABSTRACT TRUNCATED AT 400 WORDS)


Ordering Individuals with Sum Scores: The Introduction of the Nonparametric Rasch Model.
When a simple sum or number-correct score is used to evaluate the ability of individual testees, then, from an accountability perspective, the inferences based on the sum score should be the same as the inferences based on the complete response pattern. This requirement is fulfilled if the sum score is a sufficient statistic for the parameter of a unidimensional model. However, the models for which this holds true are known to be restrictive. It is shown that the less restrictive nonparametric models could result in an ordering of persons that is different from an ordering based on the sum score. To arrive at a fair evaluation of ability with a simple number-correct score, ordinal sufficiency is defined as a minimum condition for scoring. The monotone homogeneity model, together with the property of ordinal sufficiency of the sum score, is introduced as the nonparametric Rasch model. A basic outline for testable hypotheses about ordinal sufficiency, as well as illustrations with real data, is provided.


Eyeglass. 1. Very large aperture diffractive telescopes.
The Eyeglass is a very large aperture (25-100-m) space telescope consisting of two distinct spacecraft, separated in space by several kilometers. A diffractive lens provides the telescope s large aperture, and a separate, much smaller, space telescope serves as its mobile eyepiece. Use of a transmissive diffractive lens solves two basic problems associated with very large aperture space telescopes; it is inherently launchable (lightweight, packagable, and deployable) it and virtually eliminates the traditional, very tight surface shape tolerances faced by reflecting apertures. The potential drawback to use of a diffractive primary (very narrow spectral bandwidth) is eliminated by corrective optics in the telescope s eyepiece; the Eyeglass can provide diffraction-limited imaging with either single-band (Deltalambda/lambda approximately 0.1), multiband, or continuous spectral coverage.


Antioxidant activity and hepatoprotective potential of agaro-oligosaccharides in vitro and in vivo.
Agaro-oligosaccharides derived from red seaweed polysaccharide have been reported to possess antioxidant activity. In order to assess the live protective effects of agar-oligosaccharides, we did both in vitro and in vivo studies based on own-made agaro-oligosaccharides, and the structural information of this oligosaccharide was also determined. Structure of agaro-oligosaccharides prepared with acid hydrolysis on agar was confirmed by matrix-assisted ultraviolet laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) and NMR. The antioxidant effect of agaro-oligosaccharides on intracellular reactive oxygen species (ROS) was assessed by 2', 7'-dichlorofluorescin diacetate. Carbon tetrachloride was used to induce liver injury, some index including SOD, GSH-Px, MDA, AST, ALT were examined to determine the hepatoprotective effect of agaro-oligosaccharides. Agaro-oligosaccharides we got were composed of odd polymerizations with molecular weights ranged from 500 to 2500. Results from intracellular test indicated that agaro-oligosaccharides could significantly scavenge the level of oxidants in the hepatocytes, more beneficially, also associated with the improvement of cell viability In vivo studies of the antioxidant effects on tissue peroxidative damage induced by carbon tetrachloride in rat model indicated that agaro-oligosaccharides could elevate the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and decrease the level of malondialdehyde (MDA), glutamate oxaloacetate transaminase (AST), glutamic pyruvic transaminase (ALT) significantly. At 400 mg/kg, MDA level reduced 44 % and 21 % in liver and heart, SOD and GSH-Px increased to highest in liver and serum, while ALT level decreased 22.16 % in serum. Overall, the results of the present study indicate that agaro-oligosaccharides can exert their in vitro and in vivo hepatoprotective effect through scavenging oxidative damage induced by ROS.


A Fluorescence-Based High Throughput Screen for the Transporter Associated with Antigen Processing.
The transporter associated with antigen processing (TAP) is essential for antigen presentation by major histocompatibility complex (MHC) class I molecules. Traditional methods used to analyze peptide transport mediated by TAP require radioactive labeling of peptides and time-consuming manipulation of Concanavalin A-Sepharose. Drug discovery research requires rapid and reliable evaluation of large number of samples for bioactivity. To meet these requirements a nonradioactive, HTS assay for peptide transport activity of TAP has been developed. The radioactive label in the traditional assays has been replaced by a fluorescent label without compromising the transport efficiency of labeled peptide or the sensitivity of the assay. The use of multiscreen filtration plates has facilitated higher throughput and eliminated the centrifugation steps used in traditional TAP assays. The HTS assay shows similar kinetic characteristics as compared to the traditional assay. The HTS assay has been adapted on a Quadratrade mark 96-320 96-channel pipetting station (Tomtec, Hamden, CT) by optimizing time course, dose response of TAP to peptides and adenosine triphosphate (ATP), signal/noise ratio, reproducibility, and reagent stability. This HTS system has been utilized to screen a multiplexed compound library with a maximum of throughput 17,600 compounds per week.


The EnvIMS Study: Design and Methodology of an International Case-Control Study of Environmental Risk Factors in Multiple Sclerosis.
Multiple sclerosis (MS) is a chronic disease of the central nervous system, often resulting in significant neurological disability. The causes of MS are not known; however, the incidence of MS is increasing, thereby suggesting that changes in lifestyle and/or environmental factors may be responsible. On this background, the Environmental Risk Factors in MS Study or EnvIMS study was designed to further explore the etiology of MS. The design and methodology are described, providing details to enable investigators to (i) use our experiences to design their own studies; (ii) take advantage of, and build on the methodological work completed for, the EnvIMS study; (iii) become aware of this data source that is available for use by the research community. EnvIMS is a multinational case-control study, enrolling 2,800 cases with MS and 5,012 population-based controls in Canada, Italy, Norway, Serbia and Sweden. The study was designed to investigate the most commonly implicated risk factors for MS etiology using a self-report questionnaire. The use of a common methodology to study MS etiology across several countries enhances the comparability of results in different geographic regions and research settings, reduces the resources required for study design and enhances the opportunity for data harmonization.


Simple diode pumping of a power-buildup cavity.
We have developed a novel and simple passive technique to frequency lock an antireflection-coated diode laser to a power-buildup cavity. The beam reflected from a mode-mismatched cavity is spatially filtered so the maximum reflected intensity occurs at a cavity resonance. We experimentally and theoretically investigated the locking behavior and found that it is both robust and stable. In an implementation of the technique a 100-W intracavity beam can be generated by use of only a two-mirror cavity and a 15-mW 635-nm diode laser driven by a dc current.


Pollen-stigma adhesion in Arabidopsis: a species-specific interaction mediated by lipophilic molecules in the pollen exine.
To investigate the nature and role of cell adhesion in plants, we analyzed the initial step of pollination in Arabidopsis: the binding of pollen grains to female stigma cells. Here we show this interaction occurs within seconds of pollination. Because it takes place prior to pollen hydration, it also requires adhesion molecules that can act in a virtually dry environment. We developed assays that monitored adhesion of populations of pollen grains and individual cells. Adhesion between pollen and stigma cells is highly selective - Arabidopsis pollen binds with high affinity to Arabidopsis stigmas, while pollen from other species fails to adhere. Initial binding is independent of the extracellular pollen coat (tryphine), indicating that adhesion molecules reside elsewhere on the pollen surface, most likely within the exine walls. Immediately after pollination, the stigma surface becomes altered at the interface, acquiring a pattern that interlocks with the exine; this pattern is evident only with pollen from Arabidopsis and its close relatives. Purified exine fragments bind to stigma cells, and biochemical analyses indicate that this specific, rapid and anhydrous adhesion event is mediated by lipophilic interactions.


Contractile properties of inner and outer smooth muscle bundles from pig urinary detrusor.
Like in the human detrusor, the pig urinary detrusor muscle consists of two layers: compactly arranged smooth muscle bundles on the mucosal side (inner layer) and loosely arranged smooth muscle bundles on the serosal side (outer layer). The contractile properties of muscle bundles of both layers were measured using the stop test followed by an isometric contraction. Total and passive forces were measured in ten muscle bundles from the inner and outer muscle layers. Active force was defined as the difference between total and passive force. The curvature and the unloaded shortening velocity of the force-velocity relation were calculated from the shortening forces measured during the stop test. The rate of force development was calculated from the isometric contraction. Differences in contractile properties between both layers were pairwise tested using the Wilcoxon Signed Ranks test. Percentage wise, the outer layer muscle bundles produced the highest active isometric force. The shortening forces were also higher in the outer layer bundles. As a result, both the curvature and the unloaded shortening velocity, derived from the average force-velocity relations fitted to the data sets, were higher in the muscle bundles from this layer. Finally, the outer layer muscle bundles contracted significantly faster than those of the inner layer. Muscle bundles from the outer layer of pig detrusor were found to be faster and stronger (more phasic) than the weaker and slower (more tonic) bundles from the inner layer, suggesting that during bladder contraction the outer layer of the detrusor does more work than the inner layer.


Antibody-protein interactions: benchmark datasets and prediction tools evaluation.
The ability to predict antibody binding sites (aka antigenic determinants or B-cell epitopes) for a given protein is a precursor to new vaccine design and diagnostics. Among the various methods of B-cell epitope identification X-ray crystallography is one of the most reliable methods. Using these experimental data computational methods exist for B-cell epitope prediction. As the number of structures of antibody-protein complexes grows, further interest in prediction methods using 3D structure is anticipated. This work aims to establish a benchmark for 3D structure-based epitope prediction methods. Two B-cell epitope benchmark datasets inferred from the 3D structures of antibody-protein complexes were defined. The first is a dataset of 62 representative 3D structures of protein antigens with inferred structural epitopes. The second is a dataset of 82 structures of antibody-protein complexes containing different structural epitopes. Using these datasets, eight web-servers developed for antibody and protein binding sites prediction have been evaluated. In no method did performance exceed a 40% precision and 46% recall. The values of the area under the receiver operating characteristic curve for the evaluated methods were about 0.6 for ConSurf, DiscoTope, and PPI-PRED methods and above 0.65 but not exceeding 0.70 for protein-protein docking methods when the best of the top ten models for the bound docking were considered; the remaining methods performed close to random. The benchmark datasets are included as a supplement to this paper. It may be possible to improve epitope prediction methods through training on datasets which include only immune epitopes and through utilizing more features characterizing epitopes, for example, the evolutionary conservation score. Notwithstanding, overall poor performance may reflect the generality of antigenicity and hence the inability to decipher B-cell epitopes as an intrinsic feature of the protein. It is an open question as to whether ultimately discriminatory features can be found.


Broadly tunable L-band multiwavelength BEFL utilizing nonlinear amplified loop mirror filter.
We demonstrate a widely tunable L-band multiwavelength Brillouin-erbium fiber laser utilizing a nonlinear amplified fiber loop mirror filter (AFLMF). By manipulating polarization controllers placed in the fiber loop, the erbium peak gain spectrum is able to be shifted. The nonlinear AFLMF induces wavelength-dependent cavity loss and serves as an amplitude equalizer. In addition, it provides flexibility on controlling the amount of light reflected and transmitted into and out of the laser's cavity. By utilizing 100 mW 1480 nm pump and 1.1 mW Brillouin pump power, an average of 24 stable output channels are generated by the proposed structure that could all be tuned over the whole L-band window from 1570 nm to 1610 nm.


Linkage of lamins to fidelity of gene transcription.
Two major events are emerging as being involved in the fine regulatory mechanisms that control gene expression through the nuclear envelope/lamina. The first is the linkage of envelope proteins to transcription factors, most of which function as transcriptional repressors. The second is the interplay between nuclear lamina/nuclear envelope proteins and chromatin, through more or less direct interactions with DNA-binding proteins. The first mechanism appears to play a major role in tissue-specific functions, whereas the latter mechanism likely modifies higher-order chromatin organization, thus regulating differentiation and aging. This review highlights the importance of the study of a group of genetic diseases, collectively referred to as laminopathies, to understand the actual role played by lamins in ensuring the fidelity of gene expression and in modulating cell differentiation.