Miracle-dz/UniFAR-FTUs huggingface dataset:
The kinetics of competitive antagonism of nicotinic acetylcholine receptors at physiological temperature

Detailed information about the ligand‐binding site of nicotinic acetylcholine receptors has emerged from structural and mutagenesis experiments. However, these approaches provide only static images of ligand–receptor interactions. Kinetic measurements of changes in protein function are needed to develop a more dynamic picture. Previously, we measured association and dissociation rate constants for competitive inhibition of current through embryonic muscle acetylcholine receptor channels at 25°C. Little is known about competitive antagonism at physiological temperatures. Here, we performed measurements at 37°C and used thermodynamics to estimate the energetics of antagonism. We used rapid solution exchange protocols to determine equilibrium and kinetics of inhibition of acetylcholine‐activated currents in outside‐out patches by (+)‐tubocurarine, pancuronium and cisatracurium. Kinetic rates as high as 600 s−1 were resolved by this technique. Binding was primarily enthalpy driven. The 12°C increase in temperature decreased equilibrium antagonist binding by 1.7‐ to 1.9‐fold. In contrast, association and dissociation rate constants increased 1.9‐ to 6.0‐fold. Activation energies for dissociation were 90 ± 6, 106 ± 8 and 116 ± 10 kJ mol−1 for cisatracurium, (+)‐tubocurarine and pancuronium, respectively. The corresponding apparent activation energies for association were 38 ± 6, 85 ± 6 and 107 ± 13 kJ mol−1. The higher activation energy for association of (+)‐tubocurarine and pancuronium compared with cisatracurium is notable. This may arise from either a more superficial binding site for the large antagonist cisatracurium compared to the other ligands, or from a change in receptor conformation upon binding of (+)‐tubocurarine and pancuronium but not cisatracurium. Differences in ligand desolvation and ligand conformation are not likely to be important.
Hag Mediates Adherence of Moraxella catarrhalis to Ciliated Human Airway Cells

ABSTRACT Moraxella catarrhalis is a human pathogen causing otitis media in infants and respiratory infections in adults, particularly patients with chronic obstructive pulmonary disease. The surface protein Hag (also designated MID) has previously been shown to be a key adherence factor for several epithelial cell lines relevant to pathogenesis by M. catarrhalis, including NCIH292 lung cells, middle ear cells, and A549 type II pneumocytes. In this study, we demonstrate that Hag mediates adherence to air-liquid interface cultures of normal human bronchial epithelium (NHBE) exhibiting mucociliary activity. Immunofluorescent staining and laser scanning confocal microscopy experiments demonstrated that the M. catarrhalis wild-type isolates O35E, O12E, TTA37, V1171, and McGHS1 bind principally to ciliated NHBE cells and that their corresponding hag mutant strains no longer associate with cilia. The hag gene product of M. catarrhalis isolate O35E was expressed in the heterologous genetic background of a nonadherent Haemophilus influenzae strain, and quantitative assays revealed that the adherence of these recombinant bacteria to NHBE cultures was increased 27-fold. These experiments conclusively demonstrate that the hag gene product is responsible for the previously unidentified tropism of M. catarrhalis for ciliated NHBE cells.
Rostral ventrolateral medulla: Selective projections to the thoracic autonomic cell column from the region containing C1 adrenaline neurons

Anterograde, retrograde, and combined axonal transport methods were used to describe the descending efferent projections of a region of rostral ventrolateral medullary reticular formation important in cardiovascular control. We have termed this region, which contains C1 adrenaline‐synthesizing neurons, the nucleus reticularis rostroventrolateralis (RVL).
Neutralizing antibodies to interferon beta-1b multiple sclerosis: a clinico-radiographic paradox in the BEYOND trial

Background: The frequency and impact of neutralizing antibodies (NAbs) to interferon beta-1b (IFNβ-1b) on clinical and radiographic outcomes is controversial. Objective: To assess NAb impact in the BEYOND study. Methods: 2244 patients were randomized (2:2:1) to receive IFNβ-1b, either 250 or 500 µg, or glatiramer acetate, 20 mg, and observed for 2–3.5 years. NAb titers were determined every 6 months. A titer ≥20 NU/ml was considered NAb positive. Efficacy was compared between NAb-positive and NAb-negative patients, using comprehensive statistical analyses, taking into account the delayed appearance of NAbs, the time-dependent changes in the relapse rate, spontaneous reversions to NAb-negative status, NAb-titer level, and also adjusting for baseline factors. Results: In the IFNβ-1b 250 µg group, NAb-positive titers were detected (≥ once) in 319 patients (37.0%); of these, 112 (35.1%) reverted to NAb-negative status. In the IFNβ-1b 500 µg group, 340 patients (40.7%) became NAb-positive and 119 (35.0%) reverted to NAb-negative status. In both IFNβ groups, especially the 250 µg arm, NAb-positive status was not associated with a convincing impact on any clinical outcome measure by any statistical analysis. By contrast, in both IFNβ groups, NAbs were associated with a very consistent deleterious impact on most MRI outcomes. Conclusion: There was a notable dissociation between the impact of NAbs on MRI and clinical outcomes. On MRI measures, the impact was consistent and convincing, whereas on clinical measures a negative impact of NAbs was not found. The basis for this clinico-radiographic paradox is unknown but it suggests that the relationship between NAbs and the therapeutic effects of IFNβ-1b is complex.
Assignment1 of the human Stra13 gene (STRA13) to chromosome 3p26 by in situ hybridization

We have recently cloned Stra13 (Boudjelal et al., 1997), a novel retinoic acid (RA) responsive basic helix-loop-helix (bHLH) factor that shares the highest identity with the murine Hes proteins (Sasai et al., 1992; Ishibashi et al., 1993). Overexpression of Stra13 in P19 embryonal carcinoma cells results in increased proliferation, and an altered differentiation program in the presence of RA. Since dysregulated expression of bHLH factors has been reported in leukemogenesis and solid tumor development (Robb et al., 1997; Lipkowitz et al., 1992; Ball et al., 1993; Borges et al., 1997) we determined the localization of STRA13 and show that it maps to the short arm of chromosome 3 which is frequently affected in several cancers (Rostomily et al., 1997).
Studies on fate and toxicity of nanoalumina in male albino rats

The work aimed to evaluate the nanoalumina toxicity on the histological architecture, some haematological and biochemical aspects in male albino rats, during acute and sublethal experiments. Rats, in acute experiments, were injected with a single-acute dose of 3.9 g or 6.4 g or 8.5 g of aluminium oxide (Al2O3) kg− 1, whereas those of sublethal were injected with 1.3 g of Al2O3 kg− 1 2 days− 1. One-way analysis of variance indicated that injected doses and the experimental periods were significantly affected by haemoglobin (Hb) content; haematocrit value (Hct); white blood cell (WBC) count; blood platelet (Plt) count; mean corpuscular volume (MCV); mean corpuscular Hb (MCH) and MCH concentration (MCHC). In acute experiments, Hct, WBC count, MCV and Plt were significantly higher than the corresponding controls, whereas Hb, MCH and MCHC markedly decreased. In comparison with the related controls after 1, 3 and 7 days post-injection, red blood cell count, Hb, Hct, WBC count, Plt and MCV were significantly increased, but begun to decrease after 14 or/and 28 days and were associated with a marked decrease in MCH and MCHC. In serum of rats injected with acute or sublethal dose, the concentrations of total protein (TP) and total lipid (TL) were significantly lesser than the corresponding controls, whereas the levels of urea, uric acid, creatinine and the activities of aspartate aminotransferase and alanine aminotransferase were markedly increased. The injected doses were directly proportional with all the studied biochemical parameter, except the TL and TP that exhibited a negative correlation. Histologically, the highest acute and sublethal doses of nanoalumina caused hepatic irregular disarray, necrosis to the hepatic and Kupffer cells that are associated with congested blood sinusoids. The renal tissues characterized by the appearance of inter-tubular congestion that is accompanied by the dilation of the vascular glomeruli that completely occupied Bowman’s capsule and accompanied with partial disappearance of the renal tubule’s brush border. The brain showed a progressive degeneration of neurons in both the experiments.
Impact of RING and BRCT Domain Mutations on BRCA1 Protein Stability, Localization and Recruitment to DNA Damage

Abstract Mutations within the tumor suppressor BRCA1 cause the majority of hereditary breast and ovarian cancers. The BRCA1 protein is an important regulator of DNA double-strand break repair, and BRCA1-deficient cells are highly sensitive to ionizing radiation. Furthermore, BRCA1 function may contribute to enforcement of the G2 cell cycle checkpoint. E3-ubiquitin ligase activity is the only known enzymatic activity of BRCA1, which is mediated by the N-terminal RING finger domain. The C-terminal BRCT repeat domain, which mediates protein-protein interactions, is the only other identified structural domain. By investigating cancer-linked mutations within each domain, we demonstrate that truncation of the BRCT domain greatly impairs the stability and nuclear localization of BRCA1 protein. A missense mutation within the RING domain does not affect these biochemical properties. However, both mutant forms of BRCA1 fail to colocalize in nuclear foci with the known BRCA1-interacting proteins BARD1 and BACH1, which are important for DNA repair. This failure occurs despite the continued ability of the RING mutant protein to interact with BACH1 and the ability of the BRCT mutant to interact with BARD1. Furthermore, neither mutant form of BRCA1 is recruited into DNA damage-associated foci marked by γ-H2AX. Therefore, our data suggest that both the RING and BRCT domains of BRCA1 are required for an early step in the function of BRCA1 during DNA repair: recruitment to the sites of DNA damage.
Comprehensive Genome-Wide Transcriptomic Analysis of Immature Articular Cartilage following Ischemic Osteonecrosis of the Femoral Head in Piglets

Objective Ischemic osteonecrosis of the femoral head (ONFH) in piglets results in an ischemic injury to the immature articular cartilage. The molecular changes in the articular cartilage in response to ONFH have not been investigated using a transcriptomic approach. The purpose of this study was to perform a genome-wide transcriptomic analysis to identify genes that are upregulated in the immature articular cartilage following ONFH. Methods ONFH was induced in the right femoral head of 6-week old piglets. The unoperated femoral head was used as the normal control. At 24 hours (acute ischemic-hypoxic injury), 2 weeks (avascular necrosis in the femoral head) and 4 weeks (early repair) after surgery (n = 4 piglets/time point), RNA was isolated from the articular cartilage of the femoral head. A microarray analysis was performed using Affymetrix Porcine GeneChip Array. An enrichment analysis and functional clustering of the genes upregulated due to ONFH were performed using DAVID and STRING software, respectively. The increased expression of selected genes was confirmed by a real-time qRTPCR analysis. Results Induction of ONFH resulted in the upregulation of 383 genes at 24 hours, 122 genes at 2 weeks and 124 genes at 4 weeks compared to the normal controls. At 24 hours, the genes involved in oxidoreductive, cell-survival, and angiogenic responses were significantly enriched among the upregulated genes. These genes were involved in HIF-1, PI3K-Akt, and MAPK signaling pathways. At 2 weeks, secretory and signaling proteins involved in angiogenic and inflammatory responses, PI3K-Akt and matrix-remodeling pathways were significantly enriched. At 4 weeks, genes that represent inflammatory cytokines and chemokine signaling pathways were significantly enriched. Several index genes (genes that are upregulated at more than one time point following ONFH and are known to be important in various biological processes) including HIF-1A, VEGFA, IL-6, IL6R, IL-8, CCL2, FGF2, TGFB2, MMP1, MMP3, ITGA5, FN and Col6A1 were upregulated in the immature articular cartilage following ONFH. A qRTPCR analysis of selected genes confirmed the upregulated expression observed in the microarray analysis. Conclusion Immature articular cartilage responds to ONFH by the upregulation of genes involved in hypoxic stress response, angiogenesis, matrix remodeling and inflammation. This study provides novel insights into the multi-faceted role of immature articular cartilage, with inflammation as a key component, following ONFH in piglets.
Self-Management of Lymphedema: A Systematic Review of the Literature From 2004 to 2011

Background:Little is known about the effectiveness of activities used to self-manage lymphedema. Objective:The aim of this study was to evaluate the level of evidence of peer-reviewed lymphedema self-care literature published from January 2004 to May 2011. Methods:Eleven major medical databases were searched. Articles were selected for inclusion or exclusion into the category of self-management of lymphedema by topic experts. The definition of self-management included activities that individuals initiate and perform for themselves without the assistance of others. Articles were scored according to the Oncology Nursing Society’s Putting Evidence Into Practice levels of evidence. Results:Sixteen articles met criteria for inclusion in this review, with self-management as the independent variable. Ten categories of self-management were established subsequently and articles were classified by levels of evidence. In these categories, no self-management studies were rated as “recommended for practice,” 2 studies were rated “likely to be effective,” none was rated “benefits balanced with harms,” 7were rated “effectiveness not established,” and 1 was rated “effectiveness unlikely. ” Discussion:Few studies included measures of outcomes associated with lymphedema, and there was a scarcity of randomized controlled trials in lymphedema self-management. A demonstrated need exists for the design and testing of self-management interventions that include appropriate outcome measures.
Analytically monitored digestion of silver nanoparticles and their toxicity on human intestinal cells

Abstract Orally ingested nanoparticles may overcome the gastrointestinal barrier, reach the circulatory system, be distributed in the organism and cause adverse health effects. However, ingested nanoparticles have to pass through different physicochemical environments, which may alter their properties before they reach the intestinal cells. In this study, silver nanoparticles are characterised physicochemically during the course of artificial digestion to simulate the biochemical processes occurring during digestion. Their cytotoxicity on intestinal cells was investigated using the Caco-2 cell model. Using field-flow fractionation combined with dynamic light scattering and small-angle X-ray scattering, the authors found that particles only partially aggregate as a result of the digestive process. Cell viabilities were determined by means of CellTiter-Blue® assay, 4′,6-diamidino-2-phenylindole-staining and real-time impedance. These measurements reveal small differences between digested and undigested particles (1–100 µg/ml or 1–69 particles/cell). The findings suggest that silver nanoparticles may indeed overcome the gastrointestinal juices in their particulate form without forming large quantities of aggregates. Consequently, the authors presume that the particles can reach the intestinal epithelial cells after ingestion with only a slight reduction in their cytotoxic potential. The study indicates that it is important to determine the impact of body fluids on the nanoparticles of interest to provide a reliable interpretation of their nano-specific cytotoxicity testing in vivo and in vitro.
Model Averaging Using Fractional Polynomials to Estimate a Safe Level of Exposure

Quantitative risk assessment involves the determination of a safe level of exposure. Recent techniques use the estimated dose‐response curve to estimate such a safe dose level. Although such methods have attractive features, a low‐dose extrapolation is highly dependent on the model choice. Fractional polynomials,( 1) basically being a set of (generalized) linear models, are a nice extension of classical polynomials, providing the necessary flexibility to estimate the dose‐response curve. Typically, one selects the best‐fitting model in this set of polynomials and proceeds as if no model selection were carried out. We show that model averaging using a set of fractional polynomials reduces bias and has better precision in estimating a safe level of exposure (say, the benchmark dose), as compared to an estimator from the selected best model. To estimate a lower limit of this benchmark dose, an approximation of the variance of the model‐averaged estimator, as proposed by Burnham and Anderson,( 2) can be used. However, this is a conservative method, often resulting in unrealistically low safe doses. Therefore, a bootstrap‐based method to more accurately estimate the variance of the model averaged parameter is proposed.
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4 10 13 17 22 28 3 12 21 31 D e D e p a p a r r t m e n t s t m e n t s This article presents the Incremental Commitment Model which emerged from a National Research Council study and related efforts; it shows promise of improving integration of hardware, software, and human factors into the systems engineering and acquisition process. ConOps:The Cryptex to Operational System Mission Success This article covers the Concept of Operations document – what it is and what it is not – and describes the role it played in four Air Force programs. This tutorial integrates the definitions and processes of software engineering standards into the software systems engineering process developed by the Institute of Electrical and Electronics Engineers. Framework in the systems engineering of complex systems that can be used to improve fault tolerance, describes some apparent deficiencies of the framework from a fault-tolerance perspective, and provides suggestions for improvement. This article provides a framework for examining the differences between systems engineering and system of systems engineering, and notes that additional work is needed in the development of normative and prescriptive models.
Thinning intensity and growth response in SW-European Scots pine stands

The effect of different thinning intensities on growth and yield was studied in Pinus sylvestris L. stands at the south-western limit of its distribution area (Central Spain), using five long-term thinning trials. Data were analysed collectively considering several factors (trial, block, plot and period) as random effects. Total volume and volume increment decreased with thinning intensity, this loss being more significant in the case of moderate and heavy thinning. No difference was found among treatments for total basal area or the increment in basal area. The results revealed an optimum basal area (Assmann’s definition) between 85 and 100% of the basal area in unthinned plots. Volume growth loss associated with heavy thinnings (reduction of 18% in volume increment) was smaller than that reported in Central and Northern European regions (greater than 25%). Height increment was not influenced by thinning, whereas dominant and quadratic mean diameter increments increased with the thinning intensity. The response of diameter growth to thinning was greater at younger ages (less than 50 y) and in medium-sized trees. RésuméL’effet de différentes intensités d’éclaircie sur la croissance et la production de peuplements de Pinus sylvestris L. a été étudié à la limite sud-ouest de sa zone de répartition (centre de l’Espagne), à partir de cinq essais d’éclaircie suivis à long terme. Les données ont été analysées collectivement en prenant en compte différents facteurs (essai, bloc, placeau et période) comme facteurs aléatoires. Le volume total et l’accroissement en volume diminuaient avec l’intensité d’éclaircie, cette perte devenant plus significative dans le cas d’éclaircies modérées et fortes. Aucune différence n’a été trouvée entre traitements pour la surface terrière totale ni pour sa vitesse d’accroissement. Les résultats révèlent un optimum de surface terrière (selon la définition d’Assmann) compris entre 85–100 % de la surface terrière des placeaux non éclaircis. Les pertes de croissance en volume associées aux éclaircies fortes (réduction de 18 %) étaient plus faibles que celles qui sont rapportées pour les régions du Centre et du Nord de l’Europe (supérieures à 25 %). L’accroissement en hauteur n’a pas été influencé par l’éclaircie alors que l’accroissement quadratique moyen en diamètre augmentait avec l’intensité d’éclaircie. La réponse en termes d’accroissement en diamètre à l’éclaircie a été plus forte pour les jeunes arbres (moins de 50 ans) et les arbres de taille moyenne.
In vivo13C NMR measurement of neurotransmitter glutamate cycling, anaplerosis and TCA cycle flux in rat brain during [2‐13C]glucose infusion

The aims of this study were twofold: (i) to determine quantitatively the contribution of glutamate/glutamine cycling to total astrocyte/neuron substrate trafficking for the replenishment of neurotransmitter glutamate; and (ii) to determine the relative contributions of anaplerotic flux and glutamate/glutamine cycling to total glutamine synthesis. In this work in vivo and in vitro13C NMR spectroscopy were used, with a [2‐13C]glucose or [5‐13C]glucose infusion, to determine the rates of glutamate/glutamine cycling, de novo glutamine synthesis via anaplerosis, and the neuronal and astrocytic tricarboxylic acid cycles in the rat cerebral cortex. The rate of glutamate/glutamine cycling measured in this study is compared with that determined from re‐analysis of 13C NMR data acquired during a [1‐13C]glucose infusion. The excellent agreement between these rates supports the hypothesis that glutamate/glutamine cycling is a major metabolic flux (∼0.20 µmol/min/g) in the cerebral cortex of anesthetized rats and the predominant pathway of astrocyte/neuron trafficking of neurotransmitter glutamate precursors. Under normoammonemic conditions anaplerosis was found to comprise 19–26% of the total glutamine synthesis, whilst this fraction increased significantly during hyperammonemia (∼32%). These findings indicate that anaplerotic glutamine synthesis is coupled to nitrogen removal from the brain (ammonia detoxification) under hyperammonemic conditions.