Quantitative and Dynamic Catalogs of Proteins Released during Apoptotic and Necroptotic Cell Death

The inflammatory functions of the cytokine tumor necrosis factor (TNF) rely on its ability to induce cytokine production and to induce cell death. Caspase-dependent and caspase-independent pathways-apoptosis and necroptosis, respectively-regulate immunogenicity by the release of distinct sets of cellular proteins. To obtain an unbiased, systems-level understanding of this important process, we here applied mass spectrometry-based proteomics to dissect protein release during apoptosis and necroptosis. We report hundreds of proteins released from human myeloid cells in time course experiments. Both cell death types induce receptor shedding, but only apoptotic cells released nucleosome components. Conversely, necroptotic cells release lysosomal components by activating lysosomal exocytosis at early stages of necroptosis-induced membrane permeabilization and show reduced release of conventionally secreted cytokines

Interaction of Salivary alpha-Amylase and Amylase-Binding-Protein A (AbpA) of Streptococcus gordonii with Glucosyltransferase of S. gordonii and Streptococcus mutans

<p>Abstract</p> <p>Background</p> <p>Glucosyltransferases (Gtfs), enzymes that produce extracellular glucans from dietary sucrose, contribute to dental plaque formation by <it>Streptococcus gordonii </it>and <it>Streptococcus mutans</it>. The alpha-amylase-binding protein A (AbpA) of <it>S. gordonii</it>, an early colonizing bacterium in dental plaque, interacts with salivary amylase and may influence dental plaque formation by this organism. We examined the interaction of amylase and recombinant AbpA (rAbpA), together with Gtfs of <it>S. gordonii </it>and <it>S. mutans</it>.</p> <p>Results</p> <p>The addition of salivary alpha-amylase to culture supernatants of <it>S. gordonii </it>precipitated a protein complex containing amylase, AbpA, amylase-binding protein B (AbpB), and the glucosyltransferase produced by <it>S. gordonii </it>(Gtf-G). rAbpA was expressed from an inducible plasmid, purified from <it>Escherichia coli </it>and characterized. Purified rAbpA, along with purified amylase, interacted with and precipitated Gtfs from culture supernatants of both <it>S. gordonii </it>and <it>S. mutans</it>. The presence of amylase and/or rAbpA increased both the sucrase and transferase component activities of <it>S. mutans </it>Gtf-B. Enzyme-linked immunosorbent assay (ELISA) using anti-Gtf-B antibody verified the interaction of rAbpA and amylase with Gtf-B. A <it>S. gordonii abp</it>A-deficient mutant showed greater biofilm growth under static conditions than wild-type in the presence of sucrose. Interestingly, biofilm formation by every strain was inhibited in the presence of saliva.</p> <p>Conclusion</p> <p>The results suggest that an extracellular protein network of AbpA-amylase-Gtf may influence the ecology of oral biofilms, likely during initial phases of colonization.</p

Development of a general calibration model and long-term performance evaluation of low-cost sensors for air pollutant gas monitoring

Assessing the intracity spatial distribution and temporal variability in air quality can be facilitated by a dense network of monitoring stations. However, the cost of implementing such a network can be prohibitive if traditional high-quality, expensive monitoring systems are used. To this end, the Real-time Affordable Multi-Pollutant (RAMP) monitor has been developed, which can measure up to five gases including the criteria pollutant gases carbon monoxide (CO), nitrogen dioxide (NO2), and ozone (O3), along with temperature and relative humidity. This study compares various algorithms to calibrate the RAMP measurements including linear and quadratic regression, clustering, neural networks, Gaussian processes, and hybrid random forest–linear regression models. Using data collected by almost 70 RAMP monitors over periods ranging up to 18 months, we recommend the use of limited quadratic regression calibration models for CO, neural network models for NO, and hybrid models for NO2 and O3 for any low-cost monitor using electrochemical sensors similar to those of the RAMP. Furthermore, generalized calibration models may be used instead of individual models with only a small reduction in overall performance. Generalized models also transfer better when the RAMP is deployed to other locations. For long-term deployments, it is recommended that model performance be re-evaluated and new models developed periodically, due to the noticeable change in performance over periods of a year or more. This makes generalized calibration models even more useful since only a subset of deployed monitors are needed to build these new models. These results will help guide future efforts in the calibration and use of low-cost sensor systems worldwide.</p

Observation of the superconducting proximity effect in Nb/InAs and NbNx/InAs by Raman scattering

URL:http://link.aps.org/doi/10.1103/PhysRevB.66.134530 DOI:10.1103/PhysRevB.66.134530High-quality thin Nb and NbN films (60-100 Å) are grown on (100) n+-InAs (n=1019cm-3) substrates by dc-magnetron sputter deposition. Studies of the electronic properties of interfaces between the superconductor and the semiconductor are done by Raman scattering measurements. The superconducting proximity effect at superconductor-semiconductor interfaces is observed through its impact on inelastic light scattering intensities originating from the near-interface region of InAs. The InAs longitudinal optical phonon LO mode (237cm-1) and the plasmon-phonon coupled modes L- (221cm-1) and L+ (1100 to 1350cm-1), for n+=1×1019-2×1019cm-3 are measured. The intensity ratio of the LO mode (associated with the near-surface charge accumulation region, in InAs) to that of the L- mode (associated with bulk InAs), is observed to increase by up to 40% below the superconducting transition temperature. This temperature-dependent change in light scattering properties is only observed with high quality superconducting films and when the superconductor and the semiconductor are in good electrical contact. A few possible mechanisms of the observed effect are proposed.We gratefully acknowledge support from the United States Department of Energy through Materials Research Laboratory~Grant No. DEFG02-96ER45439! ~I.V.R., A.C.A., L.H.G., T.A.T., J.F.D., P.W.B., J.F.K.!, and from the United States Department of Energy through Midwest Superconductivity Consortium ~MISCON! ~Grant No. DE FG02-90ER45427! and the NSF ~Grant No. DMR 96-23827! ~S.W.H., P.F.M.!. SEM, XRD, XPS, and RBS materials characterizations were performed at the Center for Microanalysis of Materials and Microfabrication Center at Frederick Seitz Materials Research Laboratory, University of Illinois at Urbana- Champaign ~Grant No. DE FG02-96ER45439!. Sandia is a multiprogram laboratory operated by Sandia Corporation, a Lockheed Martin company, for the United States Department of Energy under Contract No. DE-AC04-94AL85000

A surrogate method for comparison analysis of salivary concentrations of Xylitol-containing products

Background: Xylitol chewing gum has been shown to reduce Streptococcus mutans levels and decay. Two studies examined the presence and time course of salivary xylitol concentrations delivered via xylitol-containing pellet gum and compared them to other xylitol-containing products. Methods: A within-subjects design was used for both studies. Study 1, adults (N = 15) received three xylitol-containing products (pellet gum (2.6 g), gummy bears (2.6 g), and commercially available stick gum (Koolerz, 3.0 g)); Study 2, a second group of adults (N = 15) received three xylitol-containing products (pellet gum, gummy bears, and a 33% xylitol syrup (2.67 g). For both studies subjects consumed one xylitol product per visit with a 7-day washout between each product. A standardized protocol was followed for each product visit. Product order was randomly determined at the initial visit. Saliva samples (0.5 mL to 1.0 mL) were collected at baseline and up to 10 time points (~16 min in length) after product consumption initiated. Concentration of xylitol in saliva samples was analyzed using high-performance liquid chromatography. Area under the curve (AUC) for determining the average xylitol concentration in saliva over the total sampling period was calculated for each product. Results: In both studies all three xylitol products (Study 1: pellet gum, gummy bears, and stick gum; Study 2: pellet gum, gummy bears, and syrup) had similar time curves with two xylitol concentration peaks during the sampling period. Study 1 had its highest mean peaks at the 4 min sampling point while Study 2 had its highest mean peaks between 13 to 16 minutes. Salivary xylitol levels returned to baseline at about 18 minutes for all forms tested. Additionally, for both studies the total AUC for the xylitol products were similar compared to the pellet gum (Study 1: pellet gum - 51.3 [micro]g.min/mL, gummy bears - 59.6 [micro]g.min/mL, and stick gum - 46.4 [micro]g.min/mL; Study 2: pellet gum - 63.0 [micro]g.min/mL, gummy bears - 55.9 [micro]g.min/mL, and syrup - 59.0 [micro]g.min/mL). Conclusion: The comparison method demonstrated high reliability and validity. In both studies other xylitol-containing products had time curves and mean xylitol concentration peaks similar to xylitol pellet gum suggesting this test may be a surrogate for longer studies comparing various products.NIDCR-NIH U54 DE14254; Head Start, HRSA 90YD0188/03; and MCHB, HRSA R40MC03622-03

Low energy electron attachment to cyanamide (NH2CN)

Cyanamide (NH2CN) is a molecule relevant for interstellar chemistry and the chemical evolution of life. In the present investigation, dissociative electron attachment to NH2CN has been studied in a crossed electronmolecular beams experiment in the electron energy range from about 0 eV to 14 eV. The following anionic species were detected: NHCN-, NCN-, CN-, NH2-, NH-, and CH2-. The anion formation proceeds within two broad electron energy regions, one between about 0.5 and 4.5 eV and a second between 4.5 and 12 eV. A discussion of possible reaction channels for all measured negative ions is provided. The experimental results are compared with calculations of the thermochemical thresholds of the anions observed. For the dehydrogenated parent anion, we explain the deviation between the experimental appearance energy of the anion with the calculated corresponding reaction threshold by electron attachment to the isomeric form of NH2CN-carbodiimide(VLID)1020340Accepted versio

Analgesia and functional outcome after total knee arthroplasty: periarticular infiltration vs continuous femoral nerve block

BACKGROUND: Capacity to ambulate represents an important milestone in the recovery process after total knee arthroplasty (TKA). The purpose of this study was to determine the analgesic effect of two analgesic techniques and their impact on functional walking capacity as a measure of surgical recovery. METHODS: Forty ASA II-III subjects undergoing TKA were enrolled in a randomized, double-blind, single-centre study receiving 48 h postoperative analgesia with either periarticular infiltration of local anaesthetic (Group I) or continuous femoral nerve block (Group F). Breakthrough pain relief was achieved with patient-controlled analgesia (PCA) morphine. The main outcome was postoperative morphine consumption. Early (postoperative days 1-3) and late (6 weeks) functional walking capacity (2 and 6 min walk tests, 2MWT and 6MWT, respectively), degree of physical activity (CHAMPS), health-related quality of life (SF-12), and clinical indicators of knee function (WOMAC, Knee Society evaluation, and range of motion) were measured. RESULTS: Patients in Group F used the PCA less (P=0.02) to achieve adequate analgesia. Postoperative 2MWT was similar in both groups (P=0.27). Six weeks after surgery, recovery of 6MWT, physical activity, and knee function were significantly improved in Group F (P<0.05). Preoperative walking capacity, physical activity and early total walking time were the independent predictors of early recovery. Distance and time spent walking were the predictors of functional walking exercise capacity at 6 weeks after surgery. CONCLUSIONS: Femoral block is associated with lower opioid consumption and a better recovery at 6 weeks than periarticular infiltration. Early postoperative activity measures (2MWT and walking time) were proved to be possible indicators of knee function recovery at 6 weeks after surgery

Effectiveness, cost-effectiveness and cost-benefit of a single annual professional intervention for the prevention of childhood dental caries in a remote rural Indigenous community

Background The aim of the study is to reduce the high prevalence of tooth decay in children in a remote, rural Indigenous community in Australia, by application of a single annual dental preventive intervention. The study seeks to (1) assess the effectiveness of an annual oral health preventive intervention in slowing the incidence of dental caries in children in this community, (2) identify the mediating role of known risk factors for dental caries and (3) assess the cost-effectiveness and cost-benefit of the intervention. Methods/design The intervention is novel in that most dental preventive interventions require regular re-application, which is not possible in resource constrained communities. While tooth decay is preventable, self-care and healthy habits are lacking in these communities, placing more emphasis on health services to deliver an effective dental preventive intervention. Importantly, the study will assess cost-benefit and cost-effectiveness for broader implementation across similar communities in Australia and internationally. Discussion There is an urgent need to reduce the burden of dental decay in these communities, by implementing effective, cost-effective, feasible and sustainable dental prevention programs. Expected outcomes of this study include improved oral and general health of children within the community; an understanding of the costs associated with the intervention provided, and its comparison with the costs of allowing new lesions to develop, with associated treatment costs. Findings should be generalisable to similar communities around the world. The research is registered with the Australian New Zealand Clinical Trials Registry (ANZCTR), registration number ACTRN12615000693527; date of registration: 3rd July 2015

Identifying adolescents at risk for suboptimal adherence to tuberculosis treatment: A prospective cohort study

Adolescents account for an estimated 800,000 incident tuberculosis (TB) cases annually and are at risk for suboptimal adherence to TB treatment. Most studies of adolescent TB treatment adherence have used surveillance data with limited psychosocial information. This prospective cohort study aimed to identify risk factors for suboptimal adherence to rifampicin-susceptible TB treatment among adolescents (10-19 years old) in Lima, Peru. We collected psychosocial data using self-administered surveys and clinical data via medical record abstraction. Applying k-means cluster analysis, we grouped participants by psychosocial characteristics hypothesized to impact adherence. Then, we conducted mixed effects regression to compare suboptimal adherence-defined as 10% of doses)-between clusters. Treatment setting (facility vs. home) and drug formulation (single drug vs. fixed dose combination) were interaction terms. Of 249 participants, 90 (36.1%) were female. Median age was 17 (IQR: 15, 16.6) years. We identified three clusters-A, B, and C-of participants based on psychosocial characteristics. Cluster C had the lowest support from caregivers, other family members, and friends; had the weakest motivation to complete TB treatment; were least likely to live with their mothers; and had experienced the most childhood adversity. Among the 118 (47.4%) participants who received facility-based treatment with single drug formulations, adherence did not differ between Clusters A and B, but Cluster C had six-fold odds of suboptimal adherence compared to Cluster A. In Clusters B and C, adherence worsened over time, but only in Cluster C did mean adherence fall below 90% within six months. Our findings have implications for the care of adolescents with TB. When caring for adolescents with low social support and other risk factors, clinicians should take extra measures to reinforce adherence, such as identifying a community health worker or peer to provide treatment support. Implementing newly recommended shorter regimens also may facilitate adherence.Revisión por pare

MicroRNAs in pulmonary arterial remodeling

Pulmonary arterial remodeling is a presently irreversible pathologic hallmark of pulmonary arterial hypertension (PAH). This complex disease involves pathogenic dysregulation of all cell types within the small pulmonary arteries contributing to vascular remodeling leading to intimal lesions, resulting in elevated pulmonary vascular resistance and right heart dysfunction. Mutations within the bone morphogenetic protein receptor 2 gene, leading to dysregulated proliferation of pulmonary artery smooth muscle cells, have been identified as being responsible for heritable PAH. Indeed, the disease is characterized by excessive cellular proliferation and resistance to apoptosis of smooth muscle and endothelial cells. Significant gene dysregulation at the transcriptional and signaling level has been identified. MicroRNAs are small non-coding RNA molecules that negatively regulate gene expression and have the ability to target numerous genes, therefore potentially controlling a host of gene regulatory and signaling pathways. The major role of miRNAs in pulmonary arterial remodeling is still relatively unknown although research data is emerging apace. Modulation of miRNAs represents a possible therapeutic target for altering the remodeling phenotype in the pulmonary vasculature. This review will focus on the role of miRNAs in regulating smooth muscle and endothelial cell phenotypes and their influence on pulmonary remodeling in the setting of PAH