Phosphorylation of Subunit Proteins of Intermediate Filaments from Chicken Muscle and Nonmuscle Cells

The phosphorylation of the subunit proteins of intermediate (10-nm) filaments has been investigated in chicken muscle and nonmuscle cells by using a two-dimensional gel electrophoresis system. Desmin, the 50,000-dalton subunit protein of the intermediate filaments of muscle, had previously been shown to exist as two major isoelectric variants--alpha and ß --in smooth, skeletal, and cardiac chicken muscle. Incubation of skeletal and smooth muscle tissue with 32PO4{}3- reveals that the acidic variant, alpha -desmin, and three other desmin variants are phosphorylated in vivo and in vitro. Under the same conditions, minor components of alpha - and ß -tropomyosin from skeletal muscle, but not smooth muscle, are also phosphorylated. Both the phosphorylated desmin variants and the nonphosphorylated ß -desmin variant remain insoluble under conditions that solubilize actin and myosin filaments, but leave Z-discs and intermediate filaments insoluble. Primary cultures of embryonic chicken muscle labeled with 32PO4{}3- possess, in addition to the desmin variants described above, a major nonphosphorylated and multiple phosphorylated variants of the 52,000-dalton, fibroblast-type intermediate filament protein (IFP). Filamentous cytoskeletons, prepared from primary myogenic cultures by Triton X-100 extraction, contain actin and all of the phosphorylated and nonphosphorylated variants of both desmin and the IFP. Similarly, these proteins are the major components of the caps of aggregated 10-nm filaments isolated from the same cell cultures previously exposed to Colcemid. These results demonstrate that a nonphosphorylated and several phosphorylated variants of desmin and IFP are present in assembled structures in muscle and nonmuscle cells

Impact of Environmental Factors on Bacteriocin Promoter Activity in Gut-Derived Lactobacillus salivarius

peer-reviewedBacteriocin production is regarded as a desirable probiotic trait that aids in colonization and persistence in the gastrointestinal tract (GIT). Strains of Lactobacillus salivarius, a species associated with the GIT, are regarded as promising probiotic candidates and have a number of associated bacteriocins documented to date. These include multiple class IIb bacteriocins (salivaricin T, salivaricin P, and ABP-118) and the class IId bacteriocin bactofencin A, which show activity against medically important pathogens. However, the production of a bacteriocin in laboratory media does not ensure production under stressful environmental conditions, such as those encountered within the GIT. To allow this issue to be addressed, the promoter regions located upstream of the structural genes encoding the L. salivarius bacteriocins mentioned above were fused to a number of reporter proteins (green fluorescent protein [GFP], red fluorescent protein [RFP], and luciferase [Lux]). Of these, only transcriptional fusions to GFP generated signals of sufficient strength to enable the study of promoter activity in L. salivarius. While analysis of the class IIb bacteriocin promoter regions indicated relatively weak GFP expression, assessment of the promoter of the antistaphylococcal bacteriocin bactofencin A revealed a strong promoter that is most active in the absence of the antimicrobial peptide and is positively induced in the presence of mild environmental stresses, including simulated gastric fluid. Taken together, these data provide information on factors that influence bacteriocin production, which will assist in the development of strategies to optimize in vivo and in vitro production of these antimicrobials.This work was funded by a SFI PI award “Obesibiotics” (11/PI/1137) to PD

Meteorological drivers and mortality associated with O3 and PM2.5 air pollution episodes in the UK in 2006

In this study we examine the meteorological drivers resulting in concurrent high levels of ozone (O3) and particulate matter smaller than 2.5 andmu;m in diameter (PM2.5) during two five-day air pollution episodes in 2006 (1st - 5th July and 18th andndash; 22nd July) using an air quality model (AQUM) at 12 km horizontal resolution to simulate air pollutant concentrations. The resultant UK health burden associated with short-term exposure to simulated maximum daily 8-h O3andnbsp;(MDA8 O3) and daily mean PM2.5andnbsp;is estimated at the national and regional level. Both episodes were found to be driven by anticyclonic conditions with light easterly and south easterly winds and high temperatures that aided pollution build up in the UK. The estimated total mortality burden associated with short-term exposure to MDA8 O3andnbsp;is similar during the chosen episodes with about 70 daily deaths brought forward (summed across the UK) during the first and second episode, respectively. The estimated health burden associated with short-term exposure to daily mean PM2.5andnbsp;concentrations differs between the first and second episode resulting in about 43 and 36 daily deaths brought forward, respectively. The corresponding percentage of all-cause mortality due to short-term exposure to MDA8 O3andnbsp;and daily mean PM2.5during these two episodes and across the UK regions, ranges from 3.4% to 5.2% and from 1.6% to 3.9%, respectively. The attributable percentage of all-cause mortality differs between the regions depending on the pollution levels in each episode, but the overall estimated health burdens are highest in regions with higher population totals. We estimate that during these episodes the short-term exposure to MDA8 O3and daily mean PM2.5andnbsp;is between 36-38% and 39andndash;56% higher, respectively, than if the pollution levels represented typical seasonal-mean concentrations. This highlights the potential of air pollution episodes to have substantial short-term impacts on public health.</p

Future air pollution related health burdens associated with RCP emission changes in the UK

Intergovernmental Panel on Climate Change (IPCC) Representative Concentration Pathways (RCPs) are used to simulate future ozone (O3), nitrogen dioxide (NO2), and fine particulate matter (PM2.5) in the United Kingdom (UK) for the 2050s relative to the 2000s with an air quality model (AQUM) at a 12 km horizontal resolution. The present-day and future attributable fractions (AF) of mortality associated with long-term exposure to annual mean O3, NO2 and PM2.5 have accordingly been estimated for the first time for regions across England, Scotland and Wales. Across the three RCPs (RCP2.6, RCP6.0 and RCP8.5), simulated annual mean of the daily maximum 8-hr mean (MDA8) O3 concentrations increase compared to present-day, likely due to decreases in NOx (nitrogen oxides) emissions, leading to less titration of O3 by NO. Annual mean NO2 and PM2.5 concentrations decrease under all RCPs for the 2050s, mostly driven by decreases in NOx and sulphur dioxide (SO2) emissions, respectively. The AF of mortality associated with long-term exposure to annual mean MDA8 O3 is estimated to increase in the future across all the regions and for all RCPs. Reductions in NO2 and PM2.5 concentrations lead to reductions in the AF estimated for future periods under all RCPs, for both pollutants. Total mortality burdens are also highly sensitive to future population projections. Accounting for population projections exacerbates differences in total UK-wide MDA8 O3-health burdens between present-day and future by up to a factor of ~3 but diminishes differences in NO2-health burdens. For PM2.5, accounting for future population projections results in additional UK-wide deaths brought forward compared to present-day under RCP2.6 and RCP6.0, even though the simulated PM2.5 concentrations for the 2050s are estimated to decrease. Thus, these results highlight the sensitivity of future health burdens in the UK to future trends in atmospheric emissions over the UK as well as future population projections

Predicting hopelessness: the interaction between optimism/pessimism and specific future expectancies

Improving our understanding of hopelessness is central to suicide prevention. This is the first study to investigate whether generalized expectancies for the future (optimism/pessimism) and specific future-oriented cognitions (future thinking) interact to predict hopelessness and dysphoria. To this end, participants completed measures of future thinking, optimism/pessimism and affect at Time 1 and measures of affect and stress at Time 2, 10-12 weeks later. Results indicated that changes in hopelessness but not dysphoria were predicted by the interaction between positive future thinking (but not negative future thinking), optimism/pessimism and stress beyond initial levels of hopelessness and dysphoria. Additional moderating analyses are also reported. These findings point to the fruits of integrating personality and cognitive processes, to better understand hopelessness

The role of IREB2 and transforming growth factor beta-1 genetic variants in COPD: a replication case-control study

<p>Abstract</p> <p>Background</p> <p>Genetic factors are known to contribute to COPD susceptibility and these factors are not fully understood. Conflicting results have been reported for many genetic studies of candidate genes based on their role in the disease. Genome-wide association studies in combination with expression profiling have identified a number of new candidates including <it>IREB2</it>. A meta-analysis has implicated transforming growth factor beta-1 (<it>TGFbeta1</it>) as a contributor to disease susceptibility.</p> <p>Methods</p> <p>We have examined previously reported associations in both genes in a collection of 1017 white COPD patients and 912 non-diseased smoking controls. Genotype information was obtained for seven SNPs in the <it>IREB2 </it>gene, and for four SNPs in the <it>TGFbeta1 </it>gene. Allele and genotype frequencies were compared between COPD cases and controls, and odds ratios were calculated. The analysis was adjusted for age, sex, smoking and centre, including interactions of age, sex and smoking with centre.</p> <p>Results</p> <p>Our data replicate the association of <it>IREB2 </it>SNPs in association with COPD for SNP rs2568494, rs2656069 and rs12593229 with respective adjusted p-values of 0.0018, 0.0039 and 0.0053. No significant associations were identified for <it>TGFbeta1</it>.</p> <p>Conclusions</p> <p>These studies have therefore confirmed that the <it>IREB2 </it>locus is a contributor to COPD susceptibility and suggests a new pathway in COPD pathogenesis invoking iron homeostasis.</p

N-terminal pro–brain natriuretic peptide and exercise capacity in chronic heart failure: Data from the Heart Failure and a Controlled Trial Investigating Outcomes of Exercise Training (HF-ACTION) study

To examine the relationship between N-terminal pro-brain natriuretic peptide (NT-proBNP) and exercise capacity in a large contemporary cohort of patients with chronic heart failure

Sub-micron magnetic patterns and local variations of adhesion force induced in non-ferromagnetic amorphous steel by femtosecond pulsed laser irradiation

Periodic ripple and nanoripple patterns are formed at the surface of amorphous steel after femtosecond pulsed laser irradiation (FSPLI). Formation of such ripples is accompanied with the emergence of a surface ferromagnetic behavior which is not initially present in the non-irradiated amorphous steel. The occurrence of ferromagnetic properties is associated with the laser-induced devitrification of the glassy structure to form ferromagnetic (α-Fe and Fe₃C) and ferrimagnetic [(Fe,Mn)₃O₄ and Fe₂CrO4] phases located in the ripples. The generation of magnetic structures by FSPLI turns out to be one of the fastest ways to induce magnetic patterning without the need of any shadow mask. Furthermore, local variations of the adhesion force, wettability and nanomechanical properties are also observed and compared to those of the as-cast amorphous alloy. These effects are of interest for applications (e.g., biological, magnetic recording, etc.) where both ferromagnetism and tribological/adhesion properties act synergistically to optimize material performance

Long-term responders on olaparib maintenance in high-grade serous ovarian cancer: Clinical and molecular characterization

Purpose: Maintenance therapy with olaparib has improved progression-free survival in women with high-grade serous ovarian cancer (HGSOC), particularly those harboring BRCA1/2 mutations. The objective of this study was to characterize long-term (LT) versus short-term (ST) responders to olaparib. Experimental Design: A comparative molecular analysis of Study 19 (NCT00753545), a randomized phase II trial assessing olaparib maintenance after response to platinum-based chemotherapy in HGSOC, was conducted. LT response was defined as response to olaparib/placebo > 2 years, ST as < 3 months. Molecular analyses included germline BRCA1/2 status, three-biomarker homologous recombination deficiency (HRD) score, BRCA1 methylation, and mutational profiling. Another olaparib maintenance study (Study 41; NCT01081951) was used as an additional cohort. Results: Thirty-seven LT (32 olaparib) and 61 ST (21 olaparib) patients were identified. Treatment was significantly associated with outcome (P < 0.0001), with more LT patients on olaparib (60.4%) than placebo (11.1%). LT sensitivity to olaparib correlated with complete response to chemotherapy (P < 0.05). In the olaparib LT group, 244 genetic alterations were detected, with TP53, BRCA1, and BRCA2 mutations being most common (90%, 25%, and 35%, respectively). BRCA2 mutations were enriched among the LT responders. BRCA methylation was not associated with response duration. High myriad HRD score (>42) and/or BRCA1/2 mutation was associated with LT response to olaparib. Study 41 confirmed the correlation of LT response with olaparib and BRCA1/2 mutation. Conclusions: Findings show that LT response to olaparib may be multifactorial and related to homologous recombination repair deficiency, particularly BRCA1/2 defects. The type of BRCA1/2 mutation warrants further investigation. (C) 2017 AACR

SPL7013 Gel (VivaGel®) Retains Potent HIV-1 and HSV-2 Inhibitory Activity following Vaginal Administration in Humans

SPL7013 Gel (VivaGel®) is a microbicide in development for prevention of HIV and HSV. This clinical study assessed retention and duration of antiviral activity following vaginal administration of 3% SPL7013 Gel in healthy women. Participants received 5 single doses of product with ≥5 days between doses. A cervicovaginal fluid (CVF) sample was collected using a SoftCup™ pre-dose, and immediately, or 1, 3, 12 or 24 h post-dose. HIV-1 and HSV-2 antiviral activities of CVF samples were determined in cell culture assays. Antiviral activity in the presence of seminal plasma was also tested. Mass and concentration of SPL7013 in CVF samples was determined. Safety was assessed by reporting of adverse events. Statistical analysis was performed using the Wilcoxon signed-rank test with Bonferroni adjustment; p≤0.003 was significant. Eleven participants completed the study. Inhibition of HIV-1 and HSV-2 by pre-dose CVF samples was negligible. CVF samples obtained immediately after dosing almost completely inhibited (median, interquartile range) HIV-1 [96% (95,97)] and HSV-2 [86% (85,94)], and activity was maintained in all women at 3 h (HIV-1 [96% (95,98), p = 0.9]; HSV-2 [94% (91,97), p = 0.005]). At 24 h, >90% of initial HIV-1 and HSV-2 inhibition was maintained in 6/11 women. SPL7013 was recovered in CVF samples obtained at baseline (46% of 105 mg dose). At 3 and 24 h, 22 mg and 4 mg SPL7013, respectively, were recovered. More than 70% inhibition of HIV-1 and HSV-2 was observed if there was >0.5 mg SPL7013 in CVF samples. High levels of antiviral activity were retained in the presence of seminal plasma. VivaGel was well tolerated with no signs or symptoms of vaginal, vulvar or cervical irritation reported. Potent antiviral activity was observed against HIV-1 and HSV-2 immediately following vaginal administration of VivaGel, with activity maintained for at least 3 h post-dose. The data provide evidence of antiviral activity in a clinical setting, and suggest VivaGel could be administered up to 3 h before coitus