Profiling SARS-CoV-2 HLA-I peptidome reveals T cell epitopes from out-of-frame ORFs

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Synapsin Isoforms and Synaptic Vesicle Trafficking

Synapsins were the first presynaptic proteins identified and have served as the flagship of the presynaptic protein field. Here we review recent studies demonstrating that different members of the synapsin family play different roles at presynaptic terminals employing different types of synaptic vesicles. The structural underpinnings for these functions are just beginning to be understood and should provide a focus for future efforts.     

Cloning and expression of the 3' portion of the T4 denV gene as a lacZ fusion gene

Polyclonal antibodies have been raised against endonuclease V from the bacteriophage T4. This rabbit serum, from which endemic E. coli antibodies have been removed, reacts with a single protein from T4-infected E. coli with a molecular weight of 16078 dalton. It was confirmed that these antibodies were directed against endonuclease V through the inhibition of the pyrimidine dimer specific nicking activity of endonuclease V in an in vitro nicking assay. A phage lambda gt11 T4 dC DNA library was screened for phage which produced a beta-galactosidase-endonuclease V fusion protein. Immunopositive clones were detected at a frequency of 0.25% of the plaques in the library. Restriction enzyme analyses of the DNA from 45 of these phage showed that all contained a 1.8 kb T4 EcoRI fragment which had been inserted within lambda gt11 in a single orientation. Western analysis of proteins which were produced from an induction of lysogens made from these phage reveals a single fusion protein band with a molecular weight slightly larger than native beta-galactosidase.     

Heavy Metals in Sediments of the Gold Mining Impacted Pra River Basin,Ghana, West Africa

Total concentrations of Hg, Al, Fe, As, Pb, Cu, Cr, Ni, Mn, Co, V, and Zn were determined in surface sediments collected from 21 locations within the gold mining impacted Pra River basin in southwestern Ghana. Samples were collected during both the rainy and dry seasons. We hypothesized that in the rural southwestern portion of Ghana, the lack of industrial activities makes artisanal gold mining (AGM) by Hg amalgamation the main source of water resource contamination with heavy metals. Therefore, metals showing concentration trends similar to that of Hg in the studied system are likely impacted by AGM. We found that total-Hg (THg) concentrations in riverine sediments are rather low as compared to other aquatic systems that are impacted by similar mining activities. Measured THg concentrations ranged from 0.018 to 2.917 mg/kg in samples collected in the rainy season and from about 0.01 to 0.043 mg/kg in those collected during the dry season. However, the determination of the enrichment factor (EF) calculated using shale data as reference background values showed signs of severe contamination in most of the sampled sites. In the dry season, THg concentrations correlated positively and significantly to the concentrations of As (r = 0.864, p < 0.01), Cu (r = 0.691, p < 0.05), and Ni (r = 0.579, p < 0.05). Based on our previously stated hypothesis, this could then be an indication of the impact of AGM on ambient levels of these 3 elements. However, the determined concentrations of Cu, and Ni co-varied significantly with Al, suggesting that natural sources do account for the observed levels. Accordingly, both AGM and metal inputs from weathered natural deposits are likely co-responsible for the observed levels of Cu and Ni. In contrast, the lack of correlation between As and Al tends to suggest an impact of AGM on As levels. Overall, our data suggest that besides Hg and to some extent As, the impact of AGM on ambient levels of investigated metals in the gold mining impacted Pra River remains negligible. Finally, the increase in metal concentrations from the dry to the rainy season underlines the impact of changes in hydrologic conditions on levels and fate of metals in this tropical aquatic system.     

Germline Mutations in NFKB2 Implicate the Noncanonical NF-κB Pathway in the Pathogenesis of Common Variable Immunodeficiency

Common variable immunodeficiency (CVID) is a heterogeneous disorder characterized by antibody deficiency, poor humoral response to antigens, and recurrent infections. To investigate the molecular cause of CVID, we carried out exome sequence analysis of a family diagnosed with CVID and identified a heterozygous frameshift mutation, c.2564delA (p.Lys855Serfs^∗7), in NFKB2 affecting the C terminus of NF-κB2 (also known as p100/p52 or p100/p49). Subsequent screening of NFKB2 in 33 unrelated CVID-affected individuals uncovered a second heterozygous nonsense mutation, c.2557C>T (p.Arg853^∗), in one simplex case. Affected individuals in both families presented with an unusual combination of childhood-onset hypogammaglobulinemia with recurrent infections, autoimmune features, and adrenal insufficiency. NF-κB2 is the principal protein involved in the noncanonical NF-κB pathway, is evolutionarily conserved, and functions in peripheral lymphoid organ development, B cell development, and antibody production. In addition, Nfkb2 mouse models demonstrate a CVID-like phenotype with hypogammaglobulinemia and poor humoral response to antigens. Immunoblot analysis and immunofluorescence microscopy of transformed B cells from affected individuals show that the NFKB2 mutations affect phosphorylation and proteasomal processing of p100 and, ultimately, p52 nuclear translocation. These findings describe germline mutations in NFKB2 and establish the noncanonical NF-κB signaling pathway as a genetic etiology for this primary immunodeficiency syndrome.     

Immunohistochemical analysis of the vitellogenin response in the liver of Atlantic salmon exposed to environmental oestrogens

Induction of vitellogenin (Vtg) in oviparous vertebrates has been used as a biomarker of response for environmental oestrogens. This study reports the cellular localization of oestrogen- and xenoestrogen-induced Vtg synthesis in the liver of juvenile Atlantic salmon (Salmo salar). Paraffin-embedded liver sections were incubated with homologous monoclonal antibody against Atlantic salmon Vtg. Following intraperitoneal (ip) exposure of fish to estradiol-17beta E₂; 5 mg kg-1 or 4-nonylphenol NP; 125 mg kg-1, Vtg induction was primarily demonstrated immunohistochemically in the cytoplasm of hepatocytes, endothelial cells and within hepatic sinusoids. Vtg staining of hepatocytes was not evenly distributed, as there was a high degree of polarization toward the sinusoid. The intensity of positive Vtg staining was stronger in the liver sections of E₂-treated fish, compared with NP-treated fish. Hepatocytes of E₂-, NP- and vehicle (control)-treated fish showed normal cellular structures, thus showing no evidence of histopathological changes. In parallel, indirect enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis of plasma Vtg levels show significant induction of Vtg in E₂- and NP-treated fish, as compared with untreated (control) fish. The present study demonstrates the applicability of immunohistochemistry in studies of cellular structures, processes and responses of fish exposure to oestrogen and oestrogen-mimicking compounds.     

Does Grazing Matter for Soil Organic Carbon Sequestration in the Western North American Great Plains?

Ecosystems - Considerable uncertainty remains regarding grazing-induced influences on soil organic carbon (SOC) sequestration in semiarid grassland ecosystems due to three important complications...