Data,Reagents, Assays and Merits of Proteomics for SARS-CoV-2 Research and Testing

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Highlights

• •In-depth proteomes of 4 SARS-CoV-2 cell line models (Vero E6, Calu-3, Caco-2, A549).
• •Proteomic evidence for thousands of Chlorocebus sabaeus proteins.
• •Proteomic response of Vero E6 cells to SARS-CoV-2 infection.
• •Synthetic peptides, spectral libraries, and targeted assays for SARS-CoV-2 proteins.

     

The time-dependent transport of chromium in adult rats from the bloodstream to the urine

While chromium was proposed to be an essential trace element over 40 years ago and if essential should possess a specific transport and distribution mechanism, the details of its transport from the bloodstream to the urine have not been elucidated. However, chromium is known to be maintained in the bloodstream bound to transferrin and to be excreted in the urine bound to the oligopeptide chromodulin or a similar chromodulin-like species. Injection of ⁵¹Cr-labeled transferrin into the bloodstream resulted in a rapid and insulin-sensitive movement of chromium into the tissues as Cr transferrin; greater than 50% of the Cr is transported to the tissues within 30 min. Tissue levels of Cr are maximal 30 min after injection; decreases in tissue Cr with time are mirrored by increases in urine Cr. Approximately 50% of the ⁵¹Cr appears in the urine within 360 min of injection in the absence of added insulin; insulin treatment concurrent with injection of ⁵¹Cr-labeled transferrin results in approximately 80% of the label appearing in the urine within 180 min. The removal of ⁵¹Cr from the blood is faster than the appearance of ⁵¹Cr in the urine; the lag in time indicates that the Cr transferrin in the blood and Cr in the urine are not in direct equilibrium and that intermediates in the transport of Cr must be involved. This establishes a clear pathway of transport of Cr starting from transport by transferrin from the bloodstream into the tissues, followed by release and processing in the tissues to form chromodulin, excretion into the bloodstream, rapid clearance of chromodulin or a similar species into the urine, and ultimately excretion as this species. Insulin stimulates the processing of Cr in the tissues.     

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.     

Selected Natural Materials in History

1 Technology of natural materials Early man used conveniently shaped stones as tools. ＂Workshop＂ areas have been found with large numbers of stones, some showing signs of being worked. However, organic materials like wood will decay under normal wet conditions in the presence of oxygen, so we won＇t find the same sort of evidence for wooden tools. It is safe to assume that early man used sticks as probes and clubs, and maybe even for making some sort of nestlike protection against the elements and predators, since we see chimpanzees and other animals doing this sort of thing. So wood, and ahnost certainly other plant materials such as fibrous leaves, and bone and other materials gleaned from dead animals, would be used from the earliest times. We need to know this in order to establish the idea that Man can be expected to have a long history of the use and manipulation of natural materials. This needs skills in choosing materials for certain uses on the basis of their mechanical properties, whether those properties are to do with the ease of shaping the material or the effectiveness of that material in use. Occasionally the material was chosen simply because it was readily available. If we find that a particular material was always used for a certain job, it＇s reasonable to deduce that Man was exerting materials selection criteria through experience.     

A novel strain of Brevibacillus laterosporus produces chitinases that contribute to its biocontrol potential

A novel strain exhibiting entomopathogenic and chitinolytic activity was isolated from mangrove marsh soil in India. The isolate was identified as Brevibacillus laterosporus by phenotypic characterization and 16S rRNA sequencing and designated Lak1210. When grown in the presence of colloidal chitin as the sole carbon source, the isolate produced extracellular chitinases. Chitinase activity was inhibited by allosamidin indicating that the enzymes belong to the family 18 chitinases. The chitinases were purified by ammonium sulfate precipitation followed by chitin affinity chromatography yielding chitinases and chitinase fragments with 90, 75, 70, 55, 45, and 25 kDa masses. Mass spectrometric analyses of tryptic fragments showed that these fragments belong to two distinct chitinases that are almost identical to two putative chitinases, a 89.6-kDa four-domain chitodextrinase and a 69.4-kDa two-domain enzyme called ChiA1, that are encoded on the recently sequenced genome of B. laterosporus LMG15441. The chitinase mixture showed two pH optima, at 6.0 and 8.0, and an optimum temperature of 70 °C. The enzymes exhibited antifungal activity against the phytopathogenic fungus Fusarium equiseti. Insect toxicity bioassays with larvae of diamondback moths (Plutella xylostella), showed that addition of chitinases reduced the time to reach 50 % mortality upon infection with non-induced B. laterosporus from 3.3 to 2.1 days. This study provides evidence for the presence of inducible, extracellular chitinolytic enzymes in B. laterosporus that contribute to the strain’s antifungal activity and insecticidal activity.     

Context-dependent role of ATG4B as target for autophagy inhibition in prostate cancer therapy

ATG4B belongs to the autophagin family of cysteine proteases required for autophagy, an emerging target of cancer therapy. Developing pharmacological ATG4B inhibitors is a very active area of research. However, detailed studies on the role of ATG4B during anticancer therapy are lacking. By analyzing PC-3 and C4-2 prostate cancer cells overexpressing dominant negative ATG4B^C74Ain vitro and in vivo, we show that the effects of ATG4B^C74A are cell type, treatment, and context-dependent. ATG4B^C74A expression can either amplify the effects of cytotoxic therapies or contribute to treatment resistance. Thus, the successful clinical application of ATG4B inhibitors will depend on finding predictive markers of response.     

Epistasis in iron metabolism: complex interactions between Cp, Mon1a, and Slc40a1 loci and tissue iron in mice

Disorders of iron metabolism are among the most common acquired and constitutive diseases. Hemochromatosis has a solid genetic basis and in Northern European populations it is usually associated with homozygosity for the C282Y mutation in the HFE protein. However, the penetrance of this mutation is incomplete and the clinical presentation is highly variable. The rare and common variants identified so far as genetic modifiers of HFE-related hemochromatosis are unable to account for the phenotypic heterogeneity of this disorder. There are wide variations in the basal iron status of common inbred mouse strains, and this diversity may reflect the genetic background of the phenotypic diversity under pathological conditions. We therefore examined the genetic basis of iron homeostasis using quantitative trait loci mapping applied to the HcB-15 recombinant congenic strains for tissue and serum iron indices. Two highly significant QTL containing either the N374S Mon1a mutation or the Ferroportin locus were found to be major determinants in spleen and liver iron loading. Interestingly, when considering possible epistatic interactions, the effects of Mon1a on macrophage iron export are conditioned by the genotype at the Slc40a1 locus. Only mice that are C57BL/10ScSnA homozygous at both loci display a lower spleen iron burden. Furthermore, the liver-iron lowering effect of the N374S Mon1a mutation is observed only in mice that display a nonsense mutation in the Ceruloplasmin (Cp) gene. This study highlights the existence of genetic interactions between Cp, Mon1a, and the Slc40a1 locus in iron metabolism, suggesting that epistasis may be a crucial determinant of the variable biological and clinical presentations in iron disorders.