USP32 is an active, membrane-bound ubiquitin protease overexpressed in breast cancers

USP32, on chromosomal band 17q23.1-17q23.2, is a highly conserved but uncharacterized gene that gave rise during evolution to a well-known hominoid-specific proto-oncogene, USP6. We investigated the expression profile of USP32 in human tissues and examined its functions to gain insight into this novel member of the well-conserved ubiquitination system. We detected ubiquitous USP32 expression across tissues and confirmed the predicted deubiquitination function owing to the presence of conserved peptidase signature aspargine, cysteine, histidine, and aspartic acid domains of ubiquitin-specific proteases. A Golgi localization of GFP-fused USP32 was detected by fluorescent protection assay and BODIPY-TR staining. In addition, stable silencing of USP32 caused a significant decrease in the proliferation and migration rate of cells. Based on these and the fact that USP32 maps to 17q23, which is commonly amplified in breast cancers, we analyzed USP32 expression in breast cancer cells. We detected high expression of USP32 in 50% (9 of 18) of breast cancer cell lines and 22% (9 of 41) of primary breast tumors compared to mammary epithelial cells. In summary, we report the preliminary characterization of this novel deubiquitinating enzyme on 17q23 and demonstrate its functional role in the ubiquitin system and its potential involvement in tumorigenesis.     

Phosphorylation-mediated signalling in flowering: prospects and retrospects of phosphoproteomics in crops

Protein phosphorylation is a major post-translational modification, regulating protein function, stability, and subcellular localization. To date, annotated phosphorylation data are available mainly for model organisms and humans, despite the economic importance of crop species and their large kinomes. Our understanding of the phospho-regulation of flowering in relation to the biology and interaction between the pollen and pistil is still significantly lagging, limiting our knowledge on kinase signalling and its potential applications to crop production. To address this gap, we bring together relevant literature that were previously disconnected to present an overview of the roles of phosphoproteomic signalling pathways in modulating molecular and cellular regulation within specific tissues at different morphological stages of flowering. This review is intended to stimulate research, with the potential to increase crop productivity by providing a platform for novel molecular tools.     

In silico evaluation of flavonoids as effective antiviral agents on the spike glycoprotein of SARS-CoV-2

The novel coronavirus pandemic has spread over in 213 countries as of July 2020. Approximately 12 million people have been infected so far according to the reports from World Health Organization (WHO). Preventive measures are being taken globally to avoid the rapid spread of virus. In the current study, an in silico approach is carried out as a means of inhibiting the spike protein of the novel coronavirus by flavonoids from natural sources that possess both antiviral and anti-inflammatory properties. The methodology is focused on molecular docking of 10 flavonoid compounds that are docked with the spike protein of SARS-CoV-2, to determine the highest binding affinity at the binding site. Molecular dynamics simulation was carried out with the flavonoid-protein complex showing the highest binding affinity and highest interactions. The flavonoid naringin showed the least binding energy of −9.8 Kcal/mol with the spike protein which was compared with the standard drug, dexamethasone which is being repurposed to treat critically ill patients. MD simulation was carried out on naringin-spike protein complex for their conformational stability in the active site of the novel coronavirus spike protein. The RMSD of the complex appeared to be more stable when compared to that of the protein from 0.2 nm to 0.4 nm. With the aid of this in silico approach further in vitro studies can be carried out on these flavonoids against the novel coronavirus as a means of viral protein inhibitors.     

A Disynaptic Circuit in the Globus Pallidus Controls Locomotion Inhibition

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Biosorption of oxybenzene using biosorbent prepared by raw wastes of Zea mays and comparative study by using commercially available activated carbon

Organic pollutants present in waste water have undesirable effect on the environment. Industry activities are the key sources of organic pollutants. Prime pollutants released from various sources react instantly with the environment and become derived (secondary) pollutants, which stay for an elongated time. The present research work has been carried out using biosorbent prepared from various Zea mays wastes for elimination of oxybenzene. Different parameters viz contact time, initial concentration; adsorbent dose, temperature and pH were optimized for the biosorption of oxybenzene on to the biosorbent samples. BCS (Baby corn silk) showed higher percentage of biosorption at optimum contact time of 3 h, pH between 5 and 6 and temperature at 25 °C. Analysis of equilibrium biosorption data in terms of several isotherm models revealed that Langmuir isotherm and Freundlich isotherm indicates better agreement with the experimental data. The kinetics of oxybenzene biosorption on to the biosorbents was described with the pseudo-first-order model. Thermodynamic parameters indicated that biosorption onto biosorbent was feasible in nature, spontaneous, and endothermic for some biosorbents, but on contrary not feasible, exothermic and non spontaneous for other biosorbents. The result of this study showed that the biosorbent derived from Zea mays can be used as a prospective biosorbent for oxybenzene in wastewater and also can be an alternative for the commercially activated carbon.     

Oxidative stress in erythrocytes: a study on the effect of antioxidant mixtures during intermittent exposures to high altitude

The aim of our study was to compare and assess the effectiveness of antioxidant mixtures on the erythrocytes (RBC) of adult male albino rats (Wister) subjected to simulated intermittent high altitudes—5,100 m (AL₁) and 6,700 m (AL₂)—to induce oxidative stress (OS). To achieve our objective, we pre-supplemented four sets of animals with different antioxidant mixtures [vitamin E (vit.E; 50 IU/kg BW), vitamin C (vit.C; 400 mg/kg) and l-carnitine (400 mg/kg)] in different combinations [M1 (vit.E+vit.C), M2 (vit.C+carnitine), M3 (vit.E+carnitine) and M4 (vit.C+vit.E+carnitine)] for 30 days prior to as well during exposure to intermittent hypobaric hypoxia (IHH). Membrane instability, in terms of osmotic fragility and hemolysis, decreased in RBCs of supplemented animals. There was a significant increase in the activity of glutathione peroxidase in the RBCs of supplemented animals. We confirmed OS imposed by IHH with assays relating to lipid [thiobarbituric acid reactive substances (TBARS) and lipofuscin (LF)] and protein (carbonyl, PrC) oxidation, and found a positive correlation between PrC and hemolysis, with a decrease in both upon supplementation with M3 and M4 mixtures. Fluorescence microscopic observation showed a maximum decrease in the LF content in rats administered M4 and M1 compared to those on M2 and M3 mixtures at both altitudes. We suggest that multiple antioxidant fortifications are effective in overcoming increased OS experienced by RBCs at high altitudes.     

Optimization of anticancer exopolysaccharide production from probiotic Lactobacillus acidophilus by response surface methodology

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths in the Western world. Recently, much attention has been focused on decreasing the risk of CRC by consuming probiotics. In the present study, exopolysaccharide (EPS) extracted from Lactobacillus acidophilus was found to inhibit the growth of CaCo2 colon cancer cell line in a dose-dependent manner. The experiment was performed in both normoxic and hypoxic conditions, and EPS was found to reduce the survival of CaCo2 cell line in both the conditions. Quantitative polymerase chain reaction (qPCR) studies demonstrated that EPS treatment upregulated the expression of peroxisome proliferator activator receptor-γ (PPAR-γ) in both normoxia and hypoxia conditions, whereas it upregulated the expression of erythropoietin (EPO) in the normoxic condition, but there was no significant expression under hypoxic conditions. Hence, the EPS production was optimized by Plackett–Burman design followed by central composite rotatory design. The optimized production of EPS at 24 hr was found to be 400 mg/L. During batch cultivation the production peaked at 21 hr, resulting in an EPS concentration of 597 mg/L.