Selenium Ameliorates Acetaminophen-Induced Oxidative Stress via MAPK and Nrf2 Pathways in Mice.

Overdose of acetaminophen (paracetamol), a widely used non-prescriptive analgesic and antipyretic medication, is one of the main causes of drug-induced acute liver failure around the world. Oxidative stress contributes to this hepatotoxicity. Antioxidants are known to protect the liver from oxidative stress. Selenium, a potent antioxidant, is a commonly used micronutrient. Here, we evaluated the protective effect of selenium on acetaminophen-induced hepatotoxicity. Treating Wistar albino mice with sodium selenite (1 mg/kg) before or after inducing hepatotoxicity with acetaminophen (150 mg/kg) significantly reduced the levels of liver injury biomarkers such as serum glutamate oxaloacetate transaminase and serum glutamate pyruvate transaminase. In addition, selenium-treated mice showed decreased levels of oxidative stress markers such as protein carbonyls and myeloperoxidase. Acetaminophen treatment stimulated all three mitogen-activated protein kinases (MAPKs) and Keap1 and decreased the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 in liver and in isolated mouse peritoneal macrophages, which was reversed by selenium treatment. Our findings suggest that the reactive oxygen species-mediated Nrf2 and MAPK pathways are critical players in acetaminophen-induced hepatotoxicity. These key findings offer an alternative therapeutic target for addressing acetaminophen-induced hepatotoxicity.

Aspartic protease-pepstatin A interactions: Structural insights on the thermal inactivation mechanism.

Aspartic proteases are the targets for structure-based drug design for their role in physiological processes and pharmaceutical applications. Structural insights into the thermal inactivation mechanism of an aspartic protease in presence and absence of bound pepstatin A have been obtained by kinetics of thermal inactivation, CD, fluorescence spectroscopy and molecular dynamic simulations. The irreversible thermal inactivation of the aspartic protease comprised of loss of tertiary and secondary structures succeeded by the loss of activity, autolysis and aggregation The enthalpy and entropy of thermal inactivation of the enzyme in presence of pepstatin A increased from 81.2 to 148.5 kcal mol-1, and from 179 to 359 kcal mol-1 K-1 respectively. Pepstatin A shifted the mid-point of thermal inactivation of the protease from 58 °C to 77 °C. The association constant (K) for pepstatin A with aspartic protease was 2.5 ± 0.3 × 10 5 M-1 and ΔGo value was -8.3 kcal mol-1. Molecular dynamic simulation studies were able to delineate the role of pepstatin A in stabilizing backbone conformation and side chain interactions. In the Cα-backbone, the short helical segments and the conserved glycines were part of the most unstable segments of the protein. Understanding the mechanism of thermal inactivation has the potential to develop re-engineered thermostable proteases.

What we know about plant arginases?

Nitrogen is one of the essential element required for plant growth and development. In plants, most of the nitrogen is stored in arginine. Hence, metabolism of arginine to urea by arginase and its further hydrolysis to ammonia by urease is involved in nitrogen recycling to meet the metabolic demands of growing plants. In this respect, plant arginases differ from that of animals. Animals excrete urea while plants recycle the urea. However, the studies on the biochemical and biophysical characteristics of plant arginase are limited when compared to animal arginase(s). In this review, the structural and biochemical characteristics of various plant arginases are discussed. Moreover, the significance of arginase in nitrogen recycling is explained and recent literature on function and activation of plant arginases in response to various environmental (biotic and abiotic) insults is also presented.

Application of Optimized and Validated Agar Overlay TLC-Bioautography Assay for Detecting the Antimicrobial Metabolites of Pharmaceutical Interest.

The agar overlay TLC-bioautography is one of the crucial methods for simultaneous in situ detection and separation of antimicrobial metabolites of pharmaceutical interest. The main focus of this research relies on the dereplication of an antimicrobial metabolite coriloxin derived from mycoendophytic Xylaria sp. NBRTSB-20 with a validation of agar overlay TLC-bioautography technique. This polyketide metabolite coriloxin was purified by column chromatography, and its purity was assessed by HPLC, UPLC-ESI-QTOF-MS, FT-IR and NMR spectral analysis. The antimicrobial capability of ethyl acetate extract and the purified compound coriloxin was determined by disc diffusion, minimal inhibitory concentration and agar overlay TLC-bioautography assay. The visible LOD of coriloxin antimicrobial activity was found at 10 µg for Escherichia coli and 20 µg for both Staphylococcus aureus and Fusarium oxysporum. Inter- and intra-day precision was determined as the relative standard deviation is less than 6.56%, which proved that this method was precise. The accuracy was expressed as recovery, and the values were found ranging from 91.18 to 108.73% with RSD values 0.94-2.30%, respectively. The overall findings of this investigation suggest that agar overlay TLC-bioautography assay is a suitable and acceptable method for the in situ determination of antimicrobial pharmaceuticals.

Biochemical and functional characterization of an atypical plant l-arginase from Cilantro (Coriandrum sativam L.).

Arginase is one of the key enzymes responsible for maintaining the essential levels of nitrogen among plants, but biochemical and functional characterization of arginase among plants is limited. While screening for stable plant arginase, we found cilantro possessing an abundant and stable arginase. We purified arginase to apparent homogeneity (3300-fold purification) with a specific activity of 81,728 nmoles of urea formed/mg of protein/min and its eight-tryptic fragments had amino acid sequences identical to Arabidopsis thaliana arginase. Cilantro arginase exhibited absolute requirement for Mn2+ (0.5 mM-1 mM). Unlike other known plant arginases, cilantro arginase did not hydrolyse d-arginine and other arginine analogues. While for sulfhydryl reagents the enzyme was sensitive, l-NOHA, an arginase inhibitor showed only moderate inhibition - a property distinct from tomato arginase. We also found arginine derived amino acids and polyamines can regulate cilantro arginase in vitro. In addition, we also noticed an increase in cilantro arginase activity to both biotic and abiotic stress. We conclude that, cilantro may be used as a model plant to study plant arginases and to delineate arginase role, beyond its classical role in nitrogen recycling and polyamine biosynthesis.