Category: Readings

In-silico epitope-based vaccine is the cornerstone of modern vaccine development and is illuminating by a growing number of experimental and computational methods. The reverse vaccinology approach uses computational screening of the whole proteome of a pathogen to identify the proteins with the attributes of potential vaccine targets. The selection of the right components [antigenic determinants, linkers, and intramolecular adjuvants] results in the development of a multi-epitope vaccine construct with maximum efficacy and minimum adverse effects. We provide insight into the recent advancement in epitope-based vaccines and the procedure to design the modern or next-generation vaccine. The synergy between the computational screening of vaccine candidates and experimental verification to assess the immunogenic potential improves decision-making and reduces the cost and time of the development of new therapeutics

Metabolic dysfunction-associated steatotic liver disease (MASLD) and metabolic dysfunction-associated steatohepatitis (MASH) are common clinico-pathological conditions that affecting over 30% of adults worldwide. Insulin resistance and hepatic lipid accumulation constitute the metabolic foundation of MASLD/MASH. Although previous studies have shown limited efficacy of activation of single PPARs (PPARα or PPARγ), ongoing clinical trials suggest that dual and pan-PPAR agonists may have a broader and more potent therapeutic effect on MASH by simultaneously targeting different inter-related mechanisms in this multisystem disease. Therefore, we hypothesized that a combination of PPARγ agonist (to enhance insulin sensitivity) with lipid lowering therapy (similar to PPARα) could have similar or better effects compared to PPARα/γ dual agonists. In the current study, we have investigated a novel combination of pioglitazone (a PPAR γ/α agonist) and ezetimibe (a cholesterol absorption inhibitor) in two different MASH animal models. We expected that anti-cholesterol absorption property of ezetimibe can augment the poor PPAR-α agonist property of pioglitazone in terms of lipid sensitivity in regulating steatosis. We tested pioglitazone at 2-3-fold reduced clinical dose (15mg/day) in combination to ezetimibe, since there are safety concerns associated with higher doses (30mg and 45mg, daily). Our results revealed that combination of low dose pioglitazone, with ezetimibe hold the ability to regulate the steatosis, hepatocyte inflammation and ballooning, which resulted in superior effects in terms of NAS as well as fibrosis score compared to pioglitazone alone (30mg/kg). Moreover, in vitro studies in human liver microsomes and mouse hepatocytes did not show any drug-drug interaction between pioglitazone and ezetimibe. Overall, this study provides a potential possibility for the clinical treatment of MASH with combination of pioglitazone and ezetimibe.

Pesticides are among the most widely used synthetic chemicals worldwide to protect crops, but they pose several environmental and health issues. The present study evaluates toxic effects of acetamiprid (ACMP) in rat liver and kidney tissues. Rats were exposed to ACMP (21.7 mg/kg b.wt; 1/10^th LD₅₀) for 21 days. Oxidative stress generation, apoptotic progression, and structural changes were evaluated via biochemical assays, semi-quantitative PCR, western blotting, and histopathology. ACMP exposure significantly decreased body weight and altered relative organ weight in hepato-renal tissues. Significant oxidative stress was evident by increased oxidative injuries to lipids and proteins and alterations of endogenous anti-oxidative enzymes. The administration of ACMP upregulated mRNA expression of Bax and caspase-3, while downregulated the mRNA expression of Bcl-2 and released cytochrome c into the cytosol of the liver and kidney tissues. Likewise, exposure to ACMP caused severe degenerative changes in the histo-architecture of hepato-renal tissues. The results of the present study confirmed the toxicity of ACMP in rats’ liver and kidney tissues and emphasized the need for strict regulation and more mechanistic understanding to delineate the toxicity of ACMP in mammals.

A novel, sensitive, portable method for detecting DNA from various human samples is most desirable. This study aims to develop a microdevice for on-chip DNA extraction and detection by PCR from various forensic samples. A microchip constructed by sandwiching a fusion-5 filter disc between PMMA layers was characterized using SEM, FTIR, and XRD. DNA capture efficiency of the microchip from human samples was quantified by Real-Time PCR. PCR products were evaluated off-chip by DNA sequencing (STR Typing). On-site detection was performed by visualizing the DNA amplicons on Fusion-5 filter paper under UV light after EtBr staining. Among all concentrations, 5% by weight PMMA membrane was found most suitable for PMMA-Fusion-5 filter disc fluidic Microchip, the best smooth cross-section morphology by SEM, strong absorption vibrations at corresponding wavelengths by FTIR, increased amorphous phase by XRD were confirmed. Using this microdevice, DNA extraction from human whole blood was, without any leakage, fast (≤7 minutes), most efficient (highest, Ct=27.22) as evaluated by real-time PCR, needs just 2µl blood sample as shown by a typical, balanced STR profile. The microdevice designed for on-chip DNA extraction has excellent potential for rapid, on-site DNA detection from various samples.

URN:NBN:sciencein.cbl.2024.v11.665