Manan Raval

Alcohol-associated liver disease (ALD) is driven by complex interactions among hepatic lipid accumulation, oxidative stress, inflammation, cell death, and disruption of the gut–liver axis. Therapeutic strategies capable of targeting multiple interconnected pathogenic pathways remain limited. In this study, we investigated the protective potential of graphene oxide nanoparticles (GNPs) in a chronic ethanol-fed rat model of ALD. Male Wistar rats were subjected to ethanol feeding and intermittently treated with GNPs (10 mg/kg) by oral gavage. Hepatic injury was assessed by biochemical parameters, histology, lipid accumulation, gene and miRNA expression, protein analysis, and gut microbiome profiling. Ethanol feeding induced hepatic steatosis, oxidative stress, apoptotic and necroptotic signaling, intestinal barrier disruption, gut dysbiosis, and activation of hepatic inflammatory pathways. GNP treatment markedly attenuated ethanol-induced lipid accumulation, normalized liver morphology, and reduced biochemical markers of liver injury. These effects were accompanied by restoration of antioxidant defenses, including Nrf2 and HO-1, and suppression of CYP2E1 expression and cell death–associated markers. In parallel, GNPs preserved intestinal architecture, maintained tight junction gene expression, and suppressed intestinal inflammatory responses. Gut microbiome analysis revealed partial restoration of ethanol-induced dysbiosis, including recovery of beneficial postbiotic-associated bacterial taxa. Improved intestinal homeostasis was associated with attenuation of hepatic TLR4-associated inflammatory signalling and modulation of macrophage-associated markers. Furthermore, GNP treatment partially normalized ethanol-induced dysregulation of miRNAs implicated in lipid metabolism, inflammation, and oxidative stress. Collectively, these findings demonstrate that GNPs exert a coordinated protective effect against ethanol-induced liver injury by modulating multiple pathological processes along the gut–liver axis. This multi-targeted activity highlights the therapeutic potential of graphene oxide nanoparticles as an intervention strategy for early-stage alcohol-associated liver disease.

Background Diabetes Mellitus (DM) impairs male reproductive functions by modulating apoptosis, oxidation, and the inflammatory pathway. The studies were planned to screen the ability of the hydro-alcoholic extract (AN) prepared using the roots of Argyreia nervosa (A. nervosa) (Burm.Fil.) Bojer. (Family: Convolvulaceae) in alleviating the DM-induced detrimental functioning of male reproductive functions in experimental rats. Method and results Diabetic conditions were induced by the combined administration of a high-fat diet and a single injection of streptozotocin (30 mg/kg). Diabetic rats treated with a higher dose of A. nervosa extract (ANH) showed decreased fasting blood glucose (0.52-fold), abnormal sperm morphology (0.50-fold), and sperm DNA fragmentation index (0.74-fold), along with improved semen quality. ANH increased testicular testosterone (1.40-fold), FSH (1.91-fold), LH (1.69-fold), and mRNA expression of StAR (1.75-fold), 17β-HSD (1.77-fold), and 3β-HSD (1.06-fold). It reduced MDA (0.72-fold) and raised SOD (1.93-fold) and GSH (2.13-fold). ANH also lowered mRNA/protein levels of RAGE (0.75-fold, 8.81-fold), TNF-α (0.79-fold, 2.11-fold), NLRP3 (0.68-fold, 8.15-fold), and Caspase-3 (0.87-fold, 8.45-fold). Conclusion AN, thus, showed the potential beneficial effects on DM-induced male infertility, probably through its ability to counter DM induced apoptosis and inflammation in testicular tissues along with supporting anti-oxidant mechanisms. The extract could upregulate testosterone biosynthesis by modulating steroidogenic enzyme activity and stimulating FSH and LH levels. This action further supported the improvement of sperm functioning parameters in diabetic rats. These results provided the potential of AN as an alternative treatment for the clinical management of DM-impaired male reproductive function, offering hope for future research and clinical applications.

Background: A reliable technique to determine stigmasterol and hecogenin simultaneously in Chlorophytum borivilianum root extract is essential for their analysis in pharmaceutical formulations. Aim: To establish and validate a High-Performance Liquid Chromatography (HPLC) technique to determine stigmasterol and hecogenin simultaneously in C. borivilianum root extracts. Methods: To accomplish chromatographic separation, a Shimadzu Prominence-I LC-2030C plus HPLC system with a Shimpak GIST C8 column (150mm × 4.6mm, 5μm) was used. The mobile phase was made up of water and acetonitrile (10:90 %v/v). The wavelength for detection was set at 210nm for Hecogenin and 240nm for Stigmasterol. Method validation followed Q2(R1) guidelines for ICH, assessing precision, accuracy, linearity, specificity and robustness. Results: Linearity was confirmed within the 10–60 μg/mL concentration range for both the analytes, yielding coefficients of correlation of 0.9981 for Hecogenin and 0.9997 for Stigmasterol. Hecogenin and stigmasterol had limits of detection (LOD) of 0.1373 μg/mL and 0.3521 μg/mL, respectively, while the limits of quantification (LOQ) were 0.4161 μg/mL and 1.0670 μg/mL. Recovery rates ranged from 98.66% to 101.73%, and both the intra-day and inter-day precision % RSD values were less than 2%. The assay of the C. borivilianum root extracts identified Hecogenin at 8.83% w/w and Stigmasterol at 2.94% w/w. Conclusion: The HPLC method is developed, validated and suitable for routine analysis of Hecogenin and Stigmasterol in plant-based formulations. Major Findings: The study developed and validated a novel HPLC method for simultaneous quantification of Hecogenin and Stigmasterol in Chlorophytum borivilianum extracts. The method demonstrated high sensitivity (LOD: 0.1373–0.3521 μg/mL), accuracy (98.66%–101.73% recovery), and precision (%RSD < 2%), confirming its reliability for routine phytochemical analysis.

The study aimed to evaluate the effect of roflumilast on modulating TNF-α/Caspase mediated cellular signals in cisplatin-induced nephrotoxicity in rats. The rats (Male Wistar) were divided into five groups: normal control, disease control (cisplatin: 7 mg/kg i.p.), and cisplatin + roflumilast (0.25, 0.5, and 1 mg/kg b.w., p.o.). Cisplatin was administrated to rats on 0 day, and roflumilast treatment was started from the 6th–15th days. Blood and tissue were collected. Tissue was used to measure oxidative stress, such as malondialdehyde, superoxide dismutase, and catalase. Gene expression study involved real-time PCR of key genes linked with inflammation and apoptosis, i.e. Tnf-α, Tnfr1, Tnfr2, Fas, Nfkb, Casp3, Casp8, and Nrf2. Cisplatin showed decreased serum creatinine and urea, high albumin, and total protein. Cisplatin elevated the malondialdehyde and reduced superoxide dismutase and catalase activity. Cisplatin also attributed an overexpression of Tnf-α, Tnfr1, Tnfr2, Nfkb, Fas, Casp3, and Casp8, and a decrease in the Nrf2 gene. Roflumilast decreased creatinine and urea and increased albumin and total protein levels. Roflumilast also downregulated the expression of Tnf-α, Tnfr1, Tnfr2, Nfkb, Fas, Casp3, and Casp8 and upregulated the Nrf2 gene expression. Roflumilast manifested as a potential reno-protective agent against cisplatin-induced nephrotoxicity.

Drug-resistant tuberculosis (DR-TB) represents a pressing global health issue, leading to heightened morbidity and mortality. Despite extensive research efforts, the escalation of DR-TB cases underscores the urgent need for enhanced prevention, diagnosis, and treatment strategies. This review delves deep into the molecular and genetic origins of different types of DR-TB, highlighting recent breakthroughs in detection and diagnosis, including Rapid Diagnostic Tests like Xpert Ultra, Whole Genome Sequencing, and AI-based tools along with latest viewpoints on diagnosis and treatment of DR-TB utilizing newer and repurposed drug molecules. Special emphasis is given to the pivotal role of novel drugs and discusses updated treatment regimens endorsed by governing bodies, alongside innovative personalized drug-delivery systems such as nano-carriers, along with an analysis of relevant patents in this area. All the compiled information highlights the inherent challenges of current DR-TB treatments, discussing their complexity, potential side effects, and the socioeconomic strain they impose, particularly in under-resourced regions, emphasizing the cost-effective and accessible solutions. By offering insights, this review aims to serve as a compass for researchers, healthcare practitioners, and policymakers, emphasizing the critical need for ongoing R&D to improve treatments and broaden access to crucial TB interventions. Graphical Abstract - The escalating burden of DR-TB represents a significant global health threat. - Effective strategies are needed for DR-TB's prevention, diagnosis, and treatment. - The review emphasizes DR-TB management with novel techniques and AI approaches. - It also focuses on relevant patents related to DR-TB management. - It underscores continuous research for effective DR-TB control challenges.

The roots of the plant Asparagus racemosus (Wild) (family: Liliaceae) are used alone as a lactogenic and general tonic or incorporated as a major ingredient in many herbal and Ayurveda formulations. Shatavarin glycosides containing sarsasapogenin as aglycone were found bioactive, and present in predominantly higher amount. The experiments were planned to develop an analytical method for estimation of total sarsasapogenin from the extract using HPTLC, through a systematic approach after identifying the potential failure modes, termed as critical process parameters to achieve the analytical target profile. The associated risks were mitigated by studying the combined effect of the critical process parameters on area, Rf, and tailing factor corresponding to the sarsasapogenin peak, assigned as critical method attributes. The optimization studies were carried out through Design of Experiment approach and process parameters were finalized to achieve analytical target profile. The developed method could resolve the peak of sarsasapogenin from the hydrolyzed methanolic extract prepared from the root powder using silica gel-backed pre-coated aluminum plates. The optimized mobile phase was Hexane:Ethyl acetate:Formic acid (7.0:1.9:0.35, v/v/v); the plates were subjected to post-chromatographic derivatization using 5% v/v hydrochloric acid solution, and heating of the plats for 3 min at 110˚C. The chromatogram was recorded by scanning the plates in TLC scanner using fluorescence mode at 366 nm. The developed analytical method was found to yield linear response in the range of 40-280 ng/spot. The limit of quantification was determined to be 35.88 ng/spot. The method was found selective, accurate, and robust. The amount of sarsasapogenin was found to be determined using the developed method and found to be 2.31% w/w on dried weight basis from the dried root powder. This is the first report of a systematic approach in estimating sarsasapogenin from Asparagus sp.

Background: Argyreia nervosa (Burm. Fil.) Bojer., a woody climber, is indicated in Ayurveda to treat debilities of the male reproductive system, diseases of the nervous system, and chronic ulcers. Objective A sensitive analytical method was developed to estimate bioactive scopoletin from methanolic extract prepared from the medicinally active dried roots of Argyreia nervosa (Burm. fil.) Bojer using HPLC equipped with a fluorescence detector. Methods: Chromatographic separation was achieved using a Luna (TM) (C-18, 250 x 4.6 mm, id: 5 mu m) column using an isocratic mobile phase comprising phosphate buffer (pH 3.5)-acetonitrile (80 + 20, by volume) at a flow rate of 1.0 mL/min. The excitation wavelength was 345 nm, and the emission wavelength was 444 nm. The chromatographic parameters were optimized using the design of the experiment approach after determining the combined effects of selected independent variables on area, retention time, and tailing factor (TF) for the peak corresponding to scopoletin, and the experimental design was validated by navigating through the design space. Results: The developed method was found linear in the range 10-140 ng/mL. The results of the studies confirmed the accuracy, precision, and robustness of the developed analytical method. The plant material was found to contain 0.0125 +/- 0.0001% w/w scopoletin on a dried weight basis when estimated using the developed method. Conclusion: The method was developed using the HPLC-fluoresence detection by adopting the design of experiment approach and simple sample preparation for the estimation of scopoletin from roots of A. nervosa.