Wastewater-based epidemiology has become an important approach for monitoring severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) circulation and understanding microbial dynamics within sewage ecosystems. In this study, we compared the microbial composition, antimicrobial resistance and virulence-associated determinants, functional profiles, and microbial network properties of SARS-CoV-2 RNA-positive and RNA-negative wastewater pools. Thirty wastewater samples collected from six sewage surveillance sites across Aizawl, India, were screened using RT-PCR targeting the SARS-CoV-2 N and E genes. Samples were grouped into SARS CoV 2 RNA positive and negative categories, pooled separately, and subjected to shotgun metagenomic sequencing followed by taxonomic, resistome, virulome, diversity, functional, and microbial network analyses. Bacterial communities dominated both wastewater groups with more than 98% classified reads. SARS CoV 2 positive wastewater showed microbial redistribution with reduced Bacillota abundance and increased Pseudomonadota and Bacteroidota representation. Bacterial and fungal diversity declined, whereas viral communities displayed greater heterogeneity. Resistome profiling identified greater antimicrobial resistance burden in SARS CoV 2 negative wastewater with 17,303 resistance associated counts compared with 11,780 in positive wastewater, dominated by antibiotic efflux mechanisms and multidrug resistance determinants including AcrAB TolC systems, soxR, soxS, and Mex regulators. Virulence profiling identified 400,576 and 151,498 virulence associated determinants in negative and positive wastewater respectively, dominated by effector delivery systems, immune modulation, adherence, and secretion associated mechanisms. Functional analyses demonstrated preservation of metabolic pathways despite taxonomic restructuring, while network analysis revealed reduced microbial connectivity in SARS CoV 2 positive wastewater. Collectively, wastewater shotgun metagenomics supports environmental surveillance and public health preparedness.
Objective Liver diseases are a global health concern, and there is a continuing need to investigate effective and accessible compounds that can target the major pathological processes involved in liver injury. Eremanthin, a sesquiterpene lactone, has been reported to possess antioxidant and anti-inflammatory properties. This study evaluated the hepatoprotective effects of Eremanthin against carbon tetrachloride (CCl₄)-induced liver injury in rats, with particular emphasis on changes in genes related to the Keap1/Nrf2/ARE and NF-κB pathways. Methods Antioxidant activity of Eremanthin was first confirmed in HepG2 cell lines and in vitro assays, followed by in vivo studies. Fifty rats were divided into five groups (n = 10): Group I (control), Group II (CCl4), Group III (CCl4 + silymarin), Groups IV and V (CCl4 + Eremanthin 20 and 40 mg/kg, respectively, for 21 days). Oxidative stress, inflammatory, and apoptotic markers, as well as liver enzymes, were analyzed. Histopathological evaluation was conducted to assess the impact of Eremanthin on liver architecture, while qRT-PCR analysis was employed to quantify the expression of genes. Results CCl4 administration markedly elevated serum ALT, AST, ALK, bilirubin, inflammatory cytokines (IL-1β, IL-6, TNF-α, NF-κB, COX-2), and pro-apoptotic markers (Bax, caspase-3, caspase-9), while decreasing Bcl-2, with histology showing hepatocellular necrosis and inflammation. Eremanthin significantly restored liver enzymes and bilirubin toward normal, reduced oxidative stress, altered the expression of inflammatory mediators, and was associated with changes in apoptotic markers, including increased Bcl-2 and reduced Bax and caspases. Eremanthin also increased the expression of NFE2L2, HO-1, and NQO1 and altered Keap1 expression. Histopathology confirmed improved hepatocyte integrity. Conclusion These findings indicate that Eremanthin has hepatoprotective effects against CCl₄-induced liver injury, associated with attenuation of oxidative stress, inflammation, and apoptotic responses and altered expression of genes related to the Keap1/Nrf2/ARE and NF-κB pathways.
Background Oxidative stress and inflammation are fundamental drivers of cardiovascular disease, yet their integrated relationship with canonical WNT/β-catenin signalling and inflammatory activation in human acute coronary syndrome (ACS) remains poorly understood. Here, we investigated whether impaired antioxidant defence is associated with coordinated dysregulation of WNT/β-catenin pathway genes and NF-κB-mediated inflammatory signalling in patients with ACS. Methods In this case–control study, patients with clinically diagnosed acute coronary syndrome (n = 100) and healthy controls (n = 50) were recruited. Serum superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) were measured spectrophotometrically. WNT pathway genes (WNT3A, CTNNB1, LEF1, AXIN2, DVL3, NXN) and inflammatory mediators (NFκB1, NFκB2, TNFα) were quantified by RT-qPCR in peripheral blood. Group comparisons used the Mann–Whitney U test with Benjamini–Hochberg false discovery rate (FDR) correction across all twelve biomarkers. Pairwise Spearman correlations were computed within each group and formally compared between groups using Fisher's r-to-z transformation with FDR correction. Ex vivo, PBMCs from two independent healthy donors were exposed to H₂O₂ with or without N-acetylcysteine (NAC) to explore redox-dependent regulation of the WNT/β-catenin and NF-κB pathways. PBMCs results are presented as mean ± SD of the two donors, without inferential statistical testing. Results Compared with controls, ACS patients exhibited lower SOD activity (49.04 ± 1.01 vs. 56.66 ± 1.17% activity/mg protein) and higher MDA (2.42 ± 0.24 vs. 1.13 ± 0.16 nmol/mg protein; both q < 0.01), while CAT activity was reduced but not significant (q = 0.30). CTNNB1, LEF1, AXIN2, and NXN were significantly downregulated (q < 0.05); DVL3 also differed significantly (q = 0.049), but inconsistent directionality between mean- and rank-based analyses precluded interpretation as confirmed downregulation, and NFκB1, NFκB2, and TNFα were significantly upregulated (q < 0.001); WNT3A did not differ significantly. Cases and controls differed substantially in age, smoking, and alcohol exposure; we did not attempt to adjust these associations for age or lifestyle factors; the differences above should be read as disease-associated rather than disease-specific or age-independent findings. Two gene pairs (WNT3A–DVL3 and CTNNB1–DVL3) exhibited significant disease-associated shifts in correlation strength (Fisher's r-to-z, BH-FDR q < 0.05). In an exploratory, two-donor ex vivo pilot, H₂O₂ exposure showed a directional pattern of WNT suppression and NF-κB induction similar to that seen in patients, with apparent attenuation after N-acetylcysteine (NAC) pretreatment. Conclusions Systemic oxidative imbalance was associated with altered expression of WNT/β-catenin and NF-κB pathway-related transcripts in this unmatched ACS–control comparison; because age, smoking, and alcohol exposure differed markedly between groups, these associations cannot be attributed specifically to ACS status. Concordant effects of ex vivo oxidative challenge and NAC rescue support a redox-sensitive WNT–NF-κB axis. These findings warrant validation in age- and lifestyle-matched, longitudinal cohorts before any diagnostic, prognostic, or therapeutic relevance can be assessed.
Saffron (Crocus sativus L.) exhibits highly temperature-sensitive flowering, posing challenges for stable reproductive production under changing climatic conditions. Functional characterization of flowering genes in saffron is constrained by its triploid and sterile nature. In this study, the Crocus FLOWERING LOCUS T-LIKE (CsFT-like) gene was constitutively expressed in Nicotiana tabacum and evaluated under four temperature regimes (18, 23, 28 and 32 °C) to investigate its effects on flowering, growth and reproductive development. Transgenic plants exhibited substantial CsFT-like transcript accumulation, reaching up to 72.49-fold relative to the calibrator. Despite high CsFT-like transcript abundance, both transgenic lines exhibited delayed flowering relative to wild-type (WT) plants, with delays ranging from 16.8 to 43.6 days across the tested temperature regimes. Capsule production was consistently reduced in transgenic plants, from 10.3 to 5.8 capsules at 18 °C and from 17.2 to 11.0 capsules at 32 °C. Plant height was also reduced at 18 °C, reaching 19.0 cm in transgenic plants compared with 40.1 cm in WT plants. These findings demonstrate that constitutive CsFT-like expression in tobacco was associated with delayed flowering and reduced reproductive output under the tested temperature conditions, highlighting the importance of genetic background and environmental conditions in determining the phenotypic effects of FT-like genes.
