The spent engine oil (SEO) is a major source of pollution and impacts the soil quality and plant growth. Phytoremediation is an environmentally friendly technique for soil remediation. The aim of this study was to assess how the phytoremediation ability and stress resistance of okra (Abelmoschus esculentus (L.) could be improved with the use of exogenous melatonin (MT). P. Moench) cultivated in a soil contaminated with SEO. Four concentrations of SEO (0, 100, 200 and 300 μM) and three MT levels (0, 100 and 200 μM) were investigated in a 4×3 factorial pot experiment. SEO stress had significant effects on plant growth, plant physiological activity and biochemical activity. MT application dose-dependently however mitigated these negative effects. Seedling exposure to MT resulted in up to 11.51%, 15.85% and 32.71% increase in shoot length, root length and number of leaves, respectively, as compared to the stressed control. Physiological parameters also enhanced, including photosynthetic rate (13.12%), chlorophyll content (26.32%), stomatal conductance (26.41%) and transpiration rate (31.62%). The maximum improvement was with the total amino acid content (107.78) in severe stress. Moreover, it is important to point out that the removal efficiency of anthracene has risen from XX% (SEO-only treatment) to XX% (SEO + 200 μM MT) showing an improvement of the phytodegradation efficiency. Two-way ANOVA showed that there were significant (p < 0.05) differences among the various SEO, MT and SEO x MT for all parameters. These results indicate that the use of exogenous melatonin in the field of phytoremediation with A. esculentus in soils contaminated with SEO could be a good option for sustainable remediation.
Background: Polymetabolic Endocrine Syndrome (PMOS) (previously known as Polycystic Ovary Syndrome PCOS) is a common endocrine-metabolic syndrome amongst reproductive aged women, particularly prevalent in South Asian women. Two of the main pathophysiological characteristics are hypoadiponectinemia and insulin resistance. Objectives: The aim of this study was to assess the therapeutic effect of administration of myo-inositol for 8-weeks (4g/day) along with life style changes on serum adiponectin and HOMA-IR in Pakistani women having PMOS. Methods: A parallel group-controlled design of 100 women (18-35 years) diagnosed with PMOS using modified NCEP ATP III criteria. After randomization 1:1 participant received myo-inositol (4g/day) and lifestyle advice (test group, n=50) or lifestyle advice alone (control group, n=50) for 8 weeks. The primary outcome was change in serum adiponectin level and the secondary outcomes were HOMA-IR, fasting glucose and fasting insulin. Results: The test group demonstrated significant adiponectin elevation (4.71±0.92 to 6.12±1.08 μg/mL, +30%, p<0.001, Cohen's d=1.42) versus controls (4.92±0.86 to 4.99±0.91 μg/mL, p=0.399). HOMA-IR reduced by 32% in the test group (4.58±1.38 to 3.12±1.29, p<0.001) versus minimal change in controls. The Inter-group effect sizes were large (adiponectin: d=2.29; HOMA-IR: d=1.64). Conclusion: Supplementation with myo-inositol in PMOS-related insulin resistance results in a dramatic improvement of adiponectin, and is an effective non-pharmacological adjunctive therapy.
Background: Plasmids are the major route of the dissemination of resistance genes between species, sectors and geographical locations, and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli are a critical health threat globally. Objective: Based on recent knowledge, this review intends to bring together the molecular epidemiology, plasmid biology, horizontal gene transfer mechanisms and therapeutic approaches in ESBL-producing E. coli, with a focus on the transmission dynamics in the realm of One Health. Methods: We searched the literature in PubMed, Scopus, and Web of Science from 2015 to 2026 using the keywords, "ESBL," "E. coli," "plasmids," "antibiotic resistance," "horizontal gene transfer," and "One Health”. Articles were selected based on their relevance to understanding the nature of plasmid mediated resistance, epidemiology, and strategies for interventions. Results: ESBL-encoding plasmids vary in different species and contain a range of resistance determinants, such as blaCTX-M, blaTEM, blaSHV genes, which are frequently found together with aminoglycoside, quinolone and colistin resistance genes, with prevalence of IncF, IncI, IncN or IncHI2 plasmid types. The pandemic ST131 clone shows good plasmid-host adaptation that has different dominant subclades in different regions. Plasmids are very flexible in terms of the presence of mobile genetic elements (ISEcp1, IS26) that allow the capture and dissemination of resistance genes. Multidrug resistance and virulence determinants converged on epidemic plasmids and resulted in hypervirulent multidrug resistant strains. Conclusions: A systems biology approach to better understand plasmid biology, together with new technologies such as long-read sequencing and artificial intelligence, and the need to support the One Health approach, are all crucial to reducing the threat of ESBL-producing E. coli. There are potential treatment options that are directed towards resistance plasmids, but they need to be developed and verified.
Background: Floods are among the most frequent climate-related disasters affecting Pakistan and often result in population displacement, disruption of health services, deterioration of water, sanitation and hygiene (WASH) conditions, and increased risk of communicable disease outbreaks. Following the July–August 2025 floods in Gilgit-Baltistan, a comprehensive assessment was required to identify priority public health and nutrition needs and inform recovery planning. Objectives: To assess the post-flood public health situation in Gilgit-Baltistan, identify major health, WASH, disease surveillance, and nutrition-related risks, and develop an operational response framework to strengthen disaster preparedness and resilient recovery. Methods: This applied public health study used secondary programmatic data extracted from the Gilgit-Baltistan post-flood field assessment conducted between September and October 2025. Descriptive analyses were performed using frequencies, proportions, and operational indicators. Health service availability, disease surveillance, WASH conditions, outbreak risks, logistics, nutrition-related information, and data quality were systematically reviewed. A risk-prioritization approach was used to identify critical operational gaps and develop an integrated intervention package. Results: All ten districts of Gilgit-Baltistan were affected by the floods, with four districts remaining affected during the assessment period. Approximately 6,000 people were affected, including 2,910 internally displaced persons residing in six temporary camps. Acute watery diarrhea, acute respiratory infections, and skin diseases were the leading morbidities. A confirmed cholera outbreak was identified in Astore, while a suspected dengue outbreak was reported in Diamer. Eleven health facilities sustained flood damage, including four completely destroyed facilities. Of 588 functional health facilities, only 372 (63.3%) were reporting through the Integrated Disease Surveillance and Response System. Major operational concerns included incomplete WASH information, open defecation practices, limited vector-control activities, insufficient nutrition screening, and inadequate data on maternal and newborn health, mental health, returnee populations, and water quality. An Integrated Flood Health and WASH Surveillance and Response Package was developed to strengthen outbreak surveillance, mobile health and nutrition services, WASH interventions, cold-chain recovery, logistics management, and emergency preparedness. Conclusion: The 2025 floods exposed important vulnerabilities in health service continuity, disease surveillance, WASH infrastructure, and nutrition-related information systems in Gilgit-Baltistan. Integrating surveillance, WASH, nutrition, and primary healthcare within a coordinated recovery framework can strengthen resilience, improve emergency preparedness, and reduce the health consequences of future flood emergencies in disaster-prone settings.
Background: Healthcare-associated (HA) infections caused by methicillin-resistant Staphylococcus aureus (MRSA) continue to be a big problem worldwide. The mecA gene is responsible for the production of PBP2a which is responsible for the resistance to all beta-lactam antibiotics. There are currently very little alternative therapeutic options as the treatment is becoming limited. The advantage of CRISPR-Cas9 technology lies in the ability to target genes with precision, however, there are technical delivery challenges in the application of this technology in bacterial pathogens. Objective: The aim of this study was to assess the efficacy of CRISPR-Cas9 delivery by liposomes with Lipofectamine and its ability to knock out the mecA gene in MRSA and make them susceptible to beta-lactam antibiotics. Methods: MO of MRSA ATCC 43300 was electroporated with a Cas9-sgRNA RNP complex using Lipofectamine CRISPRMAX with a mecA-targeting guide sgRNA designed using CHOPCHOP. An antibacterial activity was determined (CFU, growth curves, biofilm), gene targeting (PCR, Sanger sequencing), resistance reversal (MIC, disk diffusion) and safety (MTT, hemolysis). All experiments were repeated three times (n=3), data presented as mean ± SD and analyzed by One Way Analysis of Variance (ANOVA) with Tukey's post hoc test (p<0.05). Results: The treatment group showed reduction of CFU level, delay of growth and 60-70% inhibition of the development of biofilm which was significant (p=0.008). This was demonstrated by PCR with reduced intensity of mecA amplicon (1/3 of controls) and by Sanger sequencing which revealed the presence of indels in 72% of the clones. MIC dropped from 64 to 4 μg/mL (p<0.001), and inhibition zones increased from 0 to 22.3 ± 1.8 mm. Cell viability exceeded 80% (87.4 ± 3.2%), with hemolysis below 5% (3.1 ± 0.4%). Conclusion: Lipofectamine-mediated delivery of CRISPR-Cas9 effectively targets the mecA gene, reverses antibiotic resistance, and demonstrates an acceptable safety profile in vitro. These findings provide a proof-of-concept for this approach and warrant further investigation in appropriate in vivo models.
