Around the world, polycystic ovary syndrome (PCOS) is a complex endocrine-metabolic condition that typically affects 6–20% of females. Our study’s major goal was to examine how chlorogenic acid (CGA) affected mice with endocrine and metabolic problems brought on by letrozole-induced PCOS. Group I served as the control for 81 days; Group II was given Letrozole (LETZ) orally at a dose of 6 mg/kg bw for 21 days to induce PCOS; Group III was given LETZ (6 mg/kg) for 21 days, followed by treatment with CGA (50 mg/kg bw daily) for 60 days. The study indicated that LETZ-treated mice displayed symptoms of PCOS, such as dyslipidemia, hyperinsulinemia, elevated testosterone, increases in inflammatory markers and malonaldehyde, and a decline in antioxidants (Ar, lhr, fshr, and esr2) in the ovaries. These alterations were affected when the mice were given CGA and were associated with reduced levels of adiponectin. Adiponectin showed interactions with hub genes, namely MLX interacting protein like (MLXIPL), peroxisome proliferator-activated receptor gamma Coactivator 1- alpha (PPARGC1), peroxisome proliferator-activated receptor gamma (Pparg), and adiponectin receptor 1 (Adipor1). Lastly, the gene ontology of adiponectin revealed that adiponectin was highly involved in biological processes. The findings from our research suggest that adiponectin has direct impacts on metabolic and endocrine facets of PCOS.

Background and Objective: Irrational use and overuse of antibiotics is considered a major cause of antimicrobial resistance (AMR) among patients admitted to hospitals, especially in intensive care units (ICUs). ICUs are the most critical wards in healthcare settings, where the use of antibiotics is much higher compared to other wards. Therefore, the appropriate administration and monitoring of antibiotic usage in these units is a matter of concern. Materials and Methods: This retrospective study evaluated the types, utilization patterns, sensitivity, and resistance of various antibiotics used among patients admitted to the ICUs of different hospitals after the third wave of the coronavirus disease in 2019 (COVID-19) in Pakistan. Results: It was observed that more than 40% of the patients were given two antibiotics and 54.3% were given at least one antibiotic each day. A total of 768 antibiotics from different groups, based on the World Health Organization (WHO) Access, Watch, and Reserve (AWaRe) classification, were prescribed to 313 patients admitted to ICUs between April and August 2021. Among the types of antibiotics, amoxicillin/clavulanic acid was the most frequently used antibiotic (75 prescriptions). It was also observed that the majority of the bacterial isolates were more sensitive to carbapenems than the other antibiotics. The current study showed that antibiotic usage according to the AWaRe classifications was 31.8% in the Access category, 59.5% in the Watch category, and 8.7% in the Reserve category in ICUs of the studied hospitals after the third wave of COVID-19. Conclusions: The findings of the study also highlight the importance and continuous need for the implementation of antibiotic stewardship programs to optimize the appropriate utilization of antibiotics in the ICUs of the studied hospitals.

This study aimed to develop a chitosan (CHS)-coated eugenol (EUGE)-loaded nanoemulsion (CHS-EUGE-NE) to enhance brain bioavailability. A novel GC-MS/MS method was developed and validated for EUGE quantification in brain tissue and plasma alongside pharmacokinetic analysis. The nanoemulsion was optimized using a Box-Behnken design (BBD), with the final formulation containing 6.0% oil, 10% surfactant mix, and undergoing 6.5 min of ultrasonication at 30% intensity and 40 °C. The optimized CHS-EUGE-NE exhibited a globule size of 142.4 ± 11.31 nm, a polydispersity index of 0.1660 ± 0.0009, and a zeta potential of -16.20 ± 1.10 mV, which shifted to positive upon CHS coating, enhancing mucoadhesion. A linear range from 1.00 to 3000 ng/mL was established, with accuracy between 90.91% and 99.83%. Pharmacokinetic analysis demonstrated significantly increased C _max and AUC in the rat brain, correlated with behavioral improvements in the forced swimming test. No signs of toxicity were observed. These findings suggest that CHS-EUGE-NE is a promising nanoformulation for enhancing the brain-targeted delivery of eugenol for the treatment of depression and sleep disorders.

A wide-range, specific, and precise liquid chromatography tandem mass spectrometric (LC-MS/MS)technique for quantifying fluoxetine (FLX) in human plasma was developed using the RapidTrace^® automated solid-phase extraction (SPE) method; the analyte and internal standard (IS) were extricated on Oasis MCX SPE cartridges. Acetonitrile and 5 mM ammonium formate buffer (90:10 v/v) were used as mobile phase to achieve chromatographic separation on the reverse phase (C₁₈ column). The analyte and IS were ionized using +ve electrospray ionization approach which was further traced by multiple-reaction monitoring on a tandem mass spectrometer. To quantify the FLX and FLX-d5, the parent-to-daughter ion transition of m/z of 310.0/44.1 and 315.0/44.0 was used, respectively. The method demonstrated a linear active limit of 0.20–30 ng/ml with recoveries ranging from 63.04% to 79.39% for quality control samples and 61.25% for IS samples. The concentrations over the calibration range demonstrated acceptable precision and accuracy. Due to the high inconsistency of the FLX concentration data, the minimum threshold of the assay was kept at 0.20 ng/ml. The flow rate was maintained at 500 μL/min, and the time for sample analysis for each injection was 3.5 min. The method was found to be specific, sensitive, and faster with minimum utilization of organic solvents and was utilized further for metabolic and pharmacokinetic studies.

A newly UHPLC-MS/MS method development and validation for S-Allyl Cysteine was used to evaluate the comparative pharmacokinetic parameters. SC PLGA NPs (S-Allyl Cysteine Poly (D,L-lactide-co-glycolic acid) Nanoparticles) were developed by the emulsion solvent evaporation method. SC PLGA NPs showed their drug loading and encapsulation efficiency to be 5.13 ± 0.10% and 82.36 ± 4.01%, respectively. SC PLGA NPs showed a spherical morphology of an average size (134.8 ± 4.61 nm), PDI: 0.277 ± 0.004, and −25.3 ± 1.03 mV Zeta-Potential (ZP), and is suitable for oral delivery. The development and validation of the UHPLC-MS/MS bioanalytical method were performed successfully for PK-parameter examinations with 1.219 min RT, MS (162.00/73.10), and a total run-time of 2.0 min. Additionally, 1.0–1000.0 ng/mL was a linear range with inter- and intra-day accuracy of 92.55–99.40%, followed by a precision of 1.88–4.23%. SC PLGA NP’s oral bioavailability was significantly higher (** p < 0.01) in comparison to the SC-S treated groups’ (iv and oral). The antimicrobial activity of SC PLGA NPs proved to be more effective than pure S-Allyl-L-Cysteine with significant results (p < 0.01) in comparison to SC-S. SC PLGA NPs showed fitted physicochemical and enhanced antimicrobial properties, which can be helpful for oral administration. Based on the proposed research results, SC PLGA NPs were used for the improvement in oral bioavailability with a sustained and controlled release of S-Allyl-L-Cysteine delivery.

Myeloproliferative neoplasms (MPNs) present a fundamental paradox: despite sharing a small set of canonical driver mutations in JAK2, CALR, or MPL, patients exhibit striking heterogeneity in disease latency, clinical presentation, and evolutionary trajectories to myelofibrosis or secondary acute myeloid leukemia. This review synthesizes recent advances in single-cell and spatial multi-omic technologies that are resolving this paradox by moving analysis from bulk averages to individual cells and their microenvironmental ecosystems. We examine how targeted single-cell DNA sequencing reconstructs clonal architectures and phylogenies, revealing that driver mutations arise within complex mosaics where mutation order, co-mutation context, and cellular ancestry determine phenotypic outcomes. Integrated single-cell transcriptomic and epigenomic profiling exposes within-clone heterogeneity, lineage biases, and functional states that explain variable penetrance and therapy responses. Spatial transcriptomics, especially when integrated with single-cell transcriptomics, histopathology, and multiplex proteomics, further demonstrates that malignant hematopoietic stem and progenitor cells actively remodel bone marrow niches, creating localized inflammatory and fibrotic microenvironments that select for aggressive subclones. Together, these approaches support a new ecological model of MPN pathogenesis in which early epigenetic hits create permissive stem cell reservoirs, clonal competition and cooperation shape disease progression, and non-cell-autonomous niche and immune signals drive malignant metamorphosis. We discuss how this framework refines prognostication, informs rational combination therapies targeting both malignant cells and their ecosystem, and enables real-time monitoring of clonal dynamics, ultimately charting a course from descriptive atlases to actionable clinical strategies.