Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has affected billions of lives and is expected to impose a significant burden on the economy worldwide. Vaccination is the only way to prevent the infection. However, convincing people to get themselves vaccinated is challenging in developing countries such as Pakistan. Therefore, a cross-sectional questionnaire-based study was conducted (n = 982 participants) all over Pakistan to evaluate the perception, knowledge, attitude, and acceptance of the general public towards the SARS-CoV-2 vaccine, in general, and a booster dose of SARS-CoV-2, in particular. The highest number of participants were from the province of Punjab (84.5%), followed by Islamabad (3.8%), Sindh (3.7%), Khyber Pakhtunkhwa (2.7%), Baluchistan (2.6%), Gilgit Baltistan (1.4%), and Azad Jammu and Kashmir (1.4%). A total of 915 participants were vaccinated against COVID-19, out of which 62.2% received one booster dose, followed by double booster doses (25.5%) and single vaccine shots (12.3%). The highest number of vaccinated participants were from Punjab (85.8%), followed by Islamabad (3.9%), Sindh (2.8%); Khyber Pakhtunkhwa (2.6%); Baluchistan (2.3%); Gilgit-Baltistan (1.3%); and Azad, Jammu, and Kashmir (1.2%). Among the vaccinated individuals, 71.4% were unemployed, 27.4% were employed (653), and 1.2% were retired from service. However, no significant association was observed among genders and educational levels in regard to acceptance of the booster vaccine. The outcomes of the study revealed that the increased acceptance of booster doses of the SARS-CoV-2 vaccines among the public was associated with the intent of personal and family protection. Moreover, individuals with low socioeconomic status and pregnant females showed the least acceptance towards the vaccine inoculation. The study also revealed a decline trend of accepting SARS-CoV-2 vaccine among children.

Background/Objectives: Antibiotic resistance is a major public health concern, with considerable socio-economic consequences. Researchers are exploring alternative strategies, including nanotechnology, which has shown significance in targeted drug delivery. This study evaluates the synergistic antibacterial activity of azithromycin-loaded chitosan nanoparticles (AZM-CSNPs) against azithromycin-resistant clinical respiratory isolates of methicillin-resistant Staphylococcus aureus (MRSA) and Klebsiella pneumoniae (K. pneumoniae). Methods: A total of 87 sputum samples (n = 87) were collected and analyzed. The ermB gene for K. pneumoniae and the ermA gene for MRSA were used to confirm resistant isolates. Among 87 samples, 29 manifested K. pneumoniae, and 32 exhibited MRSA-positive cultures, confirmed through phenotypic and genotypic methods. The RT-PCR is performed by using a cDNA Kit to determine the gene expression. Results: The results elucidate resistance of K. pneumoniae against several antibiotics, including azithromycin (15 µg), chloramphenicol (30 µg), and amoxicillin (30 µg), while MRSA also showed resistance to cefoxitin (30 µg), azithromycin (15 µg), and gentamycin (10 µg). Reduction in the MIC value of the nanoparticle formulation showed their effectiveness. The AZM-CSNPs combined with cetirizine dihydrochloride helped to down-regulate the resistant genes. Conclusions: Notably, a strong synergistic effect was observed with AZM-CSNPs in combination with cetirizine, significantly enhancing antibacterial efficacy against resistant isolates.

The efficacy of antibiotics is endangering worldwide due to rapid development of resistant bacterial strains. It is of worth to re-evaluate previously developed antibiotics to alleviate the pressure of microbial resistance on recently developed antibiotics. In this study in vitro toxicological evaluation (cytotoxicity, mutagenicity and genotoxicity) of tilmicosin was carried out. Different concentrations (3.12, 6.25, 12.5, 25, 50, 100, 200, 400 and 800μg/mL) of tilmicosin were used. In vitro cytotoxicity was evaluated using MTT Assay. BHK-21 cells and cardiac cells from chicken embryo were used. The mutagenic potential was evaluated using Ames test. Auxotroph strains of Salmonella typhemurium (TA-98 and TA-100) were used. The genotoxicity was assessed using comet assay. Cell survival percentage of BHK-21 cells at 50μg/mL was 50.64% and IC50 was 49.64μg/mL indicating that below 49.64μg/mL all concentrations were not cytotoxic. While the cell survival percentage of chicken cardiac cells at 25μg/mL was more than 50% and at 50μg/mL was less than 50%. The IC50 calculated was 52.75μg/mL indicating that below 52.75μg/mL all concentrations are not cytotoxic for cardiac cells. The mutagenic indices of all concentrations of tilmicosin with and without S9 fraction were < 2 both for TA98 and TA100. So all the concentrations are non-mutagenic for both TA 98 and TA 100. For genotoxicity, comets were analyzed for head diameter and tail length. The damage indices were calculated. The damage was concentration dependent. The damage indices at concentrations 3.12, 6.25, 12.5 25 and 50µg/ml were not significant (P<0.01) while significant at 100, 200, 400 and 800µg/ml when compared with control (P<0.01). It may be concluded from this study that tilmicosin has not produced cytotoxicity, mutagenicity and genotoxicity at low concentrations under in vitro conditions. Key words: Tilmicosin, MTT Assay, Chicken cardiac cells, Ames test, Comet Assay

The adverse hepatotoxic effect is a great barrier for both drug development and pharmacotherapy because of its complex pathogenesis and unpredictable outcome. This research aimed to explore the hepatic toxicity and gene expression studies after the oral methotrexate administration in Wistar rats and the hepatoprotective efficacy of the metformin-loaded chitosan nanoparticles system. The nanoparticles (NPs) were synthesized by means of the ionic gelation technique with the use of the following instrumental approaches: Fourier Transform Infrared Spectroscopy, Scanning Electron Microscopy, polydispersity index, size, zeta potential analysis, drug entrapment efficiency, and in vitro drug release test. Metformin-loaded chitosan NPs exhibited a distinctive spectral peak of 3447 cm⁻¹, spherical shape, low PDI = 0.30, size of 213 nm, zeta potential of +28.1 mV, and drug entrapment efficiency of 97.34%. A total of 48 male Wistar albino rats (3–4 months old; 220 ± 25 g) were randomly allocated into eight groups (n = 6 per group) under a Completely Randomized Design (CRD) for hepatotoxicity evaluation based on biochemical, histopathological, and gene expression studies. The methotrexate treatment (Group B) caused a severe hepatotoxic and hematological effect with the increased ALT, AST, ALP, total bilirubin serum concentrations and decreased total protein, albumin, TLC, hemoglobin, RBC, HCT, and platelets count compared to the normal control (p ≤ 0.001). These alterations were markedly attenuated by metformin-loaded chitosan nanoparticles (Met-CSNPs), particularly at 75 mg/kg/day (Group H), which produced the most favorable biochemical and hematological profile among the nanoparticle-treated groups. Histopathological examination revealed Met-CSNP treatment progressively reduced hepatic injury, with the 75 mg/kg/day dose showing the greatest tissue preservation and no detectable fibrosis. Gene-expression analysis demonstrated significant suppression of hepatic TIMP1 and TGF-β expression relative to the MTX toxic-control group, with a graded dose-response across the 25, 50, and 75 mg/kg/day Met-CSNP doses. The 75 mg/kg/day formulation produced the greatest reduction in TGF-β expression (0.51-fold relative to the normal control) and the strongest overall suppression of TIMP1 among the nanoparticle-treated groups. Overall, Met-CSNPs demonstrated dose-dependent hepatoprotective and anti-fibro genic effects against MTX-induced hepatic injury, with the 75 mg/kg/day formulation showing the most pronounced protective response. Keywords: Chitosan nanoparticles, Nanoparticle drug delivery, Hepatic protection, Toxicity attenuation.