Background: Resistance to first-line anti-tuberculosis drugs in Mycobacterium tuberculosis represents a significant public health challenge, particularly in high-burden tuberculosis (TB) settings such as Pakistan, where multidrug-resistant (MDR) forms further complicate disease control efforts. Drug resistance is primarily associated with mutations in rpoB, inhA, katG, embA, embB, embC, and pncA. The emergence of novel, region-specific variants underscores the urgent need for integrating genomic surveillance into routine TB diagnostics and regional control programs. This study aimed to identify the spectrum of mutations contributing to first-line drug resistance in MDR-TB isolates from Khyber Pakhtunkhwa, Pakistan. Methods: Whole-genome sequencing was performed on 16 clinical isolates (12 MDR and 4 drug-susceptible) to identify resistance-associated mutations in rpoB, inhA, katG, embA, embB, embC, and pncA. Detected variants were interpreted using the World Health Organization (WHO) mutation catalogue to determine their association with drug resistance. Phylogenetic relationships were inferred using the Bacterial and Viral Bioinformatics Resource Center (BV-BRC) platform. Results: A total of 16 M. tuberculosis isolates were analyzed to evaluate resistance to first-line anti-tuberculosis drugs. In rpoB, 76 distinct variants were identified, including canonical mutations such as Ser450Leu and His445Arg, as well as a potentially novel substitution, Ser431Phe, predicted to confer high-level rifampicin resistance. The katG and inhA genes harbored 24 and 27 mutations, respectively, including well-characterized substitutions such as Ser315Thr and Ala114Glu, which are strongly associated with isoniazid resistance. Mutations in embA and embB were linked to ethambutol resistance, with several variants localized within conserved transmembrane domains critical for drug interaction. Phylogenetic analysis revealed substantial genetic diversity and evidence of local transmission among MDR-TB isolates. Conclusions: This study suggests that the genetic landscape of drug resistance in M. tuberculosis is highly dynamic in endemic regions. The findings highlight the importance of integrating region-specific mutation profiles into molecular diagnostic frameworks to enhance early detection, guide individualized therapeutic interventions, and strengthen strategies aimed at controlling the transmission of MDR-TB.

Purpose : Colistin-resistant Acinetobacter baumannii (CRAB) poses a growing challenge in neonatal intensive care units (NICUs) due to the emergence of plasmid-mediated Mobile Colistin Resistance (mcr) genes that can rapidly spread among neonates. This pioneering study conducted a comprehensive analysis of mcr-1, mcr-2, mcr-3, mcr-4, and mcr-5 genes in A. baumannii strains isolated from a Pakistani NICU. Notably, this investigation marks the first of its kind in this setting. In addition to genotypic analyses, the study incorporated phenotypic assays to identify the most effective method for detecting colistin resistance in these A. baumannii strains. The research also encompassed the examination of A. baumannii blood isolates exhibiting low to mid-level colistin resistance during an outbreak in a tertiary care hospital. Methods: In the genotypic investigation, DNA was extracted from strains of colistin-resistant A. baumannii, collected from the NICU of the local hospital of Islamabad, Pakistan. The specific set of primers for each mcr gene was used to detect their presence. Various phenotypic methods such as; disc diffusion, broth macro dilution, Minimum Inhibition Concentration (MIC), disc elusion and colistin agar methods were performed using predetermined calculations for phenotypic studies. Results: The study confirms the distribution of the mcr-1 gene among the eight strains, which contributes 66 % of the total number, whereas mcr-2, 3, 4 and 5 genes were not detected in all strains. Based on phenotypic analysis, A. baumannii had shown resistance against colistin at < 2µg/mL. In contrast, MIC varies from strain to strain. Conclusions: The horizontal transfer of the mcr-1 gene is responsible for developing resistance against colistin among A. baumannii in neonates. All phenotypic methods adopted in this study generated the same results, so the method selection depends upon individual comfort. However, we propose that the colistin agar method offers multiple advantages over other methods as it is more economical, easy to perform, multiple samples can be assessed simultaneously and fewer calculations are involved for colistin resistance determination and finding out clinical breakpoints.

Purpose: Colistin resistant Acinetobacter baumannii poses a growing challenge in neonatal intensive care units (NICUs) due to the emergence of plasmid-mediated Mobile Colistin Resistance (mcr) genes that can rapidly spread among neonates. This pioneering study conducted a comprehensive analysis of mcr-1, mcr-2, mcr-3, mcr-4, and mcr-5 genes in A. baumannii strains isolated from NICU. Notably, this investigation marks the first of its kind in this setting. In addition to genotypic analyses, the study incorporated phenotypic assays to identify the most effective method for detecting colistin resistance in these A. baumannii strains. Methods: In the genotypic investigation, DNA was extracted from strains of colistin-resistant A. baumannii, collected from the NICU of a local hospital of Islamabad, Pakistan. The specific set of primers for each mcr gene was used to detect their presence. Various phenotypic methods such as, disc diffusion, broth macro dilution, Minimum Inhibitory Concentration (MIC), disc elusion and colistin agar methods were performed using predetermined calculations for phenotypic studies. Results:The study confirms the distribution of the mcr-1 gene among the eight strains, which contributes 62% of the total number, whereas mcr-2, 3, 4 and 5 genes were not detected in all strains. Based on phenotypic analysis, A. baumannii had shown resistance against colistin at < 2µg/mL. In contrast, MIC varies from strain to strain. Conclusions: The horizontal transfer of the mcr-1 gene is responsible for developing resistance against colistin among A. baumannii in neonates. All phenotypic methods adopted in this study generated the same results, so the method selection depends upon individual comfort. However, we propose that the colistin agar method offers multiple advantages over other methods as it is more economical, easy to perform, multiple samples can be assessed simultaneously, and fewer calculations are involved for colistin resistance determination and finding out clinical breakpoints.

Background: Antimicrobial resistance (AMR) is one of the most rapidly growing threats to global health that is compromising the utility of currently available antibiotics and raising morbidity, mortality, and healthcare costs. Such resistant mechanisms may be overcome using innovative solutions proposed by next-generation nanotechnology, such as metallic nanoparticles, nanocarrier systems, and photothermal therapies, which enhance the outcomes of the treatments. Objective: This research paper was based on the aim to measure awareness, perceived effectiveness, challenges, and the future of nanotechnology in combating AMR among the healthcare professionals, researchers, and students, and to identify the major predictors that affect the perceived potential of nanotechnology in killing AMR-related mortality. Methods: The research involved a quantitative, purposefully sampled, cross-sectional survey study in selecting the respondents, numbering 266. The information was gathered using a structured questionnaire that consisted of six parts, namely demographics, awareness, perceived effectiveness, adoption challenges, prospects, and open-ended questions. The analysis of Likert-scale responses was done using SPSS and SmartPLS. Some statistical tools that were used were the Shapiro-Wilk test of normality, Cronbach's reliability alpha, independent samples T-test, one-way ANOVA, a Chi-Square test of independence, Pearson correlation, and multiple regression. Results: The normality assumption of all variables was met (p > 0.05), and they proved to have excellent reliability (Cronbach's Alpha = 0.87). The t-test showed that there was a significant difference between the genders as relating to the AMR awareness (p = 0.015), and ANOVA showed that there was a significant change in variations related to the child's level of education (p = 0.007). Chi-Square test demonstrated a significant relation between the gender and participation in nanotechnology training ( p = 0.032). The correlation test revealed that all the relationships that exist between awareness, perceived effectiveness, and adoption willingness are important and are either positive or negative. Regression analysis determined that positive predictive impact on the perceived potential of the nanotechnology in the reduction of the AMR-related mortality was awareness (beta = 0.42, p = 0.004), familiarity (beta = 0.36, p = 0.009), and knowledge of the concept of nanomaterials (beta = 0.28, p = 0.015). Conclusion: The research shows the importance of awareness, knowledge, and professional competence in popularizing the notion of using nanotechnology to deal with AMR. In addition to technological advancement, specific awareness and communication programs, fair access opportunities to training, and evidence-based communication will be necessary to boost the speed of use of nanotechnology-based interventions.

Purpose: Colistin resistant Acinetobacter baumannii poses a growing challenge in neonatal intensive care units (NICUs) due to the emergence of plasmid-mediated Mobile Colistin Resistance (mcr) genes that can rapidly spread among neonates. This pioneering study conducted a comprehensive analysis of mcr-1, mcr-2, mcr-3, mcr-4, and mcr-5 genes in A. baumannii strains isolated from NICU. Notably, this investigation marks the first of its kind in this setting. In addition to genotypic analyses, the study incorporated phenotypic assays to identify the most effective method for detecting colistin resistance in these A. baumannii strains. Methods: In the genotypic investigation, DNA was extracted from strains of colistin-resistant A. baumannii, collected from the NICU of a local hospital of Islamabad, Pakistan. The specific set of primers for each mcr gene was used to detect their presence. Various phenotypic methods such as, disc diffusion, broth macro dilution, Minimum Inhibitory Concentration (MIC), disc elusion and colistin agar methods were performed using predetermined calculations for phenotypic studies. Results:The study confirms the distribution of the mcr-1 gene among the eight strains, which contributes 62% of the total number, whereas mcr-2, 3, 4 and 5 genes were not detected in all strains. Based on phenotypic analysis, A. baumannii had shown resistance against colistin at < 2µg/mL. In contrast, MIC varies from strain to strain. Conclusions: The horizontal transfer of the mcr-1 gene is responsible for developing resistance against colistin among A. baumannii in neonates. All phenotypic methods adopted in this study generated the same results, so the method selection depends upon individual comfort. However, we propose that the colistin agar method offers multiple advantages over other methods as it is more economical, easy to perform, multiple samples can be assessed simultaneously, and fewer calculations are involved for colistin resistance determination and finding out clinical breakpoints.