Nadeem Ullah

Background: The emergence of colistin-resistant Acinetobacter baumannii, particularly extensively drug-resistant carbapenemresistant A. baumannii (XDR-CRAB), poses a critical global health threat; however, the molecular mechanisms underlying this resistance remain poorly characterized. Methods: This study utilized integrated tandem mass tag (TMT)-based proteomics and untargeted metabolomics to compare a representative drug-sensitive (DS) strain, a colistin-sensitive multidrug-resistant (MDR-CRAB) strain, and a colistin-resistant extensively drug-resistant (XDR-CRAB) isolate from a Guangzhou hospital, China, with a primary focus on the molecular features associated with resistance in the XDR isolate. Results: Proteomic analysis identified 260 differentially expressed proteins (DEPs) across these strains, with 98 strongly linked to the colistin-resistant phenotype. These DEPs were enriched in functions related to polysaccharide and sulfate transport, aromatic compound degradation, and cationic antimicrobial peptide (CAMP) resistance. Upregulation of the lipid A modification system, driven by increased expression of PmrA (4.2-fold), PmrB (2.9-fold), and PmrC (5.5-fold) in the XDR strain compared to the MDR strain, is consistent with this CAMP resistance. Metabolomic analysis identified 747 metabolites, with lipids and lipid-like molecules being the most altered class. The XDR strain exhibited reduced relative abundance of several glycerophospholipids and fatty acids/ conjugates, when compared to the MDR strain, indicating cell membrane remodeling. Pathway enrichment analysis further implicated purine, glycerophospholipid, starch/sucrose, and glutathione metabolism in colistin resistance. Conclusion: Colistin resistance in A. baumannii appears to involve a multifaceted strategy integrating lipid A modification, cell membrane remodeling, and metabolic reprogramming. This study provides a high-resolution molecular blueprint of colistin resistant A. baumannii and generates testable hypotheses to inform future diagnostic and therapeutic strategies against resistant infections.

Recognizing the therapeutic value of the Genus Fraxinus worldwide, this study evaluates the antioxidant potential of Fraxinus hookeri Wenz. ( F. hookeri ) against CCl ₄ ‐induced nephrotoxicity in rats. Forty‐eight rats are randomly allocated into eight groups (six rats each). Antioxidant enzymes, genotoxicity, urine and serum markers, and tissue histopathology are assessed to determine their nephroprotective effects. The Control group remains untreated, while the DMSO group receives vehicle olive oil intraperitoneally and DMSO orally (3 ml/kg). All other groups, except Control and DMSO, are given CCl ₄ (3 ml/kg, i.p., in 30% olive oil) twice weekly for 4 weeks. The CCl ₄ group receives only CCl ₄ . The Rutin group receives reference drug Rutin orally (50 mg/kg). MEFH100 and MEFH200 groups are given MEFH at 100 and 200 mg/kg, respectively, and NHFH100 and NHFH200 receive NHFH at the same doses. Rutin and F. hookeri treatment effectively ( P < 0.05) restore urine and serum markers disrupted by CCl ₄ . CCl ₄ reduced ( p < 0.05) antioxidant enzymes (CAT, SOD, POD) and increased TBAR levels and DNA damage, which are reversed by cotreatment with F. hookeri and Rutin. Histopathological improvements ( P < 0.05) are also observed with F. hookeri. The results indicate that F. hookeri enhances antioxidant defenses, supporting its potential against CCl ₄ ‐induced nephrotoxicity.

Syphilis, caused by Treponema pallidum, is resurging globally. Molecular typing allows for the investigation of its epidemiology. In Pakistan and other nations, T. pallidum subsp. pallidum has developed widespread macrolide resistance in the past decade. A study at the Peshawar Regional Blood Centre from June 2020–June 2021 analyzed serum samples from 32,812 blood donors in Khyber Pakhtunkhwa, Pakistan, to assess circulating T. pallidum strains and antibiotic resistance. Blood samples were initially screened for T. pallidum antibodies using a chemiluminescent microparticle immunoassay (CMIA). CMIA-reactive samples underwent polymerase chain reaction (PCR) targeted the polA, tpp47, bmp, and tp0319 genes. PCR-positive samples were further analyzed for molecular subtyping using a CDC-developed procedure and tp0548 gene examination. All PCR-positive samples were analyzed for the presence of point mutations A2058G and A2059G in 23S rRNA, as well as the G1058C mutation in 16S rRNA. These mutations are known to impart antimicrobial resistance to macrolides and doxycycline, respectively. Out of 32,812 serum samples, 272 (0.83%) were CMIA-reactive, with 46 being PCR-positive. Nine T. pallidum subtypes were identified, predominantly 14d/f. The A2058G mutation in 23S rRNA was found in 78% of cases, while G1058C in 16S rRNA and A2059G in 23S rRNA were absent. The research found donor blood useful for assessing T. pallidum molecular subtypes and antibiotic resistance, especially when chancres are not present. The prevalent subtype was 14d/f (51.85%), and the high macrolide resistance of 36 (78%) indicates caution in using macrolides for syphilis treatment in Khyber Pakhtunkhwa, Pakistan.

Tuberculosis (TB) is a global health problem caused by the Mycobacterium tuberculosis complex (MTBC). These bacteria secrete various proteins involved in the pathogenesis and persistence of MTBC. Among the secretory proteins, MPT64 (Rv1980C) is highly conserved and is also known as a major culture filtrate that is used in rapid diagnosis of MTBC. In the current study, we aimed to find the mutation in this highly conserved protein in isolates from the Pashtun-dominant province of Pakistan. We analyzed 470 M. tuberculosis whole-genome sequences of Khyber Pakhtunkhwa Province. Mutations in the MPT64 gene were screened through TB-Profiler and BioEdit software tools. The DynaMut web server was used to analyze the impact of the mutation on protein dynamics and stability. Among 470 MTB genomes, three non-synonymous mutations were detected in nine isolates, and one synonymous mutation (G208A) was found in four isolates. Mutation G211T (F159L), which was detected at the C-terminal domain of the protein in six isolates, was the most prominent. The second novel mutation, T480C (I70V), was detected in two isolates at the C-terminal side of the protein structure. The third novel mutation, A491C (L66R), was detected in a single isolate at the N-terminal side of the MPT64 protein. The effect of these three mutations was destabilizing on the protein structure. The molecular flexibility of the first two mutations increased, and the last one decreased. MPT64 is a highly conserved secretory protein, harboring only a few mutations. This study provides useful information for better managing the diagnosis of MTB isolates in high TB-burden countries.

The hypoxia-inducible factors (HIFs) regulate the main transcriptional pathway of response to hypoxia in T cells and are negatively regulated by von Hippel-Lindau factor (VHL). But the role of HIFs in the regulation of CD4 T cell responses during infection withM. tuberculosisisn’t well understood. Here we show that mice lacking VHL in T cells (Vhl cKO) are highly susceptible to infection withM. tuberculosis, which is associated with a low accumulation of mycobacteria-specific T cells in the lungs that display reduced proliferation, altered differentiation and enhanced expression of inhibitory receptors. In contrast, HIF-1 deficiency in T cells is redundant forM. tuberculosiscontrol.Vhl cKOmice also show reduced responses to vaccination. Further, VHL promotes proper MYC-activation, cell-growth responses, DNA synthesis, proliferation and survival of CD4 T cells after TCR activation. The VHL-deficient T cell responses are rescued by the loss of HIF-1α, indicating that the increased susceptibility toM. tuberculosisinfection and the impaired responses ofVhl-deficient T cells are HIF-1-dependent.

Background: Cancer patients, being immunocompromised, are at higher risk of coronavirus disease (COVID-19). The current study determines cancer patients’ knowledge, attitude, perception, and impact of the COVID-19 pandemic. Method: A cross-sectional online survey was conducted in Pakistan from 1 April 2020 to 1 May 2020. The study respondents were cancer patients with ages equal to or greater than 18 years. Following a request for participation, the URL for the survey was distributed on numerous channels. Other social media platforms, including WeChat, WhatsApp, Facebook, Twitter, Instagram, Messenger, and LinkedIn, were used to increase cancer patient interaction. The questionnaire comprised five different sections such as: (1) sociodemographic information, (2) knowledge, (3) attitude, (4) perception, and (5) impact of COVID-19 on cancer patients. Descriptive medical statistics such as frequency, percentage, mean, and standard deviation were used to illustrate the demographic characteristics of the study participants. To compare mean knowledge scores with selected demographic variables, independent sample t-tests and one-way analysis of variance (ANOVA) were used, which are also practical methods in epidemiological, public health and medical research. The cut-off point for statistical significance was set at a p-value of 0.05. Results: More than 300 cancer patients were invited, of which 208 agreed to take part. The response rate was 69.33% (208/300). Gender, marital status, and employment status had a significant association with knowledge scores. Of the total recruited participants, 96% (n = 200) (p < 0.01) knew about COVID-19, and 90% were aware of general symptoms of COVID-19 disease, such as route of transmission and preventive measurements. In total, 94.5% (n = 197) (p < 0.01) were willing to accept isolation if they were infected with COVID-19, and 98% (n = 204) (p < 0.01) had reduced their use of public transportation. More than 90% (n = 188) (p < 0.01) of cancer patients were found to be practicing preventative measures such as using a face mask, keeping social distance, and avoiding handshaking and hugging. Around 94.4% (n = 196) (p < 0.01) of cancer patients had been impacted by, stopped or had changed cancer treatment during this pandemic, resulting in COVID-related anxiety and depression. Conclusion: The included cancer patients exhibited a good level of COVID-19 knowledge, awareness, positive attitude, and perception. Large-scale studies and efforts are needed to raise COVID-19 awareness among less educated and high-risk populations. The present survey indicates that mass-level effective health education initiatives are required for developing countries to improve and reduce the gap between KAP and COVID-19.

Introduction: The Coronavirus disease 2019 (COVID-19) caused many problems in the health sector. Effective and safe vaccines are the only tool to combat the COVID-19 disease. Researchers and regulatory agencies have made efforts to develop such vaccines and healthcare professionals were prioritized for the vaccination program as first-line care providers. Considering this prioritization, we aimed to assess the physicians’ perceptions regarding the side effects of the COVID-19 vaccine.Methods: An interview-based study was conducted from May 5 May to November 5, 2021 (6 months) in a teaching hospital in the capital city of Islamabad, Pakistan.Results: Among the 200 physicians who agreed to participate in the interview, 74% were male. A total of 94% did not experience any side effects after receiving the COVID-19 vaccine, regardless of the type of vaccine; 6% experienced side effects. Furthermore, 90% of physicians were afraid of side effects due to the high number of vaccine doses.Conclusion: Conclusively, physicians had a positive perception regarding the COVID-19 vaccine. Healthcare authorities and related departments must take precautions to ensure the effective and safe COVID-19 vaccine to change the perceptions of physicians who had a negative perception. This will not only reduce the reluctance among physicians regarding administering COVID-19 vaccine, but will also boost and facilitate the vaccination program for the general public as well.