THE JOURNAL OF MICROBIOLOGY AND MOLECULAR GENETICS http://jmmg.wum.edu.pk/index.php/ojs <p>The Journal of Microbiology and Molecular Genetics (JMMG) will covers the latest developments in the field of Microbiology and Molecular Genetics encompassing the broad spectrum research areas including environmental microbiology, agricultural microbiology, food microbiology, medical microbiology, cancer biology, immunology, human genetics, microbial byproducts, microbial biofilms, microbial bioengineering, human genetics, molecular genetics. The journal explore the significance and interrelationships of microorganisms including bacteria, fungi, cyanobacteria, microalgae, archaea and higher eukaryotes with the advancements in the aforementioned fields. It is an open-access, multidisciplinary, double-blinded, peer-reviewed journal that publishes manuscripts across the entire spectrum of microbiology and molecular genetics.<br />JMMG will provide a multidisciplinary forum for the articles related to basic microbiology, molecular, medical and applied research. Journal of Microbiology and Molecular Genetics is a quarterly journal that publishes original research which is not under consideration for publication elsewhere. The novel and significant research will be considered for this journal.</p> <p> </p> <p><strong>PATRON IN CHIEF</strong><br /><strong>Prof. Dr. Kalsoom Pracha</strong><br />Vice Chancellor, The Women University Multan, Pakistan.<br /><strong>Email:</strong> <a href="mailto:vc@wum.edu.pk">vc@wum.edu.pk</a></p> <p> </p> <p><strong>EDITOR IN CHIEF </strong><br /><strong>Dr. Atia Iqbal</strong><br />Associate Professor, Department of Microbiology and Molecular Genetics, <br />The Women University Multan, Pakistan<br /><strong>Email: </strong>jmmgcheifeditor@wum.edu.pk</p> <p> </p> <p><strong>MANAGING EDITOR</strong><br /><strong>Dr. Sara. Janiad</strong><br />Lecturer, Department of Microbiology and Molecular Genetics,<br />The Women University Multan, Multan, Pakistan<br />Email: sara.9005@wum.edu.pk</p> The Women University Multan en-US THE JOURNAL OF MICROBIOLOGY AND MOLECULAR GENETICS 2707-6431 In-Silico analysis of the structural and functional impact of SNPs in adhesion molecules associated with cardiovascular disease and the effect of potential phytochemicals on these molecules. http://jmmg.wum.edu.pk/index.php/ojs/article/view/144 <p>Adhesion molecules are one of the most important proteins playing a major role in the regulation<br>of cell migration, apoptosis, and survival. While adhesion molecules play a pivotal role in<br>preserving cellular matrix and tissue integrity, these molecules are also involved in the<br>pathogenesis of cardiovascular diseases. The cell adhesion molecules are involved in<br>atherosclerotic plaque progression, deposition of fibrotic tissue, and thrombus formation. Selectins<br>-E and -P and members of the immunoglobulin superfamily ICAM-1 and ICAM-3 are the major<br>groups of the adhesion molecules involved in cardiovascular diseases.<br>In the current study, computational analysis was performed to understand the effects of missense<br>mutations on the structure and functions of the adhesion molecules. The computational studies<br>can lead to a better understanding of the role of single nucleotide polymorphic mutations on the<br>functions of adhesion molecules. The single nucleotide polymorphism data of adhesion molecules<br>was retrieved from gnomAD v2.1.1. Only the missense SNPs were selected, and the 3D protein<br>structures of the adhesion molecules were retrieved from AlphaFold. The 3D protein structures of<br>the adhesion molecules were mutated using USCF Chimera 1.14.<br>The comparison of structural stability, structural change, and interaction with ligands was<br>performed between the wild-type and mutant adhesion molecules by several bioinformatics tools.<br>The R458W mutant of ICAM-1 was found to be destabilizing the structure of the protein.<br>Similarly, the V88M mutant of ICAM-3 and H468Y mutant of E-selectin (SELE), along with<br>G179R and G727Q mutants of P-selectin (SELP) have also shown structural deviations as<br>compared to wild-type molecules. The R367C mutant of ICAM-1, P15L mutant of ICAM-3, and<br>T756P mutant of SELP showed a significant change in the hydrogen bonding interactions with<br>their respective ligands as compared to the wild-type protein interactions with a deviation from the<br>original binding free energies. No significant changes in the hydrogen bonding interactions were<br>observed in the G179R and G727Q mutants of SELP.<br>Moreover, phytochemicals data of several medicinal plants which were effective against<br>cardiovascular diseases was retrieved from the Indian Medicinal Plants, Photochemistry, and<br>Therapeutics database. The molecular docking was performed to analyze the effects of<br>phytochemicals on adhesion molecules. The 3,29-Dihydroxyolean-12-en-28-oic acid, a<br>phytochemical of the Panax ginseng plant was found to have an inhibitory effect on the active<br>sites of adhesion molecules ICAM-1, ICAM-3, SELE, and SELP. Furthermore, the results of this<br>study can be tested experimentally to confirm the effect of adhesion molecule mutations on the<br>onset of cardiovascular diseases and the role of 3,29-Dihydroxyolean-12-en-28-oic acid as a<br>general drug for the treatment of diseases caused by mutant adhesion molecules.</p> Mureed Hussain Muhammad Perwaiz Iqbal Koukab Imran Rana Muhammad Mateen Copyright (c) 2024 Mureed Hussain, Muhammad Perwaiz Iqbal, Koukab Imran, Rana Muhammad Mateen https://creativecommons.org/licenses/by-nc/4.0 2024-04-30 2024-04-30 5 1 1 20 10.52700/jmmg.v5i1.144 Exploring Beta-Lactamase Inhibitory Potential of Common Medicinal Plants http://jmmg.wum.edu.pk/index.php/ojs/article/view/153 <p>The use of ?-Lactam antibiotics improperly has significant negative effects on public safety and health around the world. It causes the disturbance of gut flora, the emergence of multi-drug-resistant bacteria, and environmental contamination. The focus of this investigation was the Gram-negative pathogens <em>Escherichia coli </em>(<em>E. coli</em>), and <em>Pseudomonas aeruginosa </em>(<em>P. aeruginosa</em>). Potent beta-lactamase inhibitors were found among phytochemicals from medicinal plants by <em>in-silico</em> analysis, with Gallocatechin gallate from Green tea and Glycyrrhetinic acid from Licorice being particularly effective against TEM-1 and IMP-1 ?-Lactamases, respectively. <em>In-vitro</em> analysis such as D-zone assay and time-kill assay showed that the methanolic plant extracts improved the effectiveness of antibiotics against these Gram-negative pathogens. This research emphasizes the potential of medicinal plants in combating ?-Lactam antibiotic resistance, offering alternative solutions to address this global health challenge.</p> Areej Fatima Amina Javid Mehboob Ahmed Copyright (c) 2024 Amina Javid, Areej Fatima, Mehboob Ahmed https://creativecommons.org/licenses/by-nc/4.0 2024-04-30 2024-04-30 5 1 21 35 10.52700/jmmg.v5i1.153 Production of Recombinant Human Interferon alpha 2b (rhIFN?-2b) in genetically engineered strain of Bacillus subtilis http://jmmg.wum.edu.pk/index.php/ojs/article/view/158 <p>Interferon alpha 2b (IFN?-2b) is a critical therapeutic protein with immunomodulatory properties, widely used in the treatment of various diseases, including viral infections and cancer. This study presents a comprehensive methodology for the cloning and characterization of the IFN?-2b gene, including the identification of point mutations, followed by expression in <em>Bacillus subtilis</em>. The research begins with gene isolation, primer design, RNA extraction, cDNA synthesis, and PCR amplification. Cloning into a T/A vector and subsequent sequencing reveal point mutations within the gene. The transformed construct is expressed in <em>Bacillus subtilis</em>, and the resulting protein is analyzed using SDS-PAGE. The identification of point mutations and successful protein expression in <em>Bacillus subtilis.</em> This study provides valuable insights into the IFN?-2b gene and its potential applications.</p> Saira Ashfaq Sania Mazhar Naaz Abbas Ramsha Essa Bakhtawar Bukhari Khizra Ismail Yasar Saleem Quratulain Syed Sana Riaz Copyright (c) 2024 sana riaz https://creativecommons.org/licenses/by-nc/4.0 2024-04-30 2024-04-30 5 1 37 43 10.52700/jmmg.v5i1.158 Mechanisms of Toxicity of Heavy Metals and the Microbial Strategies for their Mitigation: A Review http://jmmg.wum.edu.pk/index.php/ojs/article/view/155 <p>Heavy metals toxicity is a major concern worldwide, especially in areas where untreated industrial effluents are released in the environment. Most important heavy metals in this context are arsenic, cadmium, chromium, copper, cobalt, lead, mercury, nickel, selenium and zinc. Some of these are also essential trace elements and play crucial roles in different biological processes. Exposure to high concentration of heavy metals is hazardous for life and can lead to various medical conditions including different types of cancers. The remediation strategies to treat these contaminants involve physical, chemical as well as biological techniques, the latter of which is considered most environment friendly and inexpensive. Bioremediation is the process of removal and detoxification of toxic pollutants using microorganisms. Many microorganisms have evolved different strategies to resist and detoxify such heavy metals. This may include excluding the metals out of the cell, binding of the metals to metal binding proteins to sequester it, conversion of metals into less or non-toxic form by enzymes and alternation of cell components to reduce metal sensitivity. Different genes and proteins are involved in these processes which can be explored for biotechnological interventions and practical applications of bioremediation. This review article discusses the mechanisms of heavy metals’ toxicity, different remediation technologies available, and the various strategies that microorganisms employ to detoxify them.</p> Maria Shafiq Yasir Rehman Copyright (c) 2024 Maria, Yasir Rehman https://creativecommons.org/licenses/by-nc/4.0 2024-04-30 2024-04-30 5 1 45 63 10.52700/jmmg.v5i1.155 LDLR gene variant rs688 TT genotype; genetic association with obesity in Familial Hypercholesterolemia patients http://jmmg.wum.edu.pk/index.php/ojs/article/view/145 <p>The predominant genetic risk factor for familial hypercholesterolemia along with obesity is LDLR allele status. These receptors maintain cellular cholesterol homeostasis. For instance, a common SNP rs688 in different populations has been reported to be associated with elevated<br />plasma LDL, resulting in hypercholesterolemia. The potential role of variant rs688 with obesity in FH patients was observed. This is a case-control study with 120 blood samples of clinically diagnosed obese familial hypercholesterolemia patients by physicians and 120 healthy individuals’ blood samples were recruited for the study. Genomic DNA was extracted from blood samples. By using Tetra ARMS PCR, genotyping of LDLR rs688 (C&gt;T) exon 12 gene variation was performed. Moreover, BMI, BP, and BSL of obese FH patients were also<br />obtained; a chi-square test was performed on the obtained data to evaluate the association between rs688 and obese FH patients. Genotype CC, CT, and TT frequencies reported in the obese FH patients’ group were 0.33, 0.41, and 0.25, while in healthy individuals were 0.37, 0.41, and 0.21 respectively. Chi–square values showed a significant association with BMI, BP, and BSL. It was assessed that there is a 4% increased susceptibility of FH development along with obesity in individuals with TT genotype. So, for obese familial hypercholesterolemia, the LDLR rs688 C&gt;T gene variation can be used as a predisposing genetic marker.</p> Muneeba Afzal Hamza Altaf Tayyaba Faiz Rabia Zafar Ruqaya Shoukat Muhammad Waseem Shahid Aziz Copyright (c) 2024 Muneeba Afzal, Hamza Altaf, Tayyaba Faiz, Rabia Zafar, Ruqaya Shoukat, Muhammad Waseem https://creativecommons.org/licenses/by-nc/4.0 2024-04-30 2024-04-30 5 1 65 72 10.52700/jmmg.v5i1.145 Bacterial profile, antimicrobial susceptibility pattern and associated risk factors of urinary tract infection among pregnant women attending tertiary care hospital in Rawalpindi http://jmmg.wum.edu.pk/index.php/ojs/article/view/160 <p>The study objective to enhance treatment effectiveness by assessing risk factors, bacterial profiles, and antimicrobial susceptibility of uropathogens in pregnant women. During the period from March 2023 to August 2023, we examined 210 urine samples obtained from<br />pregnant women. The samples were subjected to uropathogens isolation, identification and susceptibility testing against thirteen antibiotic classes using the Kirby Bauer disk diffusion method, following standard procedures, to identify uropathogens in cases of significant bacteriuria. In our study there is no significant link found between age, educational level, socioeconomic status, and parity in relation to UTI during pregnancy. In 42% of the samples with significant bacteriuria, Escherichia coli emerged as the predominant uropathogens, followed by Pseudomonas aeruginosa, Staphylococcus saprophyticus, and Klebsiella pneumoniae, in that order. E. coli displayed the greatest resistance to ciprofloxacin and ceftazidime, yet demonstrated high susceptibility to meropenem, imipenem, fosfomycin,<br />and amikacin. Furthermore, a significant portion of K. pneumonia isolates exhibited resistance to ceftazidime and ciprofloxacin. For S. saprophytic, resistance was observed against penicillin and gentamicin, although they displayed 100% sensitivity to vancomycin and 81.2% sensitivity to Nitrofurantoin. The current study findings revealed that factors influencing pregnant women's UTI risk, include age, literacy, residence, multigravida, multipara, infection history, and trimester status. Access to clean water sources also played a role, highlighting the importance of addressing these variables in maternal health to<br />reduce UTI incidence. Screening antenatal patients for UTIs is imperative, and it is advisable to conduct regular screenings for pregnant women. This practice helps monitor uropathogens sensitivity patterns and facilitates the development of antibiotic protocols based on local susceptibility data.</p> Muhammad Adeel Fahad Ishaq Ameer Waqar Habib Muhammad Yousef Aizaz Ajmal Muhammad Adeel Obaid Ullah Iqbal Ahmad Alvi Copyright (c) 2024 Muhammad Adeel https://creativecommons.org/licenses/by-nc/4.0 2024-04-30 2024-04-30 5 1 73 82 10.52700/jmmg.v5i1.160 Co-Incidence of Human Papillomavirus and Epstein-Barr virus in oral cancer Patients: A pilot study. http://jmmg.wum.edu.pk/index.php/ojs/article/view/162 <p>Oral carcinoma is a major disease and leading cause of cancer-related deaths worldwide. It is the most common malignant epithelial carcinoma affecting the oral cavity, 95% of the carcinoma of the oral cavity is squamous cell type in nature. Major risk factors associated with oral carcinoma are abnormal use of alcohol, smoking, and viruses like Human Papillomavirus (HPV) and Epstein Barr Virus (EBV). The current study was designed to detect the different genes of HPV and EBV in oral carcinoma patients of Khyber Pakhtunkhwa, Pakistan. A total of 150 blood samples from oral cancer patients and 156 from healthy individuals were collected. Blood samples of patients along with control samples were screened for HPV and EBV infection for which genomic DNA was extracted by phenol/chloroform method. HPV and EBV detection was performed by polymerase chain reaction on agarose gel electrophoresis. Among 150 oral cases, 118 (79%) were males and 32 (21%) were females. HPV 56 (37%) and EBV 48 (32%) were detected in oral cancer patients, while co-infection of HPV/EBV 22 (14.6%) was detected in oral cancer cases. Taken together, it is concluded that HPV and EBV infection, poor oral hygiene and tobacco products can be the main environmental factors for the development and progression of oral carcinoma in the Pakistani population. Future research with a large sample size is needed to elucidate the mechanism involved in HPV and EBV infection and oral carcinoma.</p> Iqra Bibi Sumaira Sarwar Afsheen Ashfaq Iqbal Ahmad Alvi Afshan Saleem Maimoona Sabir Muhammad Ikram Copyright (c) 2024 Iqra Bibi, Sumaira Sarwar, Afsheen Ashfaq , Iqbal Ahmad Alvi, Afshan Saleem, Maimoona Sabir https://creativecommons.org/licenses/by-nc/4.0 2023-04-30 2023-04-30 5 1 105 117 10.52700/jmmg.v5i1.162 Retrospective Analysis of Tuberculosis Epidemiology in Okara, Pakistan http://jmmg.wum.edu.pk/index.php/ojs/article/view/150 <p>Tuberculosis, commonly called T.B., is a persistent bacterial illness attributed to <em>Mycobacterium tuberculosis</em>. Tuberculosis (T.B.) is a prevalent infectious disease that results in millions of deaths each year, particularly in low-income countries. A retrospective study was conducted between December 1, 2022, and April 30, 2023. Sputum smear microscopy was performed on all individuals suspected of having the T.B. condition. Out of the entire pool of 711 cases under suspicion, 227 cases, which accounts for 31.93% of the total, were confirmed positive for Tuberculosis. The male population is more susceptible to T.B. infection than their female counterparts, with a distribution of 59.47% and 40.53%, respectively. The age group of 30-44 years exhibited the highest rate of pulmonary tuberculosis cases at 24.67%, whereas the age group of 60-74 years recorded the highest percentage of cases at 25.56%. Based on our research, it was determined that the incidence of Tuberculosis exhibited a statistically significant increase within the overall populace of Okara District. The objective of the present study was to examine the Prevalence of Tuberculosis (T.B.) infection and raise awareness among the population regarding T.B. infection in the Okara district of Punjab, Pakistan. Effective disease control and elimination require appropriate management, diagnosis, treatment, and public consciousness.</p> Muhammad Zeeshan Ali Ali Raza Talha Javed Habib Ullah Tahir Anam Tanveer Muhammad Sharjeel Rawal Sumera Sabir Muhammad Adeel Adil Momena Habib Virdah Ijaz Hafiz Muhammad Iftekhar Hafiza Aqeela Iftikhar Copyright (c) 2024 Ali Raza, Muhammad Zeeshan Ali, Talha Javed, Habib Ullah Tahir, Anam Tanveer, Muhammad Sharjeel Rawal4, Sumera Sabir, Muhammad Adeel Adil, Virdah Ijaz , Momena Habib https://creativecommons.org/licenses/by-nc/4.0 2024-04-30 2024-04-30 5 1 97 103 10.52700/jmmg.v5i1.150