INTERNATIONAL JOURNAL OF APPLIED AND CLINICAL RESEARCH
Myopathic Disorder in an 8-Year-Old Male with Familial Predisposition: A Case Report
Myopathic disorders in children are a heterogeneous group of neuromuscular conditions characterized by progressive muscle weakness, fatigue, and impaired motor function. Early recognition is crucial for diagnosis and management. We report the case of an 8-year-old boy from Charsadda presenting with progressive weakness in the muscles of the hands and legs, accompanied by fatigue, cramps, spasms, and movement difficulties. The patient has a positive family history, including an affected brother and consanguineous marriages within the family. The patient was evaluated at Lady Reading Hospital (LRH), highlighting the importance of clinical recognition, family screening, and early supportive care in pediatric myopathies.
Copyright (c) 2025 Syed Muzammil Hussain Shah, Aqsa Ali (Author) https://creativecommons.org/licenses/by-nc-nd/4.0
Long-Standing Parkinson’s Disease with Severe Dyskinesia and Response to Clozapine
Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized by motor and non-motor symptoms, often complicated by long-term dopaminergic therapy (Rafieipour et al., 2020; Gopalakrishna & Alexander, 2015). We reported a case of early-onset PD with a strong family history, marked disease progression over three decades, and severe levodopa-induced dyskinesias showing partial response to clozapine.
Parkinson’s disease, consanguinity, dyskinesia, clozapine
Copyright (c) 2024 Aqsa Ali, Syed Muzammil Hussain Shah (Author) https://creativecommons.org/licenses/by-nc-nd/4.0
Severe Intellectual Disability in a Child Born to Consanguineous Parents
Intellectual disability (ID) is a neurodevelopmental disorder characterized by significant limitations in intellectual functioning and adaptive behavior, with onset during the developmental period (Schalock, 2011). Consanguinity is a known risk factor for inherited neurodevelopmental disorders (Mokhtari & Bagga, 2003).
Intellectual Disability, consanguinity, pediatric neurology, ASD, epilepsy
Copyright (c) 2024 Syed Muzammil Hussain Shah (Author) https://creativecommons.org/licenses/by-nc-nd/4.0
Comprehensive Review of Conjunctivitis: Etiology, Diagnosis, Clinical Differentiation, and Evidence-Based Management
One of the most common eye diseases observed in primary care and ophthalmology is conjunctivitis, which accounts for more than 1% of all primary care visits and places a significant financial strain on the healthcare system. To distinguish it from other potentially vision-threatening causes of red eye, a comprehensive clinical assessment is required due to its many etiologies, which include viral, bacterial, allergic, toxic and systematic disease-associated types. The current research on conjunctivitis’ epidemiology, risk factors, clinical manifestation, diagnostic techniques, and treatment compiles in this review. Given the clinical overlap and frequent misinterpretation, special attention is paid to differentiating infectious from non-infectious forms and viral from bacterial etiologies. The limits of laboratory diagnostics advancements, especially PCR and fast antigen detection, are examined. Conjunctivitis associated with severe mucocutaneous conditions such Stevens-Jhonson syndrome, reactive arthritis, rosacea, and graft-versus-host disease are also discussed in the review. Evidence-based recommendations and possible risks are taken into consideration when evaluating current treatment options, including the function of antibiotics, antivirals, corticosteroids, immunomodulators, and supportive medicines. Reducing needless antibiotic usage, avoiding problems, and enhancing patient outcomes all depend on an understanding of the various presentations and suitable management techniques.
Conjunctivitis, Red eye, Ocular Infection, Diagnosis, Antibiotics
Copyright (c) 2025 Syed Muzammil Hussain Shah, Aqsa Ali, Maria Nayyar (Author) https://creativecommons.org/licenses/by-nc-nd/4.0
High Prevalence of Iron Deficiency Anaemia Among Pregnant Women in District Peshawar: A Cross-Sectional Study
Anaemia among pregnant women remains a pressing public health concern, especially in tropical developing nations. Having too few healthy red blood cells can cause extreme fatigue, weakness, and shortness of breath for expectant mothers. This study aimed to determine the prevalence of and risk factors associated with iron-deficiency anaemia amidst expectant mothers in Peshawar District, Pakistan. A cross-sectional research design was employed, selecting 100 pregnant patients from Shah Medical Center in Peshawar using proportional-to-size sampling. Clinicians drew 4 ml of venous blood from participants to clinically assess anaemic status. The investigators aimed to elucidate the prevalence of deficient iron levels among local mothers-to-be and identify potential predisposing factors through statistical analysis of the collected medical data. Among 100 samples, 25 showed to be anaemic and the parameters were counted lower than normal. As per the World Health Organization, the criteria for anaemia in pregnancy were applied and serum ferritin < 20µg/l was the cut-off point for iron deficiency. Data regarding risk factors was gathered using pre-designed questionnaires. The prevalence of iron-deficiency anaemia in pregnant women was 22% while most of the cases of anaemia were mild. In this research, the risk factors accompanying anaemia included the period of gestation, poor advice, iron pills intake during antenatal, socioeconomic status, and poor hygiene condition. This study concludes that the high prevalence of iron-deficiency anaemia among pregnant women can be averted if the prevention includes giving iron supplements, proper diagnosis of gestation, and health education
Copyright (c) 2025 Hina Shaheen, Saba Mazhar Shah (Author) https://creativecommons.org/licenses/by-nc-nd/4.0
In Silico Design of a Multi-Epitope Vaccine Candidate Against Lassa Virus Using Immunoinformatics and Molecular Modeling Approaches
Lassa hemorrhagic fever is a viral acute zoonotic disease caused by Lassa mammarenavirus (LASMV) that is part of the Arenaviridae family. With rising global infection and mortality, developing an effective LV vaccine remains an urgent worldwide necessity. In this research, utilizing immunoinformatics methods, we developed a multi-epitope vaccine for the prevention and treatment of LV. Experimentally determined epitopes of the LV were retrieved from ViPR and IEDB databases and subsequently analyzed for various immunological and physiochemical properties. The finalized 12 epitopes were used for vaccine constructions [(LV-1 (300 aa) and LV-2 (353 aa)] with the help of adjuvant (beta-defensin-1 and ribosomal protein) linked to the N-terminal by an EAAK linker. To make the immunogenicity better of the vaccine, a pan-HLA DR binding epitope (13aa) was also introduced and selected epitopes were separated with GPGPG linkers. Predicted vaccine constructs were safe, antigenic, and stable with modeled secondary and 3D structures against LV. The complex between the final vaccine and immune receptors (RIG-I) was evaluated by molecular docking and showed a favorable interaction pattern, including hydrogen bonding and high binding affinity of -10.99 (LV-1) and -29.22 (LV-2) kcal/mol. Codon optimization and in-silico cloning verified the suggested vaccines’ effective expression in E.Coli. Finally, in-silico immune simulation demonstrated that LV-1 and LV-2 would also strongly stimulate the immune system against LV. The final vaccines need experimental confirmation of their safety and efficiency in the control of LV infections.
Copyright (c) 2025 Syed Muzammil Hussain Shah, Aqsa Ali, Maaz Iqbal, Aqib Iqbal, Fatima Tu Zuhra, Zainab Liaqat (Author) https://creativecommons.org/licenses/by-nc-nd/4.0
Adrenaline, aspirin and corticosteroids should be administered mindfully while treating myocardial infarction and Kounis syndrome
<a title="In the important case report by Sirsha Chatterjee et al [1], a 44-year-old Indian woman developed dizziness, widespread weakness, profuse perspiration, pressure in her chest, epigastric pain, and widespread, itchy rashes on her face one hour after ingesting oral amoxicillin clavulanate. The provisional diagnosis was Kounis syndrome. The patient began receiving, among others, medications including adrenaline nebulization, intravenous hydrocortisone and oral aspirin. The High sensitive troponin was raised but coronary angiography was not performed because the patient refused further cardiac workup. This report raises serious issues with the usage of adrenaline, aspirin, and hydrocortisonehe. These medications, which are used to treat Kounis syndrome, myocardial infarction, or thrombosis [2], paradoxically have the potential to trigger these diseases" href="https://www.ijacr.com/index.php/home/In the important case report by Sirsha Chatterjee et al [1], a 44-year-old Indian woman developed dizziness, widespread weakness, profuse perspiration, pressure in her chest, epigastric pain, and widespread, itchy rashes on her face one hour after ingesting oral amoxicillin clavulanate. The provisional diagnosis was Kounis syndrome. The patient began receiving, among others, medications including adrenaline nebulization, intravenous hydrocortisone and oral aspirin. The High sensitive troponin was raised but coronary angiography was not performed because the patient refused further cardiac workup. This report raises serious issues with the usage of adrenaline, aspirin, and hydrocortisonehe. These medications, which are used to treat Kounis syndrome, myocardial infarction, or thrombosis [2], paradoxically have the potential to trigger these diseases">In the important case report by Sirsha Chatterjee et al [1], a 44-year-old Indian woman developed dizziness, widespread weakness, profuse perspiration, pressure in her chest, epigastric pain, and widespread, itchy rashes on her face one hour after ingesting oral amoxicillin clavulanate. The provisional diagnosis was Kounis syndrome. The patient began receiving, among others, medications including adrenaline nebulization, intravenous hydrocortisone and oral aspirin. The High sensitive troponin was raised but coronary angiography was not performed because the patient refused further cardiac workup. This report raises serious issues with the usage of adrenaline, aspirin, and hydrocortisonehe. These medications, which are used to treat Kounis syndrome, myocardial infarction, or thrombosis [2], paradoxically have the potential to trigger these diseases
Adrenaline, aspirin, corticosteroids, Kounis syndromes, myocardial infarction
Copyright (c) 2025 Nicholas Kounis, Virginia Mplani, Maria Bozika, Ioanna Koniari (Author) https://creativecommons.org/licenses/by-nc-nd/4.0
REVIEW: CRISPR-Cas Methodologies in the Detection and Treatment of Infectious Illnesses
The identification of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and their corresponding Cas proteins, originally a bacterial defense mechanism, has initiated a transformative phase in genetic engineering with significant ramifications for clinical treatment. This review examines the swift advancement and many uses of CRISPR-Cas techniques in the identification and management of infectious diseases, emphasizing their capacity to revolutionize contemporary clinical practice. In diagnostics, systems such as SHERLOCK (for RNA) and DETECTR (for DNA) employ the collateral cleavage activity of Cas nucleases (e.g., Cas12 and Cas13) to enable ultra-sensitive, isothermal, and quick detection of viral and bacterial nucleic acids, including SARS-CoV-2. These field-deployable devices, in conjunction with Next Generation Sequencing augmentation methodologies such as DASH and FLASH for low-frequency targets, signify a substantial advancement beyond conventional diagnostic techniques. Therapeutically CRISPR-Cas provides a means to achieve a functional cure for chronic and latent viral infections by accurately targeting and modifying integrated viral genomes that are typically unreachable by standard antivirals. Promising pre-clinical research has shown the eradication or inactivation of viruses such as Human Immunodeficiency Virus-1 (HIV-1) and Hepatitis B Virus (HBV) through tactics like multiplexed guided RNAs and less-mutagenic base editors. Although the remarkable promise is evident, issues of delivery efficiency, off target editing, and host immunity must be resolved as these technologies go into preliminary clinical trials. Ultimately, CRISPR-Cas techniques represent a disruptive force, offering accurate, diverse, and readily changeable tools that are set to become essential in the clinical management of infectious diseases.
CRISPR, Cas 9, Cas12, Cas13, infectious diseases
Copyright (c) 2025 Aqsa Ali, Syed Muzammil Hussain Shah, Majid Khan, Zainab Liaqat, Shoaib ur Rehman https://creativecommons.org/licenses/by-nc-nd/4.0
Gene-Targeted Antimicrobial Strategies Using CRISPR-Cas Systems Against Multidrug-Resistant Bacterial Pathogens
The rapid emergence of antibiotic-resistant bacteria has significantly outpaced the development of new antimicrobial agents, posing a global health threat. This review investigates the application of CRISPR-Cas9 and CRISPR-dCas9 systems as innovative strategies to combat resistance in key bacterial pathogens. A systematic literature search identified studies focused on methicillin-resistant Staphylococcus aureus (MRSA) and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli. In MRSA, the CRISPR-dCas9 system was used to suppress mecA gene transcription, leading to a 77% reduction in gene expression without compromising bacterial viability-an approach with implications for microbiome-safe interventions. In contrast, CRISPR-Cas9 targeting of conserved ESBL gene sequences in E. coli successfully cleaved resistance determinants, resulting in >99% re-sensitization to β-lactam antibiotics. The engineered pRESAFRESBL plasmid demonstrated cross-sensitivity restoration, suggesting plasmid-wide disruption of co-transferred resistance genes. Together, these findings highlight the transformative potential of CRISPR-based technologies in reversing resistance phenotypes through precise, gene-targeted interventions. This approach paves the way for next-generation antimicrobial therapies that either neutralize resistance while preserving beneficial flora or directly eradicate resistant clone
: CRISPR-Cas9, CRISPR-dCas9, antibiotic resistance, MRSA, ESBL-producing E. coli, gene edit ing, bacterial re-sensitization.
Copyright (c) 2025 Karishma Noor, Muhammad Ibrahim Shehzad, Dewa khan, Afaq Ali khan (Author) https://creativecommons.org/licenses/by-nc-nd/4.0
Comparative In Silico Analysis of Wild-Type and Mutant-Type Akt2 Gene Mutation (C.58c>T) In Type-2 Diabetes Mellitus
Type 2 Diabetes Mellitus (T2DM) is a complex and chronic metabolic disorder that has been frequently linked to single nucleotide polymorphisms (SNPs) in the AKT2 gene. AKT2, a key isoform of the AKT kinase family, plays an essential role in metabolic regulation and has been implicated in the development and progression of several cancers, including ovarian, breast, and pancreatic malignancies. Genetic mutations in AKT2 often lead to impaired insulin signaling and secretion, positioning this gene as a crucial therapeutic target for both metabolic and oncogenic pathologies. In the present study, a computational analysis was undertaken to examine the structural and functional consequences of a specific point mutation in the AKT2 protein. Bioinformatics tools predicted the mutation to be pathogenic, resulting in notable destabilization of the protein’s native conformation. To further explore therapeutic interventions, molecular docking simulations were performed with two candidate ligands, MESOBILIVERDIN and MK-2206, against the mutant AKT2 protein. MESOBILIVERDIN exhibited superior binding affinity and enhanced structural stability, with a calculated binding energy of −13.02 kcal/mol, compared to MK-2206, which showed a binding energy of −10.5 kcal/mol. These findings suggest both compounds possess therapeutic potential; however, MESOBILIVERDIN appears to be the more promising ligand for mutation-specific targeting in T2DM.Collectively, this study offers a computational framework for assessing mutation-induced alterations in protein behavior and supports the application of structure-based drug design approaches in the development of targeted therapies for genetically defined subtypes of metabolic disorders.
Type 2 Diabetes Mellitus, AKT2 gene, SNPs, insulin signaling, molecular docking, MESOBILIVERDIN, MK-2206, protein stability, pathogenic mutation, structure-based drug design.
Copyright (c) 2025 Shamama Khan, Aizaz Ali, Areeha Nadeem, Asma Ahmed, Attiya Hameed, Rabia Hashmi, Amman Sardar, S Ashmam Ali shah, Usama, Sahar Nigar (Author) https://creativecommons.org/licenses/by-nc-nd/4.0
Computational Analysis of GRIN2A C1448R Variant in Epilepsy-Associated Intellectual Disability and their Structural Impact on NMDA Receptor Binding with Rapastinel and Dizocilpine
Epilepsy-associated intellectual disability (ID) encompasses a spectrum of neurodevelopmental disorders often caused by genetic mutations affecting synaptic transmission. This study employed a dry-lab, in silico approach to identify and characterize high-confidence candidate genes implicated in epilepsy-related ID, with a focus on GRIN2A, a subunit of the N-Methyl-D-Aspartate (NMDA) receptor complex. The pathogenic variant, c.4342T>C (p.Cys1448Arg), was subjected to computational analysis, including pathogenicity prediction, protein stability evaluation, and molecular docking. Two ligands, Rapastinel (GLYX-13) and Dizocilpine (MK-801) were selected for docking based on their clinical and pharmacological relevance to NMDA receptor modulation. Comparative docking revealed a reduction in binding affinity for Rapastinel (from −7.5 to −7.1 kcal/mol) and a slight increase for Dizocilpine (from −8.3 to −8.4 kcal/mol) in the mutant receptor. Protein–ligand interaction profiling showed that Rapastinel’s hydrogen bonding network was disrupted, while Dizocilpine retained hydrophobic interactions. Normal mode analysis indicated increased structural flexibility, with lower eigenvalues and elevated B-factors in the mutant complex, suggesting destabilized receptor conformation. These findings provide molecular insight into how the C1448R mutation may contribute to synaptic dysfunction in epilepsy-associated intellectual disability. This study underscores the utility of computational biology in exploring genotype–phenotype relationships and may guide future drug development efforts targeting GRIN2A related epileptic disorders.
Epilepsy GRIN2A, NMDA receptor, molecular docking, Rapastinel, Dizocilpine, C1448R mutation, ligand binding affinity, receptor conformation.
Copyright (c) 2025 Sumaiya Yousaf, Rimsha Khan, Uzair Malik (Author)
Computational Drug Repurposing for The Management of Pathogenicity of Beta-Thalassemia
Beta-Thalassemia is an inherited disorder caused by mutation in the HBB gene, resulting in the abnormal synthesis of beta globin protein. The c.92G>C (p.Arg31Thr) mutation of HBB gene is prevalent in South Asian population and results unstable protein. The current study uses computational approaches to analyze the pathogenicity of this mutation and the ability of three ligands, 4-Phenylbutyric Acid (4-PBA), Voxelotor, and Heminin stabilizing mutant beta-globin protein. Bioinformatics tools (MutationTaster, SIFT, PolyPhen-2, FATHMM) confirmed that the mutation is deleterious and impact the function of protein. I-Mutant and Mu-Pro suggested that the stability of protein decreases due to p.Arg31Thr mutation. Molecular docking was carried out using AutoDock Vina revealed that 4-PBA and Hemin show stronger binding affinity towards the mutant protein (−6.5 kcal/mol and −6.8 kcal/mol, respectively) as compared to wild-type protein, suggesting a stabilizing effect. Post-dock analysis employing Protein-Ligand Interaction Profiler (PLIP) showed that enhanced hydrophobic interactions and hydrogen bonding were observed in mutant-ligand complexes. In contrast, Voxelotor represented a reduction in the binding affinity to mutants, indicating that conformational changes have affected the binding. These findings indicate that 4-PBA and Hemin can be potentially used to reduce the severity of beta-thalassemia by stabilizing the mutant beta-globin. Further in vitro and in vivo validation is needed to confirm the results since in silico approaches alone cannot be relied upon. This study underlines computational drug repurpose as a promising strategy for treatment of Beta-Thalassemia.
Beta-Thalassemia, 4-Phenylbutyric Acid (4-PBA), Voxelotor, MutationTaster, SIFT, Pol-yPhen-2, FATHMM.
Copyright (c) 2025 Samrina Muskan, Sajila Mustafa, Usama, Uzair Malik (Author)
Computational Analysis of the Structural and Functional Impact of the PRKN Gene Mutation (c.1130G>A) in Parkinson’s Disease
Parkinson’s disease (PD) is a neurodegenerative disorder associated with genetic mutations, including those in the PRKN gene, which encodes the E3 ubiquitin ligase parkin. This study employed computational approaches to investigate the pathogenic effect of the PRKN G>A mutation (position 1130) and identify potential therapeutic ligands. Comprehensive Database analysis (MalaCards, DisGeNET, OMIM) identified PRKN as a key PD-associated gene with p value (P < 0.05), pathogenicity analysis (MutationTaster, FATHMM, Polyphen-2) confirmed the mutation as deleterious, while protein stability (I- I-mutant, Mu Pro) revealed protein destabilization. Molecular docking simulations demonstrated N-aryl benzimidazole enhanced binding affinity for the mutant PRKN (−7.5 kcal/mol vs. −5.5 kcal/mol in wild-type), mediated by salt bridges (GLU, GLU) and hydrophobic contacts, contrasting kinetin showed no mutation-dependent affinity changes (-5.6 kcal/mol). Protein-protein interaction networks (STRING) further implicated mutant PRKN in disrupted mitochondrial regulation. These findings highlight N-aryl benzimidazole as a promising therapeutic candidate for mutation-specific PD treatment and provide a computational framework for targeting genetic subtypes of neurodegenerative diseases.
Parkinson’s disease, PRKN, MutationTaster, FATHMM, Polyphen-2, N-aryl, Molecular docking and Simulation
Copyright (c) 2025 Attiya Hameed, Amman Sardar, Aizaz Ali, S Ashmam Ali Shah, Usama Nazif, Rabia Hashmi, Shamama Khan, Sajela Akbar (Author)
Epidemiological Study of Parkinson’s disease in the Population of Khyber Pakhtunkhwa-Pakistan
Parkinson’s disease (PD) is a neurological disorder characterized by motor impairment affecting millions of people globally. Significantly, in Pakistan approximately 450,000 PD cases have been reported previously. This study aims to ascertain the prevalence of PD in Khyber Pakhtunkhwa a province of Pakistan. Interviews were conducted with PD patients, admitted to the Department of Neurology Lady Reading Hospital, Peshawar. Already prepared questionnaires were used to record the clinical and demographic data on the basis of age, gender, comorbidities, and family history. Clinically, it was identified that Parkinson’s patients showed different features such as tremors, muscle rigidity, bradykinesia, difficulty walking, dementia, frequent falls, weakness, difficulty speaking, sleeplessness, apathy, and dyskinesia. According to the statistical analysis, PD primarily affected those between the ages of 55 and 64 and it was more common in men than women. The significance of this study was to explain the epidemiological and clinical features of PD in the KP province of Pakistan. The current study will support accurate diagnosis and treatment as well as disease management.
Parkinson’s disease, Gender, Age groups, Prevalence.
Copyright (c) 2024 Aqsa Ali, Syed Muzammil Hussain Shah, Noor, Karen Jacqueline Cloete (Author)
A Systematic Review of Genetic, Clinical, and Gender Associated Aspects of the SNCA Gene in Familial Parkinson’s Disease
Parkinson's disease (PD) presents as a progressive neurological disorder that manifests through motor symptoms such as tremors and movement stiffness, which drastically affect both mobility and quality of life. This is caused by dopamine-producing cells in the brain. The worldwide prevalence is estimated at 149 cases per 100,000 persons, indicating a rise to 25.2 million by 2050. By conducting a comprehensive PubMed database search for SNCA AND Parkinson's from 2020 to 2025, with adult populations as the focus, the researchers identified relevant studies for data extraction and analysis. The SNCA gene holds critical significance because it produces alpha-synuclein, which forms toxic Lewy bodies in brain tissue. Rare SNCA mutations that result in gene multiplication trigger severe early-onset Parkinson's disease, while common genetic variations such as REP1 polymorphism increase idiopathic PD risk. Research indicates that individuals with longer SNCA-Rep1 alleles have a greater chance of developing Parkinson's disease while also experiencing worsening cognitive impairment and non-motor symptom severity. Men exhibit a 1.5-2 fold higher prevalence of Parkinson's disease and experience symptoms earlier in life compared to women, but women display tremors more frequently when diagnosed with the disease. Female patients show elevated striatal dopamine measurements while experiencing symptoms and describe a greater intensity of pain and emotional responses. The evaluation of motor symptoms like bradykinesia through gyroscope signals demonstrates potential since it aligns closely with clinical rating scales such as the Unified Parkinson's disease rating scale (UPDRS). Many non-motor symptoms, including neuropsychiatric difficulties, sleep disorders, and autonomic dysfunction, affect up to 95% of patients with hyposmia and lower quality of life before motor symptoms appear. Analysing how genetics and gender contribute to symptom development is crucial for developing improved diagnostic tools and treatment methods for Parkinson's disease.
Systematic Review, SNCA, Parkinson’s Disease, Unified Parkinson's disease rating scale (UPDRS).
Copyright (c) 2024 Izzat Afridi, Muhammad Yaseen, Romessa Wali Khan, Aizaz Ali, Saqib Ullah (Author)
Revolutionizing Healthcare: Advances in Biotechnology for Enhanced Disease Diagnosis and Treatment
Biotechnology is changing healthcare by giving new solutions for disease diagnosis and treatment. Advances in gene editing, RNA-based therapeutics, and cell and gene therapies are innovating values in medical intervention. Techniques such as CRISPR-Cas9 enable precise genetic modifications and offer new avenues for treating genetic disorders and complex diseases. RNA-based therapeutics- mRNA rice and RNA interference are increasingly being applied in treating cancer, genetic disorders, and autoimmune disorders, expanding their scope beyond vaccines. Cell and gene therapies, like CAR-T cell therapy, are evolving to improve their effectiveness and safety over a broader range of diseases. This article describes methodologies in all these cutting-edge technologies and how they may provide targeted and personalized treatment. The discussion also brings into play artificial intelligence that further accelerates drug discovery and enhances the reliability of diagnostics. The recent results presented showcase the potential of these technologies with promising improvements in the treatment outcome of various diseases. The discussion discusses how biotechnology can revolutionize healthcare, leading to personalized medicine in which treatments are tailor-made to dose individually based on genetic profiles. This not only increases treatment efficiency but would also reduce its side effects; it can be said that it represents a significant shift towards precision medicine. Essential issues that still exist include ethics, regulations, and outreach. The biotechnology industry continues to evolve and is likely to take a front-row seat during transformation within the healthcare landscape, offering actionable and sustainable solutions. Overall, the present review gives a detailed overview of contemporary advances in biotechnology concerning changing the realm of healthcare and management of patient outcomes. Methodologies, observations, and future directions will be discussed, shedding light on how biotechnology will define the course of medicine in the future.
Gene Editing, RNA-based Therapeutics, Personalized Medicine, CRISPR-Case 9, Bio-technology, Vaccine Development
Copyright (c) 2024 Horriya Shabbir, Suneela Saeed, Maria Nayyar, Saba Mazhar Shah, Nadia Nisar, Esha Nawaz (Author)
Biochemical Analysis and Therapeutic Potential of Extract from Mushroom (Pleurotus ostreatus)
Pleurotus ostreatus is a popularly consumed species due to its taste, flavor, large values of nutrition, and pharmacological products. This research aims to investigate the nutritional, medicinal, and toxicological properties of locally cultivated oyster mushrooms in Khyber Pakhtunkhwa, Pakistan. The study consists of the evaluation of the presence of toxins using HPLC, proximate composition, nutritional profile, hemolytic activity (against human erythrocytes), cytotoxic activity (against brine shrimp), antibacterial potential, and anticancer properties of crude methanolic extract of P. ostreatus. Toxicological evaluation through HPLC revealed the presence of ochratoxin A (3.4 ppb) and Penicillic acid (2.7 ppb). Proximate analysis showed that the moisture content in 5 g of P. ostreatus was approximately 3.00±01%, 3.6±0.5% of proteins, 10.5±0.4% of crude fiber, 1.5±01% of lipids, and 89.3% of carbohydrates. Similarly, the mineral profile showed the presence of Fe, Mg, Na, Zn, Cr, Mn, and K. During hemolytic activity, the biocompatibility of extract towards human RBCs was free from the risk. The cytotoxic effects on shrimps were minimal and were found to be safe for living cells with a 64.57 LC50 value. The extract showed maximum antibacterial activity against P. aeruginosa, E. amnigenus, E. coli, E. cloacae, Citrobacter, and S. Serratia. The extract also showed noticeable cytotoxic activity on human Rhabdomyosarcoma cell lines. Overall, our results indicated that locally cultivated mushroom (P. ostreatus) is a rich source of nutrients and therapeutic agents with obscure toxicity.
Edible mushroom; hypocholesterolemia activities; polysaccharide content; Nutritional benefits; Food production
Copyright (c) 2024 Huma Gul, Sajeela Akbar, Sadaf Gohar, Syed Sohail shah, Faiza Mazhar, Sheema (Author)
Mutational Analysis of Carbapenem Resistance Genes in Citrobacter Freundii Isolated from Urinary Tract Infections (UTIs) Patients, Peshawar
In the current era, pathogens become more virulent due to the development of resistance against the major families of antibiotics. The most common infection which is both community-acquired and hospital-acquired is the Urinary tract infection (UTI). The current study is intended to determine the mutational analysis of carbapenem resistance in the isolates of Citrobacter freundii isolated from UTI patients. A total of 145 urine samples were collected from the UTI suspected patients at Khyber Teaching Hospital, (Peshawar, Pakistan). Out of 145 samples, 40 samples were detected as positive isolates of C. freundii. The phenotypic identification of isolates was done via API-10S kit. The carbapenem antibiotic resistance genes were detected via Polymerase Chain Reaction (PCR). The antibiotic susceptibility testing was done via Kirby Bauer method and the results revealed that the isolates were highly resistant towards cefepime (97.5%), amoxicillin (95%), and meropenem (92.5%). The molecular analysis revealed that the VIM gene was detected in 3 isolates, the NDM-1 gene in 6 isolates, and the OXA-48 gene was detected in 4 isolates of C. freundii. The result of Next generation sequencing revealed that no mutation was detected in the sequences of these carbapenem resistance genes. The output of the current study concluded that carbapenem resistance is emerging rapidly mainly in the uropathogens and the antibiotic resistance genes enhance the ability of uropathogens to combat against antibiotics.
Citrobacter freundii, UTIs, VIM gene, NDM-1 gene, OXA-48 gene, phenotypic identifi-cation, API-10S kit, Kirby Bauer method.
Copyright (c) 2024 Maryam Mumtaz, Rukhsar Niaz, Fareeha Khalid, Khadija Mumtaz, Kaleem Ur Rehman, Mamoon Rasheed (Author)
Molecular characterization of β-Thalassemia in Mehsud and Wazir tribes of inbreed Pashtun Population showed different patterns, indicating genetic heterogeneity for β-Thalassemia
β-Thalassemia (β-thal) is a monogenic disorder characterized by mutations in the HBB gene, which affect globin production, leading to hypochromic and microcytic anemia. This hereditary blood condition is marked by a decreased or absent synthesis of the β-globin chain of hemoglobin. It is notably prevalent in regions with a high prevalence of consanguineous marriages, such as the Pashtun population. The present study aimed to assess and compare the incidence of the most commonly occurring mutations of β-thalassemia in the Mehsud and Wazir tribes of Pashtun ethnicity. Additionally, the study examined the inheritance patterns of these mutations in patients and explored the level of consanguinity among parents. The findings hold potential significance for prenatal diagnosis (PND), genetic counseling, and carrier screening, which could help manage and reduce the occurrence of affected births within these tribes and the broader Pashtun population. This study involved the collection of 230 peripheral blood samples from affected patients, their parents, and siblings from families with at least one transfusion-dependent child, as well as sporadic patients from the Mehsud tribe. These samples were collected from various areas of the South Waziristan tribal district in the Mehsud region of Khyber Pakhtunkhwa (KP), Pakistan. A similar approach was followed for collecting 230 blood samples from the Wazir tribe, residing in different areas of the Frontier Region (FR) Bannu, KP, Pakistan. All samples were analyzed using the Amplification Refractory Mutation System Polymerase Chain Reaction (ARMS-PCR) technique to detect the six most common β-thalassemia mutations prevalent among Pashtun populations. The findings revealed some unique characteristics, particularly within the Mehsud tribe. The most prevalent mutation in this tribe was codon 41/42 (–TTCT), followed by frameshift codons FSC 8/9 (+G) (HBB: c.27_28insG), IVS-I-5 (G>C), and FSC-5 (–CT). Interestingly, the mutation spectrum in the geographically adjacent Wazir tribe differed. In this tribe, IVS-I-5 (G>C) was the most frequently detected mutation, followed by FSC-5 (–CT), among the six reported mutations. The findings of this study reveal distinct heterogeneity in the mutation patterns of β-thalassemia between the geographically adjacent Mehsud and Wazir tribes of the Pashtun ethnicity. These results highlight the importance of tailored parental counseling to address disease recurrence effectively. Furthermore, they emphasize the need for large-scale mutation screening and comprehensive prenatal diagnostic efforts within the Mehsud and Wazir tribes, as well as the broader Pashtun population.
Hemoglobin (Hb), Prenatal Diagnosis (PND), beta-thalassemia (β-thal).
Copyright (c) 2023 Mohammad Junaid, Sami Ullah (Author)
The molecular characterization of β-Thalassemia in Afridi and Wazir tribes of inbreed Pashtoon population showed different patterns, indicating genetic heterogeneity for β-Thalassemia
β-Thalassemia (β-thal) is a monogenic disease resulting from mutations in the HBB gene, leading to aberrant globin production and causing hypochromic and microcytic anemia. The current study aimed to assess and compare the incidence of the most frequently occurring mutations of beta-thalassemia in sub-tribes Afridi and Ahmadzai of the Wazir tribe belonging to the Pashtoon Ethnicity, additionally the study investigated the inheritance pattern of these mutations in affected patients and the prevalence of consanguinity among the parents. Furthermore, this study holds potential significance in the context of prenatal diagnosis (PND), Genetic counseling, and carrier screening to control the occurrence of affected births not only in these tribes but in the entire Pashtoon Population. During the current research, 300 peripheral blood samples of affected patients, their parents, and siblings were compiled both from families having at least one transfusion-dependent child and sporadic patients from Ahmadzai Wazir residing in different areas of Wazir subdivision Banu Khyber Pakhtunkhwa (KPK), Pakistan. The same procedure was followed for collecting 300 peripheral blood samples from Afridi Wazir residing in different areas of Wazir sub-division Banu and Nort Waziristan Tribal District (NWTD), KPK, Pakistan. These samples were analyzed for the six most common β-thalassemia mutations found in the Pashtun population via the amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) technique. Results obtained were a bit unique as the most common mutation detected in Afridi Wazir were Codons 41/42 ( – TTCT), interestingly followed by frameshift codons FSC 8/9 (+G) (HBB: c.27_28insG), IVS-I-5(G>C) and FSC-5 (–CT). While results obtained for Ahmadzai Wazir were different than the above, and interestingly reported IVS-I-5 (G > C) to be the most recurrent followed by FSC-5 (–CT) among the six reported mutations. The findings of the present study show differences within the geographically adjacent situated sub-branches of highly inbreed Wazir tribe of Pashtoon Ethnicity, clearly demarcating genetic heterogeneity for β-Thalassemia. These observations need serious consideration in implementing parental meetings about disease recurrence in the future, large-scale mutation screening, and PND for the population of the Wazir tribe and the whole Pashtun ethnicity as well.
β-thalassemia (β-thal); Hemoglobin(Hb), Prenatal Diagnosis (PND)
Copyright (c) 2023 Saqib Ullah, Afrasiab, Momen Khan, Hidayat Ullah, Ihsan Ullah, Moheb khan, Imran khan (Author)
Molecular Characterization of Sulfonamide and Fluoroquinolone Resistant Genes of Morganella morganii Isolates from Urinary Tract Infections (UTIs) Patients, Peshawar
Uropathogens pose a significant threat to public health, affecting both community and hospitalized patients. Among these, Gram-negative bacteria represent a predominant group. In recent years, Morganella morganii has emerged as a notable cause of urinary tract infections (UTIs) worldwide, with a concerning rise in prevalence attributed to the evolution of resistance mechanisms against multiple antibiotics. The present study aimed to investigate the prevalence of M. morganii, its associated antibiotic-resistance genes, and its correlation with demographic factors such as gender and age. The findings revealed that M. morganii was more frequently isolated from females (n = 27) than males (n = 16). The highest prevalence rate (37.2%) was observed in the age group of 21–40 years. The antibiotic susceptibility profile of M. morganii isolates indicated high resistance to cotrimoxazole (77%), followed by ciprofloxacin (67%), Cefoperazone (65%), cefotaxime (58%), gentamicin (28%), amoxicillin-clavulanate (26%), imipenem (16%), Fosfomycin (9%), meropenem (7%), piperacillin-tazobactam (7%), and amikacin (5%). PCR analysis revealed the presence of antibiotic-resistance genes in the resistant isolates. Among cotrimoxazole-resistant isolates, the sul1 and sul2 genes were detected in 23 isolates each. Similarly, in ciprofloxacin-resistant isolates, the gyrA and gyrB genes were identified in 11 and 10 isolates, respectively. Interestingly, certain isolates harbored various combinations of these resistance genes. Seven isolates contained all four resistance genes (sul1, sul2, gyrA, and gyrB), while seven isolates harbored three genes in combinations (sul1, sul2, gyrA or sul1, sul2, gyrB). Nine isolates exhibited two-gene combinations (sul1 and sul2).
uropathogen, urinary tract infections (UTIs), sul1, sul2, gyrA, gyrB, PCR analysis.
Copyright (c) 2023 Rukhsar Niaz, Ramsha Shah (Author)
Molecular Characterization and Mutational Analysis of Virulence Genes in Clinical Isolates of Klebsiella pneumoniae Obtained from Khyber Teaching Hospital, Peshawar
Klebsiella pneumoniae is a Gram-negative opportunistic pathogen implicated in various nosocomial and community-acquired infections. This study aimed to investigate key virulence factors contributing to the pathogenicity of K. pneumoniae. A total of 100 bacterial isolates were analyzed, of which K. pneumoniae was identified through 16S rRNA sequencing. The remaining isolates were classified as Escherichia coli, Citrobacter, and Shigella species. The virulence potential of K. pneumoniae was assessed by detecting the presence of the cnf1 and sfa genes, which were identified in 7% and 17% of the isolates, respectively. Despite their low prevalence, these genes were sufficient to enhance the pathogenic capacity of the bacteria. Mutational analysis revealed genetic variations in the cnf1 gene, although these mutations were pre-existing within the analyzed strains.
Klebsiella pneumoniae, virulence factors, 16S rRNA sequencing, cnf1 gene, sfa gene, muta-tional analysis.
Copyright (c) 2023 Sadaf Gohar, Sajeela Akbar, Faiza Mazhar, Syed Sohail shah, Muhammad Taha, Asmat Ullah (Author)
Molecular Detection and Characterization of Efflux Pump Genes Associated with Multidrug Resistance in Clinical Isolates of Staphylococcus aureus in Peshawar, Pakistan
Staphylococcus aureus is an opportunistic pathogen responsible for a wide range of human diseases. It is renowned for its capacity to develop resistance to multiple antibiotics, primarily due to diverse resistance mechanisms, including efflux pumps, which significantly contribute to intrinsic resistance. This study aimed to investigate the expression of efflux pump genes, namely NorA, MepA, and MdeA, in clinical isolates of S. aureus obtained from Khyber Teaching Hospital and Hayatabad Medical Complex. A total of 200 clinical samples were collected. The samples were initially cultured on Mannitol salt agar and identified based on physical characteristics and biochemical tests, including catalase and coagulase assays. Molecular confirmation of S. aureus isolates was performed using the mecA gene, which is specific to this bacterium. Antibiotic susceptibility testing was conducted following CLSI guidelines to determine resistance patterns against a panel of antibiotics. The presence of efflux pump resistance genes (norA, mgrA, mepA, mepR, and mdeA) was detected using polymerase chain reaction (PCR). Among the 17 antibiotics tested, the highest resistance was observed against ciprofloxacin, penicillin, ampicillin, and cefoxitin. Out of 200 isolates, 177 were resistant, with various efflux pump genes detected. Statistical analysis using ANOVA (Single Factor) revealed significant findings (p = 0.004). The results highlight that efflux pumps play a crucial role in intrinsic resistance mechanisms in S. aureus. Consequently, this bacterium has evolved into a multidrug-resistant pathogen, posing a significant challenge to the effective treatment of infections caused by S. aureus.
Staphylococcus aureus, antibiotic resistance, efflux pump genes, multidrug resistance, polymerase chain reaction, clinical isolates.
Copyright (c) 2023 Aqsa Zafar, Hoor Hamid, Hadiba Saadat, Talha Imtiaz, Khushal Akbar (Author)
Molecular Analysis of Consanguineous Families Having X-Linked Congenital Stationary Night Blindness in Khyber Pakhtunkhwa, Pakistan
X-linked congenital stationary night blindness (XLCSNB) is a vision disorder characterized by reduced visual acuity, myopia (short-sightedness), strabismus, and nystagmus. This condition results from mutations affecting the transmission of visual signals in the retina. Genetic studies have identified the proximal short arm of chromosome X (Xp11.4) as the region harboring the candidate gene associated with XLCSNB. Electroretinogram (ERG), a diagnostic test, plays a critical role in differentiating XLCSNB categories by evaluating the electric responses of retinal, bipolar, and ganglion cells to light and dark stimuli. To locate the candidate gene, four microsatellite markers (DXS574, DXS8012, DXS993, and DXS1201) were designed near the proximity of the target region on Xp11.4. The gene was identified as the NYX gene, encoding nyctalopia, a 481-amino-acid protein, and a member of leucine-rich repeats (LRR). Nyctalopia plays a pivotal role in signal transmission between retinal and bipolar cells. The expression of the NYX gene is low in tissues such as muscles, the brain, the retina, and the testis. Following linkage analysis, specific primers were designed to amplify both exons of the NYX gene. Gel electrophoresis was used to observe DNA bands, and subsequent sequencing of the gene was performed to identify mutations associated with XLCSNB. Three mutations were identified in the NYX gene, all of which had been reported previously. These mutations, predominantly splicing mutations, are known to be pathogenic alterations. The NYX gene encodes 13 leucine-rich repeats (LRRs), and the identified missense mutations primarily affect these LRR sites, leading to impaired retinal signal transmission and disturbed eye function. To date, 42 different mutations in the NYX gene have been reported, with splicing mutations being the most common. This study successfully identified mutations in the NYX gene associated with XLCSNB. Although previously reported, the findings provide further insight into the molecular mechanisms underlying this disorder. Future research should focus on functional analyses of the NYX genes to better understand their role in regulating retinal cell interactions and signaling pathways.
Myopia, Strabismus, Nystagmus, Electroretinography, NYX Gene, Leucine-Rich Repeat Proteins, Retinal Bipolar Cells, Pathogenic Variants
Copyright (c) 2023 Faiza Mazhar, Sajeela Akbar, Sadaf Gohar, Syed Sohail shah, Aizaz Ali, Saqib Ullah (Author)
Epidemiological Analysis of Epilepsy Prevalence in the Pashtun Population: A Comparative Study in Khyber Pakhtunkhwa, Pakistan
Epilepsy is a significant neurological disorder with a high prevalence in low- and middle-income regions, including Khyber Pakhtunkhwa (KP), Pakistan. Despite advancements over the past two decades, limited epidemiological data is available regarding the clinical and demographic features of epilepsy in this region. Socioeconomic barriers and cultural factors further impair the treatment gap. This study aimed to investigate the prevalence and clinical characteristics of epilepsy in the Pashtun population of KP, focusing on factors such as age, gender, parental consanguinity, and family history. A cross-sectional study was conducted at the Department of Neurology, Lady Reading Hospital (LRH), Peshawar. Data was collected through structured interviews and questionnaires from epileptic patients admitted to the hospital. The study analyzed demographic and clinical factors, including disease onset, age, gender, parental consanguinity, and family history. Among the surveyed population, males were found to be more affected by epilepsy compared to females. The highest prevalence was observed in individuals aged 11 to 20 years. Many patients reported parental consanguinity, indicating a possible genetic predisposition. Additionally, a family history of epilepsy was frequently observed, further emphasizing the hereditary nature of the disorder in the study population. The study highlights key epidemiological patterns of epilepsy in KP, with a higher prevalence among males and adolescents. The outcomes underscore the importance of addressing genetic factors and implementing targeted public health interventions, including early diagnosis and community education, to reduce the treatment gap and improve regional patient outcomes.
Epilepsy, prevalence, Khyber Pakhtunkhwa, gender, age groups, parental consanguinity, family history, clinical features, neurological disorder
Copyright (c) 2023 INTERNATIONAL JOURNAL OF APPLIED AND CLINICAL RESEARCH
Systematic Review of Emerging Technologies in Cystic Fibrosis Treatment: Gene Therapy and CRISPR Strategies for the Future
The current systematic review aimed to evaluate research on cystic fibrosis (CF), a genetic disease that affects multiple organs, particularly the lungs, and is associated with high morbidity and mortality. A total of 7831 relevant studies were identified from search databases, and 27 studies were ultimately considered appropriate for review after applying eligibility criteria including three longitudinal studies and the remaining cross-sectional ones. All studies included a healthy control group, with a combined total of 1839 individuals with CF and 2178 controls. The age range varied across studies, but the majority were carried out in adults. The studies had different aims, including evaluating and comparing different techniques for gene therapy and CRISPR, and assessing changes in body nutrition status. Other studies focused on the evaluation of lung function, inflammation, and clinical parameters. Animal models have played a crucial role in advancing CF gene therapy. Various animal models have been developed, including pigs, ferrets, rats, zebrafish, and sheep, each with its advantages and limitations. The CF pig model has facilitated the measurement of CFTR correction in vivo, which enhances the correlation of CFTR expression and transportation of Cl- and HCO3 as a result decreases the implication of cystic fibrosis in gene therapy. Gene editing technologies, such as CRISPR/Cas9, have emerged as promising approaches to modifying nucleic acid sequences in CF research. These tools hold the potential to repair the endogenous CFTR gene and restore its function, but efficient in vivo gene delivery remains a significant challenge. Assessing changes in body composition can provide valuable information on the effects of gene therapy or CRISPR on the overall health of CF patients. The proper evaluation of nutritional composition in the CF treatment is essential, as current therapies such as CFTR modulators primarily target the respiratory system and may not fully address the systemic effects of the disease. The CRISPR techniques and gene therapy treatment have the potential to provide more comprehensive and long-lasting treatments for CF and the assessment of body composition changes is essential for future clinical trials.
Cystic Fibrosis, genetic disease, systematic review, morbidity, mortality, gene therapy, CRISPR/Cas9, CFTR gene, CFTR modulators, systemic effects, clinical trials, nucleic acid modification, endogenous CFTR repair, health assessment, respiratory system.