INTERNATIONAL JOURNAL OF APPLIED AND CLINICAL RESEARCH
The International Journal of Applied and Clinical Research (IJACR) is a peer-reviewed, open-access journal, IJACR is committed to advancing scholarly discourse in the biomedical sciences and biological sciences. It provides a vibrant platform for the dissemination of innovative research across a broad range of Health research. From basic medical sciences to specialized areas in medicine and dentistry, rehabilitation medicine, allied health sciences, clinical psychology, public health, Neurosciences, and Antibiotic resistance, IJACR embraces the entire spectrum of medical knowledge. IJACR caters to a diverse audience, including medical Graduates, Postgraduates, postdoctoral researchers, and Healthcare professionals. The journal features a variety of scholarly works, such as original research articles, narrative and systematic reviews, case reports, short communications, special communications, thought-provoking editorials, and letters to the editor, fostering an exchange of ideas and perspectives. At the core of IJACR is an unwavering commitment to ethical medical journalism and the highest standards of research and publication integrity. It remains dedicated to safeguarding the pursuit of knowledge and maintaining the credibility of scientific inquiry.
Computational Analysis of Wild-Type and Mutant CC2D2A Highlights Differential Binding Affinity
Background: CC2D2A is a key protein implicated in ciliopathies, and mutations may alter its structural stability and ligand-binding behavior. Understanding mutation-induced changes in protein–ligand interactions is essential for elucidating its functional impact. Methods: Molecular docking analysis was performed to evaluate and compare ligand interactions with wild-type and mutant CC2D2A protein structures. Binding affinity, hydrogen bonding interactions, docking scores, and energy parameters were analyzed to assess structural and functional differences. Results: The mutant CC2D2A protein exhibited improved binding affinity (S-score: −5.0838 kcal/mol) compared with the wild type (−4.6461 kcal/mol). The mutant formed four hydrogen bonds involving Asp1576, Arg1109, and Gln1108, whereas the wild-type protein formed only one hydrogen bond with Lys1216. Additionally, the mutant showed more favorable energy parameters, indicating enhanced ligand stability and stronger intermolecular interactions. Conclusion: The mutation in CC2D2A significantly enhances ligand-binding affinity and interaction stability by increasing hydrogen bonding and improving docking energetics. These structural alterations suggest a potential functional impact of the mutation on protein activity.
CC2D2A; molecular docking; protein–ligand interaction; mutation analysis; hydrogen bonding; structural stability
Copyright (c) 2026 Khyber Ali, Saifullah, Shah Fahad Bin Ikram, Saad Ul Halim, Shahmeer Saleem, Warda Batool, Aisha Haq, Shabbir Ullah, Zain Ul Abideen , Sahar Nigar (Author) https://creativecommons.org/licenses/by-nc/4.0
A Novel GNAO1 Missense Variant Associated with Ealy-Onset Epileptic Encephalopathy
Background: GNAO1 encodes the Gαo protein, a critical mediator of GPCR signaling in the nervous system. Mutations in GNAO1 cause early-onset epileptic encephalopathy and movement disorders. This study characterized a novel missense variant, GNAO1 E20K (c.58G>A), using comprehensive in silico approaches. Methods: The variant was identified from gnomAD and analyzed using MutationTaster, BioEdit, and STRING for pathogenicity prediction, sequence alignment, and protein-protein interaction mapping. Molecular docking with N-acetylglucosamine (NAG) compared binding affinities between wild-type and mutant GNAO1. Results: The E20K variant was absent from population databases and affected a highly conserved glutamic acid residue (PhyloP: 9.527; PhastCons: 1.0). GNAO1 showed extreme constraint (LOEUF: 0.10; missense oe: 0.35). MutationTaster classified the variant as deleterious (97% damaging). STRING analysis revealed high-confidence interactions with GNB1 (0.996), RGS16 (0.993), DRD2 (0.950), and ADRA2A (0.956). Comparative docking demonstrated reduced NAG binding affinity in the mutant (−5.5913 kcal/mol) versus wild-type (−5.8479 kcal/mol), with altered hydrogen bonding patterns and a strengthened Lys46 interaction (−5.5 kcal/mol) in the mutant. Conclusions: The GNAO1 E20K variant exhibits multiple lines of in silico evidence supporting pathogenicity, including extreme evolutionary conservation, population rarity, high gene constraint, and altered ligand binding. Functional validation and clinical evaluation are warranted.
GNAO1; Epilepsy; molecular docking; protein-protein interaction; neurodevelopmental disorder; GTPase; pathogenicity prediction; N-acetylglucosamine
Copyright (c) 2026 Iffat Maab (Author) https://creativecommons.org/licenses/by-nc/4.0
Integrative Bioinformatic Analysis of TCF7L2 (Q9NQB0) Polymorphisms Associated with Type 2 Diabetes
Background: Type 2 diabetes mellitus (T2DM) is a multifactorial metabolic disorder influenced by both genetic and environmental factors. Among the susceptibility genes, TCF7L2 is one of the most significant contributors to insulin secretion and glucose metabolism. Objective: This study aimed to investigate the structural and functional impact of TCF7L2 polymorphisms associated with T2DM using an integrated bioinformatics approach. Methods: Disease-associated genes were identified using DisGeNET and MalaCards databases, and overlapping genes were analyzed using Venny 2.1. Functional enrichment analysis was performed using ShinyGO to determine the biological roles of TCF7L2. A missense variant, c.4C>G (p.Pro2Ala), was selected from the gnomAD database for pathogenicity assessment. The deleterious nature of the variant was predicted using SIFT, PolyPhen-2, and MutationTaster. Three-dimensional structures of wild-type and mutant proteins were modeled using AlphaFold. Molecular docking with resveratrol was performed using AutoDock Vina to evaluate binding interactions. Results: Functional enrichment analysis indicated that TCF7L2 is involved in insulin signaling, pancreatic β-cell development, and glucose metabolism pathways. The selected missense mutation was predicted to be deleterious by multiple in silico tools. Molecular docking revealed stable ligand–protein interactions, with altered binding affinity and interaction profiles between wild-type and mutant proteins. Conclusion: The TCF7L2 variant (p.Pro2Ala) may influence protein function and contribute to T2DM pathogenesis. Additionally, resveratrol demonstrated notable binding affinity toward TCF7L2, suggesting its potential therapeutic relevance.
Type 2 diabetes mellitus; TCF7L2 gene; missense mutation; bioinformatics analysis; molecular docking; protein structure modeling; resveratrol; insulin signaling pathway.
Copyright (c) 2026 Sahiba Rohma (Author) https://creativecommons.org/licenses/by-nc/4.0
Structural And Energetic Analysis of the p.Met1lle CFTR Variant
Background: Cystic Fibrosis is a genetic disorder caused by mutations in the CFTR gene, leading to impaired epithelial ion transport and altered protein function. Objective: This study evaluates the structural and functional impact of the p.Met1Ile (c.3G>A) CFTR variant using in-silico approaches. Methodology: Comparative homology modeling and molecular docking were performed using Molecular Operating Environment (MOE). Wild-type and mutant CFTR structures were analyzed for docking scores, conformational stability, and protein–ligand interactions. Pathogenicity prediction tools were also applied to assess functional consequences of the variant. Results: The wild-type CFTR showed higher binding affinity (S = −8.5677 kcal/mol) compared to the mutant (S = −7.8977 kcal/mol). The mutant exhibited altered conformational energy (E_conf = −541.4630 kcal/mol) and disrupted hydrogen bonding patterns, indicating reduced structural stability and ligand interaction. Conclusion: The p.Met1Ile variant likely induces structural destabilization of CFTR, reducing ligand-binding efficiency. These findings support its potential pathogenic role and demonstrate the utility of computational docking in variant analysis.
Cystic Fibrosis; CFTR; p.Met1Ile (c.3G>A); missense mutation; in-silico analysis; molecular docking; homology modeling; protein stability; pathogenicity prediction
Copyright (c) 2026 Zarlash Iftikhar Alam (Author) https://creativecommons.org/licenses/by-nc/4.0
Structural and Functional Analysis of an HBB Variant (p.Leu4Gln) in β-Thalassemia and Its Impact on Heme Binding Using Molecular Docking
Background: β -thalassemia is an inherited hemoglobin disorder caused by mutations in the HBB gene. This study investigated the structural and functional effects of the HBB p.Leu4Gln mutation on β-globin and its interaction with heme. Methodology: The pathogenic variant NC_000011.10:g.5227020A>T (p.Leu4Gln) was identified through UniProt and evaluated using MutationTaster. Wild-type and mutant β-globin structures were modeled with AlphaFold2, and protein–heme docking was performed using MOE. Results: The mutant protein exhibited a less favorable docking score (−9.8716 kcal/mol) than the wild type (−10.6160 kcal/mol) and a higher RMSD value (1.8171 vs. 1.2387 Å), indicating reduced binding affinity and stability. Interaction analysis revealed the formation of a metal coordination bond and a novel π-H interaction with His64 in the mutant complex, suggesting alteration of the heme-binding pocket. Conclusion: The p.Leu4Gln mutation disrupts heme binding and alters the structural integrity of β-globin, potentially contributing to impaired hemoglobin function in β-thalassemia. These findings provide molecular insights into disease pathogenesis and potential therapeutic targets.
β-thalassemia; HBB; β-globin; p.Leu4Gln; AlphaFold2; Molecular Docking; Heme Binding; Protein Structure; Mutation Analysis; Hemoglobin Function.
Copyright (c) 2026 Fasiha Bibi, Samrina Muskan, Muhammad Afnan (Author) https://creativecommons.org/licenses/by-nc/4.0
Structural and Functional Characterization of the Rare GNAT1 Variant p.Gly4Trp Reveals Impaired GDP Binding and Potential Pathogenic Association with Congenital Stationary Night Blindness
Background: Congenital stationary night blindness (CSNB) is a genetically heterogeneous retinal disorder associated with defects in rod phototransduction signaling. GNAT1 encodes the α-subunit of rod transducin, a G-protein essential for visual signal transduction in photoreceptor cells. In this study, we investigated the structural and functional consequences of the rare GNAT1 missense variant rs1699413898 (p.Gly4Trp), currently classified as a variant of uncertain significance. Methodology: The variant was retrieved from the gnomAD database with an extremely low allele frequency (0.000002056). Computational pathogenicity prediction tools, including PolyPhen-2, SIFT, and MutationTaster, consistently classified the variant as deleterious. Structural analysis was performed using the wild-type GNAT1 crystal structure (PDB ID: 1TAD), while the mutant model was generated using AlphaFold3. Molecular docking analysis of GDP with both wild-type and mutant proteins was performed to evaluate binding affinity and interaction changes. Results: Molecular docking analysis of GDP with both wild-type and mutant proteins revealed reduced binding affinity in the mutant structure (-9.1336 kcal/mol) compared with the wild type (-9.4581 kcal/mol). The mutant protein exhibited loss of critical interactions with ASP268, and P-loop residues involved in nucleotide stabilization, along with a reduced hydrogen-bonding network and altered binding geometry. Novel interactions with THR177 and GLN200, a residue previously associated with CSNB, were also observed. Conclusion: Collectively, these findings suggest that the p.Gly4Trp substitution may destabilize GDP binding and impair GNAT1 function, supporting its potential pathogenic role in recessive CSNB.
CSNB; GNAT1; missense variant; molecular docking; rod transducin; in silico analysis; GDP binding; retinal disorder.
Copyright (c) 2026 Fiza (Author) https://creativecommons.org/licenses/by-nc/4.0
In-silico Identification and Structural Characterization of a Missense Variant (p.Gly380Arg) in the FGFR3 Gene Associated with Achondroplasia
Background: Achondroplasia is the most common form of short-limbed dwarfism caused mainly by mutations in the FGFR3 gene. Methods: An integrative bioinformatics approach was applied to analyze the FGFR3 c.1138G>A (p.Gly380Arg) variant using gene-disease association analysis, population frequency assessment, pathogenicity prediction, protein interaction mapping, structural modeling, and molecular docking. Results: The variant showed an extremely low global frequency in gnomAD and was predicted to be pathogenic by MutationTaster, PolyPhen-2, and CADD. Protein interaction analysis revealed strong associations of FGFR3 with skeletal development and fibroblast growth factor signaling pathways. Molecular docking demonstrated reduced binding affinity in the mutant protein compared to the wild type, indicating altered structural interactions caused by the p.Gly380Arg substitution. Conclusion: The study highlights the pathogenic significance of the FGFR3 p.Gly380Arg variant and provides computational insights into its role in the molecular pathogenesis of achondroplasia.
Achondroplasia, FGFR3, p.Gly380Arg, Endochondral ossification, In silico analysis, Molecular docking
Copyright (c) 2026 Sajjal Durrani (Author) https://creativecommons.org/licenses/by-nc/4.0
In-silico Identification and Structural Characterization of a Novel Missense Variant (p.Ala2Thr) in the MCPH1 Associated with Autosomal Recessive Primary Microcephaly
Autosomal recessive primary microcephaly is a genetically diverse neurodevelopmental disorder marked by reduced brain size. This study used an integrative bioinformatics approach to identify pathogenic variants in the MCPH1 gene. Data from GeneCards and DisGeNET highlighted MCPH1 as a key candidate. A missense variant (p.Ala2Thr) was identified using gnomAD and validated via UniProt. Functional analysis showed MCPH1 involvement in cell cycle regulation and DNA damage response. In silico tools (PolyPhen-2 and SIFT) predicted the variant as deleterious. Structural and docking analyses suggested that the mutation may alter protein stability and interaction dynamics, potentially affecting its function.
Primary microcephaly, MCPH1, Autosomal recessive, Missense mutation, Neurodevelopment, Consanguinity, In silico analysis
Copyright (c) 2026 Faryal Arbab (Author) https://creativecommons.org/licenses/by-nc/4.0
Impact of Sociocultural Norms on Career Progression of Female Dentists: A Cross-Sectional Study from Khyber Pakhtunkhwa, Pakistan
Background: Female dentists face multiple challenges that may hinder their career progression, including sociocultural norms, family responsibilities, and workplace barriers. This study aimed to explore the perceptions of female dentists in Khyber Pakhtunkhwa, Pakistan, regarding factors influencing their professional advancement. Methodology: A cross-sectional survey was conducted among 213 female dentists working in both public and private dental sectors across Hayatabad, Peshawar City, Abbottabad, and Swat. A structured, validated questionnaire assessed sociodemographic characteristics, professional practice, and perceived barriers categorized into family, interpersonal, environmental, and sociocultural domains. Data were analyzed using descriptive statistics and logistic regression to examine associations between these barriers and promotion status. Results: Most participants were aged 30–39 years (53.5%), married (76.1%), and specialists (65.7%), with the majority practicing full-time (93%). Family-related challenges were the most reported barriers, with 47.9% strongly agreeing that family responsibilities and 49.2% that balancing work and family affected their careers; 59.2% identified childcare responsibilities as a significant challenge. Interpersonal barriers included male dominance (26.8%) and gender discrimination (20.2%), while environmental challenges were most notable in safe transportation (41.3%) and driving restrictions (39.9%). Sociocultural barriers were also prominent, particularly societal beliefs assigning primary child-rearing responsibilities to women (73.2% strongly agree/agree) and the lack of family-friendly policies (58.2%). Logistic regression indicated that family and sociocultural challenges were significantly associated with lower promotion rates. Conclusions: Female dentists in Khyber Pakhtunkhwa face multifaceted barriers to career progression, driven largely by family, sociocultural, and environmental factors. Addressing these challenges through mentorship, flexible work policies, and equitable promotion practices may enhance professional advancement and leadership representation for women in dentistry.
female dentists, career progression, sociocultural barriers, family responsibil-ities, Khyber Pakhtunkhwa, professional advancement.
Copyright (c) 2026 Gul Afshan Syed, Zarghona Fazal Sher, Sumaya Batool, Khushboo Rashid, Muhammad Khalid (Author) https://creativecommons.org/licenses/by-nc/4.0
Comparative Study of Cardiovascular Risk and Metabolic Syndrome in male and female patients with Rheumatoid Arthritis
Rheumatoid arthritis (RA) is associated with an increased burden of cardiovascular disease (CVD) and metabolic syndrome (MetS), though sex-specific patterns remain underexplored. This cross-sectional study aimed to compare the prevalence of MetS and estimate the 10-year cardiovascular risk between male and female RA patients. A total of 300 RA patients (150 males, 150 females) were evaluated using harmonized MetS criteria and the SCORE-2 algorithm. Results showed that MetS was significantly more prevalent in females (56.0% vs. 36.7%, p < 0.001), characterized by higher rates of central obesity and low HDL-cholesterol. In contrast, males exhibited higher 10-year cardiovascular risk (7.4% vs. 5.1%, p < 0.001). However, the presence of MetS disproportionately elevated cardiovascular risk in females, nearly equalizing risk between sexes in the MetS subgroup. Multivariate analysis confirmed MetS as a stronger predictor of high cardiovascular risk in women. These findings highlight significant sex-based disparities in metabolic and cardiovascular profiles in RA, underscoring the need for sex-stratified risk assessment and targeted management strategies to mitigate cardiovascular morbidity.
Copyright (c) 2026 Hamad Ali, Abdullah, Zain Ul Abideen, Aizaz Ul Hassan, Muhammad Sajid, Abdur Razaq, Muhammad Afaq (Author) https://creativecommons.org/licenses/by-nc/4.0
Late-Onset Alzheimer’s Disease in an Elderly Woman
Alzheimer’s disease is a progressive neurological condition and is the most prevalent cause of dementia in an elderly population. Some of the common symptoms include gradual cognitive decline, memory loss, and behavioral abnormalities that interfere with day-to-day functioning. We report the case of an 88-year-old woman from Peshawar who has had gradual memory loss, persistent questioning, and poor location and object identification for eight years. Along with this disease she was suffering from diabetes mellitus and osteoarthritis. The patient also had a positive family history of Alzheimer disease which may indicate neurocognitive issues. The clinical manifestation of late-onset Alzheimer’s disease, the significance of family history, and the difficulties in treating dementia in older individuals with several comorbidities are all highlighted in this case.
Late-onset Alzheimer disease, Dementia, cognitive decline, Family history, elderly female
Copyright (c) 2026 Aqsa Ali, Syed Muzammil Hussain Shah, Rabia Anum, Hafsa Urooj (Author) https://creativecommons.org/licenses/by-nc/4.0
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/4.0
A Case Report on Huntington’s Disease Presenting with Progressive Motor and Cognitive Decline
An uncommon, hereditary, autosomal dominant neurological condition, Huntington’s disease (HD) is marked by increasing motor dysfunction, cognitive loss, and mental symptoms (Stoker et al., 2022). A trinucleotide (CAG) repeat expansion in the HTT gene on chromosome 4 is the cause of the illness (Kumar et al., 2020). Clinical onset usually happens in mid-adulthood, and symptoms steadily get worse over time, eventually resulting in death and severe disability (Ogilvie et al., 2023). One important diagnostic characteristic is a rich family history spanning several generations (Kringlen et al., 2017). Genetic counseling, family screening, and proper symptom treatment all depend on early detection of HD.
Huntington’s disease, Chorea, Neurodegenerative disease, Family history, Case report
Copyright (c) 2026 Syed Muzammil Hussain Shah, Aqsa Ali, Hafsa Urooj, Rabia Anum, Majid Khan, Zainab Liaqat (Author) https://creativecommons.org/licenses/by-nc/4.0
Early-Onset Epilepsy with Pontocerebellar Hypoplasia in a Consanguineous Pakistani Family
Epilepsy is a prevalent neurological condition marked by frequent, spontaneous seizures and symptoms which affects individuals of all ages worldwide (Leenen et al., 2016). Early-onset epilepsy frequently indicates an underlying genetic aetiology with autosomal recessive inheritance, especially when consanguinity and a favorable family history is present (Balestrini et al., 2021). According to estimates, there are 9.99 cases of epilepsy for every 1000 people in Pakistan. It is more common among younger people (those under 30) (Deng et al., 2014). The diagnosis of severe neurodevelopmental epileptic disorders is further supported by neuroimaging findings such as anomalies in the brainstem and cerebellum (Adamczyk et al., 2021).
Epilepsy, Pontocerebellar hypoplasia, Consanguinity, Autosomal recessive inheritance, Myoclonic seizures, MRI
Copyright (c) 2026 Syed Muzammil Hussain Shah, Aqsa Ali, Hafsa Urooj, Rabia Anum (Author) https://creativecommons.org/licenses/by-nc/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/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/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/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/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/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 (Author) https://creativecommons.org/licenses/by-nc/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/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/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) https://creativecommons.org/licenses/by-nc/4.0
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) https://creativecommons.org/licenses/by-nc/4.0
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) https://creativecommons.org/licenses/by-nc/4.0
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) https://creativecommons.org/licenses/by-nc/4.0
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) https://creativecommons.org/licenses/by-nc/4.0
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) https://creativecommons.org/licenses/by-nc/4.0
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) https://creativecommons.org/licenses/by-nc/4.0
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) https://creativecommons.org/licenses/by-nc/4.0