Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 Intranasal Formulations for Organophosphorus Nerve Agent Poisoning: A Review of Recent Developments in Treatment and Prophylaxis 1784 1796 236313 10.30491/jabr.2025.549112.1921 EN Mohammad Hadi Baghersad Applied Biotechnology Research Center, New Health Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 0000-0002-1940-5388 Journal Article 2025 09 24 Organophosphorus (OP) nerve agents represent a grave global threat, encompassing both chemical warfare agents and highly toxic pesticides. The high morbidity and mortality associated with OP exposure, primarily due to the irreversible inhibition of acetylcholinesterase (AChE), underscore the critical need for rapid and effective medical countermeasures. Current standard-of-care, which relies on intramuscular or intravenous administration of anticholinergics (atropine) and cholinesterase reactivators (oximes), is limited by logistical challenges and, crucially, an inability to provide adequate protection to the central nervous system (CNS). This review examines the burgeoning field of intranasal (IN) formulations as a novel paradigm for OP antidote delivery. The IN route offers a direct nose-to-brain pathway that bypasses the restrictive blood-brain barrier (BBB), enabling CNS-targeted therapy. Preclinical research has demonstrated that IN obidoxime can completely prevent mortality and seizure-induced neuronal degeneration in animal models, an effect not observed with standard peripheral treatment. The clinical success of intranasal diazepam for seizure control provides a powerful precedent for this approach. Moreover, IN delivery holds promise for the rapid administration of prophylactic agents and next-generation bioscavengers. While the absence of human clinical trials for IN nerve agent antidotes remains a significant research gap, the well-documented advantages and efficacy of intranasal naloxone in a similar emergency setting provide a strong rationale for continued development. This report synthesizes recent findings, analyzes the comparative advantages of IN formulations, and identifies key research directions to translate these promising countermeasures into clinical practice.

https://www.biotechrep.ir/article_236313_b2d64e779b978ab650f5ac6cc997f48f.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 The Roles of miRNAs in Human Lung Cancer 1797 1803 236314 10.30491/jabr.2025.473677.1775 EN Ali Nemani Khiavi Molecular Biology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 0009-0005-3743-1124 Mahdi Fasihi-Ramandi Molecular Biology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 0000-0002-8242-4880 Journal Article 2024 09 23 Lung cancer is the most common type of cancer in the world, and smoking is the most important cause of lung cancer. Notably, small cell lung cancer accounts for about 15% of all lung cancers, and non-small cell lung cancer accounts for approximately 85% of all new lung cancer diagnoses. Most patients are diagnosed with the disease in advanced stages due to insufficient screening programs and the late onset of clinical symptoms. As a result, patients have a poor prognosis. MicroRNAs are a family of small non-coding RNAs (21-25 nucleotides) that can inhibit mRNA translation and promote mRNA degradation by base-pairing to complementary sites on the target mRNA. Through this mechanism, miRNAs alter gene expression post-transcriptionally. The first non-coding RNA, lin-4, was identified as a miRNA in Caenorhabditis elegans in 1993. The specific characteristics of miRNAs, including their tissue and even cellular specificity, stability in various biological fluids, and their dysregulation during tumorigenesis, make miRNAs potential biomarkers and therapeutic targets in cancers that should be considered. Given the high toxicity of chemotherapy drugs in human lung cancer, miRNAs could be a more suitable option with lower toxicity. However, more testing and research are needed to improve therapeutic performance and reduce adverse effects on healthy cells so that they can replace chemotherapy drugs with harmful effects on the body. This review aims to investigate the role of microRNA types in the development or prevention of human lung cancer.

https://www.biotechrep.ir/article_236314_c27983465568080ed4c8cc08bfff8ff3.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 Photoprotection of Chlorella sp. and Tetradesmus dimorphus Extracts on Ultraviolet-Irradiated Human Retinal Pigment Epithelial Cells 1804 1814 236315 10.30491/jabr.2025.495987.1824 EN Yi Teng Tang Division of Applied Biomedical Science and Biotechnology, School of Health Sciences, IMU University, 57000 Kuala Lumpur, Malaysia Yun Zhi Chin Division of Applied Biomedical Science and Biotechnology, School of Health Sciences, IMU University, 57000 Kuala Lumpur, Malaysia Chun Yik Lew Division of Applied Biomedical Science and Biotechnology, School of Health Sciences, IMU University, 57000 Kuala Lumpur, Malaysia Farhan Aisar Mes Ewan Division of Applied Biomedical Science and Biotechnology, School of Health Sciences, IMU University, 57000 Kuala Lumpur, Malaysia Kiat Fatt Chia Centre for Postgraduate Studies by Research, IMU University, 57000 Kuala Lumpur, Malaysia Rhun Yian Koh Division of Applied Biomedical Science and Biotechnology, School of Health Sciences, IMU University, 57000 Kuala Lumpur, Malaysia Chiew Yen Wong Division of Applied Biomedical Science and Biotechnology, School of Health Sciences, IMU University, 57000 Kuala Lumpur, Malaysia Centre for Environmental and Population Health, Institute for Research, Development and Innovation (IRDI), IMU University, 57000 Kuala Lumpur, Malaysia 0000-0003-0534-6206 Chun-Wai Mai Centre for Cancer and Stem Cell Research, Institute for Research, Development and Innovation (IRDI), IMU University, 57000 Kuala Lumpur, Malaysia Chooi Ling Lim Division of Applied Biomedical Science and Biotechnology, School of Health Sciences, IMU University, 57000 Kuala Lumpur, Malaysia 0000-0002-2926-892X Journal Article 2024 12 26 Introduction: Ultraviolet radiation induces retinal cell injury associated with age-related macular degeneration (AMD). Bioactive compounds from microalgae have demonstrated photoprotective properties against ultraviolet B (UVB)-induced injury, however, their protective activity on the retinal pigment epithelium (RPE) remains unexplored. In this study, the photoprotective effects of Chlorella sp. and Tetradesmus dimorphus extracts on UVB-irradiated ARPE-19 cells were investigated. Materials and Methods: ARPE-19 cells were irradiated with UVB, followed by treatment with Chlorella sp. and T. dimorphus extracts from distilled water, absolute methanol, 20% aqueous methanol (v/v), or 95% ethanol. Viability of ARPE-19 cells was assessed using 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), while oxidative stress was evaluated with diacetyldichlorofluorescein (DCFHDA). The presence of acidic vesicular organelles (AVOs) and apoptotic cells was detected through acridine orange and Hoechst 33258 staining. Results: Aqueous-methanol (20%) extracts of Chlorella sp. (1 μg/ml) improved ARPE-19 cell viability by 23.26%, while other solvent extracts of T. dimorphus did not show significant improvement compared to the control group. Besides, aqueous extracts of Chlorella sp. (1 μg/ml) showed the greatest reduction in reactive oxygen species (ROS) level by 0.46-fold, while aqueous-methanol (20%) extracts of T. dimorphus (10 μg/ml) decreased the ROS level by 0.33-fold. Moreover, treatment with aqueous-methanol (20%) extracts of Chlorella sp. and T. dimorphus reduced autophagy by 38.52% and 36.26%, respectively. Apoptotic activity was attenuated by aqueous extracts of Chlorella sp. (37.33%) and T. dimorphus (46%). Conclusions: Chlorella sp. and T. dimorphus extracts exert photoprotective effects on UVB-irradiated ARPE-19 cells, highlighting their potential as a novel approach for AMD treatment.

https://www.biotechrep.ir/article_236315_ad1b6b1c378956c8292217d2e7bc1f7e.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 Facilitating Wound Healing by Inducing M2 Macrophage Polarization Using Polymeric Nanoparticles; an Experimental and Molecular Study 1815 1827 236316 10.30491/jabr.2025.558418.1941 EN Hamid Kooshki Nanobiotechnology Research Center, New Health Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 0000-0002-8401-9918 Mehrdad Rezaeian Cancer Epidemiology Research Center, AJA University of Medical Sciences, Tehran, Iran 0000-0003-2985-8396 Reza Heidari Cancer Epidemiology Research Center, AJA University of Medical Sciences, Tehran, Iran Infectious Diseases Research Center, AJA University of Medical Sciences, Tehran, Iran Medical Biotechnology Research Center, AJA University of Medical Sciences, Tehran, Iran 0000-0001-6862-6972 Mozafar Mohammadi Applied Biotechnology Research Center, New Health Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 0000-0003-3581-658X Ali Shakerimoghaddam Noncommunicable Diseases Research Center, Neyshabur University of Medical Sciences, Neyshabur, Iran 0000-0002-8401-9918 Sakineh Hajebi Biomaterial and Medicinal Chemistry Research Center, AJA University of Medical Sciences, Tehran, Iran 0000-0002-4005-3241 Zahar Hami Toxicology Research Center, AJA University of Medical Sciences, Tehran, Iran 0000-0001-6179-2812 Ali Saberian Faculty of Agriculture, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran 0000-0002-8401-9918 Mostafa Shahrezayee Trauma Research Center, AJA University of Medical Sciences, Tehran, Iran 0000-0002-8401-9918 Mohsen Chamanara Toxicology Research Center, AJA University of Medical Sciences, Tehran, Iran 0000-0002-6201-2918 Journal Article 2025 11 09 Introduction: Targeting macrophages has garnered significant research interest due to their pivotal role in inflammation, cancer, and wound healing. This study investigates the impact of nanoparticle design on macrophage interactions and its subsequent influence on wound healing. Materials and Methods: Four nanoparticles (hyaluronic acid, chitosan, dextran, and methacrylic acid) were selected based on their biocompatibility and targeting potential. Molecular dynamics simulations revealed that hyaluronic acid exhibited the most favorable interactions with the macrophage membrane, positioning it as a promising candidate. To validate these findings, an in vivo experiment was conducted using hyaluronic acid nanoparticles on wounded rabbits. Results: The results showed that the factors involved in the stability of hyaluronic acid nanoparticles, including contact area average (40 nm2), Gyration Radius analysis (4.5 nm) and time dependent Rg (5.8 nm) were at their lowest level compared to another nanoparticle. Also the lowest entropy (700 kj/mol.k), minimum distance(1.1 nm) and energy average(-2000 kj/mol) showed the appropriate stability of hyaluronic acid nanoparticles. The results demonstrated that topical administration of hyaluronic acid nanoparticles significantly accelerated wound healing. The formulation had an average hydrodynamic diameter of approximately 104 ± 25 nm, a zeta potential of –18 ± 2 mV, and a polydispersity index (PDI) of 0.306. HA nanoparticles significantly reduced wound size compared to the control group on days 7, 14, and 21 (p < 0.01). Conclusions: This study utilized molecular dynamics simulations (MDS) to explore the interaction between four nanoparticles (hyaluronic acid, chitosan, dextran, and methacrylic acid) and the macrophage membrane, focusing on their potential to stimulate macrophages and influence wound healing.

https://www.biotechrep.ir/article_236316_cdfea0d249d7cf72ca4109e24d49ac82.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 The Storage Stability of Total DNA in Dried Blood Samples under Different Temperatures and Sunlight Exposure 1828 1839 236321 10.30491/jabr.2025.531425.1886 EN Jeanne V. Samsonova Faculty of Chemistry, Lomonosov Moscow State University, Moscow, Russia Vavilov Institute of General Genetics, Moscow, Russia 0000-0002-0670-0951 Nikolay Yu. Saushkin Faculty of Chemistry, Lomonosov Moscow State University, Moscow, Russia 0000-0002-4565-2075 Farida R. Gurgenova Vavilov Institute of General Genetics, Moscow, Russia 0009-0001-0481-906X Valery N. Voronkova Vavilov Institute of General Genetics, Moscow, Russia 0000-0002-9660-0179 Yuri A. Stolpovsky Vavilov Institute of General Genetics, Moscow, Russia 0000-0003-2537-1900 Aleksei K. Piskunov Vavilov Institute of General Genetics, Moscow, Russia 0000-0001-6439-7121 Journal Article 2025 06 30 Introduction: The collection of dried blood may be linked to certain problems during storage and transportation. The stability of DNA on supports other than traditional cellulose is not well known. This study aimed to directly compare natural (cellulose) and non-natural (glass fiber) membrane supports for short-term and long-term storage of dried blood samples under various temperature and sunlight conditions. Materials and Methods: Dried blood samples on two membrane supports (cellulose and glass fiber) were examined for both short-term and long-term storage stability across temperatures from –78 °C to +60 °C. The total DNA yield was measured, and the quality of extracted DNA was verified by PCR of mtDNA D-loop. The color change of dried blood was evaluated using RGB analysis. Results: 53-68% of total DNA was lost in frozen whole blood or dried blood stored at -20 °C and -78 °C for one year. For dried samples stored at room temperature on a shelf exposed to sunlight, the greatest total DNA degradation occurred. RGB profile analysis of dried blood showed an increase in the green component over one year. A brownish or greenish color indicates significant DNA loss. After 1.5 years, PCR for the mtDNA D-loop was successful for all samples stored at +4 °C or lower. Conclusions: The glass fiber membrane showed advantages over cellulose regarding DNA yield and storage stability. Sunlight and high temperatures are key factors to consider when collecting and transporting dried blood samples. The blood's color can serve as an indicator of possible DNA yield: the darker the blood, the lower the DNA yield.

https://www.biotechrep.ir/article_236321_486856f02b18d2305503aa43095e9e56.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 Probiotic Niche Specificity and Microbiome Analysis of Different Non-Dairy based Traditional Food Samples Using Metagenomic Approach 1840 1851 236322 10.30491/jabr.2025.494841.1821 EN Bijayanta Sircar Laboratory of Microbiology and Experimental Medicine, Department of Zoology, University of Gour Banga, Malda-732103, West Bengal, India 0000-0002-1794-0635 Manisha Mandal Department of Physiology, MGM Medical College and LSK Hospital, Kishanganj, Bihar-855107, West Bengal, India 0000-0002-9562-5534 Shyamapada Mandal Laboratory of Microbiology and Experimental Medicine, Department of Zoology, University of Gour Banga, Malda-732103, West Bengal, India 0000-0002-9488-3523 Journal Article 2024 12 19 Introduction: People consume different types of traditional dairy and non-dairy-based fermented foods worldwide. The literature review suggests that there are scientifically unexplored diverse non-dairy-based fermented foods. The present study was conducted to determine the taxonomic and metabolic status of beneficial bacteria in various non-dairy-based traditional food samples using a metagenomics approach. Materials and Methods: Metagenomic analysis of food samples may reveal the presence of many beneficial culturable or non-culturable bacteria through Next Generation Sequencing techniques. In this study, we used six non-dairy-based food products, including two types of idli batter, vegetable fermented mixture, two types of vegetable pickles, and the Indian traditional food Shinni. Phylogenetic relationships, taxonomic plot analysis, presence of enriched genera, core microbiome comparative analysis, species richness in terms of α and β diversity, and functional profiling of the microbiome were also determined in this research work. Results: All the samples showed a good abundance of probiotic bacterial presence. Specifically, Pediococcus, Weissella, Lactococcus, Acetobacter, and Lactobacillus were statistically more viable genera than others. Functional profiling of the six metagenome samples displayed the top nine KEGG Orthology metabolism and the top fourteen clusters of orthologous genes metabolism. Conclusions: Variation in the richness and diversity of bacteria in all six samples, along with variations in metabolic functions, indicates the need for further investigation to explore the nutritional value and bioactivity of beneficial bacteria in our region.

https://www.biotechrep.ir/article_236322_26bf6a73973800135f725e9211cce560.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 Combining Machine Learning Algorithms with Meta-Analysis and WGCNA to Identify Biomarker-Responsive Genes to Environmental Stresses in Thermus thermophilus HB8 1852 1864 236326 10.30491/jabr.2025.490923.1811 EN Abbas Karimi-Fard Department of Cell and Molecular Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran 0000-0002-6496-5018 Abbas Saidi Department of Cell and Molecular Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran Masoud Tohidfar Department of Cell and Molecular Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran 0000-0002-0175-7406 Seyede Noushin Emami Department of Molecular Biosciences, Wenner-Gren Institute, Stockholm University, SE 106 91 Stockholm, Sweden Journal Article 2024 11 26 Introduction: Thermus thermophilus is a thermophilic bacterium known for its resilience in extreme environments. Investigating its transcriptomic responses to environmental stresses can uncover critical adaptive mechanisms. Materials and Methods: This study analyzed transcriptomic data from 10 microarray datasets, including 63 samples (36 stress-exposed and 27 controls). Stress conditions included copper, cold, zinc, iron, heat, salt, H2O2, tetracycline, diamide, and alkylation. Differentially expressed genes (DEGs) were identified through meta-analysis, followed by Gene Ontology (GO) enrichment analysis. Weighted gene co-expression network analysis (WGCNA) was employed to detect stress-associated gene modules. Machine learning approaches—decision tree, logistic regression, random forest, adaptive boosting, SVM-RFE, and XGBoost—were used to prioritize key genes. Results: Meta-analysis revealed 54 upregulated and 196 downregulated genes under stress. GO analysis highlighted significant enrichment in ion transport, localization processes, and transmembrane transporter activity. WGCNA identified two stress-related modules, cyan and lightcyan. SVM-RFE and XGBoost outperformed other machine learning models with superior accuracy, precision, recall, and F1-scores. TTHA0798 emerged as a hub gene consistently identified across machine learning and DEG/WGCNA analyses. Conclusions: This study provides a comprehensive analysis of the stress responses of T. thermophilus, identifying TTHA0798 as a key hub gene. The integration of transcriptomic data, co-expression analysis, and machine learning offers valuable insights into the adaptive mechanisms of this extremophile, paving the way for further functional studies.

https://www.biotechrep.ir/article_236326_afb04170eae032421873dc20a3642326.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 Effect of Long-Term Exposure to Shallow Well Water on Semen Parameters and mRNA Levels of Spermatozoa Tail Formation-Responsible Genes in Rams 1865 1874 236327 10.30491/jabr.2025.548170.1914 EN Alaa Kamil Abdulla Department of Medical Biotechnology, Faculty of Biotechnology, Al-Qadisiyah University, Al-Diwaneyah, Iraq 0000-0001-8315-0116 Omar Hussein Harran Department of Agricultural Biotechnology, Faculty of Biotechnology, Al-Qadisiyah University, Al-Diwaneyah, Iraq 0000-0002-3185-4557 Qasim Zamel Bneed Department of Medical Biotechnology, Faculty of Biotechnology, Al-Qadisiyah University, Al-Diwaneyah, Iraq 0009-0005-6606-5848 Journal Article 2025 09 20 Introduction: Exposure to heavy metals is linked to impaired animal fertility through disruptions in gene expression in spermatozoa. However, the mechanisms behind these effects are not fully understood. This study aimed to assess the impact of long-term consumption of shallow well water contaminated with heavy metals on semen quality and mRNA expression of genes related to sperm tail formation in rams. Materials and Methods: Eighty sexually mature rams were enrolled, including 40 from four locations in Al-Diwaniyah governorate (Al-Shamiya, Al-Dughara, Al-Hamza, and Al-Shanafiya), which had been drinking shallow well water since birth (experimental group), and 40 drinking municipal tap water (control group). Water samples were analyzed for pH, total dissolved solids, salinity, turbidity, and heavy metal content (cadmium, lead, mercury). Semen was collected using an artificial vagina and evaluated by computer-assisted sperm analysis (CASA). Total RNA was extracted from spermatozoa, and the expression of CATSPER, AKAP4, and SPAG6 genes was quantified using one-step real-time PCR with SYBR Green. Results: Shallow well water had significantly higher turbidity, salinity, dissolved solids, and heavy metal concentrations compared to tap water (p ≤ 0.05). Rams drinking well water showed a significantly lower percentage of motile sperm (p < 0.001). Additionally, the expression levels of CATSPER, AKAP4, and SPAG6 genes were significantly reduced in the well water group compared to the control group (p < 0.001). Specifically, CATSPER expression decreased by 85.5%, AKAP4 by 82.8%, and SPAG6 by 87.4% in the well water group relative to controls. Conclusions: Chronic exposure to contaminated shallow well water markedly impairs sperm motility and downregulates key genes involved in tail formation in rams, emphasizing the reproductive risks of heavy metal pollution.

https://www.biotechrep.ir/article_236327_04ab9b8ad785bbea194b7a0ac3967d43.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 The Potential of Serum CTHRC1 Expression Changes for Early Breast Cancer Detection: A Clinical Investigation 1875 1882 236328 10.30491/jabr.2025.483014.1792 EN Mahsa Mombeyni Biology Department, Faculty of Sciences, University of Gonbad Kavous, Gonbad Kavous, Iran Mohammad-Reza Mahmoudian-Sani Thalassemia and Hemoglobinopathy Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Hossein Sabour Department of Plant Production, College of Agriculture Science and Natural Resources, University of Gonbad Kavous, Gonbad Kavous, Iran Hossein Karimpourian Thalassemia and Hemoglobinopathy Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Eisa Jorjani Biology Department, Faculty of Sciences, University of Gonbad Kavous, Gonbad Kavous, Iran Journal Article 2024 10 12 Introduction: This study aimed to evaluate the expression of Collagen Triple Helix Repeat Containing-1 Protein (CTHRC1) in the serum of breast cancer patients, addressing the need for non-invasive biomarkers and assessing its diagnostic potential for early detection of breast cancer. Materials and Methods: This case-control study involved 24 newly diagnosed breast cancer patients and 24 healthy controls. RNA was extracted from serum and converted into complementary DNA (cDNA), and the expression levels of CTHRC1 were measured. The significance of differences in gene expression was analyzed using GenEx software. Results: Most patients exhibited metastasis to the axillary lymph nodes and were classified as stage I. There was no statistically significant difference in CTHRC1 expression between the breast cancer patients and the control group. The Area Under the Receiver Operating Characteristic Curve (AUC) for serum CTHRC1 levels was 0.601. The sensitivity and specificity of CTHRC1 were 54.16% and 55.14 %, respectively. Conclusions: While the findings of this study did not align with those from studies on breast cancer tissue, it is anticipated that the sensitivity, specificity, and AUC of the ROC will improve with an increased sample size. The majority of our patients had axillary metastasis and were in stage I. Previous studies suggest that CTHRC1 expression is elevated in advanced stages of metastasis, both in serum and tissue. To establish the diagnostic value of CTHRC1, larger-scale clinical studies with more participants are warranted.

https://www.biotechrep.ir/article_236328_64ad845758d7f8f572b12800f60842ba.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 Applying the Alkaline Lysis Method for rProtein A Extraction from E. coli Bacteria: A Simple Approach for Large Protein Sample Extraction 1883 1886 236329 10.30491/jabr.2025.555733.1935 EN Hamid Kooshki Nanobiotechnology Research Center, New Health Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 0000-0002-8401-9918 Mozafar Mohammadi Applied Biotechnology Research Center, New Health Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 0000-0003-3581-658X Mehdi Ebadi Applied Biotechnology Research Center, New Health Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran Mohammad Sadegh Hashemzadeh Nanobiotechnology Research Center, New Health Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 0000-0003-3524-1798 Journal Article 2025 10 27 Introduction: The increasing demand for monoclonal antibodies in medicine requires scalable and efficient production methods. Because of its strong affinity for immunoglobulins, Staphylococcal protein A is a key tool for antibody purification. Conventional methods of extracting recombinant protein A (rProtein A), such as sonication, are costly, inefficient at high volumes, and difficult to scale up. Due to the inherent stability of rProtein A in alkaline pH, this study used NaOH to extract rProtein A from E. coli. Materials and Methods: The protein was expressed in E. coli BL21 (DE3), and then cell disruption was achieved through sonication and alkaline lysis. Ni-NTA affinity chromatography was employed to purify the isolated proteins. An ELISA-based assay evaluated the functionality and binding affinity of the purified rProtein A by determining the equilibrium dissociation constant (Kd). Results: Alkaline lysis proved more effective, releasing nearly 98% of rProtein A from the cell pellet compared to 40% with sonication. Additionally, the functionality of rProtein A remained intact, exhibiting excellent antibody-binding affinity with a Kd of 6.95 nM, slightly better than the 7.04 nM Kd of the sonication-produced protein. Conclusions: Overall, alkaline lysis is a reliable, cost-efficient, and scalable alternative to sonication for the primary recovery of rProtein A from bacterial cells.

https://www.biotechrep.ir/article_236329_05c4b001db9f7e35fad05656b209f933.pdf

Baqiyatallah University of Medical Sciences Journal of Applied Biotechnology Reports 2322-1186 12 4 2025 12 01 Statistical Optimization of CYP124 Expression: A New Benchmark for Mycobacterial Cytochrome Engineering 1887 1888 236332 10.30491/jabr.2025.554511.1932 EN Piruz Shadbash Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran 0009-0008-1328-482X Marzieh Bahari Babadi Department of Biochemistry, Medical School, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 0009-0002-9280-389X Journal Article 2025 10 20 The study by Gunawan and colleagues (J Appl Biotechnol Rep. 2025;12(3)) introduces a statistically grounded optimization strategy for the heterologous expression of Mycobacterium tuberculosis CYP124 in Escherichia coli using the Box–Behnken Design (BBD). The research identifies the optimal combination of FeCl₃, δ-aminolevulinic acid (5-ALA), and IPTG for maximizing yield and purity of CYP124, a cytochrome P450 enzyme implicated in cholesterol metabolism and drug biotransformation. This commentary highlights the scientific significance, methodological robustness, and biotechnological implications of their findings, particularly in the context of tuberculosis (TB) drug discovery and enzyme engineering.

https://www.biotechrep.ir/article_236332_761790233f9c8b6b2949120fea674c43.pdf