AJMB: Avicenna Medical Journal of Biotechnology https://www.AJMB.org/ Avicenna Medical Journal of Biotechnology The Role of Biotechnology in Latest Therapeutic Approaches for Diabetes Mellitus https://www.AJMB.org/En/Article.aspx?ID=60568 Sepideh Hajivalizadeh , Shahin Akhondzadeh Wed, 31 Jan 2024 00:00:00 GMT en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir Factor VIII as a Novel Biomarker for Diagnosis, Prognosis, and Therapy Prediction in Human Cancer and Other Disorders <p><span style="font-size:10.0pt">Coagulation factor VIII (FVIII) is an essential cofactor in the coagulation cascade, encoded by the <em>F8</em> gene on the long arm of chromosome X (Xq28). FVIII is normally circulated in complex with Von Willebrand factor (VWF) and has relevant emerging extracoagulative functions. Dysregulation of FVIII is associated with tumor progression, and could be used as a novel biomarker for tumor screening and monitoring. In breast cancer, bladder cancer, colorectal carcinoma, esophageal carcinoma, hepatocellular carcinoma and lung cancer, <em>F8</em> is regarded as an oncogene. In coronary heart disease, hemophilia A and liver disease, <em>F8</em> dysregulation has been recognized as a potential biomarker for disease diagnosis and prognosis. However, the basis of these differential expression levels remains to be understood. In this review, which is a mixture of literature review and bioinformatics analysis we described the biological functions and characteristics of FVIII, and also its expression level in non-malignant disorders and various cancers.</span></p> https://www.AJMB.org/En/Article.aspx?ID=60569 Sheyda Khalilian , Zahra Mohajer , Soudeh Ghafouri-Fard Mon, 15 Jan 2024 00:00:00 GMT en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir The Protective Effect of Crocin on Rat Bone Marrow Mesenchymal Stem Cells Exposed to Aluminum Chloride as an Endocrine Disruptor <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Background:</span></strong><span style="font-size:10.0pt"> Mesenchymal Stem Cells (MSCs) have the ability to self-renew and proliferate which gives them healing properties in various tissues. Aluminium chloride (AlCl<sub>3</sub>) is a chemical compound with harmful effects on health; oxidative stress caused by Aluminium has been reported previously. Crocin, a major component of <em>Crocus sativus </em>(saffron), has antioxidant properties and has shown therapeutic potential. Researchers have been looking for ways to reduce the harmful effects of AlCl<sub>3</sub>. </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Methods:</span></strong><span style="font-size:10.0pt"> To investigate whether crocin can reduce AlCl<sub>3</sub> cytotoxicity, rat Bone Marrow Mesenchymal Stem Cells (BM-MSCs) were isolated, cultured and divided into four experimental groups. The first group was the control, which was untreated cells. The second and third groups were treated with crocin (</span><span style="font-size:10.0pt">50, 100, 250, 500 </span><em><span style="font-size:10.0pt">&micro;M</span></em><span style="font-size:10.0pt">) and AlCl<sub>3</sub> (20, 25, 30 <em>mM</em>) for 24 <em>hr</em>. The fourth group was pre-treated with crocin (</span><span style="font-size:10.0pt">250, 500 </span><em><span style="font-size:10.0pt">&micro;M</span></em><span style="font-size:10.0pt">) for 24 <em>hr</em> and then treated with AlCl<sub>3</sub> (20 <em>mM</em>) overnight. Cytotoxicity was assessed using the MTT assay. Mineralization was evaluated by alizarin red staining. Sox-2 and E-cadherin expression were measured using real-time PCR.</span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Results:</span></strong><span style="font-size:10.0pt"> The results showed that AlCl<sub>3</sub> caused cytotoxicity on BM-MSCs and decreased the mRNA expression of Sox-2 and E-cadherin, which are important for the maintenance of self-renewal and proliferation of BM-MSCs. In contrast, crocin protected the self-renewal characteristic of BM-MSCs by increasing Sox-2 expression and also preserved the proliferative effects on BM-MSCs by upregulating E-cadherin expression (***p&le;0.001). </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Conclusion:</span></strong><span style="font-size:10.0pt"> Overall, the study suggests that crocin can protect BM-MSCs from AlCl<sub>3</sub>-induced cytotoxicity by upregulate Sox-2 expression and E-cadherin expression. This suggests that crocin may be a potential therapeutic agent for the treatment of AlCl<sub>3</sub>-induced toxicity. </span></span></p> https://www.AJMB.org/En/Article.aspx?ID=60570 Elaheh Amini, Zahra Baharvand, Azadeh Niknejad, Yasaman Tabari, Sahel Shemshadi Mon, 15 Jan 2024 00:00:00 GMT en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir The Protective Effect of N-acetylcysteine against Deltamethrin-Induced Hepatotoxicity in Mice <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Background:</span></strong><span style="font-size:10.0pt"> Exposure to pesticides is of concern to public health officials worldwide. Deltamethrin is a synthetic <span style="color:black">pyrethroid </span>pesticide which is widely used in agriculture and veterinary medicine. Deltamethrin poisoning is always one of the concerns in medical centers</span><span style="font-size:10.0pt"> due to the </span><span style="font-size:10.0pt">deltamethrin induced </span><span style="font-size:10.0pt">hepatotoxicity.</span><span style="font-size:10.0pt"> This study evaluated the hepatoprotective effects of N-acetylcysteine (NAC) against deltamethrin induced hepatotoxicity in mice</span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Methods:</span></strong><span style="font-size:10.0pt"> A total of 40 BALB/c male mice were randomly divided into four groups; the first group was used as a control (0.5 <em>ml</em> normal saline); Groups 2-4 were treated with NAC [160 <em>mg/kg</em> Body Weight (BW)], deltamethrin (50 <em>mg/kg</em> BW), and NAC plus deltamethrin. At 1 and 24 <em>hr</em> after treatment, the animals were sacrificed and blood and liver samples were obtained for analysis and the liver/body ration, hepatic enzymes as </span><span style="font-size:10.0pt">Aspartate aminotransferase (AST), Alanine Transaminase (ALT), Alkaline phosphatase (ALP), Lactate dehydrogenase (LDH), Glutathione</span><span style="font-size:10.0pt"> (GSH) content and <span style="background-color:white"><span style="color:black">Reactive Oxygen Species</span></span> (ROS) level were measured. For comparison between more than two experimental groups, one-way ANOVA following Tukey test was used by SPSS software.</span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Results:</span></strong><span style="font-size:10.0pt"> The deltamethrin significantly increased AST, ALT, ALP, and the level of ROS level at the end of 1 and 24 <em>hr</em> after treatment; while the LDH level and GSH content were decreased. Mice in the deltamethrin treated group had a higher liver/body weight ratio than in other treated groups after 24 <em>hr</em>. On the other hand, NAC in combination with deltamethrin significantly reduced the activities of AST, ALT, ALP, and increased GSH levels.</span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Conclusion:</span></strong><span style="font-size:10.0pt"> This study demonstrated that NAC has a hepatoprotective role against deltamethrin-induced toxicity.</span></span></p> https://www.AJMB.org/En/Article.aspx?ID=60571 Ali Ameri, Alireza Rahmati, Shadi Soroushfar , Mehdi Lalehzari, Tahereh Dehghani, Hamed Haghi-Aminjan, Jebreil Shamseddin, Mahmoud Omidi Mon, 15 Jan 2024 00:00:00 GMT en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir A Simple High Yield Technique for Isolation of Wharton's Jelly-derived Mesenchymal Stem Cell <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Background:</span></strong><span style="font-size:10.0pt"> The isolation of Mesenchymal Stem Cells (MSCs) from various tissues is possible, with the umbilical cord emerging as a competitive alternative to bone marrow. In order to fulfill the demands of cell therapy, it is essential to generate stem cells on a clinical scale while minimizing time, cost, and contamination. Here is a simple and effective protocol for isolating MSC from Wharton&#39;s Jelly (WJ-MSC) using the explant method with various supplements. </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Methods:</span></strong><span style="font-size:10.0pt"> Utilizing the explant method, small fragments of Wharton&#39;s jelly from the human umbilical cord were cultured in a flask. The multipotency of the isolated cells, were confirmed by their differentiation ability to osteocyte and adipocyte. Additionally, the immunophenotyping of WJ-MSCs showed positive expression of CD73, CD90, and CD105, while remaining negative for hematopoietic markers CD34 and CD45, meeting the criteria for WJ-MSC identification. Following that, to evaluate cells&#39; proliferative capacity, various supplements, including basic Fibroblast Growth Factor (bFGF), Non-Essential amino acids (NEA), and L-Glutamine (L-Gln) were added to either alpha-Minimal Essential Medium (&alpha;-MEM) or Dulbecco&#39;s Modified Eagle&#39;s Medium-F12 (DMEM-F12), as the basic culture media.</span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Results:</span></strong><span style="font-size:10.0pt"> WJ-MSCs isolated by the explant method were removed from the tissue after seven days and transferred to the culture medium. These cells differentiated into adipocyte and osteocyte lineages, expressing CD73, CD90, and CD105 positively and CD34 and CD45 negatively. The results revealed that addition of bFGF to &alpha;-MEM or DMEM-F12 media significantly increased the proliferation of MSCs when compared to the control group. However, there were no significant differences observed when NEA or L-Gln were added.</span></span></p> <p><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Conclusion:</span></strong><span style="font-size:10.0pt"> Although bFGF considerably enhances cell proliferation, our study demonstrates that MSCs can grow and expand when properly prepared Wharton&#39;s jelly tissues of the human umbilical cord.</span></span></p> https://www.AJMB.org/En/Article.aspx?ID=60572 Bahare Niknam, Arezou Azizsoltani , Neda Heidari, Samaneh Tokhanbigli, Helia Alavifard, Mahsa Haji Valili, Davar Amani, Hamid Asadzadeh Aghdaei, Seyed Mahmoud Hashemi , Kaveh Baghaei Mon, 15 Jan 2024 00:00:00 GMT en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir Simple Determination of Bosentan in Plasma Samples by Reversed-Phase High-Performance Liquid Chromatography <p style="text-align:justify"><span style="font-size:12pt"><span style="color:black"><strong><span style="font-size:10.0pt">Background:</span></strong><span style="font-size:10.0pt"> In order to measure the plasma levels of Losartan and Bosentan, a sensitive Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) technique was developed.</span></span></span></p> <p style="text-align:justify"><span style="font-size:12pt"><span style="color:black"><strong><span style="font-size:10.0pt">Methods:</span></strong><span style="font-size:10.0pt"> To compare bioavailability, the Area Under the Curve (AUC), peak plasma concentration (Cmax), and time to Cmax (Tmax) were employed. The standard curve (150-2400 <em>ng/ml</em>) was linear (R<sup>2</sup>=0.999), relative errors were between 2.4 to 10.05% and the coefficient of variation (CV%) ranged from 1.52 to 10.88. A single dosage (test and reference) was used for the <em>in vivo</em> investigation, which involved 16 healthy individuals.</span></span></span></p> <p style="text-align:justify"><span style="font-size:12pt"><span style="color:black"><strong><span style="font-size:10.0pt">Results:</span></strong><span style="font-size:10.0pt"> The AUC0-48, AUC0-, Cmax, and Tmax of the test and reference had no statistically significant differences. The C<sub>max</sub> and 95% confidence intervals of the ratio of C<sub>max</sub> of the two formulations were 0.93-0.96 and 97.6-135%, respectively. </span></span></span></p> <p style="text-align:justify"><span style="font-size:12pt"><span style="color:black"><strong><span style="font-size:10.0pt">Conclusion:</span></strong> <span style="font-size:10.0pt">Therefore, it was established that </span><span style="font-size:10.0pt">generic Bosentan was equivalent to Bosentan from Actelion and that both medications could be regarded as equally effective in clinical settings. The blood level of Bosentan could be measured using this straightforward procedure in all hospital laboratories.</span></span></span></p> https://www.AJMB.org/En/Article.aspx?ID=60573 Zahra Khalighi , Hori Ghaneialvar, Armin Soltani, Ali Khorshidi, Elahe Karimi, Ardeshir Moayeri, Naser Abbasi , Masoumeh Tahmasebi, Ali Aidy Mon, 15 Jan 2024 00:00:00 GMT en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir Poultry Gastrointestinal-derived Lactic Acid Bacteria (pGIT-d-LAB) Inhibit Multiple Antibiotics Resistance Bacterial and Fungal Pathogens <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Background:</span></strong><span style="font-size:10.0pt"> To develop a probiotic formulation for poultry feed, a few poultry gastrointestinal derived lactic acid bacteria (pGIT-d-LAB) were isolated from chicken intestinal specimens and <em>in vitro</em> experiment was performed to evaluate their efficacy as potential probiotic candidate.</span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Methods:</span></strong><span style="font-size:10.0pt"> A total of 6 strains of LAB: <em>L</em><em>actobacillus brevis</em> (<em>L. brevis</em>)<em>,</em> <em>L</em><em>actobacillus acidophilus </em>(<em>L. acidophilus</em>)<em>, Lactobacillus casei </em>(<em>L. casei</em>)<em>, Pediococci </em>spp.<em>, Lactobacillus fermentum </em>(<em>L. fermentum</em>)<em> </em>and<em> L</em><em>actobacillus plantarum </em>(<em>L. plantarum</em>)<em> </em>were isolated and cultured for collection of Cell Free Supernatant (CFS). CFS collected was tested against pathogenic bacterial isolated from chicken feces as well as prevalent fungal pathogens, utilizing agar-well diffusion techniques. A preliminary investigation into the susceptibility of the pathogens to diverse antibiotics and antifungal drugs was conducted. Bacterial pathogens exhibiting resistance to a minimum of three classes of antibiotics were subsequently identified for pGIT-d-LAB CFS screening. </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Results:</span></strong><span style="font-size:10.0pt"> The observed results revealed that the CFS derived from the isolates exhibited varying degrees of growth inhibition against different pathogens. Among the tested pGIT-d-LAB isolates, <em>L. acidophilus </em>demonstrated the most prominent zone of inhibition, measuring 18 <em>mm</em> against <em>Klebsiella pneumoniae</em> ZTAC 1233. Notably, <em>Citrobacter diversus</em> ZTAC 1255 showed resistance to all tested pGIT-d-LAB. Quantification of the metabolites produced was performed, and peak production levels was determined. <em>L.</em><em> acidophilus </em>produced the highest amount of lactic acid (1.789<em>g/l</em>), <em>Pediococci </em>spp. produced the highest amount of diacetyl and H<sub>2</sub>0<sub>2</sub> (1.918<em>g/l</em>) (0.0025<em>g/l</em>) at 48<em> hr</em> peak values respectively. </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Conclusion:</span></strong><span style="font-size:10.0pt"> The test isolates are potential probiotic candidates for controlling pathogens in poultry. </span></span></p> https://www.AJMB.org/En/Article.aspx?ID=60574 Bolanle Adeniyi, Abimbola Adesuyi , Funmilola Ayeni , Temitope Ogunbanwo , Taiwo Agidigbi Wed, 17 Jan 2024 00:00:00 GMT en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir Investigating the Effects of HMGB1 Overexpression on Colorectal Cancer Cell Migration via Oncolytic Herpes simplex Virus Type 1 (oHSV-1) <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Background:</span></strong><span style="font-size:10.0pt"> Colorectal Cancer (CRC) represents a significant global health challenge, and its progression, resistance to therapy, and metastasis are strongly influenced by the tumor microenvironment, including factors like hypoxia. This study explores the impact of High Mobility Group Box 1 (HMGB1) overexpression on CRC cell migration, while identifying potential genes associated with this process. </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Methods:</span></strong><span style="font-size:10.0pt"> To explore this, we developed oncolytic virotherapy, resulting in HSV-HMGB1, an oncolytic <em>Herpes simplex</em> virus that expresses HMGB1. HMGB1 is known its role in cancer progression, particularly in the context of cancer cell migration. </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Results:</span></strong><span style="font-size:10.0pt"> Contrary to expectations, our scratch assays indicated that HSV-HMGB1 did not significantly induce migration in CRC cells, suggesting that HMGB1 might not directly contribute to this process. Employing microarray analysis, we investigated gene expression changes linked to CRC cell migration, leading to construction of a Protein-Protein Interaction (PPI) network. This network revealed the presence of hub proteins, including as NDRG1, LGALS1, and ANGPTL4, which are recognized for their roles in cancer cell migration. The differential expression of these genes under hypoxic conditions was further validated using quantitative RT-PCR, aligning with the findings from our microarray data.&nbsp; </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Conclusion:</span></strong><span style="font-size:10.0pt"> Our findings emphasize the complex regulation of CRC cell migration, and provides valuable insights into potential molecular mechanisms and pathways. These findings have implications for further research into cancer progression and the development of therapeutic strategies.</span></span></p> https://www.AJMB.org/En/Article.aspx?ID=60575 Sara Shayan, Arash Arashkia, Golnaz Bahramali, Kayhan Azadmanesh Wed, 17 Jan 2024 00:00:00 GMT en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir CYP21A2 Gene Analysis in Southern Iranian CAH Patients and a Brief Review of the Mutation Spectrum <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Background:</span></strong><em><span style="font-size:10.0pt"> CYP21A2</span></em><span style="font-size:10.0pt"> gene mutations are responsible for more than 95% of Congenital Adrenal Hyperplasia</span><span style="font-size:10.0pt"> (CAH)</span><span style="font-size:10.0pt"> disorders with</span> <span style="font-size:10.0pt">autosomal recessive inheritance. Most of these pathogenic mutations originate from the <em>CYP21A1P</em>, a neighboring pseudogene with 98% homology, due to unequal crossing over or gene conversion events. Mutation identification of the gene could be beneficial for accurate diagnosis and outcome prediction. </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Methods:</span></strong><span style="font-size:10.0pt"> Twelve unrelated patients with CAH diagnosis</span> <span style="font-size:10.0pt">were recruited for genetic counseling. To ensure distinct amplification of the <em>CYP21A2</em> gene rather than its pseudogene, the complete sequence of the gene was amplified through two overlapping fragments by specific primers. The entire sequences were screened by direct Sanger sequencing using new sequencing primers. </span></span></p> <p style="text-align:justify"><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Results</span></strong><span style="font-size:10.0pt">: Only two pathogenic point mutations were identified. The c.293-13C&gt;G, also known as In2G, and the c.955C&gt;T mutations were found in 37.5 and 33.3% of alleles, respectively. One patient showed homozygous gene deletion. </span><span style="font-size:10.0pt">We also reviewed recent reports on <em>CYP21A2</em> gene mutations in Iran.</span> </span></p> <p><span style="font-size:11pt"><strong><span style="font-size:10.0pt">Conclusion:</span></strong><span style="font-size:10.0pt"> Evaluating the ethnicity-specific gene mutation data is significant for populations with diverse ethnic groups including the Iranian population. Although several common mutations have been reported as causative mutations among CAH patients, identifying only two common point mutations in Fars province would help prioritize exon sequencing and reduce the cost and time of genotyping.</span></span></p> https://www.AJMB.org/En/Article.aspx?ID=60576 Danial Zangene, Ali Moravvej, Homa Ilkhanipoor, Anis Amirhakimi, Zhila Afshar, Mona Entezam Wed, 17 Jan 2024 00:00:00 GMT en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir https://www.AJMB.org/En/Article.aspx?ID=60577 en eslamifar@gmail.com (Mohsen Eslamifar) ajmb@avicenna.ac.ir