Sina Gerayli; Alireza Pasdar; Sina Rostami; Samaneh Sepahi; Seyed Mousalreza Hoseini; Reza Jahanian; Aida Gholoobi; Zahra Meshkat; Mitra Ahadi
Abstract
Single nucleotide polymorphism in codon 72 of p53 gene (Arg/Pro) changes p53 protein structure and affects its activities. Hepatitis C virus (HCV) is believed to induce hepatocellular carcinoma and P53 polymorphisms have been associated with human cancers. The aim of this study was to evaluate genetic ...
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Single nucleotide polymorphism in codon 72 of p53 gene (Arg/Pro) changes p53 protein structure and affects its activities. Hepatitis C virus (HCV) is believed to induce hepatocellular carcinoma and P53 polymorphisms have been associated with human cancers. The aim of this study was to evaluate genetic variants of codon 72 of p53 gene polymorphism in HCV patients and its relationship with HCV infection.
The study was conducted on 67 HCV patients, who were referred to medical centers of Mashhad city, Iran, and 73 healthy people from the same region. Genotyping of codon 72 of p53 gene was performed by PCR-RFLP method.
The distributions of different alleles of p53 polymorphisms did not differ significantly between groups. The respective proportions of Proline homozygotes, heterozygotes, and Arginine homozygotes were 37.31%, 35.82%, 26.86% in patients and 39.72%, 27.39%, and 32.87% in the control group respectively. However, we found no significant differenece for the allelic or genotype distribution between cases and controls.
Our results indicated no strong evidence of association of the p53 polymorphism with HCV infection; however, further investigation is needed in different ethnic groups to elucidate the role of this polymorphism in HCV infection.
Samaneh Khazaei; Sedigheh Gharbi; Seyed Javad Mowla
Abstract
Esophageal squamous cell carcinoma (ESCC) is a deadly cancer with poor prognosis. In this regard, early diagnosis is of vital importance to cure the tumor in its early stages. Novel cancer diagnostic and therapeutic approaches have been recently introduced based on microRNAs (miRNAs). Also, accurate ...
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Esophageal squamous cell carcinoma (ESCC) is a deadly cancer with poor prognosis. In this regard, early diagnosis is of vital importance to cure the tumor in its early stages. Novel cancer diagnostic and therapeutic approaches have been recently introduced based on microRNAs (miRNAs). Also, accurate normalization using appropriate reference genes is a critical step in miRNA expression studies. In this study, we aimed to identify appropriate reference genes for miRNA quantification in serum samples of ESCC. In this case and control experimental study, two statistical algorithms including GeNorm and NormFinder were used to evaluate the suitability of miR-16 and 5S rRNA and their geometric mean as reference genes. Then, relative expression of miR-451 and miR-24 were evaluated while different normalizer including miR-16, 5S rRNA and their geometric mean were applied. Both GeNorm and NormFinder analyses showed that geometric mean of miR-16 and 5S rRNA is the most stable reference gene in these samples. Also, our data showed that choosing an inappropriate normalizer could change the relative expression of target genes of miR-451 and miR-24 in ESCC samples which emphasize on the importance of selecting a reliable internal control in expression analyses. We demonstrated that geometric mean of two reference genes could increase the reliability of normalizers and also by using geometric mean as reference gene, relative expression of different target is closer to reality.
Samaneh Jamshidi; Mehrdad Lahouti; Mohammad Taher Boroushaki; Ali Ganjeali; Ahmad Ghorbani; Mehdi Bihamta toosi
Abstract
Diosgenin is an important compound in pharmaceutical industry. It has various effects such as hypocholesterolemic action or antioxidant activity in HIV infected patients. Biological oxidation pathways are involved in causing or aggravating heart disease. This study investigated the potential protective ...
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Diosgenin is an important compound in pharmaceutical industry. It has various effects such as hypocholesterolemic action or antioxidant activity in HIV infected patients. Biological oxidation pathways are involved in causing or aggravating heart disease. This study investigated the potential protective effect of diosgenin on cell viability and antioxidant defenses of cultured H9C2 cells submitted to oxidative stress induced by H2O2. Viability of cells exposed to H2O2 was detected by MTT assay. The generation of ROS and hydrogen peroxide release after H2O2 were detected using the fluorescent probe H2DCF-DA. The lipid peroxidation product i.e. MDA formation was estimated by assessing the levels of thio-barbituric acid reactive substances (TBARS) using spectrophotometry. SOD activity was assayed with NWLSS (TM) Superoxide Dismutase (SOD) activity assay kit. Pretreatment of cells with 3-25 µM of diosgenin for 24 h before applying H2O2 completely prevented cell damage and significantly enhanced viability of H9C2 cells. Increased ROS induced by H2O2 was dose dependently prevented when cells were pretreated for 24 h with diosgenin. The level of the lipid peroxidation was significantly higher in H9C2 cells exposed to H2O2 as compared to the control and cells pretreated with diosgenin. SOD activity in cells treated with diosgenin significantly decreased compared with cells exposed to H2O2. These results show that treatment of H9C2 cells with diosgenin (3-25 µM) confers a significant protection against oxidative stress.
Zahra Roudbari; Mohammadreza Nassiri; Mojtaba Tahmoorespur; Aliakbar Haddad-Mashadrizeh; Ali Javadmanesh
Abstract
Human growth hormone (hGH) is a protein with multiple roles in a range of biological functions such as protein, carbohydrates and lipid metabolisms as well as immunity, tissue development and overall growth. One of the major class of biopharmaceuticals in mammalian cells is the production of recombinant ...
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Human growth hormone (hGH) is a protein with multiple roles in a range of biological functions such as protein, carbohydrates and lipid metabolisms as well as immunity, tissue development and overall growth. One of the major class of biopharmaceuticals in mammalian cells is the production of recombinant pharmaceutical proteins. In this study, we constructed a lentiviral vector carrying coding region of human GH1 (hGH) gene in order to production of recombinant hGH in mammalian cell line. hGH gene was amplified from a plasmid containing full-length hGH coding sequence and then cloned into the lentiviral vector pCDH-GFP. The HEK293T cells were transduced by the lentivirus particles as a targeted cell. hGH expression status in the recombinant cells were confirmed by RT-PCR. Additionally, western blotting analysis results showed that the recombinant cells maintained a stable hGH expression during five weeks of continuous culture. In conclusion, results of current study suggested that constructed lentiviral vector can potentially be used for a stable production of recombinant hGH protein in HEK293T cells. This methodology could be served as a foundation for further research and may open new insights toward therapeutic protein manufacturing.
Nassim Rahmani; Esmaeil Ebrahimie; Ali Niazi; Najaf Allahyari Fard; Bijan Bambai; Zarrin Minuchehr; Mansour Ebrahimi
Abstract
Allergens are proteins or glycoproteins which make widespread disorders that can lead to a systemic anaphylactic shock and even death within a short period of time. Understanding the protein features that are involved in allergenicity is important in developing future treatments as well as engineering ...
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Allergens are proteins or glycoproteins which make widespread disorders that can lead to a systemic anaphylactic shock and even death within a short period of time. Understanding the protein features that are involved in allergenicity is important in developing future treatments as well as engineering proteins in genetic transformation projects. A big dataset of 1439 protein features from 761 plant allergens and 7815 non-allergen proteins was constructed. Thereafter, 10 different attribute weighting algorithms were utilized to find the key characteristics differentiating allergens and non-allergen proteins. The frequency of Leu, Arg and Gln selected by different attribute weighting algorithms with more than 50% confidence, including attribute weighting by Weight_Info Gain, Weight Chi Squared, Weight_Gini Index and Weight_Relief. High amount of Gln and low percentage of Leu and Arg discriminate plant allergens from non-allergens
Maziar Habibi-Pirkoohi; Saeid Malekzadeh-Shafaroudi; Hasan Marashi; Saeid Zibaee; Afsaneh Mohkami; Saba NejatizaXeh
Abstract
An Agrobacterium-mediated transient gene expression assay was carried out in alfalfa (Medicago sativa) leaves for expression of a chimeric gene encoding a part of capsid protein of Foot and Mouth Disease virus called VP1. The plant leaves were transformed via agroinfiltration procedure. The presence ...
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An Agrobacterium-mediated transient gene expression assay was carried out in alfalfa (Medicago sativa) leaves for expression of a chimeric gene encoding a part of capsid protein of Foot and Mouth Disease virus called VP1. The plant leaves were transformed via agroinfiltration procedure. The presence of the foreign gene and its expression in transformed plants were evaluated by polymerase chain reaction (PCR), real time PCR, protein Dot blot and ELISA. Moreover, gene expression in the transformed leaves was quantified by ELISA method. The results obtained in this investigation indicated high level of gene expression in alfalfa leaves, showing that transient gene expression can be applied as an effective and time-saving procedure for the production of recombinant proteins. The procedures for transformation, detection of recombinant protein and its application for molecular experiments are described in the study.
Maryam Parhamfar; Arastoo Badoei-Dalfard; Milad Parhamfar; Shohreh Fahimi Rad
Abstract
Phosphorus is one of the most important nutrients for plant growth and development. Chemical Pi fertilizer is used to provide the phosphorus for the plants, but it is mostly fixed in the soil into insoluble form and become unavailable to the plants. Phosphate-solubilizing bacteria have lots of application ...
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Phosphorus is one of the most important nutrients for plant growth and development. Chemical Pi fertilizer is used to provide the phosphorus for the plants, but it is mostly fixed in the soil into insoluble form and become unavailable to the plants. Phosphate-solubilizing bacteria have lots of application in agriculture as biological fertilizer. Consumption of biofertilizers instead of chemical fertilizers can lead to environmental pollution reduction and crop production enhancement using sustainable farming. In this study, a phosphatase-producing bacterium was isolated from agricultural soil in Kerman. Screening of phosphate solubilizing bacteria was performed on the PVK medium, based on clear area diameter. The best bacterium (AG41) was identified based on 16s rDNA gene. The optimum condition for production of phosphatase was also determined and it was purified and characterized. Sequence alignment and phylogenetic tree results show that AG41 is closely related to Bacillus subtilis, with 98% homology. Phosphatase activity was determined by end point method. The best carbon, nitrogen and phosphate sources for enzyme production were 1.0% glucose, 0.5% ammonium sulfate and (0.25%) sodium phytate +(0.25%) tricalcium phosphate, respectively. Bacterial phosphatase was partially purified using ammonium sulfate fractionation followed by dialysis. Results showed that the optimum temperature for the purified enzyme activity was 40oC and it was stable at temperatures below 60°C. This enzyme was stable between pH 3.0-7.0, and the optimal pH activity was found to 5.0. These results indicated that this strain can be a notable candidate for using as biofertilizers.
