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Molecular Biology Research Communications - Volume:1 Issue: 1, Mar 2012

Molecular Biology Research Communications
Volume:1 Issue: 1, Mar 2012

  • تاریخ انتشار: 1391/07/18
  • تعداد عناوین: 6
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  • Fatemeh Khaki-Khatibi, Ali Reza Yaghoubi, Morteza Ghojazadeh, Mohammad Rahbani-Nobar Pages 1-7
    Various polymorphisms on endothelial nitric oxide synthase (eNOs) gene cause reduced production of NO, the endothelial relaxing factor, and may accelerate the process of atherosclerosis. The study designed to investigate the frequency of T-786C polymorphism of the eNOs gene in patients suffering from coronary artery disease (CAD) in north-west of Iran. One hundred twenty subjects including 60 patients with angiographically diagnosed CAD and 60 age and sex matched CAD-free subjects as control were studied. The levels of nitric oxide in the samples were measured with the Griess Method. The genotype studies were carried using allele specific PCR. Comparing with the control, reduced levels of NO were noticed in the patient group (P<0.05). Statistical analysis showed that the genotype TC associated with risk of CAD (OR=2.50, 95% CI: 1.13-5.50, P=0.023). The prevalence of C allele was significantly higher in patients compared to control group (OR=2.04, 95% CI: 1.16-3.57, P=0.013). The low levels of NO and increased frequency of T-786C polymorphism might be a risk factor in progression of coronary artery disease in the studied subjects.
    Keywords: coronary artery disease, endothelial nitric oxide synthase gene, T, 786C polymorphism, nitric oxide
  • Mansour Kharati-Koupaei, Hajar Zamani, Ali Moradshahi Pages 8-15
    In addition to biochemical, physiological and morphological analysis, molecular studies provide additional information for establishing phylogenetic relationships among different species and strains of the genus Dunaliella. In the present study, based on neighbor- joining analysis of the nuclear rDNA ITS sequence, a novel strain of the green algae Dunaliella viridis was identified from Maharlu salt lake in Shiraz, Iran. The phylogenetic tree shows that the new strain is part of a clade containing several strains of D. viridis. The new strain was designated Dunaliella viridis MSV-1 and submitted to the GenBank under the accession number HQ864830. The optimum salinity for MSV-1 growth is between 1.0 to 1.5 M NaCl and does not turn red up to 4.5 M NaCl, confirming identity of the isolated strain. With respect to growth response to copper toxicity, increase in Cu2+ concentration from 1 to 30 µM, caused progressive increase in cell number ml-1 of culture over time, whereas reduction in cell number occurred at 100 and 200 µM Cu+2. Nano copper (colloidal copper with 40 nm dimensions) showed less toxicity compared to the ionic form. Cell number ml-1 of culture did not change up to 200 µM nano copper but decreased at 500 µM. In conclusion, the analysis of the ITS sequence is a reliable basis for establishing evolutionary relationships among species and strains of the genus Dunaliella and due to rapid growth at 1.5 M NaCl and high cell density, D. viridis MSV-1 is a good candidate for biofuel production from microalgae.
    Keywords: Dunaliella viridis MSV, 1, ITS sequences, nano copper, biofuel
  • Nioosha Nekooie-Marnany, Iraj Saadat Pages 16-20
    Cyclosporine A (CsA), a cyclic polypeptide metabolite extracted from the fungus, is used clinically to combat organ graft rejection in transplant subjects. Previous studies have shown that CsA exposure enhances the production of reactive oxygen species (ROS) and lipid peroxidation, which are directly involved in CsA toxicity. To protect cells and organs against ROS, the human body has evolved a highly antioxidant protection system to neutralize free radicals. The aim of this study was to investigate the effect of CsA on mRNA expression of anti-oxidant GSTO2. To do this, Jurkat cells were incubated for 24 h with different doses of CsA, ranging from 1-80 µg/ml, and the IC50 of CsA was calculated to be 40 µg/ml. Subsequently, Jurkat cells were treated with 3 µg/ml CsA for 24 h and the gene expression of GSTO2 was quantified by quantitative Real-time PCR. Results showed that the mean (SD) expression of the GSTO2 gene in CsA treated cells was 1.10 (0.07) (when assuming an expression level in untreated cells of 1.0). However, statistical analyses showed that the alterations were not significant (t=2.29, df=2, P=0.149). These findings suggest that at this concentration of CsA, other antioxidant enzymes are up-regulated in Jurkat cell lines to detoxify free radicals induced by CsA.
    Keywords: Cyclosporine A, GSTO2, Gene expression, Real time PCR
  • Sasan Mohsenzadeh, Maryam Esmaeili, Hassan Mohabatkar Pages 21-26
    Glutathione transferases are multifunctional proteins involved in several diverse intracellular events such as primary and secondary metabolisms, signaling and stress metabolism. These enzymes have been subdivided into eight classes in plants. The Phi class, being plant specific, is the most represented. In the present study, based on the sequences available at GenBank, different primers were designed for amplifying the Phi class of glutathione transferase gene in the genome and transcriptome of Iranian barley, Karoun cultivar. After extraction of DNA and total RNA, Phi class was amplified and sequenced. Bioinformatics analysis predicted that the deduced protein sequence has two ß-sheets, eight α-helices and some intermediate loops in its secondary structure. Consequently, the sequences were submitted to NCBI GenBank with GS262333 and GW342614 accession numbers. Phylogenic relationships of the sequences were compared with existing sequences in GenBank.
    Keywords: Hordeum vulgare, Glutathione S, transferase, stress, Xenobiotics
  • Sona Talaei, Asadollah Asadi, Mojtaba Amani Pages 27-32
    Copper amine oxidases are important enzymes, which contribute to the regulation of mono- and polyamine levels. Each monomer contains one Cu(II) ion and 2,4,5 trihydroxyphenylalanine (TPQ) as cofactors. They catalyze the oxidative deamination of primary amines to aldehydes with a ping-pong mechanism consisting of a transamination. The mechanism is followed by the transfer of two electrons to molecular oxygen which is reduced to hydrogen peroxide. Inhibitors are important tools in the study of catalytic properties of copper amine oxidases and they also have a wide application in physiological research. In this study, purification of the chickpea seedling amine oxidase, was done via salting out by ammonium sulfate and dialysis, followed by DEAE-cellulose column chromatography. By using the Lineweaver - Burk plot, the Km and Vm of the enzyme were found to be 3.3 mM and 0.95 mmol/min/mg, respectively. In this study, the interaction of chickpea diamino oxidase with tetraethylene- pentamine was studied. Analysis of kinetic data indicated that tetraethylenepentamine (with Ki=0.1 mM) inhibits the enzyme by linear mixed inhibitory effect.
    Keywords: Chickpea, Copper, containing amine oxidases, Tetraethylenepentamine, Linear mixed
  • Mehdi Ghasemi, Yemen Atakishiyeva, Asadollah Asadi Pages 33-38
    Pythium irregulare oomycetes adapts with environmental changes including crude oil concentration by changing the composition of lipids in the cytoplasmic membrane and providing the required characteristics for adaptation in improper and stressful environmental situations. It was found that cultivation of Pythium irregulare LX oomycetes in the nutrient media with different concentrations of crude oil with 1.0, 2.0, 3.0, 5.0 and 10.0 (%), incubated for 5 days at 26-28°C on a rotary shaker (200 rpm) in aerobic conditions and deep culturing caused an increase in the lipid content and the unsaturation degree of fatty acids, confirming the correspondence between the increase of polar lipid/free sterol in the composition of membrane lipids’ ratio and that of polar lipids in general lipid fractions. Represented data shows that the process of adaptation of oomycetes to a stressful situation created with crude oil motivated the increase of the rate of membrane phospholipids with a high quantity of unsaturated fatty acids.
    Keywords: stress, Oil, Pythium irregulare, lipid, phospholipids