The aim of study To purify GPCR from a local strain of S. cerevisiae using Ion exchange and gel filtration chromatography techniques , by packing materials for columns which will be chosen of low cost comparing to the already used in published researches, which depend on the costly affinity chromatography and other expensive methods of purification. Local strain of S. cerevisiae chosen for extraction and purification of G-protein coupled receptor (GPCR) .The strains were obtained from biology department in Al- Mosul University, Iraq. The isolated colony was activated on Yeast Extract Pepton Dextrose Broth (YEPDB) and incubated at 30 C˚ for 24 h .Loop fully of the yeast culture was transferred to (10ml) of yeast extract peptone glucose agar (YEPGA) slant , then incubated at 30C˚for 24h , after that it was stored at 4C˚ ,the yeast cultures were reactivated and persevered after each two weeks period. S.cerevisiae was identified by morphological, microscopic characterization and biochemical test . The GPCR that extract from whole cell of S.cerevisiae was purified by ion exchange chromatography using DEAE-Sepharose ,the bound proteins (negatively charged) were then eluted using gradient concentration of NaCl ranged between( 0.1 -0.5M). Gel filtration chromatography using Sepharose 6B was applied as a second step of purification. The optical density for each fraction was measured at 280 nm by UV-VS spectrophotometer then the GPCR concentration was determined by using ELISA Kit . The fractions which gave the highest absorbance and concentration of GPCR were collected .The molecular weight of GPCR was determined by gel filtration chromatography using blue dextrin solution. Standard curve was plotted between log of molecular weight for standard protein and the ratio of Ve/Vo of GPCR . The purity of the GPCR that extracted and purified from whole cell of S, cerevisiae were carried out by using SDS-PAGE electrophoresis . In ion exchange chromatography the fraction were collected with 5 ml tube at a flow rate 0.5 ml/ min and eluted with gradient (0.1-0.5M) of sodium chloride solution. Two proteins peaks appeared after eluted by the gradient concentration of sodium chloride, while no protein peaks appeared in the washing fractions. The GPCR concentration was measured in the fractions of these two protein peaks, data indicated that GPCR located in the first protein peak (eluted at 0.1M of NaCl) at fraction numbers between 3 and 9, the maximum concentration of GPCR was 9.281 with specific activity 71.58(ng/mg)protein , 3.125 purification folds and72.9(%) yield while the second peaks (eluted at 0.4 M of NaCl) don't give any concentration for GPCR, thus its neglected. Gel filtration chromatography was used as second step of purification which applied by using sepharose 6B. Results show single active protein peaks appeared that identical with the peak of GPCR at fractions numbers(29-35). The maximum concentration of GPCR was 9.082 (ng/ml)was observed in these fractions. The specific activity for these fractions was 151.37 (ng/mg) protein with 6.608 purification folds and 39.64 (%) yield. The present study a chive a relatively high purification of GPCR from whole cell of a local strain S. cerevisiae with fold purification 6.608 and a yield of 39.64 % and molecular weight about~33KD.
This work introduces a newly synthesized triazole derivative, 5-([(2-furanyl)methyl]-thiomethyl)-4-(naphtha-2-yl)-3-hydroxyl-4H-1,2,4-triazole (FTNT), as an effective corrosion inhibitor for carbon steel A106 G/B in 0.1 M HCl solution. The structure of the novel derivative was confirmed by FTIR, 1 H NMR, and 13 C NMR spectra. Potentiodynamic polarization measurements reveal that FTNT markedly suppresses corrosion, with inhibition efficiency reaching 81.6% at 600 ppm. Thermodynamic analysis indicates spontaneous, physisorption/chemisorption-augmented adsorption of FTNT on the steel surface, best described by the Freundlich isotherm. Atomic force microscopy (AFM) surface examination demonstrated that a protective FTNT coating precipitated on
... Show MoreBackground: Red clover oil (Trifolium pratense) has isoflavones bunches which have estrogen-like exercises and may establish an option in contrast to hormone substitution treatment. The present study investigated the impact of Red clover oil on bone healing in rats by histomorphometric study. Materials and Methods: Intra bony defect was performed in right femur of thirty six healthy male albino rats. Then these rats were randomly divided into three groups (12 rats each): one control and 2 experimental groups. For control, the bony defect was left for normal healing; for the experimental (S) group, the defect was treated with hemostatic absorbable gelatin sponge; and for experimental (RS) group, the bony defect was treated with 0.2 ml red
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The definition of the role of any institution in society is achieved through its objectives, The same is true for the military and how to deal with security threats in the humanitarian field ,Terrorism, which has almost replaced the traditional pattern of war, has waged a street war and intimidated individuals, families and society ,On the other hand, he found someone to meet him from a popular crowd of volunteers to defend their homeland from different sects, sects and religions, Thus, our study will be exposed to the role of popular mobilization in human security from a sociological point of view in Samarra, a field study of 100 male and female respondents.
Microbial fuel cell is a device that uses the microorganism metabolism for the production of electricity under specific operating conditions. Double chamber microbial fuel cell was tested for the use of two cheap electrode materials copper and aluminum for the production of electricity under different operating conditions. The investigated conditions were concentration of microorganism (yeast) (0.5- 2 g/l), solutions temperature (33-45 oC) and concentration of glucose as a substrate (1.5- 6 g/l). The results demonstrated that copper electrode exhibit good performance while the performance of aluminum is poor. The electricity is generated with and without the addition of substrate. Addition of glucose substrate
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Background: Oral cancer is a highly lethal and disfiguring disease. Squamous cell carcinoma of the oral cavity constitutes about 90% of all oral malignancies. The aims of the study was to achieve an epidemiological description of the oral squamous cell carcinoma in Iraq in general and in Iraqi governorates except Kurdistan region retrospectively during period 2001-2013 Materials and Methods: Data were collected from department of oral and maxillofacial pathology, college of dentistry, university of Baghdad, Nuclear medicine and radiotherapy hospital, Iraqi cancer registry center, Main hospitals in Baghdad and Iraqi governorates, Private labs. for histopathological examinations. The descriptive and inferential statistical methods were used
... Show MoreBackground: Understanding the pathogenesis and molecular basis of Oral Squamous Cell Carcinoma (OSCC) has increased rapidly over the past few years that is essential to improve patient's prognosis and treatment modalities. The purpose of this study to evaluate the Immunohistochemical expressions of AKT, ATM, AND Cyclin E in oral squamous cell carcinoma Materials and methods: This study was performed on a forty formalin-fixed paraffin-embedded blocks which histopathologically diagnosed as Oral Squamous Cell Carcinoma. All cases were collected from the Histopathological Laboratory from patients treated surgically at Maxillofacial surgery Department at Ramadi Teaching Hospital, Iraq. Results: The immunohistochemical staining of AKT showed pos
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Liposome-mediated transfection of cancer cells provide a valuable experimental technique to study cellular gene expression and may also be adapted for gene therapy studies. However, the widely recognized advantage of liposome-mediated transfection is high efficiency. Therefore, this study were performed to optimize transfection techniques in human larynx carcinoma cell line Hep-2 using the commercial synthetic lipid TransFast™ Reagent and monitoring the expression efficiency by using the pSV-?-galactosidase Control Vector which encoded ?-galactosidase, maximum transfection efficiency were achieved with TransFast™ Reagent used at the Charge ratios of 2:1 and 0.5 µg DNA/ml, this is indicate that TransFast™ Reagent can be used as an eff
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Background: Oral squamous cell carcinoma (OSCC) is a common malignancy characterized by poor prognosis and low survival rate. The purpose of this study was to evaluate the Immunohistochemical expressions of BAD, MDM2, and P21as apoptotic markers in oral squamous cell carcinoma. Materials and methods: This study was performed on forty formalin-fixed paraffin-embedded blocks which histopathologically diagnosed as Oral Squamous Cell Carcinoma. All cases were collected from the Histopathological Laboratory from patients treated surgically at Maxillofacial surgery Department at Ramadi Teaching Hospital, Iraq. Results: The immunohistochemical staining of BAD showed positive expression in 39 (97.5%), MDM2 showed positive expression in 39(97.5%)
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Many previous investigations have found quercetin to be a powerful antioxidant and antitumor flavonoid, but its poor bioavailability has limited its use. This current study investigated the effects of two newly synthesized Quercetin Schiff bases containing 2-amino thiadiazole-5-thiol (Q1), and its benzyl derivatives (Q2) on MCF-7 human breast cancer cells. Cell viability and apoptosis were assessed to determine the toxic effects of Q1 and Q2. Cytotoxicity valuation showed that both compounds inhibited MCF-7 cell growth, and lactate dehydrogenase (LDH) activity increased in a dose-dependent aspect compared to the control group. Comet assay results observed that Q1 and Q2 induce more serious DNA damage than the control (untreated cell
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