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.
Background: Background : Patients with non-rheumatic atrial fibrillation have high risk of thromboembolism especially ischemic stroke usually arising from left atrial appendage .Transoesophageal echocardiography provides useful information for risk stratification in these patients as it detects thrombus in the left atrial or left atrial appendage. Objective : This study was conducted at Al-Kadhimiya Teaching Hospital to assess the prevalence of left atrial chamber thrombi in patients with chronic non-rheumatic atrial fibrillation using transoesophageal echocardiography and its clinical significance as well as to verify the superiority of transoesophageal over transthoracic echocardiography in the detection of these abnormalities. Type of
... Show MoreIn this work, microbubble dispersed air flotation technique was applied for cadmium ions removal from wastewater aqueous solution. Experiments parameters such as pH (3, 4, 5, and 6), initial Cd(II) ions concentration (40, 80, and 120 mg/l) contact time( 2, 5, 10 , 15, and 20min), and surfactant (10, 20and 40mg/l) were studied in order to optimize the best conditions .The experimental results indicate that microbubbles were quite effective in removing cadmium ions and the anionic surfactant SDS was found to be more efficient than cationic CTAB in flotation process. 92.3% maximum removal efficiency achieved through 15min at pH 5, SDS surfactant concentration 20mg/l, flow rate250 cm3/min and at 40mg/l Cd(II) ions initial co
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In this study, the effects of different loading doses of cerium in the prepared NaY zeolite from Iraqi kaolin were investigated. Al-Duara refinery atmospheric residue fluid catalytic cracking was selected as palpation reaction for testing the catalytic activity of cerium loading NaY zeolite. The insertion of cerium in NaY zeolites has been synthesized by simple ion exchange methods. Three samples of modified zeolite Y have been obtained by replacing the sodium ions in the original sample with cerium and the weight percent added are 0.35, 0.64, and 1.06 respectively. The effects of cerium loading to zeolite Y in different weight percent on the cracking catalysts were studied by employing a laboratory fluidized
... Show MoreBiodiesel production process was attracted more attention recently due to the surplus quantity of glycerol (G) as a byproduct from the process. Glycerol Utilization must take in to consideration to fix this issue also, to ensure biodiesel industry sustainability. Highly amount of Glycerol converted to more benefit material Glycerol carbonate (GC) was one of the most allurement compound derived from glycerol by transesterification of glycerol with dimethyl carbonate (DMC). Various parameters have highly impact on transesterification was investigated like catalyst loading (1-5) %wt., molar ratio of DMC: glycerol (5:1 – 1:1), reaction time (30 - 150) min and temperature (40 – 80) ᴼC. The Optimum glycerol carbonate yie
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Dental caries (tooth decay) is one of the most prevalent infectious disease and although of multifactorial origin, Streptococcus mutans is considered the principal pathogen in its development (i.e. bacterial processes damage hard tooth structure (enamel, dentine and cementum), producing dental cavities (holes in the teeth). The bactericidal properties of the Nd:YAG laser has been researched analyzing its use in caries prevention and bacterial reduction. One hundred twenty five samples were collected from carious teeth and isolated bacteria were diagnosed using microscopic examination, culture, biochemical tests, and Api 20 strep system. The results of this study showed that a noticeable decrease in the viability of Streptococcus mutans w
... Show MoreObjective: The current study aimed identifying the impact of rehabilitative exercises combined with ultrasonic waves on reducing pain in people with carpal tunnel compression and determining how these activities affect range of motion of the upper limb for those suffering from carpal tunnel compression. Research methodology: With pre- and post-tests, the researchers employed the experimental method in the form of two equal groups, the experimental and the control. The scientific community and sample are among the priorities that fall on the researcher, so The scientific community is determined by those suffering from carpal tunnel compression, numbering (14) patients. (12) Patients were approved and two were excluded from the resear
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In this research, a new 1, 3, 4-Thiadiazole derivatives have been synthesized by many heterocyclic reactions. Starting from (2, 5 – dimercapto -1, 3, 4-Thiadiazole) a variety of derivatives have been synthesis. Compound (1) was synthesized by the reaction of hydrazine hydrate with carbon disulphide in absolute ethanol. The compound (1) was reacted with 1, 2-dibromoethane in presence of alkali ethanol to give the compound (2). The compound (3) was formed from the reaction of compound (2) with hydrazine hydrate. Schiff base (4) was obtained by reacting of compound (3) with the compound (p-hydroxybenzaldehyde) in absolute ethanol. A variety of phenolic Schiff base (Methylolic, Etheric, and Epoxy) derivatives have been synthesized. Methylol
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