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.
To investigate the role of IL-6 and IL-8 in the immune-regulatory mechanisms involved in the recurrent spontaneous abortion of the first trimester of pregnancy. Serum level of IL-6 and IL-8 were determined in 25 women of age (20-35) years who had a spontaneous abortion of unknown aetiology during the first trimester of pregnancy .They were compared with the corresponding levels of 20 pregnant and non-pregnant women as control groups .cytokine levels were measured by (ELISA) technique .The women with spontaneous abortion had highly significant (P < 0.01) increased serum level of IL-8 and highly significant (P < 0.01 ) decreased level of IL-6 compared to those with normal pregnant and non-pregnant women. The results of this study ma
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Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen and a model bacterium for studying virulence and bacterial social traits. While it can be isolated in low numbers from a wide variety of environments including soil and water, it can readily be found in almost any human/animal-impacted environment. It is a major cause of illness and death in humans with immunosuppressive and chronic conditions, and infections in these patients are difficult to treat due to a number of antibiotic resistance mechanisms and the organism’s propensity to form multicellular biofilms. One hundred twenty clinical samples and forty hospital environmental samples (various sources) were collected from hospitals in Baghdad city during the period from Oc
... Show MoreTwo Schiff base ligands L1 and L2 have been obtained by condensation of salicylaldehyde respectively with leucylalanine and glycylglycine then their complexes with Zn(II)were prepared and characterized by elemental analyses , conductivity measurement , IR and UV-Vis .The molar conductance measurement indicated that the Zn(II) complexes are 1:1 non-electrolytes. The IR data demonstrated that the tetradentate binding of the ligands L1 and L2 . The in vitro biological screening effect of the investigated compounds have been tested against the bacterial species Staphlococcus aureus, Escherichia coil , Klebsiella pneumaniae, Proteus vulgaris and Pseudomonas aeruginosa by the disc diffusion method . A comparative study of inhibition values of
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In this study, the use of non-thermal plasma theory to remove toxic gases emitted from a vehicle was experimentally investigated. A non-thermal plasma reactor was constructed in the form of a cylindrical tube made of Pyrex glass. Two stainless steel rods were placed inside the tube to generate electric discharge and plasma condition, by connecting with a high voltage power supply (up to 40 kV). The reactor was used to remove the contaminants of a 1.25-liter 4-cylinder engine at ambient conditions. Several tests have been carried out for a ranging speed from 750 to 4,500 rpm of the engine and varying voltages from 0 to 32 kV. The gases entering the reactor were examined by a gas analyzer and the gases concentration ratio
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Iraqi crude Atmospheric residual fraction supplied from al-Dura refinery was treated to remove metals contaminants by solvent extraction method, with various hydrocarbon solvents and concentrations. The extraction method using three different type solvent (n-hexane, n-heptane, and light naphtha) were found to be effective for removal of oil-soluble metals from heavy atmospheric residual fraction. Different solvents with using three different hydrocarbon solvents (n-hexane, n-heptane, and light naphtha) .different variables were studied solvent/oil ratios (4/1, 8/1, 10/1, 12/1, and 15/1), different intervals of perceptual (15, 30-60, 90 and 120 min) and different temperature (30, 45, 60 and 90 °C) were used. The metals removal perce
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Acinetobacter baumannii received attention for its multi-drug resistant associated with many severe infections and outbreaks in clinical environment. The aims of the study are to investigate the antibiotic susceptibility profile of clinically isolated A. baumannii, biofilm production, and the efficiency of Low Frequency Ultrasound (LFU) and honey to attenuate biofilm production. A total of 100 samples were taken from different sources from Baghdad hospitals. The susceptibility patterns revealed the percentage of pan drug resistant (PDR) isolates were 1.5 %, 72.7 % were extended drug resistant (XDR), 16.7 % were multidrug resistant (MDR), and 9.1 % were non MDR and sensitive to most antibiotics used. The ability to form
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ABSTRACT
The study aims to identify the level of health services provided in private suites to government hospitals from the perspective of the recipi
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Nitrogen dioxide NO2 is one of the most dangerous contaminant in the air, its toxic gas that cause disturbing respiratory effects, most of it emitted from industrial sources especially from the stack of power plants and oil refineries. In this study Gaussian equations modelled by Matlab program to state the effect of pollutant NO2 gas on area around Durra refinery, this program also evaluate some elements such as wind and stability and its effect on stacks height. Data used in this study is the amount of fuel oil and fuel gas burn inside refinery at a year 2017. Hourly April month data chosen as a case study because it’s unsteady month. After evaluate emission rate of the all fuel and calculate exit velocity from
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