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.
The fresh water green alga Chlorella vulgaris was selected to study its ability to degrade one of polycyclic aromatic hydrocarbon (PAHs)known is anthracene (ANT). ANT is widely used in artificial products such as wood preservatives; insecticides; dye and coating materials. This algae was cultivated in CH-10 medium under constant laboratory conditions and exposed to different concentrations (1, 3, and 5 mg/l) of anthracene for 3, 5, 7, 9 and 15 days, with the concentration of ANT measured by high performance liquid chromatographic analysis (HPLC). The results showed that C.vulgaris has high ability to reduce anthracene to 80% at 1mg/Lcon centration after 3 days and 100% after 5 days, while at 3 and 5 mg / L concent
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The present study introduces description of a new species of genus Arboridia Zakhvaticin 1946, based on a large collection of Cicadellids. External morphological characters particularly male genitalia were discussed and illustrated. The genus Arboridia Zalchvatkiia (Typhlocybinae: Erythroneurini) contains small slender, fragil and attractively coloured and patterned leafhoppers. It was erected by Zakhvatkin in 1946 (Zalchvatkin, 1946). The overall length of adults ranges from 2.5 to 3.4 mm. Members of this genus can be recognized by inner apical cell of forewing which is long with oblique base; Cu confluent with this base at a point near the middle of the length of inner apical cell; two prominent circular deep brown spots on vertex (Zal
... Show MoreThe study was conducted to evaluate the anti fungal activity of
water and alcohol ic extracts (cold and hot) and the crude alkaloid
extracts of leaves, seeds and roots of Zygophyllum fabago plant against a standard isolate of C. albicans and an isolate of A. jlavus which was proved to produce aflatoxins. Investigation of presence of active antimicrobial compounds in this plant parts was carried out, crude al kaloid extract was also separated using TLC technique. The antifungal activity of all these extracts was estimated against the two fungi. Results showed variation in anti fungal
... Show MoreThe present study provides the first record species of the genus Lithobius Leach, 1814, L. ferganensis (Trotzina, 1894) which was collected from the middle of Iraq. A detailed explanation of the morphology and the diagnostic characters of specimens of both sexes is provided.
Background: Periodontal diseases are inflammatory disorders caused by the accumulation of oral biofilm and the host response to this accumulation which characterized by exaggerated leukocytes and neutrophils attraction to the sites of inflammation by chemoattractants which are a very important part of the pathogenesis of periodontal diseases. This study aimed to determine and compare the clinical periodontal parameters and the leukocyte cell types in the peripheral blood between patients with gingivitis and periodontitis with different severities compared to healthy controls. Materials and methods: This study included 150 male subjects aged between 35-50 years. They were divided into three groups: gingivitis group (n=30), periodontitis p
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The present study was conducted to determine the optimum conditions required for lipase enzyme activity extracted from germinated sunflower seeds, including temperature, pH, agitation, time of incubation, enzyme concentration, substrate type, and concentrations of mineral salts and EDTA. Optimum pH, temperature and time of incubation required for lipase stability were also determined. The results showede optimum lipase activity (3.251U/ml) wasund at 30 ÌŠC and pH 7 after 20 minutes of incubation when using 1 ml lipase enzyme with 0.02 ml of CaCl2 (10 mM) at 100 rpm of agitation and in the presence of olive oil as the substrate for enzyme reaction. EDTA appeared to have inhibitory effects, while Ca+2 and Mg+2 have stimulatory effec
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New schiff bases series (VIII) a-e and 1,3-thiazolidin-4-one derivatives (IX) a-e containing the 1,2,4-triazole and 1,3,4-thiazazole rings were synthesized and screening their biological activities. These compounds were identified via Fourier transform infrared (FT-IR) spectra, some via Proton nuclear magnetic resonance (1H-NMR) and mass spectra. The biological results indicated that all of these compounds did not reveal antibacterial effectiveness against (Escherichia coli and Klebsiella species) (G-). Some of these compounds showed moderate antibacterial activity against (Staphylococcus aureus, and Staphylococcus epidermidis) (G+), and all compounds exhibited moderate activity against Candida albicans.
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Inflammatory bowel disease (IBD) a chronic inflammatory condition of the gastrointestinal tract characterized by alternating episodes of clinical relapse and remission (Wallace, 2014), with Crohn's disease (CD) and ulcerative colitis (UC) being the two primary subtypes (Souza, 2016). The present study was achieved in Baghdad teaching hospital from November 2021 to June 2022 to prove if the diagnostic marker of Crohn’s disease (CD) (IL-6, CRP and Calprotectin( are rises in saliva as in sera and if there are significant difference in the levels of those markers between the Crohn’s disease (CD)group and control group.
The recent study was measured the levels of C-reactive protein (CRP), Interleukin-6 (IL6
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Celery and coriander are vastly applied in modern medicine and traditionally because various medicinal and nutritional benefits depend on their medicinal characteristics. The study aimed to detect, isolate and compare extracts contents of phenolic acids (caffeic and p-coumaric acids) in ethyl acetate fraction of fresh and dry aerial parts of coriander (Coriandrum sativum L.) and celery (Apium graveolens L.) of the Apiaceae family. The extraction of these constituents was carried out by maceration method using 70% ethanol and fractionation was done by using petroleum ether, and ethyl acetate. The existence of caffeic and p-coumaric acids in aerial part extracts of two plants was identified by thin-layer chromatography (TLC) and high-
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