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.
Ticks (Acari: Ixodidae) are ectoparasites that infest livestock in every geographic region of the world and are vectors of several viral, bacterial, and protozoan pathogens to both animals and humans. There is little information is available is about tick presence in Buffalo Bubalus bubalis (Linnaeus, 1758) (Artiodactyla, Bovidae) in Iraq. The current study determined the species of ticks parasitizing Buffalo in some central and southern regions included: Baghdad (Al Fathelia), Karbala (Al-Hussainia), Wasit (Kut and Al-Suwairah), Al-Qadisia (Al- Diwaniyah, Al- Saniya, Al-Mihnawea, and Afak), Thi Qar (Al-Nasiriyah and Al-chibayish), Missan (Amara and Qalaat Salih) and Basrah (Al-Haretha, Al-Madena and Al-Deer). A total of 150 Buffal
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In this work, the extraction of glycyrrhizin from Licorice using bulk liquid membrane technique was developed and optimized. The effect of various parameters such as pH of stripping and donor solutions, temperature, stirring speed and kinetic parameters were investigated. Moreover, to study the impact of the polarity of membrane solvent, two types of extraction solvents were used as a membrane solvent: n-Hexane was used as a non-polar solvent and 1-Hexanol was as a polar solvent. The optimum extraction condition was found (95.53%) using 1-Hexanol, rotating speed was 400 rpm, and pH of the acceptor and donor solutions were 8 and 5.5, respectively. The reaction kinetics constants ( and ) for the transport of glycyrrhizin from the donor pha
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Bis-anthraquinones with a unique molecular backbone, (+)-2,2’-epicytoskyrin A (epi) and (+)-1,1′-bislunatin (bis), was produced by endophytic fungi Diaporthe sp GNBP-10 associated with Gambir plant (Uncaria gambier). Epi and bis possess robust antimicrobial activity toward various pathogens. This study focus on knowing the optimum condition of epi and bis production from Diaporthe sp GNBP-10. A series of culture media with various nutrient compositions was investigated in epi and bis production. The content of epi and bis was determined by measuring the area under the curve from TLC-densitometric (scanner) experiment. The linear regression analysis was then applied to obtain the results. The optimi
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Fabrication of porous clay refractory insulating specimens from Iraqi kaolin with different percentage of Expanded Polystyrene (EPS) waste crumbs additions were investigated. After mixing and forming by hand molding, the specimens was dried and fired at 1300 oC. The structural, physical, mechanical and thermal properties of the refractory insulating products were measured. Maximum addition of EPS (1.25 wt%) lead to reduce the linear shrinkage to less than 1.7% and increased apparent porosity up to 50 %. As well as, the density, Modulus of rupture and thermal conductivity were reduced to 1.39 g/cm3, 4.1 MPa and 0.21 W/m.K, respectively. The final outcome, addition of EPS showed good results in the formation of pores without distorting the
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Background: Neonatal septicemia is a significant cause of morbidity and mortality worldwide especially so in developing countries. To reduce the mortality caused by neonatal septicemia, it became vital to diagnose it as soon as possible and treat with administration of appropriate antibiotics.Objective: To study the relationship between themicroorganisms isolated from septicemic neonates with place of delivery.Patients and Methods: Blood sample was obtained from 76 neonates (50 of them are born in Baghdad teaching hospital (Inborn), 26 of the babies are born at home or in Al-Elwya teaching hospital (out born) ,the laboratory diagnosis for the out born patients done in the same hospital(Al-Elwya teaching hospital .The aged of the neo
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Levan is an exopolysaccharide produced by various microorganisms and has a variety of applications. In this research, the aim was to demonstrate the biological activity of levan which produced from B. phenoliresistens KX139300. These were done via study the antioxidant, anti-inflammatory, anticancer and antileishmanial activities in vitro. The antioxidant levan was shown 80.9% activity at 1250 µg/mL concentration. The efficient anti-inflammatory activity of 88% protein inhibition was noticed with levan concentration at 35 µg/mL. The cytotoxic activity of levan at 2500 µg/mL concentration showed a maximum cytotoxic effect on L20B cell line and promastigotes of Leishmani tropica. Levan has dose-dependent anticancer and antileishman
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Portulacaria afra is a small succulent tree, previously belonging to the Portulacaceae family, but with further studies, the plant transferred to the Didieracea family. P. afra was used as an ornamental, vegetable, and ethnomedicinal plant. Uses of the plant by rural South Africans to treat chronic skin conditions and rashes, alleviate exhaustion, and aid in treating TB and diarrhea have been documented in folklore. According to pharmaceutical research, plant extracts off er a wide range of remedial outcomes, such as antidiabetic, antifungal, antibacterial, anticancer, antioxidant, and anti-infl ammatory. The study aims to determine some bioactive constituents responsible for pharmacological activities and traditional usefulness. Th
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The problem of water scarcity is becoming common in many parts of the world, to overcome part of this problem proper management of water and an efficient irrigation system are needed. Irrigation with a buried vertical ceramic pipe is known as a very effective in the management of irrigation water. The two- dimensional transient flow of water from a buried vertical ceramic pipe through homogenous porous media is simulated numerically using the HYDRUS/2D software. Different values of pipe lengths and hydraulic conductivity were selected. In addition, different values of initial volumetric soil water content were assumed in this simulation as initial conditions. Different value
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2-hydrazinylbenzo[d]thiazole compound [1] is produced from reaction of 2-mercapto-benzothiazole with hydrazine hydride in ethanol. Compound [1] reacted with maleic anhydride in DMF to produce (Z)-4-(2-(benzo[d] thiazol-2yl) hydrazinyl)-4-oxobut-2-enoic acid [compound (2)]. While the treatment of compound [2] with the ammonium persulfate (NH4)2S2O8 (as the initiator) in order to produce compound [3], then compound [3] reacted with thionyl chloride in benzene to produce compound [4], finally compound [4] reaction with various drugs: cephalexin, amoxicillin, sulfamethizole, elecoxib obtained polymers [5–8]. The structure of synthesized compounds identified by spectral data: fourier transform infrared (FTIR) and proton nuclear magneti
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