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.
A synthesis series of new heterocyclic derivatives (A2-A7) (pyrrole, pyridazine, oxazine and imidazol) derived from 4-acetyl-2,5-dichloro-1-(3,5-dinitrophenyl)-1H-pyrrole-3-carboxylate(A1) have been synthesised. Synthesis of compound (A2) by the reaction of starting material (A1) with hydroxyl amine hydrochloride in the presence of pyridine. Compound (A2) was reacted with hydrazine hydrate in dry benzene to give (A3) derivative. The compound )A3( deals with sodium nitrite to give diazonium salt, and the reaction diazonium salt with ethyl acetoacetate to produce compound (A4). To a mixture of compound (A4) and hydroxyl amine with sttired to yield (A5).Compound (A6) was prepared by reaction compound (A4) with thiosemicarbazide in presence
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The main aim of this study is to investigate the ability of four local entomopathogenic isolates Beauveria bassiana (Bals.) and Metarhizium anisopliae (Met.) to control the mosquito larvae in the lab. The results revealed that the isolate (MARD48) B .bassiana reduced the survival rate of the mosquito larvae to (80%) followed by the isolate M. anisopliae (MARD10) to (90%) in the first two days of treatment, and 60 and 66% respectively in the third day. The results also showed that the isolate B. bassiana (MARD48) killed 50% of the population (LC50) with the concentration 1×104 conidia/ml compared to 1×107 conidia/ml for the isolates B. bassiana (MARD14) and M. anisopliae (MARD10), and 1×108 conidia/ml for the isolate B .bassiana (MARD76).
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Background: Bone is essentially a highly vascular, living, constantly changing mineralized connective tissue. It is remarkable for its hardness, resilience and regenerative capacity, as well as its characteristic growth mechanisms. This study aimed to: 1. To evaluate the effect of bone morphogenetic protein7 (BMP7) on bone healing in artificially created intrabony defect in rabbits upper diastema, histologically. 2. To study the immunohistochemical expression of TGF-β3 and IGF-1R as bone formation markers in experimental and control groups during bone healing. Material and method: Forty male rabbits, was used in this study, 8 rabbits for each healing interval (3 days, 1,2 ,4 and 6 weeks). In each rabbit two bone holes were created on th
... Show MoreBackground: C-reactive protein (CRP) is an acute phase protein that its plasma levels increase after trauma or surgery so it is used as an indicator for the level of inflammation after surgery. The objective of this study is to investigate pre- and post-operative levels of CRP in three types of oral surgical interventions (Apicoectomy, Impaction, and Impacted teeth exposure). Materials and Methods: A total number of (48) healthy individuals aged (20-60) years who needed oral surgical intervention for either (removal of impacted third molars, exposure of an impacted canine, or Apicoectomy). A 4ml venous blood was obtained from each patient at two occasions (pre-operatively at the day of operation and post-operatively after 48 hours), then ce
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Globally, chronic kidney disease (CKD) has emerged as a significant public health concern, characterized by high rates of morbidity and mortality. To assess the risk of kidney damage, researchers have identified tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and fatty acid-binding protein-1 (FABP-1) as valuable biomarkers. This study aims to analyse the effectiveness of specific biomarkers in assessing CKD and its associated mechanisms in Iraqi patients. The study was conducted from December 2023 to May 2024. Ninety subjects, aged 48–65 years; including 60 patients with CKD (38 male and 22 female) attended the Baghdad Teaching Hospital/ Medical City/ Dialysis Unit- Baghdad, Iraq. In addition, 30 healthy people (15 male an
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Diabetes mellitus is a metabolic disorder categorized hyperglycemia resulting from defects in insulin secretion, insulin action or both. Protein tyrosine kinase (PTK) is an enzyme that catalyzes the transfer of phosphate groups from ATP to the tyrosine residues of many important proteins resulting in proteins phosphorylation. The aim of current study was to evaluate serum levels of protein tyrosine kinase enzyme and thyroid hormone (T3, T4and TSH) and to find the correlation between them in type 2 diabetes mellitus and diabetic nephropathy Iraqi patients. Methods: This study was conducted at The National Diabetes Center, Al-Mustansiriya University, Baghdad, Iraq and included 150 patients divided into three groups the first group included 50
... Show MoreBreast carcinoma is one of the greatest popular neoplasms in females. It is a major reason of demise in the world, and it is the first cancer in ranking diagnosed in Iraqi women. This study aimed to determine aminoacyltRAN-synthetase complex interacting multifunctional protein 1 and liver enzymes levels in Iraqi females with stage II breast malignance, and study the effect of chemotherapy (after surgery) on these markers. This study included 50 females patients with stage II breast malignance (before and after surgery and second dose of chemotherapy) attending the Oncology Teaching Hospital in Medical City/ Baghdad, in addition to 20 persons as controller group were chosen without any chronic diseases. Their ages ranged from (30-55) years.
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Rice is a major staple food for more than two thirds of the world population. Pathogenesis-related proteins-10 (PR10) have a range of 154 to 163 amino acid with molecular weight ~ 17 kDa. They are acidic and generally intracellular and cytosolic proteins accumulate in plants in response to biotic and abiotic stresses. In the present study, a PR10 gene and its corresponding protein were characterized in O. sativa, O. barthii, O. glaberrima, O. glumipatula, O. meridionalis, O. nivara, O. rufipogon and O. punctata. The results revealed a narrow range of variation at both DNA and protein levels in all examined species except O. glumipatula. The latter showed a relatively
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The interleukin-1 family has multifaceted roles in men٫s reproductive syste. Out of these is interleukin-1 receptor antagonist (IL-1RN) which exists in men gonads, and in case of infection and inflammatory process, its activity is increased. The current study aims to verify a possible linkage of Variable Number Tandem Repeat (VNTR) polymorphism of the IL-1RN gene with human men infertility. The study groups enrolled included 100 infertile men and 100 fertile and healthy men. Their seminal fluids were subjected to analysis. Also peripheral blood samples were collected for the assessment or detection of polymorphic Variable Number Tandem Repeats (VNTR) polymorphism of interleukin-1 receptor antagonist gene (IL-1RN). Two a
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