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.
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Pseudomonas aeruginosa gram-negative, bacilli and facultative aerobic, P. aeruginosa cause cystic fibrosis patients, wounds, burns, and immunodeficienct patients, that have many virulence factors such as pyocyanin , cytotoxic ,biofilm formation and motility, Eighty-eight isolates belonging to P. aeruginosa were collected including the 66 clinical isolates obtained from different hospitals in Baghdad and were from different sources and 22 environmental isolates from previous studies of soil near oil fields. Microscopical and cultural characteristics were studied and diagnosed using biochemical tests, VITEC device, their ability to adhere to non-living (Polystyrene), living cell line (A549) and cytotoxicity of bacterial filtrate
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protein oxidation through oxidative stress, which represents the overall status of the protein in the cell/tissue. Due to their increased levels of AOPPs were reported during T2DM. The aim of this study was to assess AOPP level in T2DM subjects with foot ulcer (DFU) and explore its correlation with infection. Type 2 diabetic patients (n=108) and healthy subjects (n=25) were enrolled in this study. The T2DM group was subdivided to diabetic patients without complications (n=25) and eighty-three (83) of them have diabetic foot. They were sub- grouped into two groups according to presence Osteomyelitis and abscess, and in reliance on medical analysis of WBC count and CRP. Group of diabetic without superficial or deep ulcer and no osteomyelitis
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Background Cadmium (Cd), one of the most abundant heavy metals, is extremely toxic to both humans and animals. hIt is well known that zinc (Zn) administration reduces Cd-induced toxicity and that metallothioneins can have a protective effect in biological systems to mitigate Cd toxicity. Objective The aim of the current study to determine if Zn administration affected the induction of MT-1 and MT-2 in the liver tissue in mice exposed to Cd. Materials and methods Metallothionein protein (MT) level in the tissue of male mice were detected using the anion -exchang high-performance liquid chromatography coupled (HPLC)assay and immunohistochemical staining. Results Single treatment to zinc or cadmium increase the level of MT in the liver, but zi
... Show MoreDuring infection, T. gondii disseminates by the circulatory system and establishes chronic infection in several organs. Almost third of humans, immunosuppressed individuals such as HIV/AIDS patients, cancer patients, and organ transplant recipients are exposed to toxoplasmosis. Therefore, the study aimed to investigate the possibility that Toxoplasma infection could be a risk factor for COVID-19 patients and its possible correlation with C-reactive protein and ferritin. Overall 220 patients referred to the Al Furat General Hospital, Baghdad, Iraq were enrolled from 2020–2021. All serum samples were tested for T. gondii immunoglobulins (IgG and IgM) antibodies, C-reactive protein and ferritin levels. In patients with COVID-19, the results
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Thyroid hormones (TH) regulate the metabolic processes required for normal development and growth; also, to organizemetabolism in adults, any defect in thyroid function leads to abnormality in thyroid hormones level. The current study hasbeen designed to find the relationship between retinol-binding protein-4 and progranulin in the serum of Iraqi women withhypothyroidism and hyperthyroidism, also, to study whether these patients are exposed to a risk of developing diabetes mellitus,and PGRN may be a biomarker in detection early stage of diabetes mellitus.Materials and Methods: in this study, serum samples were obtained from 50 Iraqis women patients, [25 patients withhypothyroidism (G2) and 25 patients with hyperthyroidism (G3)] in addition
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Background: Peripheral giant cell lesion (PGCL) and central giant cell lesion (CGCL) of the jaws have a distinct clinical behavior.Giant cell tumour (GCT) is a benign locally aggressive neoplasm affects the long bones. Both lesions are characterized histologically by multinucleated giant cells in a background of ovoid to spindle-shaped mesenchymal cells. The WW domain-containing oxidoreductase (WWOX) gene is located at 16q23.1–16q23.2, a region that spans the second most common human fragile site, FRA16D, at 16q23.2.The Ki-67 antigen is a nuclear protein that is associated with and may be necessary for cellular proliferation.Ki-67 protein is present during all active phases of the cell cycle (G1, S, G2, and mitosis), but is absent fr
... Show MoreThe purpose of this study was to know the effect of adding different percentages of soy protein a humidifier to mix Sarcoma Barker component of the beef mixture Alkhalt added Alber elderly chicken with five ratios were manufacturing Alberkr discs for sensory evaluation
Background: Oral squamous cell carcinoma (OSCC) is the most prevalent malignant neoplasm of the oral cavity and constitutes a major health problem in developing. In the last 30 years, the 5-year survival rate of patients with oral SCC has not improved despite advance in diagnostic techniques. To improve early diagnosis for this deadly disease, new biological markers are needed. HOX genes encode homeodomain-containing transcription factors involved in the regulation of cellular proliferation and differentiation during embryogenesis. HOX gene expression has been described in several adult tissues, where they performed important roles in maintaining homeostasis. Few studies have suggested that HOXA1 plays a role in tumorigenesis. Besides bein
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