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.
Background: Wound healing is a complex dynamical interaction between various cell types, the extracellular matrix, cytokines, and growth factors. osteoponetin is a substance that acts as an anti-inflammatory. Aims of study: The study was designed to identify the role of local exogenous applications of osteopontin on wound healing (in cheek skin). Materials and methods: Thirty adult male albino rats weighting an average of (250-300gm) used in this study, incisional wounds were made in the skin of the cheek of rat and they were divided into the following groups: A-Control group: 15 rats treated with 1µ l of normal saline B-Experimental groups: 15 rats treated with topical application of 1µl osteopontin. The scarification of animals we
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Background: For various reasons, inguinal hernia repair under local anaesthesia is not well accepted to both patients and surgeons. The patients fear from pain and surgeons need full relaxation and co-operation to do successful hernia repairMethods: purpose of this study is to evaluate the effectiveness of local anaesthesia in inguinal hernia repair.prospective study was made from January 2011-0ctober 2013 , on a total of 50 patients with inguinal hernia operated on under local anaesthesia. Patients were selected primarily on the basis of their willingness to accept the procedure after the technique was described to them.Results: In this study 50 patient and 58 herniorrhaphies done for them during a period of about 34months were evaluate
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In this research study theory to find the stress and emotion gases in the glass as a result of exposure to pulses of the laser beam has been the study using vehicles three major on-system axes cylindrical (r, 0, z), where I took three models of glass silica glass soda glass fused and shedtwo types of lasers where the study showed that the thermal stresses and emotions ...
Genome sequencing has significantly improved the understanding of HIV and AIDS through accurate data on viral transmission, evolution and anti-therapeutic processes. Deep learning algorithms, like the Fined-Tuned Gradient Descent Fused Multi-Kernal Convolutional Neural Network (FGD-MCNN), can predict strain behaviour and evaluate complex patterns. Using genotypic-phenotypic data obtained from the Stanford University HIV Drug Resistance Database, the FGD-MCNN created three files covering various antiretroviral medications for HIV predictions and drug resistance. These files include PIs, NRTIs and NNRTIs. FGD-MCNNs classify genetic sequences as vulnerable or resistant to antiretroviral drugs by analyzing chromosomal information and id
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Background: Pleural effusion is a common clinical
problem.
Objective: The aim of the study was to evaluate the
diagnostic utility of Carcino embryonic antigen
(CEA), CA 15- 3, and alpha-feto protein ( AFP ) as
a tumor markers in serum and pleural effusion and
evaluate the value of combining them as a diagnostic
tools that are complementary to cytology in the
diagnosis of malignancies .
Methods: Forty patients (18 malignant and 22 benign
pleural effusion) were included in this study .The
serum and effusion levels of CEA, CA 15 – 3 and
AFP were measured using immunoradiometric assay
Results: from the 40 effusions studied 26 were
exudates and 14 were transudates. The level of
pleural effusions
In this work, thermodynamic efficiency of individual cell and stack of cells (two cells) has been computed by studying the variation of voltage produced during an operation time of 30 min as a result of the affected parameters:- stoichiometric feed ratio, flow field design on single cell and feed distribution on stack of cells. The experiments were carried out by using two cells, one with serpentine flow field and the other with spiral flow field. These cells were fed with hydrogen and oxygen at low volumetric flow rates from 1 to 2 ml/sec and stoichiometric ratios of fuel (H2) to oxidant (O2) as 1:2, 1:1 and 2:1 respectively. The results showed that
... Show MoreBackground: Fertility plays great role in animal reproduction since high quality semen improves sheep industry reproduction. The current worldwide data revealed the closely related of CNP to reproductive function of rams. Aims: Evaluation the effect of CNP on cooled sperms using the traditional and molecular assays. Methods: Totally, 20 testicular samples were collected, processed to obtain the semen samples and divided into two parts; one treated with the suitable dose of CNP and the otherserved as a control. Sperm samples of both groups were cooled for 3 days and tested at 0 0h, 24h, 48h and 72h. Results: The findings revealed that the suitable dose of CNP-treated sperms was 0.0110-13. Values individual motility, live sperm
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This study was carried out to investigate the effect of dietary supplementation with different levels of parsley on semen quality of local Iraqi ganders. A total of thirty-two local ganders were used in this study during the period from the beginning of February to the end of April. The ganders were allocated for 4 treatment groups containing 8 ganders each. The treatment groups were as follows: Control diet (free from parsley); T1: Control diet + 80 g/d parsley; T2: Control diet + 160 g/d parsley; T3: Control diet + 240 g/d parsley. Semen samples were collected twice a week, fortnightly, from each gander by the dorsal-abdominal message method. The first semen collection was used to evaluate semen volume, sperm concentration, live in total
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