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.
Objectives: With the advent of ongoing novel modalities toward the treatment of human epidermal growth factor receptor 2 (HER2)/NEU - positive malignancies, the serious side effects of chemoradiotherapy have been minimized. Hence, this study was conducted to identify the patterns of immunohistochemical expression of the promising therapeutic target (HER2/NEU) among Iraqi patients with medulloblastoma in an attempt to provide basic histological information’s that would help in future clinical researches.Materials and Methods: In this retrospective study, 42 formalin - fixed paraffin - embedded tissue blocks represent cases of surgically removed medulloblastomas were retrieved from the archived materials in a specialized surgical ho
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Assessment of Salivary Macrophage Inflammatory Protein-1 Alpha Level in Different Stages of Periodontitis, Riyam Muthanna Muhammed*, Hadeel Mazin Akram
Multiple sclerosis (MS) is a neurodegenerative disorder with various clinical types. Glial fibrillary acidic protein (GFAP) is significantly elevated in the cerebrospinal fluid (CSF) of MS patients compared with that of healthy controls. The aim of this study is to evaluate serum levels of GFAP in relation to disease activity in relapsing-remitting MS patients and to compare them with those of healthy controls. Method: This study involved 58 MS patients of relapsing-remitting MS (RRMS) type, 22 in an active stage of the disease and 36 in remission, and 50 healthy individuals as age- and sex-matched controls. Blood samples were taken from the patients at the MS Clinic of the Baghdad Teaching Hospital, and the serum levels of GFAP were determ
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Background: This research identified Streptococci spp. depending on culture, biochemistry, the VITEK technique, ability to produce biofilms, and antibiotic resistance. Aim: The goal of this study was to perform microbiological procedures to evaluate the qualitative qualities of mozzarella cheese against infective Streptococci using microbiological care. Methods: Sixty (60) mozzarella cheese samples were brought from diverse markets in Baghdad from October 2023 to December 2023 at the Zoonoses Research Unit and Veterinary Public Health Department, Veterinary Medicine College, University of Baghdad. Culture of samples on agar (MacConkey and blood) and aerobically incubated at 37°C for 48 hours. Gram staining purified colonies to
... Show MoreBackground: Male infertility is a global concern and it tends to increase due to miscellaneous factors, such as environmental toxins and genetic and lifestyle choices. The aryl hydrocarbon receptor (AHR) has recently attracted attention due to its involvement in male infertility mechanisms and impact on sperm production and function. AHR, a versatile receptor expressed in various tissues, including the testes, regulates the genes involved in spermatogenesis. AHR activation is associated with cell cycle regulation and chromatin condensation during spermatogenesis. Objectives: This study aimed to investigate the influence of AHR activation on blood-testis barrier (BTB) integrity, focusing on the role of tight junction protein-1 (TJP1)
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16S rRNA gene sequence examination is an effective instrument for characterization of new pathogens in clinical specimens. Akey component of colonization, biofilm formation, and protection of the pragmatic human pathogen Pseudomonasaeruginosais the biosynthesis of the exopolysaccharide Psl.Extracellular polysaccharides,biofilm, are secreted by microorganisms into the neighboring environment and are significant for surface attachment and keeping structural safety within biofilms.Biofilm production is an important technique for the survival of P. aeruginosa,and its association with antimicrobial resistance represents a defy for patient therapeutics. The aim of the current research is to assess the antibiotic resistance manner and distribution
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Chemical analysis for evaluation of Nigella sativa L. (black cumin) seeds showed a composition of Fat 39% ; Protein 28% ; Carbohydrate 21% ; Moisture 6% and Ash 4.5% . It was found that the black seed contains the following mineral element : Magnesium 0.26 gm /100gm seed ; Calcium 0.25 gm /100gm seed and Iron 25 ?g / gm /100gm seed ; zinc 4.51?g /gm /100gm seed and Copper 3.60 ?g /gm /100gm seed. The analysis also showed that mineral element I. e. ; lead ; Cobalt ; Nickel ; Chrom ; Cadmium and Aresenic are not present . It was found that the fat of the black seed contains the following fatty acids : Myristic 2.8%; Palmtic 16.6%; Stearic 0.8 % ; Oleic 13.79% ; Linoleic 64.2% and Arachidic 1.9% .
This research of using Feldspar in the production self compacting concrete (SCC) ( 5,10,15 )% as partial replacement by weight of cement .In this research some of fresh properties of SCC ( slump flow used V-funnel test and filling ability used ( U- box test ) for concrete mixes and also some of the harden properties of SCC ( compressive and flexural tests ). The research results showed that negative effect of Feldspar on the fresh properties of self compacting concrete but the positive effect of Feldspar on the harden properties of self compacting concrete .
In this study 100 samples were collected from infected children with acute and chronic tonsillitis who attended to Al-Yarmook Teaching Hospital (ENT consultation clinic) from 5/12/2013 to 1/3/2014. The result of laboratory culture was positive in 67 samples. Depending on their cultural, morphological and biochemical characterization of bacterial isolate of them were identified as (37.31%) belonged to Streptococcus pyogenes and the diagnosis is confirmed by the use of Remel Rapid STR System, (34.32%) belonged to S.parasanguinis, (11.94%) S.mitis, (11.94%) S.oralis and (4.47%) S.thoraltensis . Results confirmed that cup assay gave highest inhibition zone after 24 hrs compare with well diffusion methods for suspension of L.
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This research paper studies the use of an environmentally and not expensive method to degrade Orange G dye (OG) from the aqueous solution, where the extract of ficus leaves has been used to fabricate the green bimetallic iron/copper nanoparticles (G-Fe/Cu-NPs). The fabricated G‑Fe/Cu-NPs were characterized utilizing scanning electron microscopy, BET, atomic force microscopy, energy dispersive spectroscopy, Fourier-transform infrared spectroscopy and zeta potential. The rounded and shaped as like spherical nanoparticles were found for G-Fe/Cu‑NPs with the size ranged 32-59 nm and the surface area was 4.452 m2/g. Then the resultant nanoparticles were utilized as a Fenton-like oxidation catalyst. The degradation efficiency of
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