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.
ABSTRACT: Pathogenic bacteria responsible for the causation of many common diseases have been identified on currency notes. The present investigation was carried out on one hundred currency notes of all the denominations (50, 100, 250, 500 and 1000RY), obtained from different occupational mainly bus drivers, hawker street, vegetable vendor, restaurants and butchers and fish seller groups in Taiz city,Yemen. Identification and characterization revealed active participation of the following species of organisms in the ascending order of percentage as E. coli(50.28 %),Staphylococci aureus(14.04 %), Klebsiellaspp(4.39 %),proteus(4.39 %), salmonella(1.25 %), shigella(0.72 %), Coagulase negative staphylococcus(0.60 %), pseudomonas(0.50 %),
... Show MoreIntroduction and Aim: The pro-inflammatory cytokine IL-39, a member of the IL-12 family plays a key role in the inflammatory response by modulating immune cell activity and inflammation. A literature search shows no study undertaken for the effect of IL-39's on arthritis so far. Hence, the purpose of this study was to investigate the role of IL-39 in rheumatoid arthritis. Materials and Methods: This study involved 80 patients with rheumatoid arthritis registered at the Rheumatology Clinic at Baghdad teaching hospital. The patients were divided into three groups based on treatments received. Group 1 included patients who were not on any treatment for arthritis, Group 2 with patients on hydroxychloroquine and or prednisone treatment,
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The electrospun nanofibers membranes have gained considerable interest in water filtration applications. In this work, the fabrication and characterization of the electrospun polyacrylonitrile-based nonwoven nanofibers membrane are reported. Then, the membrane's performance and antifouling properties were evaluated in removing emulsified oil using a cross flow filtration system. The membranes were fabricated with different polyacrylonitrile (PAN) concentrations (8, 11, and 14 wt. %) in N, N-Dimethylformamide (DMF) solvent resulted in various average fiber sizes, porosity, contact angle, permeability, oil rejection, and antifouling properties. Analyses of surface morphology of the fabricated membranes before and after oil removal revealed
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Endophytic bacteria produced analogous secondary metabolites of their hosts. Similarly, the ability to generate antioxidants is not an exception. Dragon scales (Pyrrosia piloselloides), an epiphytic plant of the Polypodiaceae family, are frequently overlooked. This research aims to isolate antioxidant-producing bacteria from dragon-scale fern leaves. The antioxidant activities were tested after the extraction procedure using ethanolic extract. Bacteria were characterized and selected as candidates for antioxidant production by screening for the production of total phenolic compounds. Antioxidant levels were determined utilizing the ABTS, FRAP, and DPPH techniques. The preliminary findings of the entire phenolic compound test rev
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The mucilage from the seeds of Lallemantia royleana family Labiatae was extracted and subjected to preformulation study for evaluation of its suitability for use as suspending agent. Furosemide suspensions were prepared using (1.5% w/v) of the extracted Lallemantia royleana mucilage, (1.5% w/v) chitosan and (0.35% w/v) xanthan gum. The mucilage was white in color and the average yield of dried mucilage obtained from L.royleana nutlets was 14 % w/w of the seeds used. It is sparingly soluble in water but swells in contact with it, giving a highly viscous solution. It is slightly acidic to neutral. It was found that the extracted natural mucilage of Lallemantia royleana exhibited a higher viscosity profil
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The present study aimed to identify the therapeutic evaluation of chitosan extracted from the fungus cushroom and pure chitosan on glucose and lipid profile in the blood of 35 male rabbits with hyperlipidemia induced experimentally by cholesterol. The tests included estimation of glucose levels, total cholesterol, triglycerides, high-density lipoproteins, low-density lipoproteins, and very low-density lipoproteins. hyperlipidemia was induced in the male rabbits used in the study which was administered orally with cholesterol 150mg/kg body weight for a week. rabbits were divided into seven groups: control, cholesterol, pure chitosan, mushroom chitosan, cholesterol and pure chitosan, cholesterol and mushroom chitosan and cholestero
... Show MoreThis study focused on the expression and regulation of BRCA1 in breast cancer cell lines compared to normal breast. BRCA1 transcript levels were assessed by real time quantitative polymerase chain reaction (RT-qPCR) in the cancer cell lines. Our data show overexpression of BRCA1 mRNA level in all the studied breast cancer cell lines: MCF-7, T47D, MDA-MB-231 and MDA-MB-468 along with Jurkat, leukemia T-lymphocyte, the positive control, relative to normal breast tissue. To investigate whether a positive or negative correlation exists between BRCA1 and the transcription factor E2F6, three different si-RNA specific for E2F6 were used to transfect the normal and cancerous breast cell lines. Interestingly, strong negative relationship was found b
... Show MoreThis study was carried out to investigate the cytogenetic effects of crude aqueous extract of Lycium barbarum on the roots tip of Allium cepa Using three concentration 125, 25, 50 mg/ ml for 2, 4, 6 hours treatment periods.This study were included some cytogenetic analysis such as mitotic index , phase index and chromosome aberration. The data showed that the treatment with 50mg/ml for 6huors led to reduce the mitotic index less than 50% . This reduction considered to have toxic and sublethal effect . These results revealed mutagenic potency by inducing differents type of chromosome aberration.
جريت دراسة مختبرية لمعرفة تأثير الزيت الطيار لقشور ثمار نبات النارنج الصفرC. aurantium تجاه النمو السطحي للفطريات Penicillium expansum، Aspergillus flavus و Fusarium oxysporum ، أظهرت نتائج الفعالية التثبيطية للزيت الطيار تأثيراً معنويا متفاوتاً في الفطريات المشمولة بالدراسة، إذ كان الزيت الطيار أكثر تأثيرأَ في الفطر P. expansum تلاه الفطر A. flavus ،في حين كان الفطر oxysporum F.أقل حساسية تجاه الزيت الطيار. بصورة عامة اظهر الزيت الطيار تأثيرا تثبيطيا
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