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.
Objective: Comprehending microbial diversity and antibiotic resistance patterns is essential for efficient treatment protocols. This study sought to determine the incidence of bacterial and fungal pathogens responsible for burn and wound infections and their antibiotic susceptibility profiles. Methods: This cross-sectional study involved 140 patients with burn or wound infections. Sterile swabs and pus aspiration were employed to collect samples, which were subsequently processed using standard microbiological procedures. Antibiotic resistance was determined using the Kirby-Bauer disc diffusion method, following Clinical and Laboratory Standards Institute (CLSI) guidelines. Data was analysed using IBM SPSS version 25.0, and the Chi-
... Show MoreBackground: Colorectal cancer (CRC) is a major killer, and nonalcoholic fatty liver disease (NAFLD) affects almost one-quarter of the global population. The aim of the study is to Investigate the correlation between nonalcoholic fatty liver disease (NAFLD) and an increased risk of colorectal cancer (CRC), as well as the relationship between liver function enzymes and specific serum biomarkers in CRC patients with NAFLD. Methods: A case-control study involving 60 participants was conducted from February to August 2022. The patients with colon cancer were examined at Baghdad Medical City's Al-Amal Hospital for Radiation and Nuclear Medicine and Oncology Teaching Hospital, and blood samples were taken. Thirty patients with NAFLD who ha
... Show MorePurpose: This study aimed to assess the thickness of alveolar bone of maxillary and mandibular incisors from orthodontics perspective. Materials and Method: A total of 73 Cone beam computed tomography for Iraqi patients (47 females and 26 males) were included in this study. The selected images were captured and imported to AutoCAD database software to perform the measurement. To measure alveolar bone thickness, a reference line was drawn through the long axis of each incisor, from the incisal edge to the root apex. Then, labial and lingual/palatal perpendicular lines were drawn to the reference line at 3, 6, and 9mm apically from the cemento-enamel junction (CEJ). Results: The buccal bone is generally thinner than the lingual/palata
... Show MoreThe science of jurisprudence is one of the legal sciences that scholars have been interested in since the first centuries of Islam, and they wrote many books about it, and the science of manuscripts is considered one of the scientific achievements in which many scholars emerged, including Imam Al-Samaani, so I chose this manuscript related to Istism to clarify its concept and meaning for all people, The student (Ali Ahmed Abdel-Aziz Sheikh Hamad) preceded me in the investigation of part of the book, and it was facilitated for me, by the grace of God Almighty, to investigate the issue (if one of the Muslim spouses apostatized and one of the infidel spouses converted to Islam until the end of the issue of if the two spouses were taken capt
... Show MoreGranular carbon can be used after conventional filtration of suspended matter or, as a combination of filtration - adsorption medium. The choice of equipment depends on the severity of the organic removal problem, the availability of existing equipment, and the desired improvement of adsorption condition.
Design calculations on dechlorination by granular - carbon filters considering the effects of flow rate, pH , contact time, head loss and bed expansion in backwashing , particle size, and physical characteristics were considered assuming the absence of bacteria or any organic interface .
CUPPER(||)AND MERCURY (||)Complexes WITH SCHIFF BASE LIGAND FROM BENZIDIN WITH ISATIN AND BENZOIN:SYNTHESIS,SPECTRAL CHARACTERIZATION, THERMAL STUDIES AND BIOLOGICAL ACTIVITIES
Background and objectives Uropathogenic E.coli (UPEC) are major causative agents of urinary tract infection (UTIs), they often possess strong biofilm-forming abilities, and capable of resisting many antibiotics, making catheter associated UTIs (CAUTIs) difficult to treat. Essential oils such as tea tree oil (TTO) have emerged as natural alternatives to antibiotics. This study aimed to evaluate the antibacterial, adhesion, and biofilm-forming efficacy of TTO against UPEC, while analyzing its effect on the gene expression of csgA gene and determining its cytotoxicity. Methods Four UPEC isolates collected from different UTIs patients from Baghdad Province. The Antibacterial activity of TTO evaluated using agar wells diffusion assay and micro
... Show More