Aim of the present study is Identification of specific gene for GPCR using specific primers .and identification of difference in PCR analysis in patients with heart thrombosis and compared with healthy, Sequencing of PCR product regarding GPCR compared for all three subject, Identification the similarity of human GPCR with local strain of yeast fifty healthy control and fifty patients with thrombosis which diagnosed medically with cardiac specific troponin t, troponin 1 levels and electro myocardiogram ECG. The aged for all subjects ranged (39-75) years patients were lying in cardiac care unit at Ibn- al- Nafees teaching hospital and Sheikh Zayed teaching hospital. Genomic DNA of whole blood was extracted from buffy coat and cell cultured handbook protocol using Bioneer- kit and Genomic DNA fungus/yeast kit was used in isolation and purification of DNA. patients divided into three groups according to their age: group A (60-75) years , group B (50-59) years , group C (39-49) years the results of genomic DNA isolation from blood cells extracted in pure form which ensured by the absorbance ratio (260/280 ) was (1.6 – 1.9 ) with a concentration of 50µg/ml and one DNA band with high resolution in gel electrophoresis. The result of genomic DNA extracted from the local strain of S. cerevisiae showed that DNA extracted with high purity because the absorbance ratio (260 /280 )was (1.7 to 2.0) with a concentration of 60 µg/ml and presence one DNA band with high resolution in gel electrophoresis. primers were designed depending on the sequence of the gene responsible for the production of GPCR on the chromosome 11 , GPCR contain three exons which covered with six primers to detect a defect in gene sequence among. Results of gel electrophoresis are showed that primer GPRX1 gave one band for (Control , A,B ) groups but absent amplified band in the patient eight and nine from group C. with molecular weight of this band is 1000 bp. The GPRX2 primer used to amplify second exon in the GPCR gene ,the molecular weight of amplified bands are 400 bp were present in all control samples and three groups of thrombosis patientsand yeast. GPRX2A primer that designed to amplify part two from second exon of GPCR gene by PCR gave one band for all samples which include control and patient, the molecular weight of this band is 500 bp. PCR analysis showed one amplify band for all control and patients group with molecular weight 500 bp for GPRX3 primer and 400 bp for GPRX3A ,300 bp for GPRX3B. The specific primers which designed to covering GPCR gene used to amplification genomic DNA of the local strain S.cerevisiae by PCR technique. Results showed all six primers which gave one band with difference molecular weight for each primer. All samples demonstrate identity planned sizes to control and local strain of S. cerevisiae samples except patient number 8 and 9 in the group(C) that showed non specialist bands in specific primer with first exon (GPRX1) .So the genetic sequence analysis of these two case based on the sequence of the remainder exons to detect the genetic defect in these case. The sequence of the first part for the second exon (X2) was identity standard sequence found on the NCBI web site for case( 8). The case (9) showed identity with the sequence present in the human gene bank but some difference in the first of sequence which neglected because it is in the place link of primer. The results for case 8 showed some mutation for Exon X2(part2). but case (9) demonstrate one deletion and one substitution. The results, also, illustrated that the ether48 thrombosis patients didn't appeared any mutation despite the positive results for ( Troponin) that gives strong indication of thrombosis. The conclusion Primer GPRX1 gave one band for (Control , A,B ) groups but absent amplified band in the patient eight and nine from group C. The molecular weight of this band is 1000 bp. The amplified band with molecular weight 400 bp were present in all control samples and three groups of thrombosis patients with primer GPRX2 and 500 bp with primer GPRX2A.PCR analysis showed one amplify band for all control and patients group with molecular weight 500 bp for GPRX3 primer.,400 bp with GPRX3A and 300 bp for primer GPRX3B.Similarity between results given by healthy group and local strain of yeast. Genetic study showed that there are only two case of patients eight and nine demonstrated mutation in nucleic location on exon two and three from GPCR gene
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
... Show MoreDespite the G protein-coupled receptors (GPCRs) being the largest family of signalling proteins at the surface of cells, their potential to be targeted in cancer therapy is still under-utilised. This review highlights the contribution of these receptors to the process of oncogenesis and points to some likely challenges that might be encountered in targeting them. GPCR-signalling pathways are often complex and can be tissue-specific. Cancer cells hijack these communication networks to their proliferative advantage. The role of selected GPCRs in the different hallmarks of cancer is examined to highlight the complexity of targeting these receptors for therapeutic benefit. Our
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Total protein and total fucose were determined in sera of thyroid
disorder patients.
Sera of (40) diagnosed by consultant hyperthyroidism, and 40 hypothyroidism were analyzed for the above parameter for control, sera of (40) normal individuals were used.
They were healthy with no appearing disorder results analysis revealed no significant differences (P<0.05) in the (mean ±SD) of total protein values in sera of hyper and hypothyroidism were compared
... Show MoreIn spite of the high rate of morbidity and mortality heart failure (HF) is common, and none of the medications are now entirely available for HF treatment. In addition to many environmental influences and clinical diseases, genetic factors may also contribute to the progression and development of HF. In the current study, samples of blood were collected from 150 heart failure patients and 130 healthy controls. We evaluated the association of four single nucleotide polymorphisms (snps) of Toll-like receptors (TLR6 and TLR5) with (HF) susceptibility in the Iraqi population. In this work, (SNP) called Toll-like receptor 5 (rs5744168, rs2072493) and Toll-like receptor 6 (rs1039559, rs5743810) were employed. (PCR-RFLP) for snps
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Visceral leishmaniasis (VL) or kala-azar is one of the worlds most neglected tropical diseases in mortality and fourth in morbidity, rK39 dipstick was used to diagnose the suspected infected patients as cheapest simple technique which can differentiate recent from chronic infection, for disease out-coming, naïve T-lymphocyte cells should be differentiated into pathogen-specific immunity responses, such as T-helper 1(Th-1) or (Th-2). HLA-G is a special protein defined as nonclassical HLA class I molecule can suppress the immune system through prevention of T-cell function by foul all T-cell mechanisms. So, this study aimed to detect and evaluate the level of sHLA-G in the sera of patients infected with VL. The results showed that there was
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This study was conducted in the specialized center of endocrinology and diabetic (AlKindy hospital from June 2004 to April 2005). Sera of 80 women (include 40 diagnosed Hyperprolactinemia (HPro) and 40 healthy women as control) were used to estimate some biochemical parameters which include prolactin (PRL),total fucose (TF), total protein (TP) and protein bound fucose (PBF), and protein bound hexose (PBHex), also TP TF , TP PBF and TP PBHex ratios. A significant elevation in TP and PBHex, in sera of HPro patients compared to control was found while PBHex/TP ratio showed a slight non significant increase in sera of patients compared to control. On the other hand a significant decrease in TF, TF/TP and PBF/TP in ser
... Show MoreAnaemia is a common extra-articular manifestation of rheumatoid arthritis (RA) where anaemia of chronic disease (ACD) and iron deficiency anaemia (IDA) are the two most frequent types. The distinction between these two types of anaemia has always been challenging requiring sophisticated techniques. Serum transferrin receptor (sTfR) a truncated soluble form of the transferrin receptor is one of the parameters that is influenced by the Iron content and supply to the erythrons and is not affected by inflammatory status and therefore the use of the sTfR/log ferritin (sTfR-F) index can be a reliable indicator of functional iron deficiency.
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The study aimed to purification of acid phosphatase (ACP) from sera of obesetype 2 diabetes mellitus patients, this study included from thirty T2DM patients and thirty control, purification process was done with several steps included precipitation with inorganic salt (NH4 ) 2SO4 30%-80%, dialysis, ion exchange chromatography by DEAE sepharose anion column and size exclusion chromatography by Sepharose 6B.ACP, BMI, FBS, HbA1c, Lipid profile, Urea, Creatinie, Insuline, Homa-IR were determined. Results showed the precipitate and concentrated protein appeared four peaks in ion exchange column. ACP located in the first and second peak with purification fold (21.1), (37.2) yield of enzyme and specific activity (173.3) IU/ml, which obtained a si
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