Gene Expression and Methylation Levels of PCSK9 Gene in Iraqi Patients with Coronary Artery Disease

يرتبط التعبير الجيني لجين  PCSK9ارتباطًا وثيقًا بمستويات الدهون وخطر الإصابة بمرض تصلب الشرايين التاجية. تهدف الدراسة الحالية إلى قياس كمية التعبير الجيني للجين PCSK9 وتأثير المثيلة على مستوى تعبيره الذي يساهم في التسبب في اضطراب الشريان التاجي الحاد. تضمنت الدراسة الحالية 150 شخصًا من المجتمع العراقي ، 100 مريض بمرض الشريان التاجي و 50 من الأصحاء. تم تحديد تركيز PCSK9 في كل عينات المصل المدروسة بواسطة تقنية ELISA ، وتم تقدير مستويات التعبير عن جين PCSK9 في الدم الكامل بطريقة qRT-PCR ، واكتشاف مستوى مثيلة الحمض النووي في جين PCSK9 باستخدام تقنية HRM-PCR. كان التعبير عن جين ب مستوى اعلى  PCSK9 6.06 ± 1.84 في مجموعة مرضى ASCAD مقارنة بمجموعة التحكم 1.00 ± 0.19. أيضا كان تركيز بروتين PCSK9 عالياً في مجموعة المرضى مقارنة بمجموعة الاصحاء P = <0.01. فضلا عن ان مستويات مثيلة PCSK9 ، اظهرت فرقاً معنوياً بين مجموعتي الدراسة بمستوى 0.002P=. في الختام ، ترتبط مستويات بروتين PCSK9 في المصل بمرض الشريان التاجي كما موضح في receiver operating characteristic (ROC) analysis ، مما يؤدي إلى اعتبار أن تركيز بروتين PCSK9  قد يكون مؤشرًا جيدًا لتطور مرض الشريان التاجي.The expression of the Proprotein Convertase Subtilisin/Kexin Type 9 gene (PCSK9) is inextricably related to lipid levels and a risk of atherosclerotic coronary artery disease (ASCAD). The present study aims to measure the quantity of PCSK9 gene expression and the effect of methylation on its expression level taking part in the pathogenesis of acute coronary artery disorder. A current study included 150 subjects from the Iraqi population, 100 ASCAD patients and 50 healthy controls. The concentration of PCSK9 in each serum sample was determined by the ELISA technique, the expression levels of the PCSK9 gene in whole blood were estimated by RT-qPCR – Quantitative Reverse Transcription PCR method, and DNA methylation level detection in PCSK9 gene by using High Resolution Melting Analysis (HRM) technique. The expression level of the PCSK9 gene was 6.06 ± 1.84 in ASCAD patients group compared with a control group 1.00 ±0.19. That indicates the up-regulation of the PCSK9 gene in patient with atherosclerosis. In addition, the PCSK9 concentration was higher in patient group in comparison to the healthy controls group P= < 0.01. However, PCSK9 methylation levels, a highly significant distinction between the two study groups p=0.002. In conclusion, serum PCSK9 levels are associated with coronary artery lesions as shown in receiver operating characteristic (ROC) analysis. Suggests that, serum PCSK9 might be a good indicator of coronary artery disease development

Food dyes as an alternative tracking dye for DNA gel electrophoresis

The chemical, physical and toxicological effects on health of synthetic dyes that used as tracking dye in the electrophoresis requires seriously search about alternative tracking dye. The present study is aimed to find an alternative dye from safe food dyes which commonly used in food coloring. Five dyes were selected depending on their chemical properties and the availability in local market: Brilliant Blue FCF, Tartrazine, Sunset Yellow FCF, Carmoisine, and green traditional, three dyes were chosen to be mixed as loading buffer: Brilliant Blue FCF, Sunset Yellow FCF as a basic because it give the whole range size of most traditional loading buffers that available in market, and adding the Carmoisine as a new indicator for the bands less than 50bp, then mixed with DNA ladder in same percentage used with traditional loading buffers to clarify the effects of dyes on DNA, migrated on 1% agarose with loading buffer promega, results showed more clarity and highly readable separation of dyes and give wide range of size in the food loading mix than promega loading dye, by viewing the gel on UV light the DNA ladder were moved smoothly, bands separated effeminately on gel and in same rate of the DNA ladder that load with promega loading buffer which indicate no interaction between the food dyes and the DNA.Our studies show that the food dye can be used as a tracking dye in place of used synthetic dye. The procedure is found to be easy, practical, safely and reliable

ApoB gene polymorphism (rs676210) and its pharmacogenetics impact on atorvastatin response among Iraqi population with coronary artery disease

Abstract Background Drug response is below genetic influence, proven by the genetic variants. Pharmacogenetics trials are performed in many diseases, including coronary artery disease. This study was designed to determine the genetic polymorphism (rs676210) Pro2739leu G > A in the lipid metabolism-related gene (ApoB gene) and its pharmacogenetic role in the response to atorvastatin drug in a sample of Iraqi population with coronary artery disease (CAD). Results Significant differences of genotype distribution in CAD patients and controls were observed in ApoB + 8216 in Iraqi population from Hardy Weinberg Analysis. It also found that dramatic difference of low-density lipoprotein (LDL-C) level in response to 40 mg/day of atorvastatin therapy, the minor allele (A) observed a greater LDL-C lowering than the wild type allele (G). In ANOVA analysis, the result showed that the rs676210, Pro2739Leu, in ApoB gene increased non significantly, but gradually in plasma level of total cholesterol (TC), triglyceride (TG), very low-density lipoprotein (VLDL), and oxidize low-density lipoprotein (oxLDL) in the order of genotype AA, GA, and GG in response to 40 mg atorvastatin. Conclusion We found the results highlighted the function of the rs676210, Pro2739Leu, in the ApoB gene in CAD etiology, and the findings support this variant’s impact in predicting the response of (LDL-C) to 40 mg of atorvastatin therapy. ApoB gene polymorphism (rs676210, Pro2739Leu), specifically the AA genotype, may help to identify individuals who will profit from atorvastatin's lowering effects

Extraction and Characterization of rapeseed oil (Brassca campestris ) and it's Effected on Microorganisms

Oil from Brassca campestris (local variety) was extracted with hexane using Soxhlet. The extracted oil was characterized and its antimicrobial activity was determined as well. The content of extracted oil was 40% with 0.5% of volatile oil .Oil was immiscible with polar solvent such as ethanol, acetone and water, while it was easily miscible with chloroform due to its hydrophobicity. The result of organoleptic tests revealed that the oil is clear yellow in color and odorless with acceptable taste. The oil was stable at 4 -25 C? for a month. Refractive index (RI) of oil was 1.4723 with density of 0.914, [both at 4-25 C?]. Boiling point 386 C?. Infra red spectroscopy (IR) indicated the presence of different chemical groups (C=C , C=O ? C-H) with different characters .Oil exhibited antimicrobial activity against Gram positive and negative bacteripa in addition to its activity against Candida albicans

Detection of Metallo-β-Lactamase Producing Pseudomonas aeruginosa Isolated from Public and Private Hospitals in Baghdad, Iraq

Metallo-β-lactamase (MBL) producing Pseudomonas aeruginosa has been reported to be an important nosocomial infection. Its intrinsic and acquired resistance to various antimicrobial agents and its ability to develop multidrug resistance imposes a serious therapeutic problem. Different clinical samples were collected from public and private hospitals in Baghdad city, Iraq. Bacterial identification was done using conventional cultural, biochemical tests, and VITEk 2 system. Minimum inhibitory concentration (MIC) testing was performed using VITEK 2 automated system. Each P. aeruginosa isolates showed resistance to Carbapenems (Imipenem and Meropenem) were subjected to Imipenem-EDTA combined disc synergy test (CDST) to investigate the production of MBL (confirmative test). The presence of bla-genes encoded IMP, VIM, and SPM-1 was detected by conventional PCR technique. A total of 75 P. aeruginosa isolates were isolated, 16 (21.3%) were able to grow on MacConkey agar supplemented with Meropenem 4mg/L (MMAC). The MIC of different antibiotics showed that 6 (37.5 %) isolates were Carbapenem resistant, MIC ≥16 µg/ml while 4 (25%) isolates appear to be MBL producer using CDST test. PCR assay revealed that 3 (50%), 1 (16.6%) of the carbapenem resistant isolates harbored blaIMP, blaSPM-1 genes, respectively. blaVIM gene was not detected in this study. The prevalence of multi-drug resistant P. aeruginosa isolates especially Carbapenem resistant bacteria was increased in Baghdad province. The blaIMP was the predominant among the MBLs genes in P. aeruginosa in this study

Genetic analysis of X-chromosomal short tandem repeat (X-STR) frequencies in Arab Iraqi male population

Abstract Background The X-chromosome short tandem repeat (STR) polymorphisms are a particular tool in the fields of human population genetics and personal identification. It was necessary in investigating complex kinship or deficiency cases in conditions where information on mitochondrial DNA (mtDNA) or Y chromosome polymorphisms have been used to explore their direct paternal line. This study aimed to investigate the allele frequency of (12X-STR) of 200 unrelated males from different region of Baghdad City to serve as a reference data base for individual identification in Iraqi population. Results Twelve X-STR loci (DXS7424, HPRTB, DXS8377, GATA31E08, DXS7423, DXS8378, DXS9895, DXS10074, DXS6809, DXS7133, DXS101, DXS6807) were successfully amplified by multiplex PCR and divided into four groups. According to measures of allele frequency, the higher alleles frequency were 16, 11, 46, 11, 14, 10, 15, 15.2, 35, 11, 25, and 11 while the lowest alleles frequency were 11, 9, 52,53, 7, 17, 14, 13, 12.2,17, 36, 15, 16, 22, 29, and 17 that observed at the 12 loci respectively. Forensic efficiency parameter for DXS8377 locus in the first group showed highest polymorphic allele in the Iraqi Arab population with the frequencies ranging from 0.005 to 0.16%. The power of discrimination (PD) value ranged from 0.663 for DXS7423 locus and 0.9066 for DXS8377 locus. In addition, the polymorphism information content (PIC) value ranged from 0.602974 for DXS7423 locus to 0.899206 for DXS8377 locus. Conclusions Overall the X-STR markers become used as an important source of information beside the autosomal and Y-STR markers, especially for kinship testing and haplotype analysis

Screening of Antimicrobial Activity from Various Pharmaceutical Companies in Iraq

An antimicrobial is an agent that kills microorganisms or stops their growth. Antimicrobial can be classified according to their function, it may be a microbicidal which is an agent that kill microbes, while those that only prevent and stops their growth are named biostatic. Antimicrobial resistance (AMR) is the ability of a microbe to resist the effects of antimicrobial medication that once could successfully treat the microbe by killing or stop its growth. So with the resistance to the antibiotics becoming more and more common and worldwide resulting in need for screening the antimicrobial activity. So in this study we used an evaluation method which is used in many laboratories to screen the activity of antimicrobial agents obtained from a local company and a foreign company on standard microorganism which includes the following (Escherichia coli, staphylococcus aurous, Pseudomonas aeruginosa, Candida albicans) and pathogenic bacteria includes (Escherichia coli, staphylococcus aurous) which obtained from renal infections and tissue burns respectively.  In conclusion the results show that the antibiotic manufactured in the local company Al-Razi in Iraq gave better effectiveness than other foreign companies and the reason for this difference may due to the active chemicals used in manufacturing the antibiotic or due to the manufacturing method

Global Incidence and Risk Factors Associated With Postoperative Urinary Retention Following Elective Inguinal Hernia Repair

Importance Postoperative urinary retention (POUR) is a well-recognized complication of inguinal hernia repair (IHR). A variable incidence of POUR has previously been reported in this context, and contradictory evidence surrounds potential risk factors.Objective To ascertain the incidence of, explore risk factors for, and determine the health service outcomes of POUR following elective IHR.Design, Setting, and Participants The Retention of Urine After Inguinal Hernia Elective Repair (RETAINER I) study, an international, prospective cohort study, recruited participants between March 1 and October 31, 2021. This study was conducted across 209 centers in 32 countries in a consecutive sample of adult patients undergoing elective IHR.Exposure Open or minimally invasive IHR by any surgical technique, under local, neuraxial regional, or general anesthesia.Main Outcomes and Measures The primary outcome was the incidence of POUR following elective IHR. Secondary outcomes were perioperative risk factors, management, clinical consequences, and health service outcomes of POUR. A preoperative International Prostate Symptom Score was measured in male patients.Results In total, 4151 patients (3882 male and 269 female; median [IQR] age, 56 [43-68] years) were studied. Inguinal hernia repair was commenced via an open surgical approach in 82.2% of patients (n = 3414) and minimally invasive surgery in 17.8% (n = 737). The primary form of anesthesia was general in 40.9% of patients (n = 1696), neuraxial regional in 45.8% (n = 1902), and local in 10.7% (n = 446). Postoperative urinary retention occurred in 5.8% of male patients (n = 224), 2.97% of female patients (n = 8), and 9.5% (119 of 1252) of male patients aged 65 years or older. Risk factors for POUR after adjusted analyses included increasing age, anticholinergic medication, history of urinary retention, constipation, out-of-hours surgery, involvement of urinary bladder within the hernia, temporary intraoperative urethral catheterization, and increasing operative duration. Postoperative urinary retention was the primary reason for 27.8% of unplanned day-case surgery admissions (n = 74) and 51.8% of 30-day readmissions (n = 72).Conclusions The findings of this cohort study suggest that 1 in 17 male patients, 1 in 11 male patients aged 65 years or older, and 1 in 34 female patients may develop POUR following IHR. These findings could inform preoperative patient counseling. In addition, awareness of modifiable risk factors may help to identify patients at increased risk of POUR who may benefit from perioperative risk mitigation strategies