Iraqi journal of biotechnology

The Effect of Genetic Polymorphism of Resistin Gene among Iraqi Breast Cancer Women

1Khadija A. Sahan, 1Ismail H. Aziz, 2Sanna N. Dawood , 3Shaymaa S. Abdul Razzaq — Mon, 03 Jul 2023 00:00:00 +0200

Breast cancer is a heterogeneous disease defined by molecular types and subtypes. It constitutes the most commonly-diagnosed cancer and the leading cause of cancer death in women, worldwide, according to the International Agency for Research on Cancer (IARC) World Cancer Reports in 2020. The study aimed to detect the relationship between Resistin Gene polymorphism (rs1862513, rs3745367) with the age and BMI of the participants diagnosed with breast cancer. A total number of 105 samples; three groups of 35 fresh blood samples were collected as malignant, benign and apparently healthy control. Conventional PCR with specific primers followed by sanger sequencing was used to characterize the two SNPs. The results revealed that (rs1862513) GG allele carriers (GG) had a breast cancer risk of (1.78) in compare with (CG) allele carriers; (OR=0.337). Genotyping and allele frequency of RETN gene at SNP (rs3745367) in three groups (control, benign and malignant) showed that AA allele with (OR=1.44) may considered as a risk factor for incidence of breast cancer. The results showed that the incidence of breast cancer was significantly higher in middle age group (40-49) year with a percentage of 42.85%. It was concluded the relation of BMI and breast cancer remains conflicting, the study show a negative correlation of BMI with tumor size, stage and grade of the tumor.

Role of miRNA-21-5p Gene Expression as a Tumor Marker and the Progression of Clinic-Pathological Characters in Breast Cancer Iraqi Female Patients

1Ruwaidah A. Abbas, 2Ismail H. Aziz — Mon, 03 Jul 2023 00:00:00 +0200

Breast cancer is a malignancy of the breast tissues that arise from uncontrolled proliferation of breast ductal and lobular epithelial cells. It’s the most prevalent malignancy among the Iraqi women. The aims of study are analyzing the expression of serum (miR-21-5p) as a molecular tumor marker and main clinic pathological characters for patients and healthy control groups. Determination of microRNA gene expression by (qRT-PCR) for both BC patients’ healthy control and statistical analysis of their demographic characteristics. The results show that high significant up regulation of serum miRNA-21 expression in BC patients (5.27) times than control group and the main clinic-pathological data show high percentage of (IDC) histopathology type, (T2) staging, the hormone receptors ER+, PR+, high metastasis LN+ and the molecular subtypes Luminal-A than other characters. In conclusions the high miRNA-21 expression levels make it act as noninvasive diagnostic tumor marker and show the aggressive phenotype progression BC diseases’ characters.

Investigation of IL-23, IL-17 and Calprotectin Levels in Blood of Iraqi Patients with Ulcerative Colitis

1Al-Hassan T. Waly, 1Abed H. Barraj, Mohammed H. Alrikabi — Mon, 03 Jul 2023 00:00:00 +0200

Ulcerative colitis (UC) is a type of inflammatory disease of the colon and it causes heavy inflammation. Knowing about cytokines is very important due to their role in the progression of the disease and in decreasing its severity. The current study aims to estimate calprotectin, IL-17, and IL-23 in the blood of Iraqi patients with ulcerative colitis. Also measuring levels of inflammation markers including ESR, WBCs, and CRP, and the reason for its height in serum or blood of ulcerative colitis patients. In this study, 30 blood samples from Iraqi patients with UC and 30 blood samples from Iraqi healthy individuals as control have been collected from 2021/11 to 2022/4, the samples were collected from Medical City Hospitals, Gastro-Enterology and Hepatology Hospital in Baghdad, Iraq. Calprotectin, IL-23, and IL-17 were measured by the enzyme-linked immune-Sorbent assay technology (sandwich-ELISA method). The Biomarkers used in clinical practice to assess disease activity are complete blood count (CBC), c-reactive protein (CRP), and erythrocyte sedimentation rate (ESR). The current study results are the incidence of ulcerative colitis between males and females is not statically significant, the mean age of diagnostic of ulcerative colitis is 33.93 ±2.59, and it has been noticed a high incidence of ulcerative colitis in Iraqi people who have medium economic status. It was concluded significant differences in levels of calprotectin, IL-17, and IL-23 compared to the control group. Also, higher levels of ESR, WBCs, and CRP in ulcerative colitis compared to the control group.

Impact of Helicobacter pylori in some Blood Parameters Change of Iraqi Patients with Gastritis Disease

1Hussein S. Qasim, 2 Rana S. Aboud — Mon, 03 Jul 2023 00:00:00 +0200

In order to find the relationship between Helicobacter pylori infection and hematological disease are disorders which primarily affect the blood and blood-forming organs. One hundred and three blood samples were taken for people aged (20-68) years for the period from 10/1/2021 to 1/3/2022, divided into three groups. The first group included 44-person H. pylori-infected with symptoms of infection, the second group had 19-person H. pylori-infected but without symptoms, and the third group included 40 people without H. pylori infection. All studied groups were carried out to measure anti-IgG Ab, Vac A and Ferritin by Enzyme Linked Immunosorbent Assay (ELISA) technique. The statistical analysis indicates a non-significant difference in Vac A (p>0.05) but a significant difference in IgG and ferritin (p < 0.05) in the positive group. It was concluded H .pylori infection may contribute to hematological disease.

Antibacterial Activity of Chitosan Nanoparticles Loaded with Syzygium aromaticum Extract against Klebsiella pneumonia

Hussein A. Shaghati , Emad H. Jassim — Mon, 03 Jul 2023 00:00:00 +0200

The present study aims to describe the antibacterial activity of chitosan nanoparticles loaded with Syzygium aromaticum extract against MDR Klebsiella pneumonia. 50 isolates of K. pneumoniae were collected from patients with urinary tract infections. The isolates were diagnosed as Klebsiella pneumoniae by classical biochemical tests and PCR technique using 16S rRNA gene as a diagnostic gene and gel electrophoresis. The sensitivity of bacteria to antibiotics was tested using the Kerby-Bauer method. Measuring the diameters of inhibition and testing the effectiveness of the chitosan nanoparticles prepared in a previous study using spared plate method and counting the colonies growth. It was concluded to demonstrate that the chitosan nanoparticles loaded with Syzygium aromaticum extract have good antibacterial efficacy against MDR Klebsiella pneumonia.

In Vivo Cytogenetic Effects of Ephedra alata L. Stems Extracts in Mitosis of Meristematic Cells in Onion Roots

Lubna Z. Mohammed , Rashad K. Mohammed — Mon, 03 Jul 2023 00:00:00 +0200

Ephedra alata. is a plant that widely available around the world and long used in folk medicine as a natural medication, was employed in the current work to prepare extracts rich in alkaloids and to test their cytotoxic potential. Alkaloids-rich and crude extracts of E.alata were compared to pure ephedrine medication for mitosis on Allium cepa. test system. Alkaloids and crude aqueous extracts of A. cepa root tips were examined for a total of five hours at five different concentrations compared to ephedrine standard. Mitotic index, phase index, and chromosomal aberration as part of the study. IC50 values of 35 mg/ml were found for each extract, indicating a sub-lethal influence on cell viability. (Toxic and sublethal effects are thought to be the result of this abnormalities, chromosome stickiness abnormalities may be caused by some faulty nucleic acid metabolic pathways and an aneugenic agents caused by the extracts with a high percentage for 75 mg/ml treatment. It was concluded the variations in the chemical constituent of the extracts represented by events caused by various chromosomal and nuclear aberrations give us promising Look of the plant as anti-tumor drug.

Evaluation of some Biochemical Parameters in Iraqi Patients with Hyperthyroidism

Noor T. Akber, Jabbar H. Yenzeel — Mon, 03 Jul 2023 00:00:00 +0200

Hyperthyroidism is autoimmune disease that is considered a thyroid gland problem. A clinical condition known as hyperthyroidism is defined by the thyroid gland producing and secreting an excessive amount of thyroid hormones. Lipid profile play an essential role in the progression and development of hyperthyroidism. The aim of this study is to evaluate the level of some biochemical parameters in Iraqi patients with hyperthyroidism. Sixty hyperthyroid patients and 30 healthy individuals with age range from (30-65) years were enrolled in this study during their attendance at the National Center for Diabetes Treatment and Research in Baghdad from the period between December 2021 to April 2022 .Blood samples were collected to evaluate the level of thyroid hormone like triiodothyronin (T3), thyroxin (T4) and thyroid stimulating hormone (TSH) using ELISA technique, and total cholesterol (TC), triglycerides (TG), high- density lipoprotein cholesterol (HDL-C) were measured spectrophotometrically by using different type of kits. The results showed highly significant (p≤0.01) increase in the level of T3, T4 in the patients with hyperthyroidism in comparison with the control group, While, TSH level showed highly significant (p≤0.01) decrease in the patients with hyperthyroidism as compared with control group. The level of TC, HDL-C and LDL-C showed a highly significant (p≤0.01) decrease in the patients with hyperthyroidism in comparison with the control group, While, the level of TG and VLDL-C showed a highly significant (p≤0.01) increase in the patients with hyperthyroidism in comparison with the control group. It can be concluded that the lipid profile play a major role in the pathogenicity of hyperthyroidism.

Study of some Hematological and Physiological Parameters and Electrolytes in Cardiomyopathy Iraqi Patients

Maryam A. Al-Khayatt , Jabbar H. Yenzeel — Mon, 03 Jul 2023 00:00:00 +0200

Cardiomyopathy (CM) is a condition that affects the heart muscle and can lead to heart failure. It can be dilated, hypertrophic, or restricted. This study was aimed to investigate the level of physiological and biochemical parameters in CM. Sixty patients with age (40-70), and thirty healthy controls with the same age range, were involved in this study during their0attendance to Iraqi Center for Heart Diseases, Ghazy Al-Hariri Hospital for Surgical Specialties in the Medical City in Baghdad. The patients were diagnosed by an expert cardiologist. The study was conducted from November 2021 to April 2022, and approved by ethical committees: Ref.:0CSEC/0121/0074 on October 20, 2021of Department of Biology, College of Science, University of0Baghdad. Blood samples and data were collected to evaluate the level of complete blood counts (CBC), some biochemical parameters like Troponin T, creatine kinase - MB (CK-MB), C-reactive protein (CRP), Lactate dehydrogenase (LDH), and blood electrolytes like (Magnesium Mg++, Sodium Na⁺, Potassium K⁺, and Calcium Ca⁺⁺) for the patients and control. Results appeared that mean total WBC count, Lymphocyte, and Monocytes percentages were significantly (P≤0.01) increased in CM patients as compared with control, while there was a highly significant (P≤0.01) decrease in RBC, Hb, and HCT in CM patients as compared with control, the levels of CK-MB, LDH, and CRP in CM patients was a significant increase as compared with the control group. Referring to electrolytes results, the levels showed that there was a significant (P≤0.01) increase in the levels of both Mg+2 and k+ in patients with CM as compared with the control group, While the level of Na+ was decreased in CM patients. It can be concluded from the current results that the wide range of biochemical abnormalities play a major role in the pathogenicity of CM patients.

Association of HOXA4 Gene Expression and Methylation with Response to Treatment in Iraqi Chronic Myeloid Leukemia Patients

1,2Noor H. Ismail, 1Abdul Hussein M. Al-Faisal, 1Ismail A. Abdulhassan — Mon, 03 Jul 2023 00:00:00 +0200

Genetic and epigenetic factors affect Chronic Myeloid Leukemia (CML) response to Imatinib mesylate (IM) therapy. This study aimed to investigate HOXA4 gene methylation and expression in CML patients and their predictive value as response markers. Blood samples were collected from fifty CML patients (25 responders and 25 non-responders to Imatinib mesylate therapy) and 50 healthy controls of same age and sex. HOXA4 gene methylation and gene expression studies were conducted by quantitative PCR (qPCR). Results revealed that CML patients had significantly higher level of HOXA4 gene demethylation and expression compared to controls group (p <0.001). The non-responders CML patients showed higher significant levels of HOXA4 gene demethylation (p<0.001) and expression (p<0.05) compared to responders CML patients. Significant risk association results of HOXA4 demethylation and expression levels (p<0.001 and p<0.05, respectively) with the development of IM resistance in CML patients according to optimal cut-off point obtained by receiver operating characteristics (ROC) analysis. In conclusion, HOXA4 gene activation due to promoter DNA hypomethylation refers to its oncogenic role in CML pathogenesis. HOXA4 gene demethylation and overexpression may serve as biomarkers for predicting IM resistance in CML patients, especially HOXA4 gene demethylation with good sensitivity and specificity.

Assessment of some Interleukins in Sample of Iraqi Patients with Ankylosing Spondylitis

Hiba H. Hashim, Reema M. Abed — Mon, 03 Jul 2023 00:00:00 +0200

Ankylosing spondylitis (AS) is a widespread persistent inflammation illness indicated by chronic inflammatory of the peripheral joints and vertebral column, which results in bone degradation consequently, has an obvious impact on the worth of the sufferers' lives. Cytokines have a key role in the pathophysiology of AS. During the progression of a disease, inflammation is likely caused by cytokines. This study included 100 AS male patients with a mean age (40.56 ± 10.19) and a disease duration of 9.04 ± 2.03 years. The age of the male control (38.78 ± 10.57) was matched (p= 0.227). The Bath AS(Ankylosing spondylitis) Disease Activity Index (BASDAI) and the Bath AS(Ankylosing spondylitis) Functional Index (BASFI) were (3.45 ± 2.03 and 3.84 ± 2.62) respectively. It was found that 57% of patients have HLA-B27 positive and the levels of erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were (25.39 ± 19.59 mm/h and 18.34 ± 3.45 mg/l) respectively. This study evaluated the levels of interleukins (IL-20, IL-22, and IL-24) with the development of Ankylosing spondylitis (AS) in the Iraqi population by using ELISA. The differences in the Interleukins concentrations in serum between patients and healthy controls were included. The results for serum levels of (IL-20, IL-22, and IL-24) appeared higher significant differences (p <0.0001) in patients as contrasted to healthy control (194.0 ± 52.90 pg/mL vs 99.52 ± 11.99 pg/mL), (373.8 ± 102.2 ng/mL vs 132.0 ± 41.63 ng/mL), and (389.6 ± 180.7 ng/mL vs 186.9 ± 29.38 ng/mL) respectively. In conclusion, higher serum concentrations of IL-20, IL-22, and IL-24 were found in male patients with AS in comparison to healthy controls. It was concluded that might be considered as biomarkers in AS development.

Gene Expression of Biofilm Formation lasB and rblB Genes of Acinetobacter baumannii Isolated from Wounds, Burns and Environmental Samples

Noor N. Abd-alkader , Wathiq A. Al-Draghi — Mon, 03 Jul 2023 00:00:00 +0200

Multiple-drug-resistant infections linked to medical devices are common due to Acinetobacter baumannii's propensity to develop biofilm. The purpose of this work is to identify the MIC of Imipenem and Chalcon and to investigate the involvement of the lasB gene in formation of biofilm by A. baumannii in order to get better understand how these bacteria can survive l in the hospital setting. As well as comparing the effect before and after of using the antibiotic and chalcone. A 250 samples were obtained from several locations in hospitals of Baghdad/Iraq and classified to 2 groups according to their source: 100 environmental and 150 clinical samples, obtained during October 2021-March 2022. Antibiotic resistance profile of A. baumannii isolates was confirmed by VITEK-2 system and in vitro biofilm-forming ability was evaluated by micro titer plate methods, respectively. Isolates were tested for the presence of lasB gene Forty (40) isolates of A. baumannii were subjected to examination under microscope and to diagnosis using biochemical tests. The (40) isolates identification was confirmed by the VITEK-2 system and a molecular detection according to the rplB gene, which is a necessary gene located in this genus. Antibiotic susceptibility testing was carried out on all (40) identified isolates, and the results detected those 36 isolates had Multi-Drug Resistance (MDR). This work indicated that 16 isolates from these 16 isolates carrying the rblB gene and 2 isolates from 6 carrying lasB that confirmed by molecular detection methods, and 38 isolates from these 40 isolates had the ability to form biofilm in the micro titer plate method, and this study show there is down expression of lasB with the antibiotic while down expression of this genes with chalcone. In conclusion, the presence of the lasB gene was significantly correlated with A. baumannii biofilm development. Biofilm production was also higher in multidrug-resistant than in other isolates. It was concluded the role of biofilm in the isolate’s resistance and the impact of the existence of lasB genes in formation of biofilm by A. baumannii strains, and lasB gene expression was reduced in those treated to imipenem and chalcone.

Gene Expression of Adenosine Deaminase Genes 1 and 2 in Female Iraqi Patients with Autoimmune Thyroid Disease

Afrah H. Abd , Maha F. Altaee — Mon, 03 Jul 2023 00:00:00 +0200

The current study was carried out to investigate the correlation of gene expressions of ADA1 and ADA2 genes with the development of autoimmune thyroid disease (AITD) in a sample of Iraqi females. One hundred patients with AITD and 80 controls were included. Quantitative real time polymerase chain reaction (qRT–PCR) was utilized for investigation of ADA1 and ADA2 gene expression among patients and controls. The correlation of age and body mass index (BMI) with AITD occurrence comparing with controls was studied. Based on the results of this study, there is high expression level of ADA1 and ADA2 genes in patients compared with healthy controls; also, the gene expression fold (2^-ΔΔCT) of ADA1 and ADA2 among AITD patients was recorded and according to results both genes showed up-regulation. It was concluded indicates both of age and BMI shows significant differences in patients compared with controls.

Evaluation of some Interleukins in Sample of Iraqi Female Patients with Rheumatoid Arthritis

1Hawraa M. Hussien, 1Reema M. Abed — Mon, 03 Jul 2023 00:00:00 +0200

Rheumatoid arthritis, commonly abbreviated as RA, an autoimmune systemic disease, is characterised by symmetric inflammatory polyarthritis. Rheumatoid arthritis mostly affects joints that are smaller, such as those in the feet and hands but it can also impact larger joints. Cytokines play an important role in the etiopathogenesis of rheumatoid arthritis during the course of the disease; inflammatory responses and joint degradation are probably caused by cytokines. The following research was carried out on 100 female RA patients and 100 female healthy control with mean ages of patients and controls (37.44±23.10 and 49.74±9.775) respectively. Erythrocytes Sedimentation Rate (ESR), Rheumatoid Factor (RF), Serum C-reactive Protein (CRP), and Anticyclic Cetrolinated Peptide (Anti-CCP) were investigated in this study with concentrations (37.44±23.10 mm/h , 22.72±1.44 IU/ml, 16.56±0.842 IU/ml and 28.09±6.68 EU/ml), respectively. This study was examined the interleukins levels (IL-34, IL-36 and IL-37) during the development of Rheumatoid Arthritis (RA) in Iraqi female. The differences in the levels of interleukins in the serum of patients as compared to healthy controls have been included in the study. The study concluded that the serum level of (IL-36 and IL-37) appeared a higher significant differences (p<0.0001) in patients compared with the healthy control, (11.45±3,3ng/mL vs 7.9±2.1 ng/mL) and (111.6±21.9 ng/mL vs 78.7±19 ng/mL) respectively. It was concluded that the rheumatoid arthritis patients' serum levels of (IL-34) were markedly lower than those of the controls (130±60.7 ng/mL vs 185.9±31.6 ng/mL) (P<0.001).

Effect of Age on Apoptosis and Necrosis of Peripheral Blood Lymphocytes in Sample of Iraqi Type 2 Diabetes Patients

Alaa N. Lateef, Bushra J. Mohammed — Mon, 03 Jul 2023 00:00:00 +0200

Type 2 diabetes (T2DM) is a metabolic disorder usually in adults, resulting from insulin insufficiency or function. Peripheral Blood Lymphocytes (PBLs) are the key components of the body's defenses which offer resistance against infective micro-organisms. Aging is an extremely intricate process related with declines in renewing potential plus adaptive reactions to stress. The aim of the study was to investigate the effect of age on apoptosis and necrosis of peripheral blood lymphocytes in a sample of T2DM Iraqi patients. Venous blood (4 ml) was collected in heparinized tubes under optimal condition and processed for peripheral blood lymphocytes separation by density-gradient centrifugation with lymphocyte separation medium. Apoptotic and necrotic lymphocytes were detected by using dual acridine orange / ethidium bromide staining and fluorescence microscopy, and the relationship between apoptosis, and necrosis with age of volunteers was analysis by using static methods. The morphological observations of lymphocytes from diabetic patients showed morphological features consistent with apoptosis more than control. Peripheral blood lymphocytes apoptosis and necrosis of T2DM patients showed significantly higher (P<0.01) when compared to control, whereas the peripheral blood lymphocytes (PBLs) apoptosis percentage reached to the maximum value (50.52 %), at age group more than 60 years of T2DM patients, however necrosis percentage reached to (21.8 %).It can be concluded that high blood glucose in poorly controlled diabetes disease is concomitant with increase rate of apoptosis and necrosis of PBLs which can be considered as a marker of severity for this disease.

Evaluation of Gene Expression of some Toll-like Receptors Genes among Iraqi Meningitis

Mahmood A. Talak, Kais K. Ghaima Institute of Genetic Eng — Mon, 03 Jul 2023 00:00:00 +0200

Meningitis is an important infectious disease with a high morbidity and mortality rate. The present study characterized the Toll-Like (receptors) (TLRs) role in the host immune response and as markers for bacterial and viral meningitis. The aim of this study evaluate the gene expression of Toll-like receptors TLR2, TLR3, TLR 4, TLR 7 and TLR9 genes the delta delta ct method among patients with meningitis infectious pathogens (Streptococcus pneumoniae, Staphylococcus spp, Herpes simplex virus1 and Epstein-Barr virus) and compared the results with the results of healthy control. The results of gene expression of TLR2 revealed that this receptor expression was increased among blood cells for infected patients with Staphylococcus spp (4.8 fold) in comparison with the control. The results of gene expression of TLR3 demonstrated that this receptor was increased among meningitis patients with Herpes simplex virus1 (5.1 fold). The receptor TLR4 was increased among patients with S. pneumoniae (6.2 fold). Also it was found that the gene expression of TLR7 and TLR9 was increased among patients through Epstein-Barr virus (6.2 fold), and Herpes simplex virus1 (4 fold), respectively. The findings of the study revealed that the mRNA expression levels of TLRs 2 and 4 had increased in infected patients with bacteria, while in the patients infected with viruses, there was increasing in the levels of TLRs 3, 7 and 9. Also thelevels of TLR2 were exhibited low increasing among patients with viruses. In conclusion, TLRs perform an important function in immune system response. to viral and bacterial meningitis and by way of genetic markers for identifying high-risk patients with meningitis and perhaps leading to novel treatment methods.

Role of IFIH1 Gene Polymorphism rs3747517 and IL-17A with its Receptor in Detection and Therapeutic approach for Type 1 Diabetes Patients

Baneen S. Mahdi, Wisal S. Abd Almohsen — Mon, 03 Jul 2023 00:00:00 +0200

Type 1 diabetes (T1D) is multifactorial autoimmune disease, overlapping factors including genetic and environmental factors contribute to the appearance of T1D in children mostly and adults. The aim of the study was to find the correlation of T1D with the chosen immune parameters and with IFIH1C polymorphisms as genetic factor for the disease and study the role of interleukin-17 in the exacerbation and evolution of T1D. This study at pediatric Teaching Hospital in the medical city between November 2021 and January 2022.The study included 100 Iraqi children divided into two groups 50 T1D patients and 50 controls. Two SNPs (rs3747517 and rs1990760) from the IFIH1 gene were carefully chosen to examine their relationship with T1D. Human IL-17A and IL-17AR enzyme linked immunosorbent assay (ELISA) kits were used to estimate the levels of interleukins mentioned above. The recognition of SNPs was achieved by using HRM (high resolution melting) real-time PCR. A Rotor gene was employed to perform qPCR-HRM, followed by an HRM analysis with ramping by 0.2 °C from 55 to 95 °C. In both selected parameters there was significant differences with average (IL-17A=260.0 , IL-17RA=238.4) of patients and an average (IL-17A=93.0, IL-17RA=90.9) in control group the p -value (0.001) , data of IFIH1 gene polymorphism rs3747517 presented as shown in wild TT, hetero TC and mutant CC genotype ,comparison between IL-17A and IL-17RA and TT,TC,CC genotype in patients group result no significant value for the IL-17A( p =0.106) but there was a significant consequence between IL-17RA and SNP data (p =0.018). The IL-17A and IL-17R levels is a key indicator of damaged β cell function and progression to explicit T1D with it is receptor. It was concluded that IL-17A involvement in T1D increasing and a possible therapeutic approach for T1D directing IL-17A with IL-17RA besides highlight on status of IFIH1 as significant genetic factor and indicator to T1D.

The Role of Long Non Coding RNA ANRIL Gene Expression and Serum Interleukin-27 Level in Metastasis of Breast Cancer Patients

1Ali K. Ibrahim, 2Mohammed I. Nader, 3Sana A. Abbood — Mon, 03 Jul 2023 00:00:00 +0200

Breast cancer is a heterogeneous disease and is the most common and prevalent form of malignancy diagnosed in women. The antisense non-coding RNA in the INK4 locus (ANRIL) belongs to long non-coding RNA family is overexpressed in several cancers including breast cancer. The study aims to understand the role of ANRIL in breast cancer this study investigated ANRIL expression in breast tumors using RT-qPCR. Additional assay for IL-27 were used in this study to evaluate the role of IL-27 in breast cancer metastasis. The study included 100 histopathologically confirmed with breast cancer. After blood collection, the serum was separated and divided into two aliquots, for total RNA extraction; cDNA synthesis and serum Interleukin-27 measurment. The ANRIL over expressed in patients with breast cancer (7.779940883) fold times compared to the healthy controls group (1.038805018). The fold change for the stages I, II, III and IV were (4.297018327, 5.553563985, 6.757504959 and 11.63189562) respectively. Twenty five cases with metastasis and fourty nine cases with lymph node involvement showed positive correlation with ANRIL expression. The levels of serum IL-27 were significantly increased in patients with breast cancer than in the control group (P=0.004). In addition, an elevation in serum IL-27 levels noticed in patients with metastasis (P=0.0001). Moreover, the serum levels of IL-27 was elevated in stage IV patients compared to stage I, II and III (P= 0.002, 0.0021 and 0.002 respectively). In conclusion these data indicate that ANRIL is highly expressed in malignant breast cells, and it may function in breast cancer prognosis, and the IL-27 may be a new prognostic biomarker of metastatic breast cancer.

Role of BIRC5 Gene Polymorphisms with Serum Level of Survivin in Iraqi Patients with Multiple Sclerosis

Masra M. Abdulkarim, Da’ad A. Hussain — Mon, 03 Jul 2023 00:00:00 +0200

Multiple sclerosis, often known as MS, is a disease of the central nervous system that is brought on by damage to the myelin sheath, which is a protective covering that surrounds and shields nerve cells in the brain and spinal cord. Undoubtedly, the immunological mechanisms are involved in the pathogenesis of this disease, but the initiating factors remain obscure. This study aimed to role the baculoviral inhibitor apoptosis protein (IAP) repeat containing 5 BIRC5) rs17878467 C/T) gene to regulate survivin levels in Iraqi patients diagnosed with multiple sclerosis. This study included hundred volunteers: fifty patients with multiple sclerosis, 18 (36.00%) males and 32 (64.00%) females. The age mean of MS patients was (38.14) years. While the fifty remaining samples were taken from healthy people 27 (54.00%) males and 23 (46.00%) females, the age mean was (41.04) years. Molecular study was performed in order to investigate the two single nucleotide polymorphisms (SNPs) for BIRC5 gene in chromosome 17 (rs9904341 and rs17878467) for studied groups which have determined using high resolution method (HRM) genotyping assay by real time polymerase chain reaction (RT-PCR) using the Qiagen rotor gene Q real-time PCR system. This study concluded that the rs17878467 C/T genotype was associated with increased risk for MS in Iraqi patients with highly significant difference. It was concluded that serum level of the survivin protein was higher in multiple sclerosis patients than the control group with statistically significant differences that can be considered a diagnostic marker.

Molecular Study of Carbapenem Resistance Genes in Proteus mirabilis Isolated from Clinical Samples in Baghdad Hospitals

Noor A. Al-Nabhani, AbdulMuhsin M. Shami — Mon, 03 Jul 2023 00:00:00 +0200

The majority of Proteus species are opportunistic pathogens that infect humans with a variety of illnesses including, infections of the respiratory system, nose, eye, ear, skin, and throat, as well as wounds and urinary tract infections. The aim of this study to isolate and identify of Proteus mirabilis; molecular screening of carbapenems Resistance genes (blaOXA-48, blaOXA-58 and blaOXA-23) of Proteus mirabilis and analysis its prevalence among local isolates in Baghdad. Two hundred and fifty samples have been collected from urine, ear swabs, burn and wound swabs of patients of both sexes and different age from several hospitals in Baghdad. The time between October 2022 to until the end of March 2023, has been used for the collecting of samples. From inpatients and outpatients in four hospitals in Baghdad. Gram's staining, microscopic examinations, cultural on MacConkey agar and blood agar plates and biochemical properties were used to identify all isolates. The diagnosis was confirmed through VITEK-2 system. According to the diagnostic results, out of 250 clinical samples, 225 were found to be positive, with 65 (26%) of the 250 isolates being Proteus mirabilis, while the remaining 160 (64%) belonged to other bacterial species. Antibiotic sensitivity test against 16 different antibiotics showed that Proteus mirabilis was highest resistance Piperacillin (89.2%), Cefotaxime (87.6%), Ceftazidime (86.1%), Cefepime (81.5%), Trimethoprim / Sulfamethoxazole (78.4%), Gentamicin (76.9%), Tetracycline (75.3%), While there is less resistance at Doxycycline (69.2%). On the other hand, moderate resistance Ceftriaxone (58.4%), Amoxicillin-clavulanic acid (56.9%), Amikacin (53.8%), Aztreonam (41.5%), and Ciprofloxacin (40%). Based on the results of the current study, P. mirabilis had only low levels of resistance to Levofloxacina (33.8%), Imipenem (15.3%) and Meropenem (1.5%). Conventional PCR using 16S rRNA housekeeping gene to Detection all isolates were P. mirabilis. In conclusion, Carbapneam resistance genes were identified by PCR. It was concluded that the existence of the OXA-23 gene gave (15, 61, 40, 43, 31, 21) isolates to be 60% resistant to carbapenem, whereas the presence of the OXA-58 gene in (6) isolates to be 60% resistant.

Prevalence of some Virulence Genes in Staphylococcus aureus Isolated from Iraqi Infertile Male Patients and its Effects on Sperm Parameters

1Haider K. Abbas, 2Abdulameer M. Ghareeb — Mon, 03 Jul 2023 00:00:00 +0200

Staphylococcus aureus is one of the most effective bacteria related with infertility and capable of immobilizing and agglutinating sperm cells. The aim of this study to identify of prevalence rate selected pathogenicity genes (can, clfa, scn and hlg) in S. aureus. This pathogen isolated from spermatozoa specimens of infertile males were attending Kamal Al-Sammrai Hospital / Baghdad during the period April 2020 until April 2021.Out of 120 cultures,25(20.83%)cultures utilized biochemical tests and microbiological assays to identify S.aureus isolates and confirmation by the Vitek 2 system. In addition, the molecular technique was based on The polymerase chain reaction assay was applied to identify according to the particular gene gmk encoding for (Guanylate Kinase).The results showed presence the gmk gene in all S. aureus isolates. The prevalence rate of examined virulence genes revealed that clfa, cna, scn and hlg genes present in 14 (56%), 14 (56%),13 (52%) and 7 (28%) among S.aureus isolates respectively. It is concluded that in general. S. aureus bacteria had negative effects on sperm parameters while in particular no effects of virulence genes in this bacterium on both sperm motility and sperm count.

Antibacterial and Cytotoxic Effect of Synthesized CuoNPs from Staphylococcus epidermidis

1Zahraa H. Kadhim, 2Mais E. Ahmed2, 1Ilker Şimşek — Mon, 03 Jul 2023 00:00:00 +0200

The risky of major concern resistance antibiotics become for public health. The alternative treatment metallic nanoparticles such heavy metals effects on antibiotic resistance bacteria different type antibiotic- impossible to treat using novel eco-friendly synthesis technique nanoparticles. The aim of the study to search for alternative antimicrobial medicines has been prompted by growing worries about multidrug resistance bacteria by synthetize novel nanoparticles. This study was to explore whether green synthesized copper nanoparticles (INPs) by multi drug resistance bacteria Staphylococcus epidermidis isolation from wound and burn infection. The results showed remarkable antimicrobial activity against Staphylococcus. aureus Minimum inhibitory Concentration range (16, 32, 64,128, 256, 512) µg/ml via well diffusion method in vitro, discover those concentrations effected in those bacteria and the best concentration is 64 µg/ml, characterization CuO NPs to prove this included atomic force microscope, Ultraviolet-visible spectrophotometer, X-ray Diffraction and Transmitted electrons microscopy, field emission scanning electron microscopy and anticancer activity was tested against cell membrane A375. The cells viability was decreased with increasing the copper nanoparticles and displayed a dose-dependent sequence of progressive cytotoxicity beginning at lower concentration to its maximum inhibition, (22) % inhibition human dermal fibroblast neonatal (HdFn) cells and (66) % inhibition of A375cells. It was concluded CuONPs were successfully synthesized friendly environment, few cost, highly CuO NPs product showed exceptional activity against bacterial strains and activity against cell line A375cells invivo.

Molecular Study of Siderophore Genes in Carbapenems Resistant Klebsiella pneumoniae Isolated from Urinary Tract Infection Patients

Zinah Sh. Ali, AbdulMuhsin M. Shami — Mon, 03 Jul 2023 00:00:00 +0200

Urinary tract infections (UTIs) frequently result from gram-negative, bacillus bacteria one of them is called Klebsiella pneumoniae .These pathogens are capable of acquiring genes that code for a variety of antimicrobial resistance mechanisms, such as carbapenem resistance. The goal of the presented work was to identify siderophore genes in carbapenemase K. pneumoniae bacterial isolates from urine samples collected from patients with complicated UTIs at five hospitals in Baghdad, Iraq. From November 2022 until the end of February 2023, samples have been collected,from five hospitals in Baghdad.A total of 180 urine samples from outpatients and inpatients who attended were taken In contrast to the ratio of male patients (38.33%), the majority of patients (61.67%) were female. In sterile containers, midstream urine (MSU) samples have been taken from patients exhibiting UTI symptoms. The samples have been incubated at of 37° C for 24 hrs while being grown on MacConkey agar, blood agar, and Hi Chrome UTI Agar. Through watching grown colonies and isolating Gram-negative bacteria, the outcomes were documented. Out of (180) clinical samples ,n=50 (27.77 %) isolates were recognized as K.pneumoniae ,then selected (10) resistance isolates to made the detdction of entB and ybtS genes .In this study, all n=10 (100%) isolates were determined positive for 16srRNA housekeeping gene, n = 10 (100%) of isolates were determined positive for the ybtS gene, and n = 9 (90%) of isolates were positive for entB gene. The current study was concluded, the high virulence factor regarding carbapenems CRKP could be due to large part to high rate of the detection of ybtS and entB genes in highly virulent CRKP.

The Effect of Antimicrobial Peptide LL-37 on Biofilm Formation of Klebsiella pneumoniae Isolated from Catheter-Associated Urinary Tract Infection Patients

Reyam M. Rifaat, Kais K. Ghaima — Mon, 03 Jul 2023 00:00:00 +0200

Opportunistic pathogen Klebsiella pneumoniae Most commonly causes nosocomial infections and particularly affects persons with weakened immune systems. For survival and immune evasion during infection, K. pneumoniae makes use of a variety of virulence factors, and caused many infections include pneumonia, bacteremia, UTI. The aim of this research was to assess the antimicrobial peptide's effectiveness against K. pneumoniae isolated from patients with UTI as an antibiofilm agent. This study included 170 urine samples which collected from October 2022 to March 2023 from patients with urinary catheters from Baghdad hospitals. The biofilm formation test was carried out for 28 strong biofilm isolates after treatment with LL-37 peptide. The results of biofilm formation after treatment with LL-37 peptide showed an obvious inhibitory activity where the most of isolates under test became ranged from non-biofilm to moderate. Results of five mdr K. pneumoniae isolates' lowest inhibitory concentrations (MICs) against LL-37 revealed that the MIC range of LL-37 were (1.9-500 µg/ml), in conclusion, LL-37 has a significant effect on K. pneumoniae growth and biofilm formation at very low concentrations.

Gas Chromatography-Mass Analysis and Evaluation of Antibacterial Activity of Mint and Basil Essential Oil

1Namareq F. Ahmed, 2ShurooK M . K. Saadedin, 1Hadeel Q . Rustam — Mon, 03 Jul 2023 00:00:00 +0200

Essential oils have grown in popularity in recent years due to their antimicrobial qualities. The aim of study to analyses the chemical component evaluate the antibacterial activities Mentha arvensis, Mint essential oil (MEO) and Ocimum basilicum, Basil essential oil (BEO), and compares the essential oil's effects in inhibition of Staphylococcus. aureus bacteria that responsible for enterotoxin production. Water distillation method used for MEO and BEO extraction, then analyzed secondary metabolites using Gas Chromatography-Mass Spectroscopy (GC-MS) techniques sixty-four compounds were identified from MEO (11.52%) 2-ispropyl-5-methylcyclohexanone(menthol), (11.33%) D-limonene, (9.92%) 2-cyclohexen-1-methylethylidene, (6.28%) Cyclohexanone and (3.71%) Cyclohexanol also fifty-three compounds were identified in BEO estragole (55.44%), linalool (18.41%), propanal,2-methyl-3phenyl (4.67%),(cyclopropylmethy) 4methyloxy benzene(1.99%), and 2(10) pinene (1.89%). This Investigation used an antibacterial disk diffusion assay and the MEO, BEO have antibacterial activities against S.aureus but MEO exceeds BEO as it more antibcterial activity against S. aureus. It was concluded that the diameter of the inhibition zones of MEO was greater than BEO.

Molecular Identification of Virulence Factors and Genotyping of Acinetobacter baumannii Isolated from Clinical Samples by ERIC-PCR

Narjis M. Abdulhussein, Mayaada, S. Mahdi — Mon, 03 Jul 2023 00:00:00 +0200

An opportunistic bacterium called Acinetobacter baumannii has significantly increased the frequency of infections in recent years. With only a limited number of “common” virulence factors, it is infections have spread rapidly through hospitals across the globe. The aim of the study a total of 150 samples were collected from various clinical sources from different age groups and gender patients in Ghazi AL Hariri Hospital and Baghdad Teaching laboratories in Medical City in Baghdad /Iraq during the period from December 2021 to March 2022. Bacterial isolates were identified and the Identification was confirmed by using Vitek-2 system. The result revealed that only (33.3%) isolates were given identical morphological belong to A. baumannii isolates. In all fifty clinical isolates of A. baumannii, a biofilm production profile was completed. The results showed that in 40 (80.0%) of the tested isolates strong biofilm was detected, while 10 (20.0%) of them were able to form moderate biofilms with an OD of 630 nm and mean ELISA reader values were between (0.1 and 0. 2). According to biofilm production tests, thirty bacterial isolates were selected, which are found to be strong biofilm production. The thirty selected isolates were introduced to molecular assay. The results revealed that all selected isolates (100%) have virulence genes (CsuE1 and OmpA genes). Depending on the thirty isolates, the ERIC-PCR banding patterns have indicated 2 to 6 bands with a total similarity level of about (74.1%). It was concluded.

Molecular Identification of Tinea spp. Causing Tinea Disease using ITS Sequencing Analysis

Qadisiyah H. Hashoosh, Alaa M. AL-Araji — Mon, 03 Jul 2023 00:00:00 +0200

Tinea is a group of keratinophilic fungal infections caused by dermatophytes that invade the skin, hair, and nails. As the conventional methods for dermatophytes detection are time-consuming or lack enough specificity, accurate diagnostic methods such as molecular techniques are required for precise identification and differentiate between two closely related species of dermatophytes. To disclose the useful role of DNA sequencing in the recognition of dermatophytes using the ITS region and comparing the current local isolates with those provided by the NCBI. The study aims to diagnose the dermatophyte isolates causing ring worm diseases using the inter transcribed spacer (ITS) region and find its phylogenetic analysis. one hundred ten (skin scraping, hair clippings and nails) samples were gathered from patients with dermatophytosis. All specimens were being microscopically inspected and then cultured on sabouraud's dextrose agar (SDA) with cycloheximide, PCR with ITS1 and ITS4 primers was used to confirm the positive dermatophytes' cultures. The PCR results were sequenced, and the rRNA database was used to do analysis on the sequences and verify the homogenic data of the microorganisms (NCBI). Substitutions in ribosomal RNA gene were found 37 isolates from dermatophytes and 8 isolates of them contained variations of the percentage of conformity with the gene bank 99% and they were registered with the gene bank and wrote down the new code that was registered in the table and in front of the heterogeneous sample. Trichophyton mentagraphytes recorded the highest frequency from cases. Trichophyton simii recorded the highest genetic several transitions, transversion, deletion, and insertion mutation when compared to other globally known. The phylogenic trees showed clear genetic interfere. It was concluded, demonstrate that the ITS region developed in this study can be used for detecting the Microsporum, Trichophyton, and Epidermophyton.

Assessment of HNF4A Gene Expression as a Potential Biomarker for T2DM Predicting and its Correlation with Biochemical Parameters in Iraqi Population

1Alaa H. Younus, 2Abdul Hussein M. Al-Faisal — Mon, 03 Jul 2023 00:00:00 +0200

Type 2 diabetes mellitus (T2DM) is a complicated disease that is caused by a combination of genetics and environment. The hepatocyte nuclear factor (HNF4A) is responsible for regulating hepatic gluconeogenesis and insulin production. Several studies have connected the corresponding gene to type 2 diabetes. This study involved the collection of 150 blood samples, which were divided into three groups: fifty youthful patients (10-35 years) and fifty elderly patients (40-85 years) with a previous diagnosis of T2DM. Additionally, fifty blood samples from healthy, typical individuals have been collected. All of the participants had their complete medical histories taken, received a comprehensive clinical examination, and had standard laboratory tests performed, such as blood sugar levels while fasting, hemoglobin A1c levels, and total cholesterol. Also, the gene expression of HNF4A in comparison with housekeeping gene GAPDH has been done. The results showed that levels of fasting serum glucose, hemoglobin A1c and total cholesterol had a significant increase in T2DM patients as compared to control group. The analysis of HNF4A gene expression revealed that there was no statistically significant difference for gene expression of the HNF4A in compression with control. The study conclude that a significant correlation existed between HNF4A gene expression with the biochemical tests.

Molecular Detection of Candida spp. Isolated from Female Patients Infected with COVID-19 in Baghdad City

Farah, H. Al- Qursh, Israa A. Ali — Mon, 03 Jul 2023 00:00:00 +0200

Coronavirus disease 2019 (COVID-19) is an infectious disease with severe acute respiratory syndrome and first recognized in Wuhan, China, and it has since spread to the world, resulting in the coronavirus pandemic to 2020. The present study aimed to evaluate Molecular study of some types of vaginal fungi isolated from recovered women from Covid-19 in Baghdad governorate. The study was conducted on 213 samples collected between December 2021 and March 2022, where the number of positive samples reached 188 with percentage 88.26%, while the number of negative samples reached 25 with percentage 11.73% by taking vaginal swabs from various female patients in Al- Kadhimiya Teaching Hospital. Three of Candida spp. were isolated: Candida albicans, which appeared in 60 samples with a percentage 41.37%, 50 isolates from Candida glabrata with a percentage 34.48% , 58 isolates from Candida pichia with a percentage 24.13% , all Candida spp_. It was conclude that molecular diagnosis results of fungal isolates using the polymerase chain reaction technique

Detection of FsrB Quorum-Sensing Gene and Biofilm Production in Enterococcus faecalis Isolated from UTI Women

1Hawraa A. Hamel, 1 Abdulameer M. Ghareeb, 2 Saja A. Shareef — Mon, 03 Jul 2023 00:00:00 +0200

Enterococcus faecalis are Gram-positive responsible for a variety of infections in susceptible hosts, including endocarditis, bacteremia, urinary tract infections (UTI), and mouth infections, The adhesion of E. faecalis to host tissues, notably urinary tract infections, is a crucial phase in the pathogenesis of the organism. The aim of the study Detection of a FsrB Gene and Biofilm Production in Enterococcus faecalis Isolated. The patients with various age groups who had urinary tract infections were included in this study for collection 150 urine samples from (Baghdad Hospital of Medical City and teaching laboratories of Medical City) in Baghdad Between November 2022 and February 2023. Urine samples were cultured on differential culture media and the isolated bacteria were identical according to cultural characteristics, biochemical test and VITEK- 2 system also molecular method was used in the identification of bacterial growth. The results revealed 32 isolates were belonging to E. faecalis and this was confirmed through amplification of species specific gene (23SrRNA). All 32 E. faecalis isolates were subjected to further molecular analysis to check for the presence of the diagnostic biomarker FsrB gene, which is linked to the quorum-sensing system that regulates the production of gelatinas to control biofilm growth. The results showed that 26 (81.2%) E. faecalis possess fsrB gene. Biofilm producers was detected in E. faecalis isolates using microtiter plate method (MIC) which that the results indicated that 9 (28.12%) of bacterial isolates were strong biofilm production, 11 (34.375%) as moderate and 12 (37.5%) as week biofilm production. Antibiotic susceptibility of the bacterial isolates was examined against 22 types of antibiotics. Isolates showed resistance to Cefpodoxime, Eryrthomycin and Ceftriaxone (96.875%), while the lowest resistance (12.5%) was against Amikacin. According to our study, the spread of Enterococcus faecalis began to increase significantly compared to previous years. The study proved that Enterococcus faecalis were highly resistant to Cefpodoxime, Eryrthomycin and Ceftriaxone and highly sensitive to Amikacin. The study proved that all isolates were biofilm producers. Also The study proved that all isolates contain the 23SrRNA gene The current in vitro study demonstrats the presence of FsrB quorum-sensing gene.

Prevalence of Multi-Antibiotic Resistance marA and Quorum Sensing luxS Genes and Evaluation of Biofilm Formation in Uropathogenic Escherichia coli

1Asmaa A. Muhaimeed, 2Abdulameer M. Ghareeb — Mon, 03 Jul 2023 00:00:00 +0200

Uropathogenic Escherichia coli (UPEC) are one of the main organisms that cause 80–90% of community-acquired urinary tract infections (UTIs) and catheter-associated urinary tract infections (CAUTIs). It is essential that UPEC have the ability to colonize the bladder by producing phenotypic virulence factors such flagella, pili, curli, and non-pilus adhesion. Pathogens growing on indwelling medical devices can communicate with one another using quorum sensing (QS) signals. The direction of the formation of biofilms, which result in the encasing of bacterial cells in extracellular polymeric substances (EPS), is crucially influenced by QS. The aim of this study was to evaluate the pathogenicity of the main pathogenic E. coli isolated from UTI patients and determine antibiotic resistance pattern and biofilm production. From December 2022 to February 2023, Out of 150 urine samples taken from UTI patients in various hospitals in Baghdad/Iraq, 25 UPEC isolates with various clinical UTI symptoms were found. These isolates were recognized by conventional and rapid techniques, and their identification was later verified by a molecular strategy based on the 16S rRNA gene. All 25 E. coli isolates were tested for antibiotic sensitivity using the Kirby-Bauer disk diffusion method on Muller-Hinton agar, and it was the highest resistance toward cefotaxime and amoxicillin-clavulanate, while the lowest toward meropenem, nitrofurantoin, and imipenem. Using traditional PCR techniques and specially designed primers, 25 UPEC isolates were screened for the presence of the multi-antibiotic resistance marA gene and the quorum sensing luxS gene. According to the results, both investigated genes were present in high percentages (92%). It concludes that this high percentage of marA is correlated with a high rate of antibiotic resistance and luxS, which indicate a high ability of biofilm formation; thus, the marA and luxS percent indicate the level of pathogenicity among multidrug-resistant UPEC.