Iraqi journal of biotechnology

Impact of Polyethylene Glycol 6000 and Sodium Chloride Stresses Combined and Single on Morpho-Physiological Traits and Expression of CAT Gene in Wheat (Triticum aestivum L)

1Muhammaed J. Husain, 1Kamil M. AL-Jobori — 2024-11-27

In some arid and semiarid places like Iraq, wheat growing may experience challenges with both drought and soil salinity. The goal of the present study was aimed to evaluate how drought and salt stress alone and combined affects growth and physiological characteristics and fresh weights of wheat plants to identify their response mechanisms. Two wheat genotypes (Latifya and Iba99) were treated with drought stress of-0.6Mpa (induced by PEG6000 solutions) ), Salt stress of -0.6Mpa (induced by NaCl solutions) and combined stress (0, -0.5, -1.0, -1.5, -2.0Mpa). Results revealed there was a marked variation in studied traits in these two genotypes under single and combined (D+S) stress. Stress treatment significantly decrease fresh weights of shoot, root and whole plant, leaf water content (LWC), relative water content(RWC),, chlorophyll (a), chlorophyll (b) , total chlorophyll, and Carotenoids ,but it increase water saturation deficit (WSD).electrolyte leakage(EL) , expression of CAT gene leaf content of Na⁺ and Cl^- . Interestingly, increased K⁺ at most stress treatments. Together, the findings showed that the combined effects of salt and drought stress could result in increased WSD, Na⁺ and Cl- buildup, and membrane degradation. It was concluded the Latifya genotype demonstrated excellent performance for all examined traits. It is clear that the Latifya genotype is more tolerant to stress than the Iba99 genotype.

Isolation and Identification of Staphylococcus aureus from Iraqi Patient with Atopic Dermatitis in Baghdad Hospitals

1Shaimaa A. Yassin, 1Zainab H. Abood — 2024-11-27

Atopic dermatitis (AD) is one of the most chronic skin diseases that designated that redness, irritation and sever lesion. S. aureus has been tandemly detected in those lesions. The goal of the study was to characterize the aerobic and anaerobic microbiology of AD that had been secondary infected. Swab samples have been collected from patients diagnosed with AD. Specimens were examined microscopically and aerobically during cultivation using suitable culture media. The antibiotic susceptibility and the diagnosis of the bacterial isolates were confirmed using the VITEK 2 system. The results were interpreted according to the Clinical and Laboratory Standards Institute (CLSI) guidelines for 2020. The PCR technique was applied to characterize the Methicillin-resistant Staphylococcus aureus (MRSA) pathogen and determine the prevalence of its most important virulence genes, including 16S rRNA and the SPA gene. Eight different bacterial strains were tested from 200 AD patient swaps; all bacterial types isolated from eczematous lesions and nearly healthy areas were considered 80% S. aureus isolates and 20% other strains from eczematous lesions, while 40% S. aureus isolates and 60% other bacteria from nearly healthy areas. It was concluded all isolated S. aureus showed high resistance to the B-lactam group but showed variable susceptibility to other antibacterials used.

The Role of miRNA-9 as a Predisposing Factor for Metastasis in Breast Cancer among Iraqi Patients from Localized to Locally Advanced and Metastatic Stages

1Mays T. Abdullah, 1 Ismail H. Aziz, 2 Ahmed Z. Alsammarraie — 2024-11-27

Breast cancer (BC) is the highly prevalent malignancy afflicting women worldwide, with an alarming 2 million new cases diagnosed in 2020. The escalating incidence and mortality rates of this disease have demonstrated a consistent upward trend over the past three decades. These trends are primarily attributed to modifications in risk factor profiles, advancements in cancer registration methodologies, and enhanced capabilities in cancer detection techniques. This study was aimed at the Oncology Teaching Hospital/Baghdad Medical City and the Oncology Unit at Al-Yarmouk Teaching Hospital in Baghdad, encompassing a cohort of 150 samples divided into two groups: a blood group comprising 90 samples (control, localized, locally advanced, and metastatic BC patients) and a tissue group comprising 60 samples (benign and malignant BC). The study spanned from March 2022 to January 2023, involving patients aged 24 to 75 years. The primary objective of this investigation was to assess miRNA-9 gene expression across all sample types, with gene expression levels normalized to the housekeeping gene U6 and quantified using the ∆Ct value and the fold change (2-∆∆Ct) method. The results concluded an upregulated fold expression of miRNA-9, with the highest expression observed in locally advanced and metastatic BC (fold expression 2.404±0.1364 compared to other groups. In localized breast cancer, the fold expression was 1.795±0.092, and in malignant tissue, it was 1.972±0.119, both compared to the apparently healthy control group.

Study Association between DRD2 Gene Expression and Infertility among Some of Iraqi Females

1Eman A. Faris AL-bandawi, Essam F. AL-Jumaili, 2Zaynab K. Alhusni — 2024-11-27

Infertility is commonest endocrine disease occurring in women of reproductive age. The aim of study to determine the D2 dopamine receptors gene expression and clarify altered concentrations of dopamine in infertile women and controls. Also, to assess their role in infertility and their correlation with measured biochemical parameters .This study was carried out in the Institute of Genetic Engineering and Biotechnology for Postgraduate Studies - University of Baghdad through the period from January 2023- April 2023, The patients were taken from the Kamal Al-Samarrai Infertility Treatment Hospital in Baghdad the study was included 100 individuals (50 controls and 50 infertile females). Biochemical examinations were done include serum concentrations of dopamine (DA), luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin were measured by enzyme-linked immuno sorbent assay (ELISA) and enzyme linked fluorescent assay (ELFA) techniques. The RNA was extracted from whole fresh blood the concentration and purity of RNA Based on nanodrop device was performed, then converted to cDNA to measure the level of gene expression for DRD2 by using Real Time PCR technique then converted to cDNA to measure the level of gene expression for DRD2 by using Real Time PCR technique. In this study, the results showed a significant decreases in dopamine level in infertile Women in comparison to controls group (P< 0.01). In addition The analysis of hormones level (LH), (FSH) and (PRL) infertile female showed a highly significant increase in PRL and LH compared to controls group (P≤0.01). The results of the fold expression of DRD2 was showed down-regulated in patient, whereas it was up-regulated in control group. In conclusion gene expression of DRD2 gene and Dopamine can be utilized as biomarkers for early diagnosis of female infertility in Iraqi femals as one of the laboratory diagnostic methods.

Linezolid Resistance and Biofilm Formation in Invasive and Commensalism Staphylococcus Epidermidis

1Hind H. Muneam, 1May T. Flayyih — 2024-11-27

Staphylococcus epidermidis is a fundamental reason of catheter-related infections septicemia condition in immunocompromised individuals. To determine if skin-related S. epidermidis isolates are distinct from those that cause septicemic illnesses. This study aim to comparative the ability of bacterial to produce biofilm and linezolid resistance genes S. epidermidis. Methods 150 specimens were isolated from Urine, wound and blood from different hospitals and 150 specimens from urine and fingerprintf healthy individual in Baghdad city from July 2021 to January 2022. The bacterial isolates were recognized with biochemical test and vitek 2 system. for their potential role in biofilms formed the polystyrene micro-titer plate (MTP) estimate using to measure the capacity to biofilm producer. However, the MTP method capably to categorized the bacterial isolates to strong, moderate, weak, and non-biofilm producers. The biofilm genes (icaA and icaD) and Linezolid resistance gene cfr were detection by PCR method. The results of this study showed 30 (10%) Staphylococcal epidermidis strains were isolated and identified from 300 samples. The MTP method was classified the isolates to 11/30(36.6%) strong, 6/30(20%) moderate and13/30 (43.3%) weak biofilm formation. The PCR results revealed that 11/15(73.3%) and 7/15(28.5%) from healthy and clinical Staphylococcus epidermidis isolates respectively were complete icaABCD Operon, 4/11(36.3%) and 2/7(14%) were cfr gene positive. While 3/15 (20%) and 8/15(53.3%) were defect icaADBC operon, which contain 3/8 (37.5%) cfr gene positive and no one from healthy samples. Finally, 1/15 (6.6%) lacking to icaABCD operon and cfr positive strains. It was concluded, icaA, icaD and cfr genes have a significant role in staphylococcal infections caused as a virulence marker.

Molecular Detection of adeL, adeR and adeS Genes of Acintobacter baumannii Isolated from Different Clinical Samples

1Ahmed A. Faraj, 1Wathiq A. AL-Draghi — 2024-11-27

The emergence of multidrug-resistant Acinetobacter baumannii has become a global concern, particularly for patients susceptible to burn, blood bacteremia, and wound infections. In this research endeavor, was collected and analyzed 150 clinical samples from various sites within Baghdad/Iraq hospitals, specifically the Laboratory of Medical City, spanning from October 2022 to March 2023. Through meticulous culturing procedures, A. baumannii isolates were identified via microscopic examination and biochemical assays and Vitek-2 tests. To further validate the identification, a molecular approach was employed, confirming the presence of the 16S-rRNA gene in (n=32) of the A. baumannii isolates using PCR analysis. This molecular verification reinforced outcomes obtained through conventional diagnostic techniques including culture, biochemical tests, chromagar selective media and Vitek-2 tests. An exhaustive antibiotic susceptibility evaluation was performed, revealing multi-drug resistance in 32 isolates. Significantly, all isolates exhibited the presence of the adeL gene, while 31 and 22 isolates carried the adeR and adeS genes respectively. This study was concluded comprehensive insights into the prevalence of multidrug-resistant A. baumannii in Baghdad hospitals. It was concluded highlighting the significance of molecular techniques in accurate identification and understanding antibiotic resistance patterns.

Impact of siRNA-Mediated Gene Silencing on IntII Gene Expression for Clinical Isolated Pseudomonas aeruginosa

1Albab F. Alfarras , 1Wathiq A. Hatite Al-Daraghi — 2024-11-27

Integrons hold significant importance in gram-negative bacteria, which are known for being susceptible to acquiring antibiotic resistance. The aim of this study was to identify Integron class II (intII) genes and induce silencing in the intII gene by designing small interfering RNA (siRNA) molecules specifically targeting this gene in Pseudomonas aeruginosa and demonstrating a silencing effect on antibiotic resistance. The DNA was extracted from 30 clinical isolates of P. aeruginosa to identify the intII gene, and a specific siRNA was designed and attached to gold nanoparticles (AuNPs) for delivery into bacterial cells. The replica plate method and RT-PCR techniques were applied to assess the impact of silencing. The findings indicated the presence of the intII gene in 10% of the isolates, and the process of joining siRNA and AuNPs was very effective. Phenotypically silencing that was induced by siRNA showed significant differences between transformed (sensitive) and non-transformed (resistant) colonies, but in contrast, gene expression of the intII gene was increased after being treated with siRNA. The study concluded that siRNA-AuNPs could be a potential therapeutic agent for P. aeruginosa infections. Based on the findings, treatment with siRNA showed a very high significant difference between transformed (susceptible) and non-transformed colonies (resistance), and the pre-treatment with siRNA dramatically reduced the bacteria load.

Study the Expression of miRNA34a-5p Gene among Iraqi Breast Cancer Patients

1Sama S. Majeed, 1Ismail H. Aziz — 2024-11-27

MicroRNAs (miRNAs) are short non-coding RNA sequences (approximately 22 nucleotides) that play an important role in gene regulation. This research is to evaluate the serum expression (miR34a-5p) in the Iraqi population as important biomarkers. Circulating serum miRNA34a-5p expression was measured using RT-qPCR in 50 patients women suffering from breast cancer compared to 50 apparently healthy controls. The significant increases in groups (41-50) years old (p=0.016) and over 60 years old (p=0.02) compared to control groups. 28 years old (46%) of these 50 breast cancer women were under 50 years old, and 22 years old (44%) were over 50 years old. According to body mass index, it is a significant difference in the frequency distribution of patients and control subjects according to BMI (P = 0.020). Where high frequency distribution of patients with breast cancer occurs in the obesity group 23 patients (47%) compared with other groups. The results showed the expression of miR34-5p is high1y significant (p=0.004) in the patients compared with contro1groupe.

Antibacterial Activity and Molecular Detection of Plantaricin

1Aula H. Obaid , 1Ali H. Saliem — 2024-11-27

Escherichia coli O157:H7 is the cause of more people and animals getting sick with diseases like hemolytic uremic syndrome (HUS) and enterohemorrhagic diseases (EHEC).This study was conducted for molecular detection and evaluate the antibacterial activity and mechanism of action of plantaricin against E. coli O157:H7 in comparison with ciprofloxacin. This experiment was carried out through obtaining plantaricin from Lactobacillus plantarum and study the sensitivity of the E.coli O157H7 by MIC and MBC and identify its mechanism of action by Fe-Scanning electron microscope (Fe-SEM). The plantaricin showed pronounced concentration dependent antibacterial activity. The results of the presence study activity suggest plantaricin may have the perfect to be choice in clinical control. The result of inhibition of plantaricin was ( 25.72 mm to 30.83 mm) while the result of MIC was (0. 625mg/ ml) and MBC was (1.25 mg/ ml) , while for ciprofloxacin, the result was( 16.75 mm to 22.04 mm ) , at (20 and 40µg/ ml ) respectively . I was concluded The Fe-SEM identification indicated that DNA leaked from cells due to the cell lysis of E.coli O 157:H7 and plantaricin interacts with the target cell membrane.

Evaluation of IL27 Gene Expression Related with Treatments in Samples of Multiple Sclerosis Iraqi Patients

1Zahraa K. Lafi, 1Bushra J. Mohammed — 2024-11-27

Background: Multiple sclerosis (MS) is a neurological and demyelinating illness that causes persistent nerves inflammation. The causes of this illness are varied and complex. The goal of the current research was to evaluate Interleukin-27 (IL27) gene expression in a sample of Iraqi patients with MS. Methods: Blood samples were collected from 75 Iraqi patents suffered from MS, who taken various treatments as Interferon-ß, Fingolimod, Retuxan and Natalizumab. Also, the study included 75 apparently healthy volunteers as a control group, all subjects with age ranged between (22-58 years). IL27 gene expression was estimated by using Real-time Polymerase Chain Reaction (RT-PCR). The statistical analysis was done by Statistical Analysis System (SAS) to identify the impact of different factors on research parameters Results: The results found that the most commonly medication used in treatment MS Iraqi patients was Betaferon 30 (40%) followed by Gilenya 23(32%) with highly statistical significant (P≤0.01).The results of estimation IL27 gene expression showed noteworthy decrease in patients’ group (1.25+0.08 fold) when compared with control group (3.24+0.09 fold) at highly significant difference (P≤0.01), also the outcome of treatment effect on IL27 gene expression in MS patients revealed that no significant differences between medication types in spite of the patients who taken Betaferon had the highest level of IL27 gene expression (1.40+ 0.09 fold) among other patients . Conclusion: It was concluded that a significant decrease of IL27 gene expression levels in MS patients as compared with control, and these levels were not affected by the different of treatments types.

Demographic and Biochemical Study of Beta Thalassemia Major Associated with Liver and Pancreas Disorders in Adult Sample of Patients from Baghdad province

1*Hawraa A. Luaibi , 1Bushra J. Mohammed — 2024-11-27

β-thalassemia is an inherited autosomal recessive blood disorder results from decreased or missing synthesis of beta globin chains in hemoglobin. This study was aimed to examine the relationship between liver and pancreas disorders of beta thalassemia major with demographic and biochemical aspects, in adult Iraqi patients. Blood samples were collected from 40 patients suffered from beta thalassemia with pancreas disorder as group A, along with 40 patients suffered from thalassemia with liver disorder as group B, and 40 patients suffered from thalassemia without pancreas or liver disorders as group C, from Ibn Al-Baladi Hospital, Baghdad, and 40 samples from age and gender-matched apparently healthy individuals as a negative control (Group D), all subjects with age more than 18 years. Ferritin serum level was determined by using automated immunocheiluminometric analyzer. ALT level was estimated through Atellica™ CH Analyzer, and amylase was evaluated via ARCHITECT c4000 device. The results revealed that the highest number of thalassemia patients located at the age group (18-28) years with high significant difference (P≤0.01), which was 91 (75.83%), and higher incidence was for male patients 72 (60%) with significant difference (P< 0.05). Also results showed elevation of ferritin level in all patient groups when compared with control, and the highest value appeared in liver disorder patients (4014.9 ng/ml) with high significant difference (P<0.01). The results revealed that ALT level was significantly higher (76.430 U/l in liver disorder patients, as well as results showed that the level of amylase was lower in all patient groups as compared to the control group (84.150 U/l) with high significant difference (P<0.01). This study concluded that there was relationship between liver and pancreas disorders of beta thalassemia major with ferritin, ALT and amylase in adult Iraqi patients.

Pin1 Gene Expression and Some Biochemical Parameters in Iraqi Population with Chronic Kidney Disease

1Safa S. Mahdi Al-Shattawi, 1Essam F. Al-Jumili, 2Ula M. Ridha AL-Kawaz — 2024-11-27

A widespread general medical problem is chronic kidney disease (CKD). It is a primary source of illness and mortality globally and is becoming more acknowledged as a global public health concern, particularly in developing nation. The purpose of this study was to assess the clinical importance of Pin1 gene mRNA expression in samples taken from CKD patients in Iraq and some biochemical parameters. Blood samples from 120 individuals were collected and sorted into the following three groups: Group 1 consists of 40 samples from CKD patients on dialysis, Group 2 of 40 samples from CKD patients who are not on dialysis, and Group 3 of 40 samples from controls who appear to be in good condition. Reverse transcription polymerase chain reaction (RT- PCR) was used to evaluate the messenger RNA (mRNA) expression levels of Pin1 in peripheral blood, and GAPDH was used as a housekeeping gene. Serum parathyroid hormone (PTH) was determined by using ELISA Kit assay. The renal routine test includes serum urea and serum creatinines were detected using an automatic biochemical analyzer (SIMENS ATELLICA, USA). Results revealed that non-Dialysis CKD patients had significantly higher level of Pin1 gene expression (P >0.01) than Dialysis CKD patient and control. The level of serum parathyroid hormone (S.PTH) was show highly significant increase with (p>0.01) in non-dialysis CKD patient than other group (dialysis and control group) while serum urea (S.Uea) and serum creatinin (S.cr) were show high significant increase with (p>0.01) in dialysis group compare to other study groups. It was concluded a high level of expression of Pin1 gene in chronic kidney disease with SHP and high protein level of PTH.

The Role of Parathyroid Hormone Receptor 1 Gene Polymorphism at rs1138518 SNP in the Incidence of Osteoporosis in a Sample of Iraqi Women

1Bushra A. Jassam Al-Juaifari, 2Essam F. Alwan Al-Jumaili, 3Ali A. Mahdi — 2024-11-27

Osteoporosis is a medical condition characterized by the weakening of bones, which makes them fragile and more prone to fractures. It is often associated with bone mineral density (BMD) reduction and changes in bone microstructure. Osteoporosis is influenced by a combination of genetic, diet and lifestyle factors, specific genetic variants can contribute to an individual's risk of developing the condition. The PTHR1 gene encodes the parathyroid hormone receptor1, which plays a role in calcium homeostasis and bone remodeling. Genetic variations within this gene, including rs1138518, have been studied in relation to bone health. The study aimed to detect the specific relationship between the rs1138518 variant on the PTHR1 gene and osteoporosis in a sample of Iraqi women their ages between (20-50) years old. A total number of 120 samples: two group of (60) women diagnosed with osteoporosis and (60) apparently healthy control. The DNA was extracted for both groups and the PTH1R SNP (rs1138518) gene were detected by qRT-PCR. The result showed there was high significant difference between patient and control in genotypes of mutant allele (AA) (51.67% vs. 13.33%, respectively; odds ratio =1.376; p-value =0.0001**, and that A allele frequency was significantly increased in patients compared to control (0.67 vs. 0.33) and the wild allele T showed decreased frequency in patients (0.22 vs. 0.78). The genotype of rs1138518SNP was showed significant difference between patient and control in PO4 (p-value= 0.05*). The TA genotype of rs1138518 SNP was effective in increasing the serum level of MMP-9 in osteoporosis patients (4006.13 ±131.30) compared TT or AA genotype (3495.12 ±174.01 and 3581.09 ±95.10, respectively; p=0.017*). This study was concluded there is an A allele – related risk factor for osteoporosis in Iraqi women, while the wild allele T might have a protective effect.

Study the Association of mi 423-5p Expression with Coronavirus-19 Infection in Samples of Iraqi Patients

1Israa H. AL-zubaidy, 1 Saife D. AL-Ahmer , 2 Mohammed I. Mohsin Aldafaee — 2024-11-27

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the agent responsible for the extremely contagious sickness known as coronavirus disease 2019 (COVID-19). Non-coding RNAs known as microRNAs (miRNAs) have a vital role in controlling the expression of genes. This study aims to evaluate the role of mi423-5p as a biomarker for the detection and monitoring of COVID-19, it was planned to achieve that through the following steps: a collection of 50 nasopharyngeal swabs and 50 blood samples from the COVID-19 Patient group, and control group (health), detection of gene expression of miR-423-5p by using qRT-PCR.The statistical analysis data of mi 423-5p indicates that there were statistically significant differences between the infected patients and control and patients had the vaccination or not, mi 423-5p was significantly upregulated in patients, that the P value =0.012, and there were significant differences whether the patient had the vaccination or not, the p-value was = 0.028. The study was concluded that mi423-5p could be useful as a biomarker for COVID-19 infection and it also helps evaluate the effectiveness of vaccines used against COVID-19.

Detection of Biofilm Operon, Some Virulence Factors, and Antibiotics Susceptibility of S. aureus Isolated from Patients in Holly Karbala City

1Zahraa M. Wannas, 1 Hayfa H. — 2024-11-27

The formation of biofilms in Staphylococcus aureus has the potential to impede the effectiveness of antibiotics, leading to challenges in infection treatment. The aim of this research is to determine the prevalence rate of some virulence genes in S. aureus isolate and learn more about the virulence components of S. aureus. The S. aureus was clinically isolated from different 250 samples in Karbala City (al-Husain Medical City and al-Hassan al Mujtaba Teaching Hospital). Isolate biofilm formation was examined using a microtiter plate assay. The biofilm operon and some of the MSCRAMMs (microbial surface components recognizing adhesive matrix molecules) genes were detected by PCR assay. The disk methods and the VITEK 2 compact were utilized to determine antibiotic susceptibility. The sixty S. aureus isolates were isolated and identified. The microtiter plate method divided the isolates into (25%) strong, (66.6%) moderate, and (8.3%) weak biofilm. PCR assay results showed that 100% of isolates have ica operon, 100% have clumping factors genes, and 0% of (bap). This research proved the association between the icaADBC operon and biofilm production in S. aureus.

Gold Nanoparticles Biosynthesized from Synephrine Extracted from Citrus aurantium Peels and its Cytotoxicity Assay

1Ayat S. Jadou, 1 Rusol AL-Bahrani — 2024-11-27

Nanotechnology is one of the most productive fields of study in current science.Nanoparticles (NPs) and nanomaterials are finding new uses at a rapid rate. The aim of the study that synthesis of nanoparticles from peels, which are considered Citrus auriantum. In this study synephrine was extracted from C. auriantum peels and biosynthesized gold nanoparticles from it, then characterization of synthesized gold nanoparticles (AuNps) by UV-visible spectroscopy Fourier Transform Infrared (FTIR) and FESEM which confirm the presence of AuNps in diameter size range to (20.8-34.3) nm and determined their toxicity assay on lymphoid human cell in 24 ,48 and 72 hours least cytotoxicity value was in 3.013 in 10 mg/ml for Citrus auriantum extract in 24 hours. It was concluded that synephrine extracted has no toxic effect inhibition rate of this maximum 5.698 in AuNp biosynthesized in 20 mg/ml concentration in 72 hours.

Evaluation of IL10 Gene Expression in Samples of Systemic Lupus Erythematous Iraqi Patients

1Sajood H. Abd allah, 1Wiaam A. AL-Amili — 2024-11-27

Systemic Lupus Erythematous (SLE) is a type of chronic multisystemic autoimmune disease which impacts women more frequently than males. The antibodies produced by the immune system attack healthy cells The goals of the current research were to evaluate IL10 gene expression in a sample of Iraqi women patients with Systemic Lupus Erythematous and determine the influence of IL10 gene expression level on SLE disease. A total of 54 women blood samples were obtained from individuals diagnosed with Systemic Lupus Erythematous (SLE) in Iraq. Additionally, 46 seemingly healthy women were included as a control group. The age range of all participants was between 15 and 51 years. The assessment of IL10 gene expression was conducted using Real-time Polymerase Chain Reaction (RT-PCR). The results of IL10 gene expression detection showed that no differences between the control group (1 fold change) and the patient group(1.043 fold change). It was concluded no effect of IL10 gene expression in development SLE Iraqi patients.

Detection of Some Capsule Genes (ugE, wabG and ycfM) in Carbapenem Resistance Klebsiella pneumoniae Clinical Isolates from Baghdad Hospitals

1Dalal A. Salman, 1 Luma S. Mohammed — 2024-11-27

The rising prevalence of carbapenem-resistant K. pneumoniae has emerged as a significant concern in the field of public health. Consequently, it is imperative to conduct an in-depth study of this issue, particularly in relation to the potential association between capsule genes and the development of carbapenem resistance. The aim of the study identifying effective strategies to reduce the dissemination of carbapenem antibiotic resistance. The 120 samples were collected from different clinical specimens (burns, sputum, urine, and wound) between October 2022 and January 2023.), according to culturing , microscopic examination, biochemical tests and vitek 2 system , only 50 isolate (41.6%) isolates were found to be Klebsiella pneumoniae. The percentages of isolation were found to be (46%) by sputum (22%), (20%), and (12%) in urine, sputum, wounds and burns respectively. Antibiotic sensitivity testing on three types of carbapenem discs, namely imipenem, meropenem, and doripenem, revealed that meropenem was the most effective antibiotic, followed by imipenem, and finally doripenem, with resistance percentages of (30%, 40%, and 56%, respectively). By using PCR specific primers designed particularly for this study, capsule associated genes were detected in twenty-five resistant and sensitive isolates. cfm was found to be the most predominant gene, followed by wabG and uge, with percentages of (88%, 84%, and 72%, respectively). It was concluded there is an association between capsule associated genes and carbapenem resistance in K. pneumoniae.

Evaluation the Serum Level and Gene Expression of IL-6 in Iraqi Systemic Lupus Erythematous Patients

1Amina S. Jalal, 1Wiaam A. AL-Amili — 2024-11-27

The chronic systemic autoimmune disorder known as systemic lupus erythematous (SLE) can be distinguished serologically by both cellular and humeral immunologic abnormalities such as direction of many autoantibodies against non-organ specific intracellular antigens. The SLE is created by a combination of several environmental and genetic factors. The current study included 54 Iraqi females with SLE who were diagnosed by specialists in Baghdad Teaching Hospital / Medical City and Kadhimiya Teaching Hospital from June 2022 until September 2023 and 46 healthy women with an age range of 10-50 years old. This study aimed to evaluate the serum level and gene expression of interleukin-6 (IL-6) in SLE patients using ELISA technique and real time-PCR in addition to the possibility of their relationship with the disease pathogenesis. The demographic study showed that the highest age group of patients was 40.74% (21-30). Results show that the level of IL-6 serum were highly significant (p< 0.01) increased in patients group compared to control group. Additionally, IL-6 gene expression was increased about eight folds compared to control group. These results pointing to the association of IL-6 with SLE. The present study recommend to use IL-6 as a biomarker for SLE.

Evaluation of miRNA-126 Expression and Some Immunological Parameters as Diagnostic Biomarker in Chronic Myeloid Leukemia Iraqi Patients

1Aya K.. Mohammed, 1Zahraa K. Zedan — 2024-11-27

Chronic Myeloid Leukemia (CML) is a hematologic malignancy characterized by the uncontrolled proliferation of myeloid cells in the bone marrow. Recent advancements in cancer research have highlighted the crucial role of cancer stem cells (CSCs) in tumor initiation, progression, and treatment resistance. The aim of this research is to investigate the potential of miRNA-126, CD33, and TNF-alpha as biomarkers for identifying and characterizing CSCs in a cohort of Iraqi patients with CML.The research methodology involves the collection of bone marrow samples from a cohort of Iraqi CML patients. Subsequently, various molecular techniques, including polymerase chain reaction (PCR) and flow cytometry, are employed to analyze the expression levels of miRNA-126 and CD33, in these samples. The choice of these biomarkers is based on their known associations with cancer stem cells and their relevance to CML.The results of this study showed that there were significant increases in miRNA-126 and CD33 (p<0.05) in a sample of Iraqi patients with CML compared to healthy people. It was concluded the dramatic increases in the expression of each parameter as shared above.

Investigation of IL-4 (rs2243250) Gene Polymorphism in Sample of Iraqi Allergic Patients

1Mohammed S. Jebur, 1Suham A. Jaber, 1Assad A. Muhsen, 1Ahmed S. Mohammed — 2024-11-27

Allergy is the reaction by your immune-system. Substances that cause reactions include food and pollen. The study was aimed to reveal circulating T-lymphocyte-subsets and related cytokines during asthmatic-attacks is still unclear. This study included 89 subjects total; 31 controls, 23 Allergy Food and 35 Allergic Rhinitis individuals. The aim of this study is to estimate the Total IgE levels in allergic patients and healthy control subjects. Also, investigates about single nucleotide polymorphisms (rs2243250) of IL-4 genes in allergic diseases such as, allergic rhinitis and allergic food. Patients' serum IgE levels were noticeably greater than those of the controls (AF: 234.4 ± 35.8, AR: 253.4± 29.1, control: 48.0 ± 5.7 IU/ml, P<0.001). The CC genotype in the IL-4 gene was not related with AR patients. Additionally, the polymorphism (rs2243250 = C-590T) in the IL-4 gene had no correlation with AR in the sample being examined due to the sample size of the subject was small. It was concluded the sample being examined and IgE is a unique immunoglobulin that plays a central role in the pathophysiology of allergic disorders.

The Association of Paraoxonase 1 (PON-1) Gene Polymorphism at rs854560 SNP (L55M) and Some Oxidative Stress Parameters in Serum with the Incidence of Recurrent Spontaneous Abortion in Iraqi Women

1Zainab T. Hussein, 2Ismail A. Abdul-Hassan — 2024-11-27

Recurrent spontaneous abortion (RSA) is characterized by the occurrence of two or more spontaneous pregnancy losses before reaching the midpoint of gestation. Oxidative stress assumes a central role in the development of RSA, serving as a key factor in its pathogenesis. The objective of this study is to explore the link between the L55M polymorphisms found in the PON1 gene, which codes for an antioxidant enzyme, and the susceptibility to RSA.Paraoxonase 1 (PON1) is a critical enzyme in the body's antioxidant defense system, safeguarding cells against damage caused by reactive oxygen species. The most prevalent PON1 polymorphism, L55M (rs854560), is known to influence PON1 activity. In this research, blood samples were collected from two groups: 50 women with a history of RSA and 50 women who were apparently healthy. Following DNA extraction, Restriction Fragment Length Polymorphism (RFLP) analysis was employed in conjunction with PCR amplification to identify single-nucleotide polymorphisms (SNPs) and ELISA for determining PON1 activity, total antioxidant capacity (TAC) and HDL concentration in serum. The result of this study showed decreasing in PON1 activity (8.15 ± 2.15 versus 9.66 ± 3.75 , respectively) and TAC (2.80 ± 0.10 versus 3.53 ± 1.52, respectively) in patients with RSA than apparently healthy women, while HDL concentrations higher in women with RSA than apparently healthy women (12.68 ± 5.53versus 9.01 ± 5.30, respectively) which are lead to increase oxidative stress and related with RSA. The genotype results showed that the percentage of TA genotype was in women with recurrent spontaneous abortion significantly higher than that of apparently healthy women (60% versus 32 %, respectively, OR=3.5) and this represent a risk factor for recurrent spontaneous abortion incidence. Also, the percentage of AA genotype in women with recurrent spontaneous abortion was higher than that of apparently healthy women but non-significant (12% and 4% 68%, respectively). The values of allele frequency for T allele were 0.8 and 0.6 for apparently healthy women and women with recurrent spontaneous abortion, respectively. The values of allele frequency for A allele were 0.2 and 0.4 for apparently healthy women and women with recurrent spontaneous abortion, respectively. It was concluded the heterozygous TA genotype is related with the risk of RSA and the minor allele (A allele) increased in women with RSA.

Association of TGF-B1 Protein Level with Breast Cancer Risk in a Sample of Iraqi Women

1Muhammad M. Yasser, 1Fadi S. Hekmet, 1Muhammad A. Ali AL-Bedhawi — 2024-11-27

Breast carcinoma is the most prevalent cancer-related cause of death in women, and metastasis is the main factor in morbidity. The aim of the study level of TGF-B1 between the age groups. The total number of new cases of cancer in Iraq during the year 2019 was 35,864. Transforming growth factor (TGF) is a multifunctional cytokine whose abnormal expression is linked to cancer development and metastasis. The tumor growth factor TGF-β is a major component of epithelial–mesenchymal transition (EMT), immune evasion, and metastasis during cancer progression. It is produced by a variety of cells within the tumor microenvironment (TME), and it is responsible for regulating the activity of cells in this milieu. TGF-b is also central to immune suppression within the tumor microenvironment, and recent studies have revealed roles in tumor immune evasion and poor responses to cancer immunotherapy. Then, focusing on cancer, we discuss the roles of TGF-b signaling in distinct immune cell types and how this knowledge is being leveraged to unleash the immune system against the tumor. The aim of this study to investigate the role of TGF-B1 in Breast Cancer risk Method: measure the protein level by ELISA Sandwich method. It was concluded that found significant increase in cancer patients compare with controls, according to age group that was no- significant association and the result show significant increase in metastasis patients compare with non-metastasis group.

Assessment of MicroRNA (miR-146a) as Potential Biomarkers and Disease Activity in Iraqi Patients with Rheumatoid Arthritis after Biological Treatment

1Aseel A. H. Hussien, 2Mohammed I. Nader, 3Waseem K. Kaith — 2024-11-27

Rheumatoid arthritis (RA) is a debilitating form of persistent autoimmune inflammation that affects the joints. Rheumatoid arthritis's primary treatment is disease-modifying anti-rheumatic agents (DMARDs), such as methotrexate (MTX) and Enbrel. The current study aimed to measure the gene expression level of MiR146a as well as demographic data and clinical presentations for patients with RA compare with control models after biological treatment. The study includes 70 patients with rheumatoid arthritis and 30 healthy individuals as a control group, with an age range between 20 to 60 years. Blood was drawn from everyone who participated in this study. In this study, ESR, HB, and WBC were evaluated in rheumatoid arthritis patients. The findings showed that the majority of age groups were in the third group (40-49 years), where there was a 26.1% percentage, and that the lowest number was in the first group, which included ages 29, where it was 8.7 percent. Erythrocytes Sedimentation Rate (ESR), C-reactive Protein (CRP), CDAI and smoking were examined in this study with concentrations (8.0±2.7 × 10⁹/L, 10.56±1.40, 13.95± 9.2, 16(23.2%)) respectively. It was concluded that MiR-146a transcript levels were higher in RA patient whole blood samples than in healthy control samples.

Molecular Detection of MexA, MexB Efflux Pump Genes and MexR Regulatory Gene in Clinical Isolates of Pseudomonas aeruginosa

1,2Farah M. Al-Qurashi, 2 Wathiq A. Al-Draghi — 2024-11-27

Pseudomonas.aeruginosa, possess the capacity to acquire level of resistance to many drugs. The MexAB-OprM system, is considered one of the most prominent efflux pumps associated with multi-drug resistance. The aims of the study to detect the appearance of MexA ,MexB efflux pump genes and MexR regulatory gene in Pseudomonas. aeruginosa . In this study, we examined seven clinical isolates of P. aeruginosa that were isolated from patient’s attending (Al-Kindi Teaching Hospital, Shaikh Zayed Hospital, and Imam Ali Hospital).; identified by Vitek 2 compact , Detection of MexA, MexB efflux pump genes and MexR regulator gene by PCR technique , primers are designed in the college of science / biotechnology department/Iraq. All three of the genes, MexA, MexB, and MexR, were existing in all seven clinical isolates of P. aeruginosa, according to conventional PCR results. It was concluded that carrying efflux pump genes can make P. aeruginosa more resistant to several drugs.

Detection of CTLA4 Gene Polymorphism and Inflammatory Cytokine Profile Among Inflammation Bowel Disease Patients

1Rand M. Abd AL-Rhman, 1Jinan M. Jawad AL-SAffar — 2024-11-27

Inflammatory bowel diseases (IBD) are chronic inflammatory conditions of the gastrointestinal tract driven by unsuitable immune responses to an changed gut microbiome in heritably susceptible individuals. The present study was designed to determined the CTLA4 (rs231775) single nucleotide polymorphisms (SNPs) using sequencing analysis and assessment the serum levels of interleukin 11,15,18 and CTLA4 level using ELISA technique , that are related to etiology and pathogenesis of inflammation bowel disease (IBD) in two groups of Iraqi patients crohn’s disease (CD) and ulcerative colitis (UC). When comparing IBD patients to controls the results of CTLA4 (rs231775) SNP revealed some variations, the G allele was demonstrated increased incidence in IBD patients than controls. The IL11 levels (mean ± SD; pg/mL) were significantly(p≤0.05) increased (472.61±162.89 and 460.43 ± 144.65) in CD and UC patients, respectively compared with healthy control (160.15±160.56) pg/ml. Similarly, serum level of IL15 was significantly increased in crohn’s disease patients and ulcerative colitis patients (451.18±197.33vs. 395.20 ± 183.54 pg/ml; P > 0.05) , respectively compared to controls (112.15±129.22 pg/ml; p ≤0.05). Whereas, the IL18 levels (mean ± SD; pg/mL) were increased (43.68 ±16.52 and 41.56 ± 10.79) in CD and UC patients, respectively compared with healthy control (40.76 ± 13.27) pg/ml, though, the variation was not statistically significant. The CTLA4 levels (mean ± SD; pg/mL) were show significantly( p ≤0.05)decreased (0.64 ± 0.28 and 0.61 ± 0.29) in CD and UC patients, respectively compared with healthy control show a significant (0.80 ± 0.40) pg/ml .It was concluded that CTLA4 SNP effected on CTLA4 protein expression, and cytokines 11,15 were appeared role in IBD than interleukins 18 and these results were indicated not significantly variation between CD and UC groups (P>0.05).

Isolation, Identification of Escherichia coli and Haematological Study for Iraqi Woman with Acute UTI

1Ruqya I. Abdulwahhab, 1Sahar M. Hussein — 2024-11-27

Urinary tract infection is one of the most common infections worldwide, especially in women, caused by different types of microbial agents. The purpose of this study was to focusing on infection of gram negative bacteria , haematological parameter and renal function in Iraqi women with acute UTI. Setting and design: 1-Sample collection from women with uti infection, 2- determine acute uti cases, 3-isoulation and identification of bacterial infection, 4- investigation for complete blood picture, 5- study renal function. A total of 220 samples (blood, serum and urine from the same case) were collected which included 120 samples that infected with UTI with mean age of (30-60) years old, in addition a 100 samples as controls were collected from healthy individuals. Morphological characteristics and biochemical tests were determined for acute women samples closely related to gram-negative bacteria that included microbiology that cultured on MaConky and EMB agar and examined by microscopic. The biochemical test included in this study were (Indole, Methyl-red, Voges-Proskauer, Citrate, Oxidase, Catalase), then used Api and Vitik for detected gram negative bacteria that presented. Finally used 16s rRNA for detection of E.coli bacteria. Evaluation clinical biomarkers were determined for women samples includes WBC,TYM, platelets, MPV, CRP, Urea, creatinin and BUN. Statistical analysis used: The statistical package for social sciences (SPSS) application was used to observe the effects of various factors in research parameters. Least significant difference (LSD) test (ANOVA) or T-test was rummage-sale to significant compare among means. The results showed that 100 isolates gave typical phenotypic characteristics and biochemical tests as gram negative bacteria, that divided to the genus Escherichia coli was 96(80%), Klepsiala was 15(13%), Enterobacterasea was 15(13%) and Protease was 4(3%). Also confirmed the result for E.coli bacteria by using 16srRNA. Clinical biomarkers were determined for all women samples that infected with UTI, the most common hematology and renal tests that studied in UTI infection women were WBC, MPV, LYM, PLT, CRP, Urea, BUN and Creatinine then compared with their levels in healthy control group. The results showed that there were significant increasing (p-value ≤ 0.05) in WBC, MPV and BUN, while there were non-significant increasing (p-value ≥ 0.05) in LYM, PLT, CRP, urea, Creatinine. It was concluded that many etiology that lead to UTI infection in women, it may be includes gram-negetive bacteria, gram-positive bacteria and fungi. The most causative agent of UTI infection was gram-negative bacteria, also haematological and renal function test may be good tool for diagnosis of acute UTI infection.

Effects of Thiopurine Methyle Transferase (TPMT) Gene Polymorphism on Serum TPMT Levels in Iraqi Patients Diagnosed with Ulcerative Colitis

1Sawsan A. Yusif, 1Rafid A. Abdulkareem — 2024-11-27

Ulcerative colitis is a chronic inflammatory bowel disease. Thiopurine methyltransferase (TPMT) is an enzyme that metabolizes a class of drugs called thiopurines. The aim of this study was to examine the impact of a polymorphism (rs2842934) in TPMT gene on TPMT level. Genotyping for TPMT gene (rs2842934) polymorphism was determined by the tetra-primer amplification refractory mutation system-polymerase chain reaction (TARMS-PCR). Serum TPMT level was estimated by ELIZA Technique .The TPMT gene (rs2842934) polymorphism demonstrated a significant association with UC risk. The highest prevalence of UC was seen among patients who had heterozygous GA genotype [Odd Ratio (OR): 17.51; 95% Confidence Interval (CI): (6.109-50.199); P-value <0.001]. The homozygous genotype (GG) showed the lowest risk with only (8%) in UC patients compared to the controls (72.5%) [OR: (0.03); 95%CI. (0.011-0.100); P-value < 0.001]. The highest mean TPMT enzyme level was ±196 pg/mL in UC patients who were homozygous for A allele compared to ±0.50 pg/mL in controls; P-value <0.001. Conclusions: The TPMT gene (rs2842934) polymorphism showed that the G allele was beneficial for protection from UC. In contrast, the “A” allele of rs6721961 significantly increases susceptibility to UC.

MicroRNA-146a Gene Expression as a Potential Biomarker for Rheumatoid Arthritis in a Sample of Iraqi Patients

1Luma Y. Aldouri, 1Mohammed I. Nader, 2Mohammed H. Munshed Al-Osami — 2024-11-27

MicroRNAs (miRNAs), a type of small non-coding RNAs, play a crucial role in controlling post-transcriptional gene expression, suggesting their potential involvement in autoimmune disorders. The objective of the current study was to assess the expression levels of miR-146a as potential indicators for diagnosing rheumatoid arthritis (RA) and to examine their correlation with disease activity. A total of one hundred blood samples divided into 50 RA patients and 50 healthy individuals were obtained. The medical histories of all participants were collected, and they were provided with a comprehensive clinicopathological assessment. Disease activity for the group of patients was evaluated using the Disease Activity Score 28 (DAS28). The RNA content of blood samples was extracted, followed by cDNA synthesis. Subsequently, the levels of miR-146a transcripts were measured using Real-time PCR. Significant statistical differences were seen between patients and healthy controls in terms of miR-146a relative expression, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and anti-cyclic citrullinated peptide (anti-CCP) levels (p < 0.01). The miR-146a expression level, with a fold change of 4.87, exhibited a substantial upregulation in the whole blood samples obtained from patients diagnosed with RA compared to those obtained from healthy controls. Furthermore, a positive correlation was seen between the miR-146a expression level and the levels of ESR, DAS28, CRP, and anti-CCP in the RA patients. In diagnosing RA, MiR-146a had superior diagnostic capabilities, exhibiting the most significant levels of sensitivity and specificity at 74% and 87%, respectively. This was further supported by an area under the curve (AUC) value of 0.90, obtained at a cut-off value of P2.55. In comparison, the diagnostic performance of anti-CCP was not as favorable. The diagnostic test demonstrated a sensitivity of 52%, specificity of 100%, and AUC of 0.81 when using a cut-off value of P 122.5U/ml. In summary, the increased expression of miR-146a in the entirety of blood samples from individuals with RA may serve as a possible indicator of disease activity and severity in those with established RA.

Cytotoxicity of Newcastle Disease Virus (NDV) against cultured breast cancer cell lines

Fatima A. Karim1, Saife D. Al-Ahmer2, Ahmed M. Al Shammari3 — 2024-11-27

The second leading cause of death for women is breast cancer. The disease has been successfully managed by conventional therapies. The aim of study to found novel strategies, such as oncolytic virotherapy, were required to fight it. x given by ICCMGR, was cultivated using chicken eggs from commercial hatcheries in Al-Hur area, a suburb of Holy Kerbala province. Propagated NDV induced considerable hemorrhaging in infected embryos compared to unaffected control embryos and destroyed all embryos within 48 hours. For additional testing, allantoic fluid containing the virus was collected, purified, and kept at -20°C. Hemagglutination (HA) assay was employed to quantify the NDV titer which show positive findings as a typical haemagglutination mesh pattern of chicken red blood cells at 2⁸, or 256 HAU. Using Vero cells and Flint's algorithm to calculate the Tissue Culture Infection Dose 50 (TCID50) test, the viral dilution and syncytia growth in the seeded wells were linked, and the result was found to be 3.5 * 10⁶ viral unit/ml. The multiplicity of the viral infection (MOI) 1,2 and 3 was prepared and tested on Amj-13 and MCF-7 of which cell viability and growth inhibition rate using MTT assay was examined. Results of Optical Density (OD) and Inhibition Rate Growth Percentage (IR%) were calculated using Statistical Analysis System (SAS) to determine Least Square Means (LSM) to compare the significance difference between the mean values at p ≤ 0.05. There were proportional significant increases, (P ≤ 0.05), in inhibitory rate of both cancer cell lines treated with all concentrations of NDV alone compared with control of which the highest inhibitory rate was recorded at concentration (3 MOI) suggesting the more viral particles, the greatest cytotoxicity caused that reached 73.14% and 67.7 % respectively. The cytopathic effects (CPE) caused to cell lines due to the exposure NDV was observed under inverted microscope. Cells suffer morphological changes including granulation and shrinkage of cytoplasm, which lead eventually to separation and floating of the infected cells in the culture media, after 72 hrs. large round empty spaces were so apparent. These observations were not detected in control cell culture within the same time of examination of infected cells. The results were concluded promising results in not just eradicating cancerous cells, but as potential cancer vaccine.