مهدى حسين محيل العمّار

ABSTRACT Eight distinct phylogenetic groups of Escherichia coli strains have been identified through a novel technique called quadruplex PCR. The exact evolutionary relationships among these bacterial groups remain ambiguous. The Clermont phylotyping technique has been employed in this study to elucidate the phylogenetic groups of E. coli. This study aimed to establish the connections between isolates derived from clinical and environmental samples, isolate them, diagnose them, and subsequently classify E. coli bacteria into phylogroups and strains. Five hundred samples were collected from Karbala, subsequently cultured, isolated, and diagnosed biochemically, with confirmation via PCR and according to the Clermont classification. In the 201 positive isolates, two pathotype groups (DEC/EXPEC) and their associated subpathotype groups (APEC, UPEC, SEPEC, MPEC) were identified, alongside seven phyllocrops (A, B1, B2, C, D, E, and F), as well as the Unknown group and clade (I, II, V). Commensal E. coli was additionally identified. For this investigation, a novel strain classification was established, and these strains were initially catalogued in the NCBI gene library under the designation "Hetero-hybrid pathogenic," which is the plural of "EXPEC + DEC + DEC." An extension number was obtained upon the initial registration of ten new E. coli isolates with the NCBI. A novel category was established specifically for this study; E. coli bacteria were identified utilising specific diagnostic primers, and pathogenic strains initially discovered in Iraq were diagnosed

Abstract. The study aimed to evaluate the gene expression of genes (TLR-4,NOD-2) in patients of Otitis media and healthy persons. This finding included 50 samples that collected from healthy subject and 100 samples from a patients suffering from otitis media who attended Al-Sadr Medical City (ENT Department) in Al-Najaf Governorate during the period from February 2022 to June 2022. The samples had an average age ranging from 5 to 70 years. The gene expression of these genes among those suffering from Otitis media and healthy individuals have been investigated in this case-control research. Using a PCR technology. polymerase chain reactions were carried out to amplify each sample for the patient and control groups. The results of the molecular study (gene expression) showed a high significant increase in the level of gene expression in patients for the two genes NOD-2, TLR-4 genes (14.78 ± 2.369, 16.42 ± 3.158), respectively, with a significant difference at P≤0.05. TLR-4, NOD-2 as used as a molecular diagnosis Otitis Media patients

Background: Our aging domestic population is disproportionately affected by senile dementia and neurological impairment caused by Alzheimer's disease (AD), a complex and progressive age-related disorder. In the field of human pathogenic neurobiology, as crucial regulators of gene expression, microRNAs (miRNAs). Brain cells are known to have a high transcriptome, which means they are actively involved in gene transcription and mRNA generation. Objectives: This research aimed to examine the correlation between the expression of miRNAs-155 and patients diagnosed with Alzheimer's disease. Methodology: The study enrolled 100 participants: 50 healthy controls and 50 AD patients diagnosed according to DSM-V criteria, the quantification of miRNA-155 expression was performed; through the use of real-time PCR. Results: The expression of miRNA-155 is significantly greater in the Alzheimer's disease group (9.94±3.134-fold change) than in the control group (1.07±0.52) (P= 0.000). In order to evaluate miR-155's expression efficiency, a receiver operating characteristic (ROC) curve was constructed. Both the sensitivity and specificity were found to be quite high in the study, measuring 1.000 for each. The area under the curve (AUC) was also 1.000, indicating excellent performance. The 95% confidence interval for the AUC was from 0.1.000 to 1.000. The cut-off value used in the analysis was 3.88. Conclusion: Ultimately, miRNA-155 might potentially be used as a diagnostic tool to differentiate individuals with Alzheimer's disease from those who are healthy. According to the findings, that miRNA-155 has the potential to function as a genetic indicator for Alzheimer's disease. This work offers significant insights for future investigations on miRNAs as molecular indicators in medical domains and as promising targets for therapeutic interventions.

Abstract Objective: To discuss the association between single nucleotide polymorphism (SNP) of rs1929992 in IL- 33gene and IL-33 serum level in rheumatoid arthritis (RA) susceptibility among Iraq population. Methods: A total of 50 samples were collected from 35 RA patients from November 2018 untill end of January 2019 together with 15 healthy physical examines in the same period were chosen as the subjects. The serum IL-33 levels measured by commercial ELISA kits . Erythrocyte sedimentation rate ,white blood cell were measured by standard laboratory techniques The RFLP-PCR reaction technique was used to detect the genotype distributions for rs1929992 in IL-33gene was carried out by using restriction enzyme. The frequency of each allele and genotypes distribution was calculated so as to evaluate the association between genotype distribution and RA susceptibility. Results: Serum IL-33concentration was significantly higher in patients with RA than in control groups. The homozygous genotype AA recorded higher frequency in RA patients (42.9%) than controls (6.7%) with a significant difference (P-value 0.001). Homozygote genotype GG frequency (45.7%) was a significant in patients compared to controls subject (33.3%) with a significant difference (P-value 0.016), and the genotype heterozygous GA frequency (11.4%) were non-significant in patients compared to controls (60.6%). The allele frequency for allele G was (51.4%)in patients compared with controls ( 63.3%) with a significant difference (P-value 0.022)while for the allele A was (48.6%) in patients compared with controls( 36.7% )with a significant difference (P- value 0.001). Conclusions: significant correlation between RA patients susceptibility and genotype AA and alleles at rs1929992in IL-33 gene is observed. From this study showed that the IL-33 levels were influenced by genetic variation at SNPs rs5743708and rs1929992, respectively.

ABSTRACT Burning is damage to the skin and loss of the primary barrier of infection. Burned skin is at risk of infection as long as the barrier is absent. If not treated, infection caused by serious burns can be life-threatening and lead to sepsis. Sepsis is a life-threatening organ dysfunction due to the anti-infection host not responding; early treatment is critical. Sepsis is the leading cause of death, resulting in up to 50 to 60% of burn injury deaths. Therefore, the present study aimed to diagnose sepsis in burn patients and to study the relationship between TNF-α (-308G →A) gene polymorphism and burn patient with sepsis. Recorded results showed that procalcitonin (ng/mL) (PCT) significantly higher in sepsis than in without sepsis burn patients group (p < 0.05), 9.16 ± 5.07 versus 0.57 ± 1.27 respectively. The genotypes relative frequency in burn patient with sepsis were as follow : GG ( 40 % ), GA ( 45%) and AA (15%); while in burn patient without sepsis subjects : GG (53.3%), GA (33.3%) and AA ( 13.3%). G allele is higher frequency (62.5%) in burn patients with sepsis than A allele (37.5%). The presence of heterozygous GA genotype in the TNF-α-308 was associated with sepsis of burn patient

and sustainable. This study aimed to biosynthesize Zinc oxide nanoparticles by using Bacillus spp and evaluate their activity exposure to some environmental factors. The Zinc oxide nanoparticles was evidenced by trans-coloration of the reaction medium from yellowish into turbid yellow. Biogenic nanoparticles have been characterized using Atomic force microscope (AFM), X-ray Diffraction (XRD), Scanning electron microscope (SEM), and Energy dispersive spectroscopy (EDS). It is found that Zinc-oxide nanoparticles was homogenous nanorods, with size distribution (20–80nm). The average size of biogenic nanoparticles was 26 nm.

Summary This study aimed to isolate and diagnose Burkholderia cepetia from clinical specimens of diabetic foot and 2019, which includes 120 clinical specimens for both sexes with an age ranged between ) old. The diagnosis of bacteria isolates was based on microscopy, as well as the culture and biochemical Burkholderia cepacia 8(6.6%). Antibiotic sensitivity test was examined by dick diffution method, Burkholderia cepacia under study which included;, ceftriaxone, cefoxitin and Cefepime, with percentage of (100%); ticarcllin of (87.5%); aztreonam and amikacine with percentage(62.5); meropenem , imipenem and gentiamycin with percentage of (37.5%). Burkholderia cepacia isolates were with blaOXA and (6/8) of isolates carry blaCTX-M gene, all isolates of Burkholderia cepacia gave negative result of blaIMP and blaKPC gene. Finally PCR analysis showed that the integron gene was (3/8 %).

Abstract Renal failure results in the abrupt loss of kidney function, leading to the retention of waste products, electrolyte disturbances, and volume status changes. The estimation of glomerular filtration rate (eGFR) has been recognized as the best standard screening for kidney disease. .the study aimed to estimate the level of HCMV IgM/ IgG antibody in renal failure patients and the use of the RFLP-PCR technique in detecting the polymorphism of IL-1B. The study conducted on fifty renal failure patients (34 males and 16 females) and 20 randomly healthy subjects as a control group. The study demonstrated that 22(44%) of patients’ blood samples were positive for Cytomegalovirus antibody rapid test. plasma concentration IL-1 was estimated in all study groups, the level of IL-1 was significantly elevated in all renal failure patients compared with healthy persons. Digested DNA bands at 190 bp and 115 bp were produced by homozygote genotype of IL-1 -511(C/C), digested DNA band at 305bp, 190bp, and 115bp were produced by heterozygous genotype of IL-1 -511 (C/T) whereas homozygote genotype of IL-1-511 (T/T) was produced one fragment 305bp and remained undigested. Conclusion. The renal failure patients are highly susceptible to infection with CMV. CMV IgM/IgG antibody test is an important test in the diagnosis of CMV. The severity of this disease may be associated with the presence of CT genotype and T allele of IL-1 in renal failure patients.

ABSTRACT The biosynthesis of nanoparticles by using microorganisms is developing as an ecofriendly method for nanoparticle synthesis because of its cheap, simple and non-toxic. Bacillus sp. can be used for producing iron oxide nanoparticles. In addition, it has the ability for the biosynthesis of Fe3O4 nanoparticles. The nanoparticles producing was evaluated by using ultra violate-visible (UV-visible) and Fourier-transform infrared spectroscopy (FT-IR) methods also the production and size of the nanoparticle was confirmed by X-ray Diffraction and Field Emission Scanning Electron Microscope (FESEM) to confirm the accuracy of iron oxide nanoparticles. pH, Temperature, and Incubation time of production of iron oxide nano-particle also studied. Keywords: Bacteria, Drug delivery, Iron Oxide, Nanoparticles. International Journal of Pharmaceutical Quality Assurance (2020): DOI: 10.25258/ijpqa.11.1.13 How to cite this article: Jubran AS, Al-Zamely OM, Al-Ammar MH. A Study of Iron Oxide Nanoparticles Synthesis by Using Bacteria. International Journal of Pharmaceutical Quality Assurance. 2020; 11(1): 88-92

Objective: To discuss the association between single nucleotide polymorphism (SNP) of rs1929992 in IL- 33gene and IL-33 serum level in rheumatoid arthritis (RA) susceptibility among Iraq population. Methods: A total of 50 samples were collected from 35 RA patients from November 2018 untill end of January 2019 together with 15 healthy physical examines in the same period were chosen as the subjects. The serum IL-33 levels measured by commercial ELISA kits . Erythrocyte sedimentation rate ,white blood cell were measured by standard laboratory techniques The RFLP-PCR reaction technique was used to detect the genotype distributions for rs1929992 in IL-33gene was carried out by using restriction enzyme. The frequency of each allele and genotypes distribution was calculated so as to evaluate the association between genotype distribution and RA susceptibility

ABSTRACT : The study was conducted on 50 patients infected with peptic ulcers and 20 blood samples from apparently healthy people. H. pyloriwas investigated by using the Stool antigen rapid test for stool samples and in the blood samples by using Serum antibody rapid test. Restriction fragment length polymorphism PCR (RFLP-PCR) REFLP-PCR technique was used to determine polymorphism in the promoter region of the IL-1b gene in patients and non-infected patients. After using the stool antigen rapid test for stool samples and the serum antibody rapid test to confirm the H. pylori infection, the results were 50 (100%) positive and 20 (100%) negative. The genotype distributions results of the IL-1b -511 (C/T) single nucleotide polymorphism (SNP) showed that the frequency of the CC genotype between H. pylori positive group and H. pylori negative group was 34%, 30% respectively with a significant difference between patients and non-infected patients. Also for CT genotype distributions, there was a significant difference between H. pylori positive group and H. pylori negative group (52%, 45% respectively, while the frequency of the TT genotype between H. pylori positive group and H. pylori negative group was not significant between the two groups. More than 66% of infected patients with H. pylori have T allele in the two genotypes: CT and TT for - 511 (C/T) SNP at promoter region of IL-1B gene.These findings referred that there was an association between thispolymorphism and H. pylori infection in peptic ulcer patients; therefore this gene is considered as a risk factor and may be predict the probability of developing of this disease in Iraqi population.

ABSTRACT : Ventilator associated pneumonia is one of the most important types of pneumonia that infect persons, who are recumbent under mechanical ventilation in the hospitals worldwide. This study has been to investigate some of causative microorganisms at molecular and immunological levels, as well as other features such as seasonal variations, age distribution and detect the most suitable methods for diagnoses. The study has been accomplished in Al-Najaf Province, in Al-Zahraa Teaching Hospital, Al-Sadr Teaching Hospital and Al-Hakeem General Hospital during the period from September 2017 to December 2018. Blood and nasal swabs samples have been from 80 patients with ventilator-associated pneumonia, their age ranged from (few days to 80 years). Immunochromtographic assay for RSV has been used to detect their antigens in nasal swab samples after that the positive results have been confirmed by reverse transcriptase real time PCR technique, were showed incidence in 25% of the infections with RSV. This study has shown that the molecular method is more suitable for the diagnosis of the cases than the other methods.

Background: The biosynthesis of nanoparticles (NPs) using microorganisms is developing as an eco-friendly method for NP synthesis due to it’s cheap, simple, and non-toxic. Iron supplement drugs such as ferrous sulfate are the most used drug for the treatment of iron deficiency anemia, but this supplement reported that can yield undesirable side effects. Methodology: This study estimates the effects of iron oxide and chitosan iron oxide NPs (IONPs) on serum iron, ferritin, and transferrin markers in comparison to ferrous sulfate in iron deficiency anemic albino white mice. The biosynthesis of iron oxide and chitosan IONPs and the mice were divided randomly into two major (A and B) groups and four minor groups (B1, B2, B3, and B4), each of four mice. No material was added to the nipple of Group A (control). Group B1 was a positive control group, where mice were let without treatment during the completely of experimental period. Group B2 was treated with one daily dose (4 mg/kg) of commercially available iron supplement product containing ferrous sulfate, Group B3 was treated with one daily dose (4 mg/kg) uncoated IONPs, and Group B4 was treated with one daily dose (4 mg/kg) chitosan-coated IONPs. Results: Serum iron and ferritin in Groups B3 and B4 were significantly higher than Group B2 (P < 0.05). In addition, transferrin was lower. The average level of serum iron in the same groups was remarkably more than the groups received ferrous sulfate with similar doses (P < 0.05). Conclusion: The freshly Bacillus sp. has the capability to biosynthesized Fe3O4 NPs with faster rate and slight toxicity.

Abstract This study aimed to determine the P.aeruginosa that causes skin infection by detecting of pathogenisity markers. Bacterial isolated and identificated phenotypically and genotypically, as well as detects the pathogenicity markers encoding genes producing from P.aeruginosa by using PCR techniques. One hundred twenty clinical specimens were collected from patients suffering from different clinical infections during the period from September 2014 to January 2015 in AL-Sadder Medical city. The identification of the P.aeruginosa isolates depended on colonial morphology, microscopic examination and biochemical tests. P.aeruginosa had the ability to produce capsule, biofilm , adhesion, protease, bacteriocin, haemolysin, -lactamase and gelatinase. Regard to molecular study the outcome showed that 24 (100%) of P. aeruginosa isolates carrying lasB gene, 24(100%) carrying aprA, 23(95%) carrying plcH while 13(54%) carrying algD. To conclusion that P. aeruginosa able to produce sever pathogenicity markers which responsible for pathogenicity of P.aeruginosa infections.

Abstract A total of (50) clinical specimens were collected from patients suffering from gall bladder attending to AL-Sadder Medical City and AL-Furat General Hospital include (gall bladder tissue, stool and blood) during the period from December 2016 to September 2017 from different age and sex. The identification of Salmonella typhi isolates were depended on colonial morphology and biochemical tests as a primary identification. The final identification was performed with the automated VITEK-2 compact system and PCR technique. According to the results obtained by the VITEK tests and PCR technique, thirty five clinical isolates of Salmonella typhi were obtained. This study revealed that totally 35 (70%) positive result include that 23 (65.7%) isolates was positive from gall bladder tissue , 11 (31.4%) from stool and 1 (2.8%) from blood. The study investigated the virulence factors of Salmonella typhi, which play a major role in enterococcus pathogenicity. Salmonella typhi had the ability to produce colonization factor antigen, biofilm L. form and flagellar swarming variation while did not produce hemolysin. The study showed that the virulence factors of Salmonella typhi , which play a major role in their pathogenicity among gall bladder chronic infection.

This study aims to detection antibiotic resistance gene in Acenitobacter bumannii isolates collected from different diseases. It was included 276 specimens collected from different sources including patient suffer from burn, wound, upper respiratory tract inflammation, urinary tract inflammation, osteocytes, endocarditis and blood infection. 52 of these isolates belonged to italic pleas which have been identified through several laboratory tests including the morphology characterizes, biochemical, API 20 E and VITK -2 compact system technique. The distribution of isolates was burn (34.6 %), wound (28.8 %), urine (19.2), blood (9.6%), sputum (5.7%), plural fluid and CSF (1.9 %). The molecular study revealed that A. baumannii have the gene of ESBL, oxacillance and MBLs bla OXA like 51, bla OXA like 24, bla PER, bla TEM is (94%, 89%, 84%, 96%) respectively also the gene aadA and aadB is (76% and 84%) respectively fluroquinolone gyrA (66%) and finally all isolates have the gene bla TEM.

This study aimed to detection the intI gene and its relationship with multi-drug resistance of Salmonella typhi. isolates. The results of the antibiotic sensitivity test showed that the S.typhi isolates were highly resistant to the antibiotic; the results of PCR technique for intIgene amplification show that there are where 14/30 S. typhi isolated had positive results of amplification.

ABSTRACT : Pseudomonas aeruginosa is an opportunistic pathogen particularly affects those with impaired host or mucosal immunity and has a broad range causes impaired host or mucosal immunity and has a broad range of presentations, including respiratory infections in cystic fibrosis and mechanically ventilated patients, blood stream infections in premature neonates and wounds in burns injuries in the healthcare setting. This study aimed to determine the P.aeruginosa that causes skin infection by detecting of pathogenisity markers. Bacterial isolated and identificated phenotypically and genotypically, as well as detects the pathogenicity markers encoding genes producing from P.aeruginosa by PCR techniques. About one hundred twenty clinical specimens were collected from different clinical infections P.aeruginosaidentification according to colonial morphology, microscopic examination and biochemical tests. Results show that P. aeruginosa had ability to produce capsule, biofilm, adhesion ,protease, bacteriocin ,haemolysin, b-lactamase and gelatinase. Regard to molecular study the outcome showed that 24 (100%) of P. aeruginosa isolates carrying lasB gene, 24(100%) carrying aprA, 23(95%) carrying plcH while 13(54%) carrying algD. Present results concluded that P. aeruginosa able to produce sever pathogenicity markers which responsible for pathogenicity in any patients infected.