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الكشف عن الرز المحور وراثيا باستخدام انواع مختلفة من التفاعل الانزيمي المتسلسل PCR == Detection of Genetically Modified Rice By Different Type of PCR

Author name: ياسمين ابراهيم فرحان
Supervisor name: امنة نعمة الثويني
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: In recent years, foods produced by genetic engineering technology have been on the world food markets. The biosafety aspects, regulations, and labeling for these foods are still contentious issues in most countries. Thus detection and quantificationof GMOs play crucial role for developing regulations on GM foods.In this study, eighty six non - labeled rice samples from different locals and exported market were analyzed to detect the genetic modification using a DNA based detectionvmethods as, conventional Polymerase chain reaction (PCR), and Real time PCR (RTPCR).The DNA rice samples were extracted by manual C - hexadecyl - Trimethyl - Ammonium - Bromide (CTAB) method and wizard kit method. The result revealed that DNA yield by the two methods is comparable. Rice DNA tends to be of a higher concentration when purified with the CTAB method; however, this particular DNA is more easily to amplify, the optical density (OD) was recorded 1.70 - 1.98 and the concentration of DNA quantified by fluorometer DNA rice samples, ranged from 11 to 50.5 ?g/?l. The DNA rice sample has also been used successfully with the Wizard Genomic DNA Purification Kit, and showed varieties in quality, the OD was recorded 1.65 - 1.95, and the concentration between 4.7 - 43.8 ?g/?l.The rice specific gene (sps gene) was detected by PCR. The results demonstrate that the purity of the extracted DNA in all tested rice samples was sufficiently high for a sensitive PCR analysis and the primer of detected gene appeared clearly at 251pb.Three genes; CaMV 35S promoter, NOS terminator, and insecticide resistant gene Cry1Ac were used to detect of GM rice by PCR, and Real time PCR using oligonucleotide sets targeting to novel genes. The result showed that there was no positive result reaction with conventional PCR, while the outcome of gradient PCR revealed a positive reaction in one sample (Uncle Bens brown) for CaMV35S promoter only. Gradient PCR with 12 replicons for each sample was used for qualitative detection of CaMV35S promoter gene, after optimization of melting temperature and cycles run (45 cycles) , the results appeared positive in the last three grades (63.9, 64.6, 64.9) for CaMV35S promoter, but NOS terminator, and CryIAc were recorded negative results.The result of Real - Time PCR clarified that the CaMV35S promoter specific primer showed strong amplification with Ct, and Tm values were reached into 33.73, 38.63 and 61.55, 62.92 in two samples Uncle Bens brown and Himalayan brown, respectively, whereas NOS terminator gave positive results in four samples Maxims, Laasturiana, Carolin white and Mahatma, and the values Ct and Tm reached to30.87, 30.31, 30.54, 33.75 and 64.53, 64.61, 62.62, 63.87 respectively in comparison with the positive control, while CryI Ac which did not show any positive signal.It was concluded that using molecular methods like Real - time PCR will be useful tool for detecting GM rice such as a part of the approval detection processes because of the rarity of data concerning consumption of GM rice in Iraq.

تاثير الاصابة بداء المقوسات الكوندية على المستويات الهورمونية والمدورات الخلوية خلال فترة بلوغ الانسان في محافظة بغداد == The Effect of Toxoplasmosis On Hormonal And Cytokines Levels During Human Maturity In Baghdad Province

Author name: ياسمين رياض عبد الكريم الخناق
Supervisor name: صباح ناصر العلوجي | خولة حوري زغير
General topic: Biology
Specific topic: Microbiology - Parasites
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: داء المقوسات الكونديه مرض عالمي الانتشار حيث يصاب معظم الاشخاص ذوي المناعة الجيدة بطفيلي المقوسات الكونديه وغالبا دون ظهور اعراض. ان الهدف الرئيسي لهذه الدراسه هو فهم الفروقات الجنسيه, الهرمونيه والمناعيه في سن البلوغ في الاشخاص الذين لديهم اجسام مضاده نو | Toxoplasmosis is a worldwide disease where most healthy, immunocompetent individuals infected by Toxoplasma are almost asymptomatic.The primary goal of this study is to perceive the hormonal and immunological sex - differences in puberty age who have positive anti - Toxoplasma IgG specific antibodies. The secondary goal is to inspect the endocrine - immune interaction in these persons by detecting the effect of testosterone and oestradiol hormones level on cellular immune response namely, IL - 4, IL - 12. From the first of November 2012 till the end of April 2013, 303 blood samples were collected from apparently healthy male and female students of Al - Erfan, Ignadeen and Algawahery schools and Baghdad University, Both sexes where divided into two age groups : group (A) which included subjects with age range (12 - 15) years old and group (B) which included subjects with age range (16 - 19) years old. All serum samples were tested for toxoplasmosis by using Latex agglutination test and ELISA anti - Toxoplasma IgG antibodies test. As well as, all serum samples were tested by using ELISA technique for detection of serum mean concentration of testosterone, oestradiol hormones, IL - 12 and IL - 4. The results revealed that 107/ 303 (35.31%) of the studied subjects showed seropositive toxoplasmosis, 60 males and 47 females of 107 positive samples showed high significant (p<0.01) differences in comparison to uninfected subjects. Males group B have recorded the highest percentage 34(41.46%) of the infection. Positive association was found between toxoplasmosis and testosterone level in asymptomatic toxoplasmosis cases compared to uninfected group. high mean concentration of testosterone in toxoplasmosis infected males recorded (15.03± 1.04 ng/ml) and (12.4± 0.91 ng/ml) in groups A and B respectively, in comparison to control group which recorded (8.03± 0.78) and (9.86± 0.83) in groups A and B respectively. Also toxoplasmosis infected females revealed high levels of testosterone hormone which represented (4.83± 0.06 ng/ml) and (2.55± 0.03 ng/ml) in groups A and B respectively, with a significant (p?0.05) differences between them, while the control group recorded (0.10± 0.02 ng/ml) and (0.90±0.03 ng/ml) in group A and B respectively.The present study showed a significant (P?0.05) decrease in the mean concentration of oestradiol E2 hormone in toxoplasmosis infected males and females in comparison with uninfected ones. E2 mean concentration was (41± 2.48 ng/ml) and (56± 2.91 ng/ml) for male groups A and B, respectively, in comparison to control group which recorded (67± 2.08 ng/ml) and (74± 2.42 ng/ml) in group A and B respectively, while it was (188 ± 12.48 ng/ml) and (196 ± 16.52 ng/ml) for female group A and B respectively, in comparison to control group which recorded (221±12.09 ng/ml) and (233± 15.63 ng/ml) for group A and B, respectively.The mean concentration of E2 hormone in toxoplasmosis infected females according to their menstrual cycle showed low levels in ovulation, late follicular and luteal phases, which represent (37.5 ± 2.59 ng/ml), (131 ± 16.7 ng/ml) and (76± 3.92 ng/ml) respectively, while the mean concentration of this hormone in uninfected females was (52.4± 2.88 ng/ml), (271.6± 21.04 ng/ml) and (196.2± 12.37 ng/ml) in the three phases of menstrual cycle. This study showed high significant (p?0.05) level of IL - 12 in both males and females with latent toxoplasmosis in comparison with free - toxoplasmosis groups. The mean concentration of IL - 12 in infected males was (4.75 ± 0.88 pg/ml) and (4.12 ± 0.69 pg/ml) in male groups A and B respectively, in comparison to control group which was (2.86± 0.53 pg/ml) and (2.46± 0.62 pg/ml) in groups A and B respectively, while it was (5.60 ± 0.12 pg/ml) and (6.04 ± 0.26 pg/ml) in infected female groups A and B respectively, in comparison to control group which recorded (3.32± 0.89 pg/ml) and (4.27± 0.15 pg/ml) in group A and B respectively. IL - 4 recorded quite elevated level in toxoplasmosis infected males (groups A and B) which was (15.09 ± 0.92 pg/ml) and (17.67 ± 0.78 pg/ml) respectively, in comparison to control group which recorded (13.89± 0.84 pg/ml) and (14.92± 0.69 pg/ml) in groups A and B respectively, Meanwhile the mean concentration of IL - 4 in toxoplasmosis infected females showed mild elevation in both groups A and B which was (15.14 ± 0.84 pg/ml) and (16.06 ± 1.13 pg/ml) respectively, in comparison with toxoplasmosis free subjects which recorded (16.53± 1.22 pg/ml) and (15.15 ± 0.97 pg/ml) with no significant differences between them. Interactions between the endocrine and immune systems may mediate sex differences in response to toxoplasmosis infection.

تحديد الصفات المظهرية والوراثية لانواع بكتريا المكورات المعوية المعزولة من المرضى في محافظة النجف الاشرف == Phenotypic And Genotypic Detection of Enterococcus Sp. Isolated From Patients In Al - Najaf Al - Ashraf Governorate

Author name: زهراء حميد عودة القريشي
Supervisor name: مهدي حسين محيل العمار
General topic: Biology
Specific topic: Microbiology - Bacteria
Degree: Master
Language: English
University location: Najaf
First pages:
Abstract: The study aimed to isolate and identify the Enterococcus spp. from different clinical specimens and study the virulence factors of predominant species, as well as detected the virulence factors encoding genes such as efaA (endocarditis - associated antigen), esp (enterococcal surface protein), eep (stimulating of pheromones expression) and enlA (enterolysin A) genes by PCR techniques.There were three hundred clinical specimens collected from patients suffering from different clinical infections during the period from September 2013 to January 2014 in AL - Sadder Medical City and AL - Hakem General Hospital. The identification of the Enterococcus spp. isolates were depended on colonial morphology, microscopic examination and biochemical tests as a primary identification. The final identification was performed with the automated VITEK - 2 compact system using Gram positive - Identification (GP - ID) cards.According to the results obtained by the VITEK tests forty two clinical isolates of Enterococcus were detected, which distributed into : (22) isolates from urine, (8) vaginal swabs, (6) seminal fluid, (4) throat swabs and two isolates from wound swabs with no isolates from cerebral spinal fluid, stool and blood specimens.This study revealed that the E. faecalis is more distributed with 25(59.52%), followed by E. faecium with 10 (23.80%), E.avium with 5 (11.90 %), E. durans and E. raffinosus with 1(2.39%) for each.The study investigated the virulence factors of E. faecalis, E.faecium and E.avium, which play a major role in enterococcus pathogenicity. E. faecalis and E.faecium had the ability to producecapsule, gelatinase, biofilm, adhesion, protease, bacteriocin, haemolysin and cytolysin except ? - lactamase produced only by E. faecalis while E.avium produced all these virulence factors except gelatinase, bacteriocin and cytolysin.The results revealed variation in the resistance of bacteria to antibiotics, E.faecalis express absolute resistance (100%) to Erythromycin, high resistance against Gentamycin, Tetracyclin and Vancomycin but high susceptibility to Ciproflaxacin and Penicillin and moderate susceptibility to Chloramphenicol. E.faecium exhibited absolute resistance (100%) to Erythromycin, Gentamycin and Tetracyclin, high resistance against Chloramphenicol, Penicillin and Vancomycin but high susceptibility to Ciproflaxacin while E.avium exhibited susceptibility (100%) to all these antibiotics except Tetracyclin showed absolute resistance.Then detected the virulence factors encoding genes : efaA, esp, eep and enlA genes by using PCR techniques and Electrophoresis Systems.Finally, The genotypic method. Regard to genotypic study the outcome showed that 21(84%) isolates of E.faecalis, 4(40%) of E.faecium and 5(100%) of E.avium were carrying efaA gene and 17(68%) of E.faecalis, 5(50%) of E.faecium, 5(100%) of E.avium were carrying esp gene while eep gene was carrying only by E.faecalis and E.faecium ; 12(48%) and 3(30%) respectively. Also the results revealed that only 1(4%) isolates of E.faecalis have enlA gene.

التحري عن التعبير الجيني لل FOXP3 وTGF - ?1 باستخدام الطرائق الجزيئية والمناعية في سرطان الرئة اللاصغير الخلية == Detection of FOXP3 Gene Expression And TGF - ?1 Using Molecular And Immunological Methods In Non - Small Cell Lung Carcinoma

Author name: سهاد فيصل حاتم المقدادي
Supervisor name: امنة نصيف جاسم | بان عباس عبد المجيد
General topic: Biology
Specific topic: Microbiology
Degree: Doctorate
Language: English
University location: Baghdad
First pages:
Abstract: Studies have linked FOXP3 and TGF - ? expression to the outcome of certain cancers. FOXP3 is a marker known to be expression in T - regulatory cells while TGF - ? is a secreted protein usually detected in the extra cellular matrix.The present study aimed at focusing on the identification of immune markers namely FOXP3 and TGF - ? with their expression patterns in lung cancer patients as a useful tool to predict disease progression.Also it is aimed to design molecular evaluation of m RNA expression of both FOXP3 and TGF - ? in peripheral blood mononuclear cells and bronchial (brush) cells of patients with lung cancer and benign lesions, using qRT PCR; determining the T - reg level in the peripheral blood employing the High Rsolution Melting (HRM) as a novel method to detect Treg - specific demethylated region (TSDR); molecular DNA analysis of somatic mutation of exons 3, 6, 7 of FOXP3 in patients with lung cancer tissue and benign lesions and immunohistochemical (IHC) estimation of FOXP3 and TGF - ?1 in T - reg and cancer cells in formalin fixed paraffin embedded(FFPE)lung cancer tissue and benign lesions.Blood samples were collected from 30 patients with newly diagnosed, non small cell lung carcinoma and 30 patients with benign lesions.Patients were recruited at The Specialized Surgery Hospital and Oncology Teaching Hospital/Baghdad.Samples from 16 apparently healthy donors were used as control during the period from June 2012 to June 2013. The samples preservation with TRIzol reagent were subjected to molecular study including RNA and DNA extraction; reverse transcription; RT - PCR; HRM assay and DNA sequencing were done in the Molecular Oncology Unit/Guys and ST Thomas? s hospital/ King College/London/UK.The expression level of FOXP3 was high in 16(61.5%)in lung cancer cases.A significant difference was noticed between cancer cases from one side and benign lesions or healthy control on the other side p<0.05.Mean of FOXP3 expression(fold change)was significantly high(2.64±0.09)in cancer cases than in benign cases(1.32±0.04)and healthy control(1.38±0.06)with p<0.05.A significant association between high expression level and >60 age and squamous cell carcinoma in cancer cases P<0.05.The expression level of TGF - ? was high in 16(61.5%)in lung cancer cases.A significant difference was noticed between cancer cases from one side and benign lesions or healthy control on the other side p<0.05.Mean of TGF - ? expression (fold change) was significantly high (6.27±0.56) in cancer cases more than healthy control (2.87±0.09) with p<0.05.The association was significant between TGF - ? expression level (high and low) and age>60in cancer and benign groups (p<0.05), while no significant association with gender and cancer types were noticed.For FOXP3 mRNA expression in bronchial brush cells, the result showed no significant difference between the mean fold change of malignant(3.57 ± 0.06) and benign(4.02 ± 0.06) patients. The low expression was predominated both in cancer and benign cases. No significant differences were found between FOXP3 expression (high and low) and age; gender ; cancer types..According to FOXP3 T - reg specific demethylated region detection, results showed that the mean percentage of FOXP3demethylation in lung cancer patients (4.32 ± 0.04) was significantly higher than in benign lesions (3.22 ± 0.02) patients andhealthy controls(3.33±0.04). A positive correlation coefficient with high significant, was found in the group of cancer samples (R² = 0.6653;r = 0.69;P : 0.0017)on correlating percentage of Treg and demethylation of FOXP3 from one side with its m RNA expression on the other side.In benign lesion group was(R² = 0.5334;r= 0.59;p= 0.0027), While in the control group a positive correlation but a weak significance was found (R²= 0.2383;r=0.28;P=0.0437).FOXP3 gene sequencing revealed high frequency of missense mutations c.715 GTA>CTA : V 239 L in 17 (94.44%) in malignant sample and non cancerous cases7 (87.5%)without statistical differences. Missence mutations were also detected in exone 3 in 3(16.67%) cancer cases and in 1(12.5 %) benign lesion.No missense mutations could be detected in exon 6. Intronic mutations and silent mutation were variable in three exons without statistical differences. Many cases of adenocarcinoma have shown multiple mutations either of missense or Intronic types. Missense mutations of exon 7 were correlated significantly with an age of 60?years. Exon 3 mutations were significantly associated with adenocarcinoma. Positive FOXP3 Immunohistochemistry (IHC) staining in tumor cells was associated with high missense mutations frequency 10(55.55%) in exon7, while in exone 3 was 2(11.11%). Negative FOXP3 IHC staining in the tumor cells was associated with seven missense mutations in exon7 and one (5.55%) in exon 3, in addition 4(23.53%) cases of the exon 7 missense mutations were associated with negative FOXP3 expression in lymphocytes.The result showed that FOXP3 by using IHC staining was positive in 21(70%) of nuclei of cancer cells, and 22(73.3%)in Treg infiltrates.The positive cancer cells and Treg infiltrates associated significantly with age>60 (p<0.05).No significant association was found withgender, cancer type, while there is association with moderate differentiation compared to poorly differentiation (p<0.05). High frequence of FOXP3 expression score 3 and high intensity were appeared in nuclear cancer cells compared to benign lesions cells, while Treg infiltrates with score 1 and high intensity was high frequency in malignant and benign.The result showed that TGF - ?1 by using IHC staining was positive in 25(83.3%) in the cancer cells and 21(70%) in stromal cells. No significant difference was noted between positive expression in malignant and benign lesions p>0.05.No significant association was noticed between positive cells expression and age, gender, cancer type and differentiation p>0.05. High frequency of TGF - ?1 expression score 3 and high intensity in malignant cells and benign. Also stromal cell expression score 3 and high intensity were predominated in malignant and benign lesions. The high and moderate intensity expression was more frequent in matrix surrounding cancer cells compared to non cancerous.Total agreement and kappa coefficient between FOXP3 and TGF - ?1were poor in malignant and benign epithelial cells and stroma, while the perfect agreement was between expression of TGF - ?1in stromal cells

تعدد الاشكال الوراثي للحركيات الخلوية وHLA - DQB1 في مرضى السل الرئوي == Genetic Polymorphisms of Cytokines And HLA - DQB1 In Pulmonary Tuberculosis Patients

Author name: خلود كريم حسن
Supervisor name: علي حسين ادحية
General topic: Biology
Specific topic: Microbiology
Degree: Doctorate
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: The present study aimed to understand the correlation between serum level of nine cytokines (IL - 1?, IL - 1RA, IL - 2, IL - 4, IL - 6, IL - 10, IL - 12, IFN - ? and TNF - ?) and their genetic polymorphisms at 16 gene positions defined by sequence specific primer - polymerase chain reaction (SSP - PCR) in pulmonary tuberculosis (PTB) patients, and in addition HLA - DQB1 gene polymorphism was also defined by SSP - PCR to determine their role in susceptibility or resistance to M. tuberculosis. Finally, serum level of cortisol was also determined in the patients.Ninety four Iraqi Arabs PTB patients (70 males and 24 females) were enrolled in the study. They were referred to the Institute of Chest and Respiratory Diseases in Baghdad for diagnosis and treatment during the period May - October 2012. A control sample of 80 apparently healthy individuals was also included and matched patients for gender (60 males and 20 females) and ethnicity. The results are summarized in the following : 1. A significant increased serum level of IL - 1? (24.16 ± 8.82 vs. 3.20 ± 1.18 pg/ml), IL - 1RA (41.31 ± 6.64 vs. 16.85 ± 5.50 pg/ml), IL - 2 (17.63 ± 3.53 vs. 7.80 ± 1.10 pg/ml), IL - 4 (9.56 ± 2.60 vs. 3.81 ± 1.70 pg/ml), IL - 10 (34.49 ± 4.60 vs. 7.61 ± 1.70 pg/ml), IL - 12 (25.16 ± 5.85 vs. 7.70 ± 1.12 pg/ml) and TNF - ? (22.52 ± 4.41 vs. 4.97 ± 1.15 pg/ml) was recorded in PTB patients compared to controls. Also, Cortisol serum level was significantly increased in patients (215.47 ± 1.33 vs. 38.63 ± 1.74 ng/ml).2. Cytokine gene polymorphism analysis revealed that neither genotypes nor alleles of IL1A - 889, IL2 - 330, IL2+166, IL4 - 590, IL4 - 33, IL6+565, IL10 - 819, IL10 - 592, IL12B - 1188 and TNF - 238 genes showed a significant variation between PTB patients and controls. In contrast, the frequency of TT genotype of IL1RN gene at position mspal 11100 showed a significant (P = 0.004) increase in PTB patients compared to controls (65.9 vs. 43.7%). For IL4 - 1098, the frequency of TT genotype was also significantly (P = 0.048) increased inpatients (82.9 vs. 70.0%). At position - 174 of IL6 gene, a significant (P = 0.002) increased frequency of GG genotype was observed in patients (55.3 vs. 31.2%). For IL10 gene, only GG genotype at position IL10 - 1082 was observed with a significant (P = 0.045) increased frequency in patients (18.1 vs. 7.5%). At position - 308 of TNF gene, a significant (P = 6.9 x 10 - 5) decreased frequency of GG genotype was observed in patients (60.6 vs. 87.5%), while GA genotype was significantly (P = 1.3 x 10 - 4) increased (38.2 vs. 12.5%). Finally, the frequency of AA genotype of IFNG gene at position +874 demonstrated a significant (P = 0.006) increase in PTB patients (55.3%) compared to controls (33.7%).3. To determine the impact of cytokine genotypes on cytokines serum level, PTB patients and controls were distributed according to their serum level in the three genotypes of each cytokine. It was found that CC genotype of IL1RNmspal 11100 in patients was observed with the highest IL - 1RA level (52.16 ± 5.81 pg/ml) compared to TT (41.39 ± 3.23 pg/ml) or TC (38.10 ± 4.54 pg/ml) genotype. The TT genotype of IL2 at position - 330 also showed the highest level of IL - 2 (22.16 ± 4.31 pg/ml) compared to TG (17.59 ± 3.40 pg/ml) or GG (13.68 ± 3.53 pg/ml) genotype in patients. The IL4 - 1098 TT genotype showed the highest level of IL - 4 in patients (10.38 ± 2.21 pg/ml) compared to TG (6.09 ± 1.20 pg/ml) or GG (3.93 ± 0.80 pg/ml) genotype. For IL10 gene, the GG genotype of IL10 - 1082 recorded the highest level of IL - 10 (40.67 ± 2.96 pg/ml), which was significantly different from AA genotype (26.66 ± 5.65 pg/ml). At position - 308 of TNF gene, serum level of TNF - ? in GG genotype of patients demonstrated a significant increased mean compared to genotype GA (24.76 ± 1.30 vs. 19.15 ± 1.12 pg/ml). At position - 238, TNF GG genotype showed a significant increase level of TNF - ? (23.02 ± 2.91 pg/ml) in patients compared to AA genotype (17.18 ± 1.53 pg/ml) of patients. Finally, IFNG+874 AA genotype was observed with the highest IFN - ? level in patients (11.07 ± 1.12 pg/ml) compared to AT (7.97 ± 1.81 pg/ml)or TT (6.10 ± 2.20 pg/ml) genotype. In contrast, no such differences were observed in controls.4. Out of the five encountered HLA - DQB1 alleles, DQB1*03 showed a significant (P = 0.005) increased frequency in PTB patients compared to controls (71.3 vs. 50.0%). It was also observed that heterozygosity at such gene locus was significantly (P = 0.03) more frequent in patients than in controls (93.6 vs. 82.5%), while homozygosity was observed with a less percentage frequency in patients compared to controls (6.4 vs. 17.5%) and the difference was also significant (P = 0.03).Accordingly, it is possible to conclude that the cytokine profile was deviated in PTB patients, and such deviation was correlated with the genotypes of some cytokines, which might also together with HLA - DQB1polymorphism confer the individual an immunogenetic predisposition to develop M. tuberculosis infection.

التحري عن بعض المؤشرات المناعية وارتفاع نسبة وجود الفيروس HCMV في المرضى المصابين بالفشل الكلوي == Assessment of Some Immunological Markers And Viral Load For Hcmv In Patients With Renal Failure

Author name: احمد جاسم شوالة الخويلدي
Supervisor name: مهدي حسين محيل العمار | زياد متعب الخزاعي
General topic: Biology
Specific topic: Microbiology - Viruses
Degree: Doctorate
Language: English
University location: Najaf
First pages:
Abstract: اجريت هذه الدراسة على 150 مريضا الراقدين في مستشفى الصدرالتعليمي ومستشفى الحكيم (قسم الكلى) في محافظة النجف خلال الفترة الممتدة من كانون الاول 2012 الى شهر اب 2013. وكان الغرض من هذه الدراسة بيان علاقة الفيروس المضخم للخلايا بمرض الفشل الكلوي، تراوحت اعما | This study was carried out on 150 renal failure patients, who were admitted to the kidney department in AL - Sadder Medical City and Al - Hakeem hospital in AL - Najaf governorate during the period from December 2012 to August, 2013. Subjects of this study were Investigated for the role of Cytomegalovirus among them, their age ranged between (1 - 88) years. Twenty four (age - and sex - matched) healthy individuals without any evidence of chronic inflammatory disease depended as control. All patients and control divided in four age groups. Blood and urine samples were collected from patients and control for immunological (IgM, IgG, C3, C4, IL - 6, IL - 10, IL - 12 & IFN - ?) by using ELISA and molecular study by RT - PCR, respectively. The obtained results showed that HCMV - IgG antibody was 100% for all cases, while IgM was 18.66% compared with that of control. Real time - PCR amplification for presence of HCMV DNA in urine samples revealed that HCMV genome were detected in 22(14.66%) of the 150 urine samples in all age groups, that distributed into 12(21.05%) with viral load ranged (20 - 543840) Copy/ml in females and 10 (10.75%) with viral load ranged (40 - 28050) Copy/ml for males. The results of cytokines profile showed a highly significant(P<0.05) elevation in patients than control. According to sex females appeared higher IFN - ?, IL - 6, IL - 10 ( 83.86, 82.67, 9.06 pg/ml, respetively) than males were (76.57 , 79.36, 6.51 pg/ml, respectively). Where's IL - 12 were recorded higher elevation in male (28.83 pg/ml ) than females (27.35 pg/ml). According to age groups 41 - 60 age group showed high level in IFN - ?, IL - 6, IL - 10 were (96.45, 90.40, 9.62) pg/ml , respectively.While IL - 12 appeared high level (37.43 pg/ml ) in age group 1 - 20 years Complement fractions C3, C4 decreased in all groups of patients compared to those of a healthy control. C3 was recorded (64.37 mg/dl) in females, and (70.24 mg/dl) in males and decreased to (63.41 mg/dl) in age group (1 - 20) years, while C4 decreased significantly among sex it was (21.06 mg/dl) in females and(24.22 mg/dl) in males, and down to (21.65 mg/dl) in age group (21 - 40) years.The overall finding results showed that HCMV are more prevalence among chronic renal failure patients and cause immune suppressor for them by increase and decrease many immune factors.

دراسة جزيئية لعوامل ضراوة المكورات النعقودية السالبة لانزيم التجليط والمعزولة من اصابات مختلفة == Molecular Study of Virulence Factors of Some Coagulase Negative Staphylococci Isolated From Different Infections

Author name: سعاد عبد الهادي عبد الرزاق الحلو
Supervisor name: عباس شاكر جواد المحنة
General topic: Biology
Specific topic: Microbiology - Enzymes
Degree: Doctorate
Language: English
University location: Najaf
First pages:
Abstract: The study aimed to investigating the role of coagulase - negative staphylococci in human infections, and determining the predominance genes of the virulence factors. Three hundred clinical specimens were collected from out and inpatients undergoing catheter related infections and twenty specimens were collected from healthy hospital staff as a control from January 2013 to July 2013 of Al - Zahraa Teaching Hospital, Al - Sader Teaching Hospital and Al - Hakeem Hospital in Al - Najaf Al - Ashraf province. The specimens were included urine, blood, vaginal swabs, seminal fluid and wound swabs. The specimens were cultured on mannitol salt agar and the primary identification was depended on Gram stained and biochemical tests. Then finally identification with Vitek 2 system is done.One hundred isolates were identified as coagulase - negative staphylococci (CoNS), Staphylococcus haemolyticus was identified as the most frequently isolated species in (53%), followed by Staphylococcus epidermidis (26%) and Staphylococcus hominis were recorded in (21%). Most of CoNS isolates were highly resistance to penicillin G (benzylpenicillin), oxacillin, cefoxitin and erythromycin; and low resistance to rifampicin, levofloxacin and others. While, control isolates results showed moderate resistance to penicillin G and erythromycin; low - level of resistance to cefoxitin, oxacillin and other antibiotics.The investigation of virulence factors revealed that 93% of coagulase - negative staphylococci isolates were production of slime layer, DNase 58%, protease 29% and hemolysin 88%. But the results gave negative result for TNase and lipase enzymes.Monoplex and multiplex PCR were used to explore the MecA, aap, icaA, icaD, atlE, sea, seb, sec, sed, hla, hlb, sspA, sspB, geh, nuc genes. The results showed that all CoNS isolates (100%) had mecA and atlEgenes, but 98% of isolates had aap, 93% icaA and icaD genes. PCR revealed that only (14%) of isolates had genes for enterotoxins expression. (92.86%) and (7.14%), sea and seb respectively, in contrast, the sec and sed genes were not be recorded.The result showed that 47% of CoNS isolates had hla gene and 41% contain hlb gene, 29% were positive for the sspA gene whereas the sspB gene and geh and nuc2 genes not found in any of staphylococcal isolates. Finally, the result indicated that 58% of CoNS isolates were expressed the nuc1 gene.Plasmid curing was carried out in order to determine the origin of resistance and some virulence factor genes (chromosomal or plasmid - borne gene). The curing (elimination) of the plasmids of coagulase - negative staphylococci isolated was catalyzed using ethidium bromide in different concentration and high temperature (44?C). The results showed that the oxacillin resistant coagulase - negative staphylococci were plasmid mediated since 93% of the isolates showed negative result on oxacillin resistance screening agar, and absence of mecA gene from all isolates. Also, 41 of coagulase - negative staphylococci isolates that showed ? - hemolysin became non - hemolysin after manipulated with ethidium bromide.Finelly, taking into consideration the etiological importance of CoNS has often been neglected, the present investigation confirmed that these microorganisms should not be ignored or classified as mere contaminant.

تقييم اختبارات PCR وطرق الزرع الاعتيادية في التشخيص المبكر لتجرثم الدم لدى الاطفال في مستشفى حماية الاطفال التعليمي في مدينة الطب / بغداد == Evaluation of PCR And Culture Methods For The Early Diagnosis of Bacteremia In Children From Welfare Teaching Hospital In Medicine City /Baghdad

Author name: زينب صالح هادي الزبيدي
Supervisor name: محمد ابراهيم نادر
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: استهدفت الدراسة الحالية تقييم الفحص المعتمد على تقنية PCR (polymerase chain reaction) (وطرق الزرع الاعتيادية في التشخيص المبكر لتسمم او تجرثم الدم في الاطفال.531 نماذج دم تم جمعها من الاطفال المرضى الذين اعمارهم اقل من 51 سنة ومشتبه بان لديهم اعراض تجرثم | The present study has been undertaken to evaluate polymerase chain reaction (PCR) technique in the diagnosis of bacteremia in comparison with the conventional blood culture techniques in children (infant and newborn).Blood specimens were collected from 135 children under 12 years of age suspected with fever and sepsis, obtained from Welfare Teaching Hospital/Medical City/Baghdad, for the period from April/ 2013 till January/ 2014.Blood specimens were collected and processed for Blood culture and PCR. Blood culture was performed using blood culture bottles contain brain heart infusion broth and positive results were subcultured using three media (macConkey - , chocolate - and blood agar), Gram stain, biochemical tests and conformational test (Api staph and Api 20E). Polymerase chain reaction was done using the universal primer, gram positive specific primer, gram negative specific primer, 16s rRNA primer for coagulase negative staphylococci and LacZ primer for Enterobacteriaceae.Optimization trials was carried out to increase the sensitivity of the PCR by applying 57°C in the annealing step for Gram positive specific primer and Gram negative specific primer to detect Gram positive and negative bacteria in blood respectively.Blood specimens were positive for bacteria in 69 cases (51.1%) by blood culture and 74 cases (54.8%) by PCR out of a total of 135 specimens analyzed. PCR showed more sensitive results compared to blood culture for detection of neonatal bacteremia. current results were revealed the ability of PCR to recognize five pathogens which have been negative by culture, all have been coagulase negative Staphylococci.The most frequent bacteria isolated and detected by PCR and Blood culture methods were Coagulase negative staphylococci (CoNS) (n = 60) followed by Enterobacter spp. (n = 8), E.coli (n = 5) and K.pneumoniae (n= 1). Interestingly, higher incidence rate (81.1%) were documented for the late onset sepsis (LOS) in our study compared to the early onset sepsis (EOS) (18.9%) for all bacteria. LacZ PCR efficiency have been 100% for detection of Enterobacteriaceae in blood.

تاثير انزيم الكلوكوسيل ترانسفيريز المنقى من العزلة المحلية Streptococcus mutans النمط C في انتاج الاضداد (IgY) من صفار بيض طيور الدجاج == The Effect of Glucosyltransferase Purified From Local Isolate Streptococcus Mutans (Serotype C) On Egg Yolk Antibodies (IgY) Generation In Layer Hens

Author name: هاشم محمد زهراو الصبيحاوي
Supervisor name: عصام فاضل علوان الجمیلي | فارس عبد الكريم
General topic: Biology
Specific topic: Biotechnologies
Degree: Doctorate
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: استهدفت الدراسة الحالية عزل وتشخيص بكتيريا Streptococcus mutans المسؤولة عن تنخر الاسنان البشري والتكلسات (plaque) واستخراج اضداد لها من صفار بيض الدجاج Yolk Immunoglobullin (IgY) لغرض استخدامها مستقبلا كمثبطات لنمو هذه البكتيريا الخطيرة ويمكن مزجها مع مع | The presented study aimed to isolate the main agent for dental caries and teeth plaque, Streptococcus mutans bacteria, and then production of specific antibodies against these harmful bacteria by the use of chicken egg yolk immunoglobulin (IgY). S. mutans had been proposed as the main etiological agents of dental caries and high levels of mutans streptococci in the plaque is correlated with a higher risk for dental caries. Seventy five plaque samples were collected from human teeth. Forty two samples were considered to be positive bacterial isolates using MS - agar (Mitist Salivares agar). Thirty five isolates were considered belonging to the group Streptococci; among these isolates 29 isolates were expected to be belonging to mutans streptococci group according to ability of producing special kind of exopolysaccharides. Ten isolates were considered as S. mutans with a percentage of 41% depending on staining with triphenyltetrazolium chloride and tolerance with NaCl 4%, 6 isolates were classified as serotype C by using Lancefield grouping identification. These isolates were tested for production of extracellular Glucosyltransferase (GTF) through determination of their enzyme specific activity. All isolates were able to produce the enzyme; Streptococci isolate (H5) identified as Streptococcus mutans serotype C was selected as the best producible isolate for GTF with a specific activity of 2.6 U/mg. It was found that GTF of the chosen isolate (H5) was produced during the middle stationary phase (18 - 35 hr.) and its maximal productivity was reached at 22 hr. Purification of S. mutans serotype (C) H5 GTF were done by ammonium sulfate, ion - exchange chromatography (DEAE - Sephacel column), and gel - filtration chromatography using Sepharose 6B column. The best percent saturation use for precipitating GTF by ammonium sulfate was 20 - 40% with specific activity 3.4 U/mg. Two purified GTF enzymes (GTF - I and GTF - II) were detected with specific activity 8.3 U/mg, 35.5 U/mg after 22.6, and 96.1 fold of purification respectively with yield 17.2%. Purification S. mutans CA - GTF (H5) were done by 8M urea, ammonium sulfate, DEAE - Sephacel column and gel - filtration (sepharose 6B) column chromatography. The purified CA - GTF was detected with specific activity 18.1 U/mg after 24.5 fold of purification with yield 20.2%. Determination of purified GTF (GTF - I, GTF - II) and CA - GTF molecular weight was done by using gel - filtration chromatography (sepharose 6B) column with presence of standards proteins. It was found that the molecular weight of GTF - I, GTF - II and CA - GTF was 125.819, 112.201 and 84.139 dalton, respectively. The ability of GTF, CA - GTF and whole cell of S. mutans to stimulate the immune system of avian hens was tested. The intramuscular rout injection of three purified antigens (GTF, CA - GTF and whole cell) in the chest of experimental hens was done. IgG from egg yolk hens (IgY) was purified through the post immunization period (9 weeks) by using polyethylene glycol (PEG) precipitation and protein content of IgY antibodies was estimated from egg yolk and serum. Each one milliliter of purified IgY egg yolk samples GTF, CA - GTF and Whole cell, protein contained 7.06, 6.97, 3.9 mg/ml, respectively while in serum protein content about2.6, 3.1 and 3.25 mg/ml, respectively. The Sodium dodecyle sulfate polyacrylamide gel electrophoresis (SDS - PAGE) of anti - GTF (IgY) indicated that purified IgY gave two bands; 47.863 and 34.673dalton which were considered to be IgY heavy and light chains respectively. the IgY - CA - GTF sample is the best in terms IgY specificity 34.07% while the two samples (GTF, Whole cell) performed 30.5% and 29.3% respectively, Igy - GTF the best in terms purity 49% followed IgY - CA - GTF 47% and IgY - whole cell 46.3%. The immunological specificity of the three IgY samples preparations was assessed by ELISA test and the best sample that produced high titration was IgY - GTF with concentration 3.5 mg/ml, followed by the IgY - CA - GTF and IgY - whole cell with concentration 3.28 and 3.1 mg/ml respectively. The IgY - GTF inhibited approximate 75% of the specific activity GTF, while IgY - CA - GTF inhibited 50% of the specific activity CA - GTF. A double immunodiffusion test for detection of the immune response between anti - GTF IgY and purified GTF, CA - GTF and Whole cell antigens were recorded. The immunological response of anti - GTF and anti - CA - GTF was indicated by the appearance of precipitation lines on the surface gel between anti - GTF and two antigens GTF and CA - GTF while in the anti - CA - GTF and anti whole cell only with homologues antigen. The effect of different concentration of inhibitor (Amoxicillin) and anti - GTF, anti - CA - GTF and anti - whole cell on the growth of S. mutans bacteria were tested using broth dilution method and diffusion method on solid medium. Anti - GTF and anti - CA - GTF had no effect on the growth of S. mutans(H5) serotype C, while anti - whole - cell and Amoxicillin were capable to inhibit the growth of bacteria at concentration 20µg/ml and 15µg/ml respectively. The minimal inhibitory concentrations in which these concentrations were noticed at 35µg/ml and 30 µg/ml respectively. The highest zone of inhibition (40 mm) was recognized with Amoxicillin at concentration of 50 µg/ml, followed by anti - whole cell with a zone of inhibition of 34 mm at concentration of 70 µg/ml.

التشخيص الجزيئي لبكتريا Streptococci الفمويه باستخدام جينات gtfs في بعض مرضى السكري المصابين بتسوس الاسنان == Molecular Identification of Oral Streptococci Using Gtfs Genes In Some Iraqi Diabetic Dental Caries Patients

Author name: هالة كمال محسن القزاز
Supervisor name: نورية عبد الحسين علي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: اجريت هذة الدراسة في معهد الهندسة الوراثية والتقنيات الاحيائية في جامعة بغداد خلال الفترة من تشرين الثاني 2012 الى ايار 2013، للكشف عن علاقة تسوس الاسنان بين مرضى السكري والمرضى غير المصابين بالسكري DDCP) و(NDCP اعتمادا على تسوس الاسنان من انواع Streptoco | The present study was carried out in Genetic Engineering and biotechnology Institute / University of Baghdad during the period from November, 2012 to May, 2013 to detect the relationship between diabetic and non - diabetic patients according to the dental caries occurrence and its causes by Streptococcus spp. (S. mutans, S. salivarius and S. oralis (which are isolated from oral cavity, In addition, this study was carried out to study the comparison between the traditional (bacterial culturing) and molecular diagnosis methods. The total number of the studied groups was 95 Iraqi patients (45 diabetic dental caries patients (DDCP) and 50 non - diabetic dental caries patients (NDCP) of both genders who their ages ranged from 18 - 65 years old. The patients, samples including saliva and buccal swabs that randomly collected from DDCP and NDCP who were reviewing Al - Alweyia Centers of Dental Caries and Diabetic Diseases in Al - Yarmook hospital in Baghdad city. The distribution of patients who have dental caries according to genders showed high significant differences at (p<0.01) between two genders (30 females, 15 males) for diabetic dental caries patients, while in non diabetic dental caries patients, there were significant differences at (p<0.05) between two genders (29 females, 21 males). The distribution of diabetic dental caries patients according to age showed high significant differences at (p<0.01) between two genders in age group of 36 - 50 years old, While low significant differences at (p<0.05) between two genders in age group of 20 - 35 years old and no significant differences between two genders in age group more than 50 years old, In another hand in non diabetic dental caries patients, there were no significant differences between two genders in all age groups. The results of samples (saliva and buccal swabs) culturing on mitis salivarius bacitracin agar media (MSBA) appeared that out of 95 bacterial cultures, 67 bacterial cultures were grown (32 bacterial cultures for diabetic dental caries patients and 35 bacterial cultures for non diabetic dental caries patients); S. mutans, S. salivarius, and S. oralis species were identified according to the results of microscopic examination, API kit 20 - strep, hemolysis on blood agar, motility test and catalase test. The molecular study focused on the analysis of DNA which extracted directly from saliva, buccal swabs and from the bacterial culture cells of S. mutans, S. salivares and S. oralis from both diabetic dental caries patients and non diabetic dental caries patients. Polymerase chain reaction (PCR) results revealed the presence of the product with 433, 544, and 374 bp which were related to gtfD (S. mutans), gtfK (S. salivarius) and gtfR (S. oralis) respectively in all samples (saliva, buccal swabs and bacterial culture). According to the presence of these three genes, there were high significant differences at (p<0.01) between diabetic dental caries patients and non diabetic dental caries patients, while there were no significant differences according to the percentage of presence of each gene between the three species of bacteria. Sequencing of the PCR products of the gtfs (gtfD, gtfK, and gtfR) genes region showed that nine samples gave acceptable results according to National center for Biotechnology Information (NCBI) matching, while 3 samples gave no results; this may be due to an error in sequencing system. The sequencing analysis of gtfs gene (gtfD, gtfK and gtfR) revealed that in DDCP the highest percentage of recorded mutations was in the gtfR gene.While in non diabetic dental caries patients, the highest percentage of recorded mutations in the gtfK gene than gtfR genes. In gtfR gene all mutations were substitution for diabetic dental caries patients and non diabetic dental caries patients. Nevertheless, in diabetic dental caries patients the mutations in gtfK and gtfD genes distributed between substitution and deletion mutations without recording any type of insertion mutation. But, in non diabetic dental caries patients, in gtfD all mutations were distributed between three types of mutations (substitution, insertion and deletion). The highest percentage of the effect of mutations in gtfs genes (gtfD, gtfR and gtfK) in diabetic dental caries patients were silent and missense mutation's than the frameshift mutations. on the other hand, the highest percentage of the effect of mutations in gtfs genes (gtfD, gtfR and gtfK) in non diabetic dental caries patients was missense mutations as compared with the other two types of silent and frameshift mutations

دراسة تاثير ضوء الليزر الثنائي الصمام (632 نانومتر) على بكتريا المكورات العنقودية الذهبية بوجـود المثلين الازرق كمتحسس ضوئي == Study of Photodynamic Effect of (632 nm) Laser Diode Light On Staphylococcus Aureus Using Methylene Blue As A Photosensitizer

Author name: ضياء خليل اسماعيل
Supervisor name: نورية عبد الحسين علي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: شملت الدراسة ثمان وسبعين عزلة من بكتريا المكورات العنقودية الذهبية عزلت من مئة وعشرين عينة تم الحصول عليها من عينات حروق وجروح قيحية وتجرثم الدم ومن مرضى يعانون من التهابات المجاري البولية, ومن اكثر من مستشفى للفترة من تشرين الثاني‏ / لغاية اذار/ 2014.تم | Seventy eight isolates of S. aureus bacteria were obtained from one hundred and twenty samples which were collected from different body sites and lesions (urine, blood and purulent wounds and burns) of patients from both sexes during the period November - 2013 to March - 2014. Methicilline sensitivity test (5µg MET disc) showed the appearance of methicillin - resistant in thirty seven Staphylococcus aureus isolates. Following exposure to laser light with a wavelength of 632 nanometer in the presence of Methylene blue at a concentration of 300µM at various exposure times (2, 4, 6, 8, 10, 12 and 15min), the results showed that the maximum decrease in viable colony counts ranging approximately from (6.9 to 3.8) log10 CFU /ml. Highly significant reduction in the viable count was achieved at 10, 12, 15 min exposure times, and 99% killing of cells were obtained when Photosensitisation of S. aureus using diode laser light at an energy density at 458.6 watt/cm2 for 15 mints. While in their exposure to the laser light in the absence of the dye or the dye in the absence of the laser light presented no significant effect on the viability of the S. aureus isolates. Both of phenotypic and genotypic investigation of the changes in virulence factors and the antibiotic - resistance were evaluated before and after irradiation with laser light.Results of photosensitization susceptibility tests showed large variations in the susceptibility, the isolates with resistant to methicillin before laser irradiation, become sensitive to it with percentage of 21.6%; in contrast the isolates with sensitive to vancomycine become resistant to it with percentage of 32.43%.On the other hand, the isolates that were resistant to Cefotaxime before laser irradiation become within the sensitivity range after laser irradiation with percentage of 51.35%, and also there were isolates within the sensitivity range before laser, become sensitive to Ciprofloxacin with percentage of 27.02% after irradiation. And the isolates of the S. aureus with resistant to Norfloxacin before laser irradiation, become sensitive to it with percentage of 16.2%. Also resulted in decrease the activity of ? - haemolysis, with 33 (90.3%) isolates of S. aureus in comparison to control as shown in blood agar method assay. In contrast had no effect on thermonuclease enzyme after irradiation.Detection of three genes represented in MRSA isolates by a confirmatory test was carried out using Polymerase chain reaction (PCR) technique. The results of the PCR amplification of mecA gene noted that it was present in 27 (72.2%) S. aureus isolates, While hlb gene detected 17(46%) isolates of 37 samples of S. aureus isolates, 14 of 17 hlb - positive S. aureus isolates(82.3%) were showed reduction in toxin production after exposure to laser light, whereas no altered or deficiency in thermonuclease gene (nuc).

علاقه بعض طرز الجين كالبين - 10 مع حدوث مرض السكري من النوع الثاني في العراق == Association of Some Calpain - 10 Gene Polymorphisms With The Incidence of Type 2 Diabetes Mellitus In Iraq

Author name: مياسة مثنى خالد
Supervisor name: اسماعيل عبد الرضا عبد الحسن
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: اجريت الدراسه الحاليه في معهد الهندسه الوراثيه والتقنيات الاحيائيه - جامعة بغداد خلال الفتره من كانون الثاني ولغاية حزيران - 2013 للكشف عن علاقة بعض الطرز الجين كالبين - 10 بحدوث مرض السكري من النوع الثاني في العراق. تم استخلاص الدنا من الدم الكلي باستخد | The present study was carried out in Genetic Engineering and biotechnology Institute - Baghdad University during a period from January to June, 2013, for detecting the association of some calpain - 10 gene polymorphisms with the incidence of type 2 diabetes mellitus in Iraq. Genomic DNA was isolated by using Geneaid DNA extraction kit from the whole blood; conventional PCR (SNP - 44 and Del/Ins - 19) and PCR - RFLP (SNP - 43 and SNP - 63) were used to detect the calpain10 variants by using specific primers and restriction enzymes. The study population consisted of 50 subjects with type 2 diabetes and 50 with normal fasting blood glucose (80 - 110 mg/dl). The type 2 diabetic subjects were recruited from the National Center for Diabetes treatment and Research. The non - diabetic control subjects were recruited from the same area as the comprising blood donors, healthy volunteers, or hospital /university staff members. Previous studies have detected a role for Calpain - 10 (CAPN10) polymorphisms in susceptibility to Type 2 diabetes mellitus (T2DM) in many populations. This study aimed to evaluate possible associations between these polymorphisms in the CAPN10 gene (SNP - 44, SNP - 43, Del/Ins - 19, and SNP - 63) and T2DM incidence in Iraqi population. Enrichment of allele 1(2R) in Del/Ins - 19 and 2R/2R genotype were found in T2DM patients. While the alleles and genotypes distribution of SNP - 44, SNP - 43 and SNP - 63 were not significantly different between patient groups and non - diabetic control subjects. The genotype AA in SNP - 43 and genotype TT in SNP - 63 were not found neither in T2DM nor in control subjects. of the eight haplotypes detected, enrichment of both haplotype 112 defined by variants of SNP - 43, Del/Ins - 19, and SNP - 63 and haplotype 2112 defined by variants of SNP - 44, SNP - 43, Del/Ins - 19, and SNP - 63 were seen in patients. The distribution of the other haplotypes was comparable between patients and control subjects. The calpain10 haplotype combinations were also obtained, and the haplotype combinations 111/111 and 111/112; which are created by variants of SNP - 43, del/ins - 19 and SNP - 63 and; haplotype combinations 1111/2111, 1111/2112 and 1121 / 2222; created by SNP - 44, SNP - 43, del/ins - 19 and SNP - 63; were associated with increasing the risk of T2DM.

التاثيرات المضادة للاكسدة والسمية الخلوية لمركب اللكنان المنقى من بذور نبات جوزة الطيب == Antioxidant And Cytotoxic Effects of Lignan Purified From Myristica Fragrans Seeds

Author name: شیماء عصام عبد الوهاب البرزنجي
Supervisor name: عصام فاضل علوان الجمیلي
General topic: Biology
Specific topic: Biotechnologies
Degree: Doctorate
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: In this study, natural lignan dimer was isolated from nutmeg seeds (Myristica fragrans) using organic solvent, partially purified using liquid/liquid partiation, purified using anion exchanger and chemically characterized using Benedict’s Reagent, Fehling’s Reagent and Molish’s Reagent. Then, by the aid of UPLC - PDA - IT - TOF - MS System, the molecular weight (626.221 Dalton) and the molecular formula (C39H45O7) of this dimer were determined. After that, the free radical scavenging activities were studied using stable free radical compound 1, 1 - Diphenyl - 2 - Picryl - hydrazil (DPPH). Results showed that 100, 10, 1 and 0.1 ?g/ml of purified lignan had 76.7 %, 65%, 28% and 8% scavenging activity respectively, while the same concentrations of partial purified lignan had 44.3%, 18.5%, 11% and 0% scavenging activity respectively.MTT(3 - (dimethylthiazol - 2 - yl) - 2, 5 - diphenyl tetrazolium bromide) assay was conducted to determined the IC50 for both purified and partial purified lignan using 4 different cell lines A549 (human lung adenocarcinoma epithelial cells), MCF7 (breast cancer cells), PC3(human prostate cancer cells), and HepG2(liver hepatocellular cancer cells), and to determine which cells type were be affected more by this natural lignan dimmer.The IC50 values for purified lignan were 85.17, 51.16, 108.4 and 60.21 ?g/ml while the IC50 values for partial purified one were 170.1, 84.14, 154.4 and 151.3 ?g/ml using A549, MCF - 7, PC - 3 and HePG2, respectively.The high content screening analysis (HCSA) and Cellomics Thermo Scientific maltiparametric Kits were used for the evaluation of cell - lignan interaction; 100, 50 and 25 ?g/ml of purified lignan caused 87.22, 69 and 53.36% reduction in MCF - 7cell count respectively and the same concentrations caused 98.1, 97.5 and 98.55% nuclear morphology changes. Results also revealed that these concentrations caused 7.7, 7.0 and 5.83% increase in MCF - 7 cells permeability respectively and they also caused 12.22, 11.15 and 0.2%decrease in mitochondrial membrane potential respectively, while these concentrations caused 11.12, 10.1 and 10% increase in Cytochrome C releasing from mitochondria to cytoplasim respectively.Reactive oxygen species (ROS) induction in MCF - 7 cells in the presence of 200, 100 and 50 ?g/ml of purified lignan caused 20, 11.48 and 9.61% ROS reduction respectively.MCF - 7 cell cycle was studied in the presence of 100, 50 and 25 ?g/ml of purified lignan, and results revealed that this compound blocked cell cycle at Sphase and the percentages of S - phase cells reduction were 74.33, 52.4 and 67%, respectively. This reduction was dose dependent while the same concentrations had no effect on MCF - 7 mitotic cells. Cell cycle arrest was detected immunofluorescently using BrdU antibodies (S - phase cell staining) and phosphor - Histone H3 antibodies (M - phase cells staining

التشخيس المبكر لداء السكري النوع الاول باستخدام مضاد حمض الكلوتاميك منزوع الكاربوكسيل ومضاد البيروكسيديز الدرقي == Early Detection of Type 1 Diabetes Mellitus Using Anti - Glutamic Acid Decarboxylase And Anti - Thyroid Peroxidase

Author name: ريم محمد عبيد
Supervisor name: منذر مصطفى فتحي
General topic: Biology
Specific topic: Zoology - Physiology
Degree: Doctorate
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: Markers have been described in type 1 diabetes mellitus (T1DM), There is a number of specific and non specific antigens have been identified. The major autoantigens involved in the destructive process of beta - cells leading to the development of type 1 diabetes are insulin hormone, glutamic acid decarboxylase (GAD), tyrosine phosphatase enzyme or Insulinoma - associated Antigen - 2(IA - 2).This study was conducted to find the relationship between antibodies for this antigens (insulin, GAD and IA - 2) and T1DM which could be used for the early detection of T1DM in normal Iraqi population. To study the importance of anti - thyroid peroxidase (anti - TPO) as a marker for autoimmune thyroid disease (ATD) in T1DM patients, and to find the relationship between T1DM and ATD.The study was carried out on 50 blood samples of men and 30 blood samples of women with age ranged from (20 - 60 years old), they were divided in to three groups : 1. Group 1 (20 men and 10 women) whom have fasting plasma glucose (FPG) above 180 mg/dL.2. Group 2 (20 men and 10 women) whom have FPG ranged from 120 - 180 mg/dL.3. Group 3 (10 men and 10 women) whom have FPG below 120 mg/dL.Blood samples were collected from all subjects, FPG and glycated haemoglobin (HbA1c) were measured. The levels of insulin, anti - insulin, anti - GAD, anti - IA - 2 and anti - TPO antibodies were measured in the serum.The statistical analysis results showed no significant difference in the presence of antibodies, HbA1c and FPG between men and women groups, inspite of some simple differences between the two groups.A significant (p<0.05) elevation in the level of FPG and HbA1c was observed in diabetic patients group compared withnon diabetic group. Significant (p<0.05) decrease in the level of insulin of T1DM patients was noticed compared to non diabetic group. Significant (p<0.05) elevation in the level of (anti - insulin, anti - TPO) in T1DM patients compared with non diabetic group. Elevation in the level of (anti - GAD, anti - IA - 2) in the T1DM patients compared with non diabetic group.The results also showed that no positive results for (anti - insulin, anti - TPO) present in the non diabetic group. One positive result for anti - GAD and one positive result for anti - IA - 2 present in the non diabetic group, which indicate the importance of anti - GAD and anti - IA - 2 antibodies ssay in normal population that could be used as early detection of T1DM.

دراسة فسلجية لمرضى الفشل الكلوي قبل وبعد الديلزة في محافظة واسط == A Physiological Study of Renal Failure Patients Pre And Post Dialysis In Wasit Province

Author name: تمارة حمد احمد العقبي
Supervisor name: كاظم جهيد كاطع الطائي | هيثم قاسم محمد
General topic: Biology
Specific topic: Zoology - Physiology
Degree: Master
Language: English
University location: Wasit
First pages:
Abstract: تم اجراء هذه الدراسة في كلية العلوم/ قسم علوم الحياة في جامعة واسط وبالتعاون مع مركز الكلية الصناعية في محافظة واسط للفترة من 1/10/2012 ولغاية 1/5/ 2013 لغرض دراسة التغيرات الفسلجية التي تحدث لمرضى الفشل الكلوي بنوعيه الغسيل الدموي والغسيل البريتوني قبل و| This study was conducted in the Department of Biology, College of Science, University of Wasit. The samples were collected in cooperation with artificial kidney center in AL - Kut Hospital, AL - Kut city, Wasit Province, during the period from 1 /10/2012 to 1/5/2013. In order to study the physiological changes of renal failure patients before and after hemodialysis and peritoneal dialysis. The blood samples were taken from one hundered thirty four patients for this study.The patients in this study were divided into two groups according to the type of dialysis : 94 hemodialysis patients, 52 males with age range (21 - 69) years and 42 females with age range (20 - 70) years, 40 peritonial dialysis patients, 24 males with age range (21 - 60) years and 16 females with age range (23 - 63) years. All results were compared with the results of 57 healthy person who were selected from both sexes (29 males and 28 female). Eight milliliters of venous blood were collected from control subjects and patients with renal failure by using disposable syringe of 10 ml before and after dialysis process.The blood analyses was conducted for evaluating the hematological parameters which are included (Hb, PCV, WBCs and PLT ) biochemical parameters which include (Blood Urea, Serum Creatinine, Total Protein and Calicum) electrolyte parameters which include sodium and potassium, measured concentration of (Erythropoietin and Aldosterone) hormone in patients with renal failure before and after dialysis and control groups and measure the effectiveness of phagocytic cells in patients with renal failure before dialysis by using Nitroblue Tetrazolium stain (NBT). The study revealed the following results : 1 - When comparing the results of parameters between before and after hemodialysis and peritoneal dialysis with healthy control show the following : a - Significant decrease (P < 0.01) in the levels of (Hb and PCV) while significant increase (P < 0.01) in PLT in renal failure patients before and after dialysis compared to healthy control and significant increase (P < 0.01) in the numbers of WBCs in male peritoneal dialysis patients compared to healthy control.b - Significant increase (P < 0.01) in urea, creatinine, sodium, potassium and in the aldosterone hormone concentration in renal failure patients before and after dialysis compared to healthy control.c - Significant decrease (P < 0.01) in the erythropoietin hormone concentration, total protein, calcium and phagocytic cells activity in renal failure patients compared to healthy control.2 - When comparing the results of parameters between before and after dialysis for each type of dialysis showing the following : - a - All hematological parameters and the concentration of hormones (Erythropoietin and Aldosterone) observe no significant differences between before and after dialysis.b - Significant decrease in the levels of urea, creatinine and there is a significant increase in the levels of total protein and calcium after dialysis comparing with before dialysis.c - Significant decrease in the levels of potassium in hemodialysis patients after dialysis comparing to before dialysis and there is a significant decrease in the levels of sodium and potassium after peritoneal dialysis comparing to before dialysis. 3 - When comparing the results of males and females hemodialysis with males and females peritoneal dialysis shows the following : a - Significant increases in numbers of WBCs before and after dialysis in the male and female peritoneal dialysis comparing with hemodialysis b - Significant decreases in the levels of urea and creatinine after dialysis in the male and female peritoneal dialysis comparing with hemodialysis.c - No significant difference in the levels of Hb, PCV, PLT, total protein, calcium, sodium, potassium, the phagocytic activity and the concentration of hormones (Erythropoietin and Aldosterone) before and after dialysis in the male and female hemodialysis comparing with peritoneal dialysis. We conclude that renal failure patients in Wasit Province suffer from severe anemia, high blood pressure, increase in the levels of urea and creatinine, disturbance in the levels of potassium, sodium and calcium with deficiency in the body's immunity.

دراسة تشريحية مقارنة لبعض الانواع البرية من ذوات الفلقتين النامية في محافظة بغداد == Comparative Anatomy of Some Wild Dicots Spp. Grown In Baghdad Province

Author name: زبيدة عبد اللطيف اسماعيل
Supervisor name: علي حسين الموسوي
General topic: Biology
Specific topic: Plant
Degree: Doctorate
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: The present investigation dealt with comparative anatomical characters of 29 selected wild species that belongs to different families growing in different parts of Baghdad Province. These species are : 1. Brassica deflexa Boiss.2. Brassica rapa L.3. Cardaria draba (L.) Desv.4. Sinapis arvensis L.5. Sisymbrium irio L.6. Strigosella africana (L.) Boch.7. Stellaria media (L.)Vill.8. Atriplex nitens Schkuhr.9. Chenopodium album L.10. Chrysanthemum coronarium L.11. Lactuca serriola L.12. Sonchus oleraceus L.13. Medicago polymorpha L.14. Melilotus indicus (L.) ALL.15. Vicia sativa L.16. Erodium cicutarium(L.) L'H17. Geranium rotundifolium L.18. Lamium amplexicaule L.19. Malva parviflora L.20. Plantago lanceolata L.21. Polygonum aviculare L.22. Rumex dentatus L.23. Ranunculus muricatus L.II24. Veronica polita Fries.25. Astrodaucus leptocarpus (Hoghst.) H. Riedl26. Urtica urens L.27. Verbena officinalis L.28. Lippia nodiflora (L.) Rich.29. Tribulus terrestris L.Some of these species have been anatomically investigated for the first time.Anatomical characters might be useful for plant biologist for the identification of important wild plants as an additional character at global level. Comparative anatomical characters were used; such as characteristics of ordinary epidermal cells of stems and leaves and stomatal complexes. The epidermis possesses number of important diagnostic character that offer valuable clues for identification, like size, shape, in addition to indumentum of both stems and leaves. Venation system was investigated and compared. Cross sections of root were useful taxonomically especially the thickness of epidermis, cortex, phloem and xylem. The study observed presence of sclerenchyma tissues in the root cortex of some species. Cross sectioning of stems and petioles were also studied. Epidermis, cortex, pericycle, vascular bundle shapes and numbers, pith characters, presence of crystals and tannin filled cells were useful aid in distinguishing species. Vertical sections of leaf blades, shape and number of vascular bundles, thickness of palisade and spongy layers and other mesophyll characters were important taxonomically.ccording to some of these characters, species were divided into groups. This study concluded the presence of variations in characters and these were presented for the first time comparatively, so that, the anatomical characters were a good support to the exomorphological characters in the studied species. Field photographs of the different studied plants and sections of organs were put in addition to numerous tables in this work. The above results were discussed scientifically regarding mainly the environmental factors.

فعالية مستخلصات بعض الطحالب الكبيرة للفطريات الممرضة للنبات == Activity of Some Macro - Algae Extracts Against Phytopathogenic Fungi

Author name: دنيا يوسف محمد يوسف
Supervisor name: عبد اللطيف محمد جواد
General topic: Biology
Specific topic: Plant
Degree: Doctorate
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
Abstract: تضمنت الدراسة الحالية عزل وتشخيص طحالب كبيرة من بيئتين مائيتين محليتين مختلفتين. تم عزل ثلاث طحالب كبيرة هي Cladophora glomerata (N) وEnteromorpha rafsii من منطقه بحر النجف في محافظة النجف, في حين عزل Cladophora glomerata (R) من احد مبازل منطقة الراشدية ش | The present study includes isolation and identification of Macro - algae from two different environmental water bodies. Three Macro - algae were isolated, Cladophora glomerata (N) and Enteromorpha ralfsii from Baher Al - Najaf region in Holy Najaf city. In addition, C. glomerata (R) was isolated from Al - Rashdiya, north of Baghdad.Phytopathogenic fungi from soil and some infected fruits were also isolated. They were identified as Pythium ultimum and Rhizoctonia solani.Water and ethanol were used to extract each algae to evaluate their antifungal activity against isolated phytopathogenic fungi. Different concentrations of these algal extracts had been prepared which are (10, 25 and 50) mg/ml and the antagonistic activity against the isolated phytopathogenic fungi were evaluated, the hot and cold water extracts did not show any antagonistic activity against fungi. However, all these extracts were ignored in the future work for this study. Furthermore, the hot ethanolic extract was more efficient than these of cold ethanolic extracts. Results have been shown that there were significant differences when macro - algal ethanolic extract were used.C. glomerata (R), which isolated from Al - Rashdiya, was more efficient against tested fungi than the same alga what isolated from Baher Al - Najaf region. However, E. ralfsii extract was more efficient against tested fungi than C. glomerata (N). Percentage of inhibition against P. ultimum when hot ethanolic extract of E. ralfsii, C. glomerata(R) and C. glomerata(N) were (88.8, 83.3 and 63.3) respectively.However, results for cold ethanolic extract of the same algae were (63.3, 52.2 and 32.2) respectively when 50 mg/ml of algal extracts were used.Percentage of inhibition against R. solani when hot ethanolic extract of E. ralfsii, C.glomerata (R) and C.glomerata(N) were (94.4, 100 and 78.8) respectively.However, results for cold ethanolic extract of the same algae were (66.6, 64.4 and 48.8) respectively when 50 mg/ml was used. According to the difference in biological activities against those phytopathogenic fungi caused by the same species of macro - algae or different genera from different environments. Due to difference in biological activity of the same macro - algae species Physiochemical parameter for each environments obtained such as air and water temperature, Turbidity (NTU), Electrical Conductivity(EC)and Salinity%, Water pH, CA+2 concentration, Mg+2 concentration, Total Hardness and Total Alkalinity. Results indicated that soaked Cucumis sativus and Capsicum annum seeds in cold and hot ethanolic macro - algae extracts of (E. ralfsii, C. glomerata (R) and C.glomerata (N) for 24 hours were able to protect seed germination that have been grown in a Petri dish contaminated with P. ultimum and R. solani comparing with control. In addition, results indicated that spraying C. sativus and C. annum seedling with these algal extracts within two weeks old again protects these seedlings from the same phytopathogenic fungi either before or after 24 hr. However, results indicated that adding the algal extracts within two months age to the C. sativus and C. annum plants again protects these plants from the same phytopathogenic fungi comparing with control and fungicide treatment. Moreover, results indicated that the active chemical compounds in E. ralfsii was Tannins, Saponins and Flavonoids. While C. glomerata was contained Tannins, Saponins, Alkaloid and Phenols. Results indicated that both hot crud methanolic extract of C. glomerata (R) and E. ralfsii have a lot of active chemical compounds against micro - organisms by using the GC - Mass Spectrometry technology.

استخدام انزيم اللاكيز المنقى جزئيا و المنتج من بعض العزلات الفطرية في تحطيم الافلاتوكسين B1 == The Use Of Partially Purified Laccase Produced By Some Fungal Isolates In Aflatoxin B1 Degradation

Author name: امنة رامي عبد الله
Supervisor name: عبد الكريم جاسم هاشم
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:

كفاءة مستخلص الكحول الاثيلي لاوراق نبات الاس Myrtuscommuins (L.) واوراق وبذور نبات الدودونيا Dodonaea viscosa(L.في السيطرة على بعض الجوانب الحياتية لحشرة خنفساء اللوبياء Callosobruchus maculatus (Fab.) Coleoptera : Bruchidae) == The Efficiency Of Ethyl Alcohol Extracts Of Myrtus Communis L. Leaves And Dodonaea Viscose (L.) Leaves And Seeds In Control Of Some Biological Aspects Of Pea Weevil Callosobruchus Maculatus (Fab.) (Coleoptera : Bruchidae)

Author name: زينب كريم جواد القزاز
Supervisor name: محمد عمار الراوي
General topic: Biology
Specific topic: Environment
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:

تاثير تركيز محلول الفضة الغروي و النحاس المعدني في بعض انواع البكتريا المعزولة من المستشفيات == The Effect Of Colloidal Silver Concentration And Metallic Copper Against Some Hospital Acquired Infections Isolates

Author name: علياء محمد هادي
Supervisor name: اليس كيكور ملكونيان
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:

دراسة في الوبائية والتحري بتقنية لداء (PCR) سلسلة تفاعل البلمره الابواغ الخبيئة في شمال بغداد == Study In Epidemiology And PCR Detection Of Cryptosporidiosis In North Of Baghdad

Author name: حارث سعيد جعفر الورد
Supervisor name: احسان مهدي الصقر | سهيلة حياوي محمود
General topic: Biology
Specific topic: Microbiology - Parasites
Degree: Doctorate
University: University of Baghdad
Language: English
University location: Baghdad
First pages:

تاثير بكتريا الحليب العصوية كمفرز حيوي لتثبيط بعض عوامل الضراوة لبكتريا القولون == The Effect Of Lactobacillus Species As Probiotic On Some Virulence Factor Of Escherichia Coli

Author name: علياء عبد الحسين المالكي
Supervisor name: صبحي جواد حمزة
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:

تاثير المركبات القلوانية والفينولية والتربينية لنبات اللبخ Albizzia lebbeck L. Benth. في الاداء الحياتي للذبابة المنزلية Diptera : Muscidae) Musca domestica L. == The Effect Of Extracted Alkaloids, Phenols, And Terpenoids Of Albizzia Lebbeck (L.) Benth. On The Biological Performance Of House Fly Musca Domestica L. (Diptera : Muscidae)

Author name: ايناس سلطان جابر الزبيدي
Supervisor name: فوزي شناوة الزبيدي
General topic: Biology
Specific topic: Environment
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:

تاثير المستخلص الكحولي لرايزومات نبات الكركم على خطين خلويين Curcuma longa من مرض ابيضاض الدم وطبعة النواة للخلايا المعاملة بالنيكل == The Effect Of Ethanolic Extract Of Curcuma Longa Rhizomes On Two Types Of Leukemic Cell Lines And Karyotyping Of Human Lymphocytes Treated By Nickel

Author name: فرح جبار هاشم
Supervisor name: مؤيد صبري شوكت | عبد الامير ناصر الركابي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:

دراسة بعض الخواص الفيزيائية و الكيميائية والبكتريولوجية و التنوع الوراثي لبكتريا E. coli المنتجة لانزيمات بيتا لاكتاميز من مياه الصرف الصحي في مدينة اربيل == Studying Some Physical, Chemical, Bacteriological Characteristics, And Genetic Diversity Of Beta Lactamase Producing E. Coli Of The Sewage From Erbil City

Author name: رنا جبوري عزيز
Supervisor name: فوزي شناوة الزبيدي | حارث جبار فهد المذخوري
General topic: Biology
Specific topic: Environment
Degree: Doctorate
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
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