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التحري الجزيئي عن النمط الثالث لداء السكري البادئ عند النضج (MODY 3) في مرضى السكري العراقيين Molecular Investigation of Maturity Onset Diabetes of The Young Type 3 (MODY 3) In Iraqi Diabetic Patients

اسم المؤلف: اسراء عدنان ابراهيم البغدادي
اسم المشرف: نورية عبد الحسين علي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
الجامعة: جامعة بغداد - معهد الهندسة الوراثية والتقنيات الاحيائية - قسم التقانة الاحيائية
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: شملت الدراسة 63 مريضا مصابا بداء السكري غير المناعي المنشا في اعمار مبكرة من ذوي التاريخ العائلي بالاصابة بمرض السكري ممن لايعانون البدانة الذين تم اختيارهم من مرضى السكري من مراجعي المركز التخصصي لامراض الغدد الصم والسكري في الرصافة/ بغداد للفترة من ايار | This study included 63 non obese patients having early onset of non immunogenic diabetes with strong family history of diabetes selected from diabetic patients attended the specialized center of endocrinology and diabetes at Alrusafa/ Baghdad during period from the begging of May 2011 till the end of February 2013. The demographic study revealed that there is no association between the disease incidence with neither the gender nor the blood group. But there is a significant difference in the distribution of the patients according to age of the first diagnosis and gender, as the study showed that majority of male patients (53.846%) were first diagnosed with hyperglycemia below the age of 20 while the majority of females (72.972%) were first diagnosed with hyperglycemia after the age of 20.The distribution of patients according to the type of treatment also reveled association between the gender and both age of diagnosis and type of treatment of the hyperglycemia as the study showed that there is a higher percent of male patients (80.768%) using insulin at the beginning of their diagnosis with hyperglycemia or transferred later to insulin than those of females (67.565%).The present study reported for the first time in Iraq the presence of monogenic diabetes (maturity onset diabetes of the young type 3) as major cause of diabetes within non obese diabetic patients' early onset of non immunogenic diabetes with strong family history of diabetes. The sequence analysis of the hepatocyte nuclear factor 1? gene showed that Iraqis have mutational hot spot at exons 3 and 4 of this gene responsible for maturity onset diabetes of the young type 3, and that mutations in the promoter region or exons 5, 6, 8 and 9 are a rare cause of diabetes.

التنميط الوراثي لمستضدات التطابق النسيجي في عينة عراقية من مرضى السكري النوع الثاني Hla Genotyping In A Sample of Iraqi Type 2 Diabetes Mellitus Patients

اسم المؤلف: احمد كاظم محمد
اسم المشرف: محمد ابراهيم نادر بتول حسن الغرابي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
الجامعة: جامعة بغداد - معهد الهندسة الوراثية والتقنيات الاحيائية - قسم التقانة الاحيائية
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: داء السكري مرض واسع الانتشارعالميا تختلف نسبة الاصابة بين البلدان المتطورة والنامية ويعتبر المسبب الرئيسي للاعاقة والموت في العالم.داء السكري النوع الثاني هو الاكثر حدوثا حيث تبلغ نسبة الاصابة (90 - 95%) من مجموع المصابين بالسكري للانواع الثلاثة الرئيسية. | Sixty of non insulin dependent diabetes mellitus (NIDDM) patients who were diagnosed according to American Diabetes Association criteria (ADA) 2007 were selected from the specialized center of endocrinology and diabetes (Baghdad AL - Russafa Health Directorate) during the period between first of May 2013 to last of October 2013.Their age ranged 35 - 70 years. Fourty apparently healthy individuals their age ranged (35 - 70) years were considered as control. Enzymatic colorimetric methods used for measured FBS (fasting blood sugar) and HbA1c (glycohemoglobin) and enzyme linked immunosorbent assay (ELISA) for hormones and enzyme markers. Fasting blood sugar revealed high significant in patients with median (11.6 mmol/L vs. 5.9 mmol/L) and (P<0.001) in comparison to control groups.Elevation of glycated hemoglobin (HbA1c) with mean (9% vs. 5.5%) in comparison to control groups. Another reliable marker are the concentration Adpionectine hormone, Insulin hormone and adenosine deaminase activity the results of those estimated significantly difference between levels mean (20.7 vs. 34 ng /ml) in patients compared to healthy subjects (P<0.001) for adiponectin ; mean (106.6 vs. 59.3 ng/ml) for ADA (adenosine deaminase) with (P<0.001) and the median (12.1 vs. 16 uIU/ml) for insulin hormone with (P 0.001). In order to investigate the accuracy of previously mentioned tests, a statistical analysis [Receiver - Operator Characteristics (ROC)] has been applied to show the accuracy, specificity and sensitivity of the tests under test.This analysis revealed that serum ADA activity is the best marker with highly specificity 100%, sensitivity 100%, and accuracy 100% while; FBS was the best test with highly specificity 100%, sensitivity 100% and 100% accuracy in comparison with other tests. It was denoted that type2 DM was associated with certain HLA class II alleles were analyzed for their genotyping by Polymerase Chain Reaction - Sequences Specific Olegionucleotide (PCR - SSO) technique. The present study revealed that diabetic patients were positively associated with HLA - DQB1*0201 (83% vs. 5.0%) which is the most prevalent in patients followed by DRB1*1137 (46.7% vs. 0.0%); DRB1*0401 (41.7% vs. 2.5%), and DRB1*1306(15% vs.0.0%) while HLA, A*0201;B*3559;Cw*0410 and DQB1*0501 is negatively associated in type 2DM in comparison with healthy control groups.This study has shown that there is no significant association between FBS, HbA1c, serum insulin, HOMA2(Homeostatic Model Assessment2) ? - function, HOMA2 - IR, serum adiponectin, serum ADA and HLA alleles(DQB1*0201, DRB1*1137, DRB1*0401, DQB1*0501, DRB1*1306) in spite the significant associated between FBS and serum ADA and HLA - DRB1*0701 allele with (P 0.021, P 0.008) respectively.The current result concluded that there may be an important role for HLA genotyping in arising the chance for enhancing the susceptibility for either disease development or protection against its initiation.

التقييم الجزيئي لنسخ جين المقاومة الدوائية MDR1 في بعض المرضى العراقيين البالغين المصابين بسرطان ابيضاض الدم الحاد Molecullar Assessment of Multidrug Resistance Gene (MDR1) Transcript In Some Adult Iraqi Patients With Acute Leukemia

اسم المؤلف: كفاح جبار شاكر اليعقوبي
اسم المشرف: عبد الحسين الفيصل
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
الجامعة: جامعة بغداد - معهد الهندسة الوراثية والتقنيات الاحيائية - قسم التقانة الاحيائية
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: The present study aims to shed light on the follow up of acute leukemic (AL) patients at initial diagnosis and after treatment to assess the response and early relapse through evaluating the gene expression level of one of the major multidrug resistance genes which is the multidrug resistance 1 (MDR1) to investigate the possible association between level of MDR1 gene expression and the clinical outcomes and this may be considered as a potential marker for response to chemotherapy of acute leukemic patients. Furthermore, the current study correlates between the MDR1 gene phenotype and MDR1 genotype in three important coding regions (C1236T, G2677T/A, and C3435T considering the potential influence of altering MDR1 activity and its effect on therapy outcome as well as susceptibility to develop cancer.White blood cells (WBCs) isolated from 106 blood sample of acute leukemic patients were provided by Iraqi hospitals in Medical City. These samples were distributed as follows : 46 newly diagnosed patients with acute leukemia who had not received chemotherapy and follow - up 25 after 1st induction, 17 after 2nd induction and 8 at consolidation, with 10 blood samples of healthy voluntaries. Two comparative groups were taken included 46 sample of peripheral blood (PB) and 26 sample of bone marrow biopsy (BMB) in paraffin blocks to validate the level of gene expression compare to WBCs. For genotyping analysis, 56 of blood sample were taken to study genetic variation of MDR1 gene polymorphism. The samples preservation with TRIzol was done. Samples subjected to total RNA and DNA extraction, then molecular study by using reverse transcription, Quantitative Real Time - polymerase chain reaction (qRT - PCR) and direct sequencing, at Molecular Oncology Unit in Guy´s Hospital - Kings College / London.The study reached at the following results : 1 - The results showed age groups (20 - 39 years) were associated with acute myeloid leukemia (AML), while (13 - 19 years) with acute lymphoblastic leukemia (ALL).2 - The level of MDR1 gene expression showed high significant differences with WBCs compared to PB and BMB.3 - The clinical outcomes indicated that the rate of complete response (CR) of newly diagnosed acute leukemic patients was 19(41%), while 27(58.7%) was non - responder (NR).4 - Statistical analysis showed significant differences with NR at initial diagnosis in acute myeloid leukemia, while appeared after 1st induction in lymphoid type.5 - The results of positivity MDR1 gene expression were 10(21.7%) out of 46 newly diagnosed in acute leukemia, while 36(78.3%) were MDR1 - negative depend on (1.1±0.03) cutoff value.6 - The positivity MDR1 gene expression appeared mainly in non - responders patients at initial diagnosis, and with early relapse patients, after complete remission, in consolidation.7 - The MDR1 mRNA expression showed significant differences with high level in NR compared to CR patients at initial diagnosis. During treatment follow up the increased level of MDR1 gene expression in CR patients and appeared non - significantly with NR.8 - The results of MDR1 C1236T genotype and allele frequency showed that 1236CC wild type genotype and C allele were significantly frequent in healthy control. While CT heterozygous genotype frequency was highly significant in AML and no significant difference in allele frequency. ALL showed non - significant difference in genotype and allele frequency of MDR1 C1236T.9 - Odds ratio and 95% confidence interval (ORs and 95%CI) analysis showed no evidence associated with risk factor in MDR1 C1236T ALL carriers. While risk factor observed in AML with MDR1 1236CT carriers.10 - The results of MDR1 phenotype - genotype association indicate that MDR1 1236CC wild type was significantly high expression among healthy and it was aprotective genotype. While the MDR1 1236CT showed significant differences with high level of MDR1 gene expression in AML patients. Whereas ALL revealed significant differences in high level of MDR1 gene expression with MDR1 1236TT genotype. Both CT and TT were affected genotypes.11 - The results of MDR1 G2677T genotype and allele frequency indicated that 2677GA genotype significantly appeared with low frequency in healthy control with no significant difference in allele frequency. Both ALL and AML showed high significant frequency in 2677GT genotype. G allele frequency was showed significant differences in AML while non - significant in with ALL.12 - Odds ratio and 95% confidence interval (ORs and 95%CI) analysis showed the MDR1 2677GT genotype was associated with risk factor to developing ALL and AML. Whereas the GG appeared associated with AML only.13 - MDR1 phenotype - genotype association, indicate that MDR1 2677GA genotype was significantly high expression in healthy individual. While AML patients showed significant differences with high level of MDR1gene expression in 2677GT genotype. ALL showed significant differences with high level of MDR1 gene expression in MDR1 2677TT genotype.14 - The results of MDR1 C3435T genotype and allele frequency showed significant difference in genotype and allele frequency with heterozygous CT in both control and AML patients and mutant T allele. Whereas non - significant genotype and allele frequency with ALL.15 - Odds ratio and 95% confidence interval (ORs and 95%CI) analysis showed that the MDR1 3435CC genotype carriers associated with risk to developing ALL. While no risk factor associate with MDR1 C3435T variants to develop AML.16 - MDR1 phenotype - genotype association, indicate that the wild type 3435CC genotype was significantly high expression in healthy control. The MDR1 3453CT genotype showed high significance with high level of MDR1 gene expression inAML. While ALL showed significantly high level of MDR1 gene expression in 3435TT genotype.17 - The results of MDR1 genotype - phenotype association showed similar impact of MDR1C1236T, G2677T/A and C3435T genotypes in AML clinical outcomes. The MDR1 CT/GT/TT genotypes were associated in NR AML with high level of expression at presentation, compared to significant low level in CC/GG genotype. In contrast, CR patients were observed non - significant with MDR1 gene expression at presentation and significant with low MDR1CC/GG genotypes in post treatment. In regards to ALL patients the MDR1 TT genotype showed significant differences with high level of MDR1 gene expression in NR and CR ALL at presentation and significant only with NR at post treatment. So there was no clear evidence between MDR1 genotypes and clinical outcome with ALL.18 - The haplotype results showed that the three MDR1 C1236T, G2677T/A and C3435T genotype were linkage disequilibrium significantly with heterozygous haplotype B (CT - GT - CT) compared to A(CGC) and C(TTT). Also B haplotype appeared significantly with high level of MDR1 gene expression compared to A and C. According to the clinical outcome, haplotype B was observed significant differences in NR AML patients while other haplotypes were non - significant

تقييم التعبير الجيني للجينات CK19, MGB, MUC1 microRNA - 195 and microRNA - let 7a في نساء عراقيات مصابات بسرطان الثدي Evaluation of MGB1, CK19, MUC1, microRNA - 195 and microRNA - let - 7a Expression In Iraqi Women With Breast Cancer

اسم المؤلف: جودت نوري غائب
اسم المشرف: عبد الحسين مويت الفيصل
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
الجامعة: جامعة بغداد - معهد الهندسة الوراثية والتقنيات الاحيائية - قسم التقانة الاحيائية
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: The present study aimed to shed light on the identification a panel of genes with distinct expression patterns in breast cancer patients as a useful tool for breast cancer early detection and progression. The present study designed to investigate the levels of genes expression of five genes panel (MGB1, CK19, MUC1, miR - let7a, and miR - 195) in circulating free mRNA and miRNA from blood of breast cancer patients versus noncancerous samples (benign tumor and healthy controls) to establish a biomarker panel potentially useful for early detection and progression of disease. The expression patterns of the identified genes were then compared with certain clinical features (age, lymph node status, and tumour size).Blood samples from 55 patients with different stages of newly diagnosed invasive ductal carcinoma were provided by certain Iraqi hospitals. Two control groups were used in this study; 10 samples of patients with benign breast tumors, and 20 samples from healthy donors. The samples preservation with TRIzol was done. Samples subjected to total RNA extraction, and then molecular study by using reverse transcription and real time PCR at Molecular Oncology Unit in Guy´s Hospital - Kings College / London.The study reached the following results : 1. The patients’ age range was 24 - 70 years and the median was 49 years with high frequency of patients in the range of 40 - 59 years. According to the family history, 50(90.91%) of patients were have negative family history. According to the clincopathological features (lymph node status and tumor size) the percentages of patients with multiple lymph nodes and tumor size 2.0 - 2.9 cm were the highest groups, which showed statistically highsignificant differences.2. For MGB1 gene expression, the result showed that 30(54.5%) patients were MGB1 - positive while 25(45.4%) patients were MGB1 - negative.According to malignancy status the percentage of patients with high level of MGB1 gene expression 22(40%) was significantly high. In correlation with age groups there was statistically no significant differences in the levels of gene expression with age. In correlation with clincopathological features, lymph node status showed that the highest percentage of MGB1 positive patients 18(66%) were multiple for lymph node status, and the tumor size results showed that there was decreasing in the MGB1 geneexpression with increasing of tumor size. 3. For CK19 the results of present study showed that 41(74.54%) patients were CK19 - positive, while 14(25.46%) patients were CK19 - negative.According to malignancy status the percentage of patients with high level of CK19 gene expression 30(54.45%) was significantly higher in compare with benign tumor patients and healthy controls. In correlation with age groups there was statistically no significant differences in the levels of gene expression with age. In correlation to the clincopathological features, lymph node status results showed that the highest percentage of CK19 positive patients 24(88.89%) were multiple for lymph node status, and there was increasing in the CK19 gene expression with increasing of tumor size.4. Mucin 1 (MUC1) gene expression results showed that the percentage of MUC1 - positive breast cancer patients 72.73%(n=40) was significantly higher when compared with benign tumor patients and healthy controls. According to the age groups the results showed no significant correlation with patients age groups. The clincopathological features results showed that the highest percentage of MUC1 positive patients 84.21%(n=16) have few lymph node status, and there was statistically significant association between the increasing of MUC1 gene expression and tumor size. 5. The miR - 195 gene expression results showed that the percentage of patients with positive miR - 195 gene expression 83.64%(n=46) was significantly higher than patients with negative miR - 195 expression 16.36%, the study also showed that the percentage of high miR - 195 expression samples 69.09% (n = 38) was significantly higher in compare with benign tumor patients and healthy controls. According to the clincopathological features, patients with multiple and few lymph node metastasis were found to have significantly the highest percentages of miR - 195 expression, while the tumor size results showed that there was increasing in the miR - 195 gene expression with increasing of tumor size. 6. The percentage of miR - let 7a - positive breast cancer patients 81.82% was significantly higher, when compared with miR - let 7a - negative patients 18.18%. In correlation to the clincopathological features, results showed no significant correlation in miR - let7a gene expression levels with patients age groups, for lymph node status, the results showed that the highest percentages of let 7a positive patients were those with multiple lymph node and few lymph node metastasis. The tumor size results showed that there was increasing in miR - let 7a gene expression with increasing of tumor size.7. According to genes combinations, three genes combination (CK19, miR - 195 and miR - let 7a) was significantly positively expressed with percentage of 60%(33/55), which reflect their potential diagnostic and prognostic value.8. The study concluded that the three genes combination (CK19, miR - 195 and miR - let 7a) may have potential applications as a diagnostic and prognostic markers for breast cancer.