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الكشف الجزيئي عن انواع البكتريا وفايروس التهاب الكبد المرتبطين مع بيتا - ثلاسيميا Molecular Detection of Bacterial and Hepatitis Virus Types Associated with ? - Thalassemia

اسم المؤلف: شـهـد رعـد مزعـل
اسم المشرف: اياد محمد علي فاضل
الموضوع العام: علوم الحياة
السنة: 2015
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: تضمنت هذه الدراسة التحري عن انواع الاصابات البكتيرية والالتهاب الكبد الفايروسي المرتبط بمرض الثلاسيميا نوع الكبير ومقارنتها مع الاشخاص المرضى غير مصابين بثلاسيميا نوع الكبير.لغرض التشخيص انواع البكتيريا جمعت نوعين من عينات من مرضى الذين راجعوا مستشفى ابن بلدي.عينات الادرار تضمنت (41عينة من المرضى مصابين بثلاسيميا نوع الكبير و18من المرضى غير مصابين بثلاسيميا نوع الكبير) وعينات الدم تضمنت (25عينة من المرضى مصابين بثلاسيميا نوع الكبير و25عينة من المرضى غيرمصابين بثلاسيميا نوع الكبير). اعتمادا على تحديد الصفات الزرعية للمستعمرات والصفات المورفولوجية للخلايا البكتيرية ومن ثم الصفات الكميوحيوية كما امكن تشخيص العينات باستخدام .VEKIK2systemاظهرت النتائج التشخيص في عينات الادرار في المرضى مصابين بثلاسيميا نوع الكبير ان بكتيريا الاكثر تواجدا كانت Escherichia coli بنسبة 25% بينما انواع اخرى من بكتيريا ظهرت بنسبة 6% ,5% ,3% ,3% ,2% ,2% ,2% ,2%, 1.3% لبكتيريا klibseilla pneumonia, Chlamydia pneumonia Pseudomonas aeruginosa, Enterobacter colaca, proteus. spp, Salmonella typhi, Pantoea spp, sterptococcus pneumonia على التوالي. بينما من جانب اخر كانت نتائج التشخيص عينات الادرار لمرضى غير مصابين بثلاسيميا النوع الكبير حيث كانت بكتيريا الاكثر تواجدا كانت Escherichia coli بنسبة 28% بينما ظهرت نتائج الانواع بكتيريا الاخرى بنسبة 12% ,7% ,5% ,3% ,4% ,4% ,4% ,4% ,4% على التوالي لبكترياklibseilla pneumonia, sterptococcus pneumonia, Chlamydia pneumonia Pseudomonas aeruginosa, Enterobacter colaca, proteus. spp, Salmonella typhi, Pantoea. spp ..اظهرت نتائج تشخيص عينات الدم ان بكتيريا الاكثر تواجدا في المرضى مصابين بثلاسيميا النوع الكبير والمرضى غيرمصابين بثلاسيمميا النوع الكبير كانتEscherichia coli 24% ,15% على التوالي. بينما ظهرت الانواع بكتيريا الاخرى لمرضى مصابين بثلاسيميا النوع الكبير بنسبة 6% ,3% ,5%, 5%,5% ,3% ,2% ,2%, 2% على التوالي لبكتيريا streptococcus pneumonia ,klibseilla pneumonia, Pseudomonas aeruginosa, Chlamydia pneumonia, Enterobacter colaca, proteus. spp, Salmonella typhi, Pantoea. spp. بينما ظهرت الانواع بكتيريا الاخرى لمرضى غير مصابين بثلاسيميا النوع الكبير بنسبة 7%, 5%, 3.2% ,3.2% ,3.2%3.2%, 3.2%, 3.2%,3.2% على التوالي لبكتريا sterptococcus pneumonia, Chlamydia pneumonia ,klibseilla pneumonia, Pseudomonas aeruginosa, Enterobacter colaca,proteus. spp, Salmonella typhi, Pantoea.spp. اما بالنسبة لتاثير بعض المضادات الحيوية على بكتيريا Escherichia coli فقد اظهرت النتائج اختبار الحساسية لمضادات باستخدام VITKE 2 systemمقاومة هذه العزلات امبسلين (98%) وسيفاتريكسون (95%) السبروفلوكساسين (46%) السيفوتازيديم (44 (%والاموكسيسيلين (43% (والسيفوتاكزيم (40%) وليفووكساسين) 78%) والنالدكسك اسد (50%) ميثوبريم (27%) سيفابيم (40%) اكستابنيم (20%).استعملت طريقة التحليل الجزيئي PCR analysis للكشف عن عوامل الفوعة الجينية والمقاومة والتقارب الوراثي التي تمتلكها بكتيريا Escherichiacolii ، وباستخدام بادئات متخصصة لكل من الجين الخاص بالاهداب من النوع الاول (fimH) والجين الخاص بالمحفظة (kpsMTII). وجين خاص بتحلل الدم (hly) وجين خاص بالالتصاق (eae) وجين خاص ب لمقاومه مضاد حيوي ((bla - cmy2 كانت النتائج ان جين (fimH) بنسبة (58%) ، فيما كانت نسبة الجين الخاص بالمحفظة (kpsMTII) (58%) بينما جين خاص بالالتصاق (eae) كانت نسبة (20%). ونسبة جين خاص بتحلل الدم (hly) (28%), ونسبة الجين خاص بمقاومة المضاد الحيوي ((bla - cmy2 كانت (83%) بينما كانت نسبة ظهور كل جينات) 5.7%), وكانت نتائج الجين الخاص بتقارب الوراثي (Box) (58%).ولغرض دراسة الاصابات الفايروسية مرتبطة بمرض ثلاسيميا النوع الكبير وباستخدام تقنية ELSLA جمعت عينات الدم (88 من المرضى مصابين بثلاسيميا النوع الكبير و62 من مرضى غير مصابين بثلاسيميا النوع الكبير) تم جمع العينات من مركز الوطني لنقل الدم لتحري عن وجود الالتهاب الكبد الفايروسي نوع ( CوB) حيث كانت نسبة الالتهاب الكبد الفايروسي نوع (B) لدى المرضى مصابين مصابين بثلاسيميا النوع الكبير (17%) بينما كانت نسبة لدى المرضى غير مصابين بثلاسيميا النوع الكبير (16%).بينما كانت نسبة الالتهاب الكبد الفايروسي نوع (C) (82.9 %) ,(8 ,83%) لدى مرضى مصابين بثلاسيميا النوع الكبيرو غير مصابين بثلاسيمياالنوع الكبير على التوالي . | This study was carried out to detect the types of bacterial and viral infections which associated with β thalassemia major comparison to non thalassemic patients group.For Bacterial types identification two different kinds of samples were collected from Ibin ALBalady hospital. Urine samples which included (41from thalassemic patients and 18 from non thalassemic patients) and blood sample which included (25 from thalassemic patients and 25 from non thalassemic patients) depending on their cultural, morphological as well as biochemical characterization. The result identification that in urine sample showed that was most prevalent kind was Escherichia coli with percentage 25%. In addition, other bacteria were also obtained with a percentage 6%, 5%, 3%, 3%, 2%, 2%, 2%, 2%, and 1.3%. Klibseilla pneumonia, Chlamydia pneumonia, Pseudomonas aeruginosa, Enterobacter colaca, proteus. spp, Salmonella typhi, Pantoea spp, sterptococcus pneumonia respectively. While, on other hand, in non thalassemic patients the result identification that in urine sample the most prevalent kind was Escherichiacoli with percentage 28%. In addition, other bacteria were also obtained with a percentage 12%, 7%, 5%, 4%, 4%, 4%, 4%, 4%, 4% for klibseilla pneumonia, sterptococcus pneumonia, Chlamydia pneumonia, Pseudomonas aeruginosa, Enterobacter colaca, proteus. spp, Salmonella typhi, Pantoea spp, respectively. The result for blood sample identification also showed that the Escherichia coli was most common in thalassemic and non thalassemic patients it with a percentage 15%, 24% respectively, and other types bacteria were also obtain with a percentage 6%, 3%, 5%, 5%, 5.3%, 2%, 2%, 2% for , sterptococcus pneumonia, klibseilla pneumonia, Pseudomonas aeruginosa Chlamydia pneumonia, Enterobacter colaca, proteus. spp, Salmonella typhi, Pantoea spp, respectively from thalassemic patients and other types bacteria were obtain from non thalassemic with a percentage 7%, 5%, 2.3%, 2.3%, 2.3%, 2.3%, 2.3%, 2.3%, 2.3% for sterptococcus pneumonia, Chlamydia pneumonia, klibseilla pneumonia, Pseudomonas aeruginosa, Enterobacter colaca, proteus. spp, Salmonella typhi, Pantoea spp receptivity, and the results confirmed by identification VITEK 2 system kit. The VITEK 2 SYSTEM Antibiotics sensitivity test kits results revealed that Escherichiacoli were resistance to following antibioses Ampcilin (98%), Cefatriaxone (95%), Ciprofloxacin (46%), Cefipime (74%) Liovfaxacin (78%), Tetracylin (70%), Cefotaxim (40%), Aomxcilin (43%), Nalidexic acid (50%), Extapenem (20%), ceftazidime (44%). Moreover, low percentage of resistance was shown to, extapenem (20%) Trimethoprim (27%). PCR Methods used of molecular diagnosis of virulence genes, resistance, genotyping genes of E. coli by using specific primers to type 1 fimbria gene (fimH) and capsule gene (kpsMTII), hemolysin (hly), intimin (eae), ceftriaxone (bla - cmy2). PCR results the presence fimH (58%) and kpsMTII (40%) while hly (28%), eae 25%, (bla - cmy2) (83%). While the prevalence of all genes together with a percentage (7.5%) The results of box - PCR genotyping was 80%. To study the viral infection which associated with β thalassemia major by used ELSLA technique sample (88 from thalassemic and 62 from non thalassemic patients) were collected from Iraq National blood bank. For detection of HBS and HCV the result showed that HBS17% in thalassemic patients while in non thalassemic patients was16% while HCV were (82.95%), (83.87%) is percentage thalassemic patients receptivity

تعدد الاشكال الوراثية لبعض انظمة مجاميع الدم في مرضى ابيضاض الدم النيقياني Genetic Polymorphism of Some Blood Group Systems in Myelogenous Leukemic Patients

اسم المؤلف: رشا حسن صدام السوداني
اسم المشرف: نصر فرحان عبد الله
الموضوع العام: علوم الحياة
السنة: 2015
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: Assay has been made for ABO, Rh, Lewis, MNS and Duffy blood groups in a sample consisted of 80 affected individuals with chronic myelogenous leukemia (CML) and other sample consisted of 65 individuals affected with acute myelogenous leukemia (AML). Comparison have been made between each of the Leukemia sample with the control samples which consisted of 100 Healthy individuals.The results of the various comparisons were as follows : 1 - There is insignificant difference in frequency of ABO blood group in chronic and acute leukemia as compared with the control sample.2 - There are significant differences for Rh system patterns due to diminish of CcDee and ccDEe phenotypic patterns in both chronic and acute myelogenous leukemia, Moreover, there is an increase in CCDee pattern in CML and also an increase in CcDEe pattern in AML.3 - There are significant differences (P <0.001) for Lewis group blood in CML as compared with the control samples, also there is significant difference (P <0.05) in AML due to decrease in Le (a - b +) phenotype which resistant to the disease, and an increase in Le (a - b - ) phenotype.4 - There are significant differences (P <0.001) for MN blood group in both types of the disease (CML and AML), due to an increase in M + N + phenotype and decrease in M + N - phenotype. Also there is significant difference for Ss blood group in (AML), because of the increase in S + s + phenotype and decrease in S - s+ phenotype.5 - There is significant difference for Duffy group blood in CML, since the Fy (a + b - ) phenotype was decreased and the Fy (a - b +) was increased. But there is no significant difference in (AML).6 - There is no existence of disturbance or deviation in Hardy - Weinberg equilibrium for ABO, Rh and Lewis.7 - There is disturbance in Hardy - Weinberg equilibrium apparently existing for the MN blood group in both types of Leukemia as compared with the control sample . There is an increase in frequency of (N) allele in the patients and a decrease in frequency of (M) allele. There is also deviation (P <0.001) for Ss blood group in AML since allele S was increased ,but allele s was decreased in the patients as compared with the control sample .Moreover , there is deviation from this equilibrium for Duffy group blood in the overall Leukemia sample. The higher frequency of Fy ͣ allele was found in AML, while the higher frequency of Fyb and Fy alleles were found in CML . Such deviations from Hardy - Weinberg equilibrium in hematological malignancies and not deviated in the control sample may give attention about the etiology of the disease concerned.

انتاج بروتين cry1I من بكتريا Bacillus thuringiensis بواسطة التنسل الجيني Production of cry1I Protein from Bacillus thuringiensis by Gene Cloning

اسم المؤلف: نسمة طالب وناس علي
اسم المشرف: محمد الحجاج
الموضوع العام: علوم الحياة
السنة: 2015
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: البصرة
الصفحات الاولى:
المستخلص: Bacillus thuringiensis (commonly known Bt) is a ubiquitous, gram - positive and spore - forming bacterium. The organism produces intracellular crystal proteins, which are toxic to insects, during the stationary phase of its growth cycle. Because of its insecticidal activity, B. thuringiensis has been used as a biopesticide. However, it is still necessary to search for more toxins to control insect orders, which have the ability to develop resistance against such pesticides, and also to provide alternatives for chemical insecticides. The purpose of this study is to isolate B. thuringiensis strains that are collected from Basrah, and to identify the cry genes content of these isolations.In this study, 344 Bacillus species were isolated from 22 soil samples collected from different area from Basrah, fifty six of them (16,2%) were identified as B. thuringiensis strains based on colony morphology , microscopic observation of spore position in the cell and genetic analysis . Most isolations were examined by multiplex PCR using for cry 1, cry 2, and cry 9 universal primers in order to identify the type of cry gene content of these isolations. 82% of the isolations amplified cry1 gene, 76% amplified cry9 and 21% amplified cry2 genes.In this study cloned cry1I gene is using specific primer to amplify full length of gene. The cry 1I gene (2169 bp) amplified product was inserted in to the Pst1 and BamH1 sites of pdrive cloning vector joining technique to produce the recombinant vector. The cloning vector then transformed in to E.coli HB101 and the transformant cells colonies were selected by ampicillin sensitive phenotype, the efficiency of transformation was also determined to be 7,8 ×105cfu/μg. After that the cry1I protein is purified from LB broth media supported with ampicillinand used this protein against Tuta absoluta larva within concentrations (100 μg/ml, 150μg/ml, 200μg/ml) .The effectiveness of the toxin is to kill the larvae were in the concentration of (200μg/ml) in the 24 hours after treatment, while least toxicity in the concentration (100μg/ml) need three days to kill all larva.

دراسة جزيئية لتعدد الاشكال الوراثية لجين TAS2R38 (جين تذوق مادة PTC) بين سكان محافظة البصرة وعلاقتها بمرض السكري A Molecular Study of TAS2R38 (PTC Tasting Gene) Polymorphism Among Basrah Population and It's Association With Diabetes Mellitus

اسم المؤلف: حسنة عامر مهوس
اسم المشرف: فائزة عبد الوهاب احمد اسعد يحيى عايد
الموضوع العام: علوم الحياة
السنة: 2015
الدرجة: دكتوراه
اللغة: العربية
مكان الجامعة: البصرة
الصفحات الاولى:
المستخلص: This study dealt with the genetic polymorphism of Phenylthiocarbamide (PTC) tasting from phenotypic and genotypic (Molecular of TAS2R38(P49A) gene ) prospective,among sample of Basrah Population from 2014 - 2012. Also, a comparative Molecular study for the gene TAS2R38(P49A) and some biochemical parameters was made in a group of Diabetic patients for the first time in Iraq /Basrah and the world .Phenotypic study using the gradient dilutions of PTC substance method, was established among 317 individual : 96Males and 221 Females from different regions of Basrah. Increased sensitivity with low threshold 6 of Males than Females 5 was observed which indicate that the Males were more sensitive than Females in this region from Iraq .However, the tasters percent among Females still higher73.3% than Males72.9% ,and the whole population 73.2% as well .This study recorded different tasters percent in different regions : 62.1%,71.8% ,60.6%, 82.3% for the North ,AL - Zubair and Safwan, Abu - ALChaseab and AL - Fao and the center of the city respectively, although ,the whole population of Basrah found to be under Hardy - Weinberg Equilibrium . Allelic frequencies according to phenotypic analysis showed : recessive allele t had higher frequency (0.53) than dominant allele T (0.47), with elevated Heterozygous frequency Tt among all studied regions and the whole population(0.53) .The results recorded also , high BMI for non tasters tt as a whole and for both sexes in this phenotype : 28.32 ,28.26 for males and females respectively. While it had the lowest value among heterozygous 23.40 ,Statistical analysis for phenotypic study of this locus reveald an additive effect of the two alleles T,t by 81.33% with responsibility of 83.10% for the phenotypic variation found in this trait . Genetic (Molecular) analysis was done for comparative individuals and diabetic patients by sequencing the hTAS2R38(P49A) the PTC gene, among 127 individual : 59 healthy individual as a comparative group and 68 diabetic patients from both sexes and same age ranges. The results recorded a C/G allele in the position of the first SNP (145 C/G) which encode for Proline amino acid (C) or Alanine amino acid (G) : CC , CG refer to Tasters genotypes while GG refer to Non - tasters genotype. High frequency of C allele as compare to G among both healthy people and diabetic patients : 0.55, 0.53 for both respectively . Frequency of heterozygous genotype CG recorded a significant access as 0.76 among the comparative group which may implies a selective heterozygosity advantage among Iraqi population at this region. A significant differences in genotypic distributions was recorded between comparative and diabetic groups with high incidence of non tasters phenotypically tt 41% and genotypically GG 20.5 % among diabetic patients compare to healthy people : 25 % , 7% respectively . Interestingly non taster GG genotype had the lowest BMI among diabetic patients compare to othertwo taster genotypes CC, CG respectively. . Study of Genotype - Phenotype Association revealed significant correlation (p<0.0001) between them among comparative group as 86.9% and diabetic group as 48.8%. A comparative study of Biochemical analysis betweendiabetic and non diabetic individuals from different genotypes for TAS2R38 (P49A) gene showed significant elevated value of cholesterol among diabetic from the GG genotype 191.85 ± 6.57 mg/dl with non significant elevated values of Hb1c 8.93 ±1.77 and triglyceride 149.57±6.53 mg/dl. However, all these parameters were significantly more higher among diabetic than non diabetic . Significant differences(P<0.05) were recorded between diabetic and non diabetic according to their thyroid hormones levels with low values of T3 , T4 and high values of TSH among diabetic than non - diabetic. TSH hormone recorded its highest significant values among diabetic of CC genotype with 4.39 ± 0.16 μUI/ml as compare to the other two genotypes heterozygous CG and recessive homozygous GG . Although non significant , T4 recorded its lowest values among diabetic of the CC genotype .However, significantly (P <0.05 ),T3 hormone found to have lower values in the two homozygous genotypes; CC genotype for diabetic 0.54± 0.03 ng/ml and non diabetic 0.53±1.64 ng/ml ; GG genotype : 0.72± 0.04 ng/ml ; 1.82± 0.59 ng/ml for diabetic and non diabetic respectively . while individuals with CG genotype found to be non significantly different .Lastly it could be concluded that this trait may have some effect on BMI and may play a role the variation found among peoples in biochemical parameters (cholesterol, thyroid hormones ) changing's especially among diabetic patients. Ministry of High Education and Scientific Research Basrah University/ College of Education for Pure Science Department of Biology A Molecular Study of TAS2R38 (PTC Tasting Gene) Polymorphism Among Basrah Population and It's Association With Diabetes MellitusA Thesis submitted to the Council of the College of Education for Pure Science/ University of Basrah as a partial fulfillment of the requirements for the degree of Philosophy of Doctor of Science in Biology / Genetics By Hasna Amir Mohaus BSc. Biology / 1994MSc. Biology / 1998 Supervision by Assist.Prof.Dr. Prof.Dr.Faizah A.