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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

Author name: احمد كاظم محمد
Supervisor name: محمد ابراهيم نادر | بتول حسن الغرابي
General topic: Biology
Specific topic: Biotechnologies
Degree: Doctorate
Language: English
University location: Baghdad
First pages:
Abstract: 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.

انشاء وتوصيف لخط سرطان الثدي الخلوي العراقي == Establishment And Characterization of Iraqi Breast Cancer Cell Line

Author name: مرتضى عادل الشامي
Supervisor name: محفوظة عباس عمران | احمد مجيد الشمري
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: Several primary cultures were initiated from different samples obtained from Iraqi female patients of breast tumor, one sample was successful, and this sample was histological diagnosed as breast cancer infiltrating ductal carcinoma.The cell suspension was cultured in tissue culture flask and confluent monolayer achieved after 16 days from primary culture. The continuous subculture was done in grown cells in tissue culture flask each 48 - 96 hrs. Between subculture to other until across 50 passages through11months.In our current study different experiments were done to characterize the cultured continuous cells, which are studying the growth curve of the new established cell line and calculating the population doubling time that have been 22 hours.Furthermore, a morphological study was carried out by staining the cells with hematoxilin and eosin dyes. The cells were elongated multi - polar epithelial like cells with nuclear polymorphism and multi - nuclei, in addition to high nuclear to cytoplasm ratio, all these characters of the malignant tumor cells.The Cytogenetic study showed chromosomal aberrations with many numerical changes among the tumor cells and abnormal structure gives chromosomes with unknown origin called marker chromosome. In furthermore the G - band stained of normal 46XX chromosome was done to facilities the comparisons between chromosomes of the new established breast cancer cell line and normal chromosomes aberrationsImmunocytochemistry examination was done for the tumor cells grown in multi well tissue culture slide chamber to detect the presence of some hormonal receptors (ER and PR) gives negative result, and to test Her - neu2 gene that gives week positive result.

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

Author name: كفاح جبار شاكر اليعقوبي
Supervisor name: عبد الحسين الفيصل
General topic: Biology
Specific topic: Biotechnologies
Degree: Doctorate
Language: English
University location: Baghdad
First pages:
Abstract: 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

دراسة جزيئية عن جين المقاومة mecA في بكتريا العنقودية الذهبية المقاومة للمشيسلين والمعزولة من بعض مستشفيات بغداد == Molecular Study For Detection of Meca Gene In Methicillin - Resistant Staphylococcus Aureus Isolated From Some Hospital In Baghdad City

Author name: لمى ياسين موسى
Supervisor name: محمد ابراهيم نادر
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: Two hundred and thirty clinical specimens were collected from two different hospitals in Baghdad during the period (December 2012 to April 2013). These specimens were collected from visitors, hospitalized patients and the health care workers in these hospitals. The specimens included nasal swab, wound swab, burn swab, abscess and pus, sputum, ear swab, urine and blood culture diagnostic results show that 150 out of the (230) specimens gave positive bacterial cultures and (100) isolates are characterized as Staphylococcus aureus depending on the cultural and biochemical examinations.the coagulase test was performed and the results showed that from total 150 isolates of Staphylococci, 100 isolates (61%) were coagulase - positive (COPS), while only 50 isolates (39%) were coagulase negative (CONS). In addition, the distribution of methicillin resistance among Staphylococcus spp. was investigated.The use of the antibiotic susceptibility profile for these isolates was examined against methicillin resistance. Using disk diffusion method revealed that (13) isolates were proved to be methicillin resistant Staphylococcus aureus (MRSA), While (87) isolates of S. aureus showed sensitivity to methicillin (MSSA) and there was no intermediate resistance among these isolates.The ability of MRSA isolates to produce some virulence factors were investigated and the results showed that MRSA isolates produce many enzymes and toxins that contributed in their virulence such as protease, urease, dnase and gelatinase, and also produce a beta types of haemolysins.The ability to produce slime layer by MRSA isolates was also investigated and the results showed that all isolates of MRSA were produced slime layer when theytested by Congo red agar method and the results showed that all of MRSA isolates produced strong slime layer.One of the aims of this study was to adopted a accurate diagnostic method to detect S.aureus by its genetic material contents through extracting of DNA and gel electrophoresis of the PCR product for the specific gene.Detection of methicilline - resistance gene represented by A confirmatory test was carried out for the selected isolates using Polymerase chain reaction (PCR) technique for further characterization up to the species level by the amplification of (mecA) gene.This is Staphylococcus aureus specific gene that encodes the extra Penicillin Binding Protein, which is unique to methicillin - resistant staphylococci. All the (13) positive isolates by disk diffusion test are found to be positive for the presence of (mecA) gene as their agarose gel revealed the presence of DNA band of mecA gene with a molecular size about (200 bp.).Results of the detecting (femB) gene showed that it was positive in all of MRSA isolates as they appear to have a band with a molecular size of about (651 bp). The genetic determinants of methicillin resistance mecA and femB genes were amplified using multiplex PCR technique in order to identify methicillin resistant (mecA+) and susceptible (lacking mecA) staphylococci and to differentiate S. aureus (femB+) from coagulase negative staphylococci (lacking femB). All of the S. aureus isolates (100%) were found to harbor femA gene, it is species specific marker for S. aureus.

عزل وتشخيص الجين lipA المنتج من بكتريا الزائفة الزنجارية من مياه الصرف الصناعي == Isolation And Identification of lipA Gene Producing Pseudomonas Aeruginosa From Industrial Wastewater

Author name: انتصار طه لفتة
Supervisor name: واثق عباس الدراغي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: Arabic
University location: Baghdad
First pages:
Abstract: توضح هذه الدراسة عزل lipA gene من الزوائف الزنجارية من مياه الصرف الصناعية للزيوت النباتية وتشخيصها اعتمادا على تفاعل البلمرة المتسلسل (PCR). استغرقت هذه الدراسة تسعة اشهر, من شهر شرين الاول 2014 لغاية حزيران 2015. تم جمع خمسين عينة من مياه الصرف الصناع | This study clarify the isolation and identification of lipA gene in Pseudomonas aeruginosa from industerial wastewater of vegetable oils factories depending on the polymerase chain reaction (PCR) technique. The present study had taken nine monthes starting from October 2014, till the mid of June 2015. Fifty samples of industrial wastewater were collected from the factories of the general company of vegetable oils, fourty from AL Rasheed factory and ten from AL ameen factory, the samples collected from the physical, chemical and biological treatment units, and other different wastewater tanks departments. While the four sewage samples were collected from sewer service Baghdad /Alrustumaiya. For Identification of Pseudomonas aeruginosa, two types of tests were used in this study. The first type was the routine tests, which include selective cultures, bacteriological and biochemical tests. Thirty four of fifty samples (68%) gave a positive growth and results for tests that were used to confirm the presence of P. aeruginosa. In addition to diagnosis P. aeruginosa in sewage sample which prepare the biological treatment units in vegetable oils factories as active sludge, other bacterial types have been diagnosed by using traditional methods and API 20 E system. For detection of bacterial lipolytic activity, two methods were used for this purpose, the first was the screening of bacterial lipolytic activity which was based on values of clear zones diameter around bacterial colony. The best values were between (1.9 _ 2.7) cm. The second test was carried by the Gas - Chromatography - Flame ionization detector, fatty acids solutions that were produced from hydrolysis by lipase enzyme were extracted by Petrolium ether solvent then analyzed by gas chromatograph apparatus. The second type of test was the Molecular diagnosis by using the Polymerase Chain Reaction (PCR) to detect lipA gene in Pseudomonas aeruginosa by two primers. Through two primers used, lipA 948 was the best and more specialized primer to isolation lipA gene of P. aeruginosa. It gave (100%) a positive result. While the second primer lipA 558, gave (66.66%) a positive result and that may be due to the design of this oligonucleotides was not specific for lipA of P.aeruginosa. This is so as was based on highly preserved region of 12 bacterial lipA - homologous genes for many genus and many species belong to Pseudomonas. DNA sequencing done for amplicon generated using lipA 948, this sequence aligned by using BLASTn software against NCBI database to validate the results and investigate the similarity degree with other corresponding strains.

انتاج انزيم السيليلز من عزلة محلية لبكتريا B167 Streptomyces sp. واستخدامه في انتاج الوقود الحيوي == Cellulase Production From Local Isolate of Streptomyces Sp.B167 And Its Application In Biofuel Production

Author name: بنان محمود سليمان
Supervisor name: ناظم حسن حيدر
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: هدفت الدراسة الحالية الى انتاج انزيم السليليز من بكتريا الستربتومايسس ودراسة تاثير بعض الظروف الزرعية على انتاج الانزيم; انتاج الوقود الحيوي من المخلفات السليلوزية من خلال التحلل المائي للمخلفات بالاحماض والانزيمات المايكروبية. تم غربلة 74 عزلة لبكتريا St | The current study was aimed to produce biofuel from cellulosic waste material degraded by local isolate Streptomyces. Seventy four isolates of Streptomyces were screened for cellulase production in solid and liquid media. The results showed higher capability of isolate Streptomyces sp. B 167 for cellulase production and bioconversion of cellulose. Therefore, it was selected for further studies. The results of optimization revealed that the cellulase enzyme productivity by the isolate Streptomyces sp. B 167 reached to 2.1 and 2.28 U/ml after 48 h of incubation time and pH 7 respectively. Cellulase productions in tested isolate improved (2.57 U/ml) by supplementation of cellulose liquid medium with 1 % of yeast extract as nitrogen source. Additives of carbon sources like (manitol, glucose, maltose, sucrose and starch) to the process of saccharification did not improve the cellulase productivity. The bioconversion of cellulosic waste to reducing sugar was maximum with Banana peels (77.78 %) followed by the rice husk (75.56 %), orange peels (71.11 %), corn steep peels (60.0 %) and lowest bioconversions (53.33 %) were recorded with sawdust. The degradation of cellulosic waste increased with increasing substrate concentration. Maximum cellulase productivity (3.18 U/ml) and bioconversion (86.1 %) was obtained at 3 % (w/v) of cellulosic waste (Banana peels). Saccharification of cellulosic waste with different treatment methods was studied. The pretreatment of cellulosic waste with 1 % HCl and H2SO4 produces 21 and 15.8 g of reducing sugar / 100 g of cellulosic waste. In comparison, hydrolysis with Streptomyces sp. B 167 enzymes resulted a significantly higher amount of reducing sugar yield (25 g / 100 g cellulosic waste). Further fermentation of cellulosic hydrolysates was performed using Saccharomyces cerevisiae using stationary fermentation condition. Maximum yield of ethanol were (0.30, 0.19 and 0.10 g ethanol / g glucose) observed with Streptomyces sp. B 167 enzymes, HCl and H2SO4 hydrolysates respectively after 48 h of fermentation

التاثير التثبيطي لبعض المستخلصات على فعالية انزيم Angiotensin converting enzyme وبعض المؤشرات الحيوية المساهمة في ارتفاع ضغط الدم == Inhibitory Effect of Some Plant Extracts On Angiotensin Converting Enzyme Activity And Some Biochemical Marker That Associated With Hypertension

Author name: رؤى اياد يوسف
Supervisor name: غازي منعم عزيز | حسن فياض
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: The present study was sought to investigate the inhibitory effect of four crude aqueous plant extracts : Zingiber officinale Roscoe (ginger), Camellia sinensis (Green tea), Olea europaea (Olive) and Hibiscus sabdariffa (Rosella) on key enzyme linked to hypertension, Angiontensin - I Converting enzyme (ACE), and on the oxidant/antioxidants status, lipid profile in vitro and in vivo studies. Study of some biochemical biomarker demonstrated that antioxidant enzyme, oxidant enzyme, liped profile and ACE level for 75 hypertension patients. Antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) were shown to have cross relationship with ACE level in hypertension groups, while lipid profile have positive relationship with ACE level in hypertension groups. ACE activity for the four groups G1, G2, G3, G4 were 19.61±3.97, 21.3 ± 1.95, 28.06 ± 5.34 and 32.74±8.19 ng/ml respectively. From these results we concluded that ACE was associated with hypertension and its modulated by drug or herbal extracts. Angiotensin - I Converting enzyme was extracted from sheep lung with specific activity 0.08 U/mg, then the crude ACE extract was concentrated with sucrose by dialysis with specific activity 0.1 U/mg, purification fold was 1.25.The enzyme was purified partly by ion - exchange chromatography using DEAE - cellulose with specific activity 0.5U/mg, yield 30% and purification fold 6.25. ACE activity was determined using N - [3 - (2 - furyl) acryloyl]L - phenylalanyl glycyl glycine (FA - PGG) as a substrate. Results for in vitro ACE inhibitory activity using plant extracts (ginger, green tea, roselle and olive) showed that the all four crude aqueous plant extracts had inhibitory activity in different values when used in the same concentrations about (1 mg/ml), and ginger extract possessed higher inhibitory activity than other three extracts. The ACE inhibitory potency of the ginger extract was found to be significant (P<0.001) when compared with the standard anti - ACE inhibitor drug (Captopril) at the same concentration. The inhibitory activity of ginger extract with different concentration (25, 50, 100 mg/kg) in L - N - ? - nitro - L - arginine methyl ester (L - NAME) induced hypertensive mice was evaluated. Acute oral administration with L - NAME 50mg/kg.b.w causes a rise in blood pressure in normal mice. Administration of aqueous ginger extract (25, 50 and 100 mg/kg) for 4 weeks in L - NAME treated mice significantly (P<0.05) reduced the mean arterial blood pressure compared with L - NAME animals without treatment, with decreasing the serum levels of ACE; while the activity of superoxide dismutase (SOD) and glutathione peroxidase(GPx) showed a significant elevation in ginger treated L - NAME induced hypertensive mice. The results suggest that ginger extract could prevent the development of high blood pressure induced by L - NAME probably can be attributed to prevent or reduce the oxidation process and the inhibition of physiological processes of a substance L - NAME and so as it contains ginger compounds of polyphenols, which inhibits the activity of the ACE and prevent oxidation of fats and repair System Antioxident. Our study concluded that ginger might act as a natural alternative to better and safer in the prevention of negative impacts and risk factors such as high blood pressure and lipids.

تحديد تعبير الجين المتحمل للملوحة TaGSK1 في عدد من اصناف الحنـطـة == Determination of Gene Expression of Salt Tolerant Gene Tagsk1 In Wheat Cultivars

Author name: ايمان نعمان اسماعيل
Supervisor name: نورية عبد الحسين علي | مجيد ارشيد سباح
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: Arabic
University location: Baghdad
First pages:
Abstract: نفذت الدراسة الحالية في مركز بحوث التقنيات الاحيائية/ جامعة النهرين للمدة 2011 - 2012 لغرض الكشف عن جين الملوحة TaGSK1 ودراسة التعبير الجيني له في صنفين من الحنطة فرات ودجلة والتركيب الوراثي 2H المنتخبة لصفة تحمل الملوحة مقارنة مع الصنف تموز2 الحساس للملو | This study was conducted in biotechnology research center, Al - Nahrain university in 2011 - 2012 to detect the gene responsible for salt tolerant (TaGSKI) and study its expression in two wheat (Tigris and Euphrates) cultivars and the genotype (2H) were selected for salt tolerance through improvement and breeding programs as compared with to the sensitive local wheat cultivar (Tamooze 2). Seeds of the these cultivars were sown in flasks that contained Ms media in three salt levels (0, 15, 25) ds/m with three replication for each. Five seeds from each cultivars were sown in each replicate. After 10 - 15 days from the sowing, the percentage of germination was estimated, and samples of leaves were collected for RNA extraction and then changed to the cDNA. B - actin gene that consider endogenous control and salt tolerant gene TaGSK1 were amplificated by using PCR technique to discover the gene and by QPCR to estimate gene expression by determine the CT (Cycle Threshold) value for B - actin and TaGSK1 genes in wheat plants of the studying cultivars. Number of DNA dilutions of 189bp molecular weight that extracted from agarose gel were used to determing CT value for each dilution. Standard curves were drawn to find out value of PCR Efficiency which was used for gene expression for the salt tolerant gene TaGSK1.The results of germination percentage (%) showed that there were high differences between the two salt tolerant cultivars (Tigris and Euphrates) and 2H genotype and local cultivar (Tamooze 2). Euphrates cultivar gave 100% germination percentage, and the Tigris cultivar and 2H genotype gave 89, 86% germination percentage respectively at 25ds/m. whilst the local cultivar gave zero germination percentage at the same level of salinity. At 15ds/m level, also the Euphrates cultivar gave 100% germination percentage while the Tigris cultivar and 2H genotype gave 96, 94% germination percentage respectively as compared to Tamooze cultivar that gave 13% germination percentage. All the cultivars have 100% germination percentage at 0.0ds/m level. The conclusion of this result is the two cultivars (Tigris and Euphrates) and 2H genotype more salt tolerance than local cultivar at this growth stage which more salt sensitive than others growth stages. The results of PCR reaction were also indicated that Tigris and Euphrates cultivars and 2H genotype have salt tolerant gene TaGSK1, while this gene did not exist in the local cultivar. There were two bands of TaGSK1 gene that have 189bp and 404bp molecular weight in the Tigris and Euphrates cultivar and the genotype 2H, while the local cultivars have only one band that have 404bp molecular weight. These results were indicated that 189bp molecular weight of this gene is responsible for salt tolerance character in these cultivars.The results of QPCR reaction also were revealed that there is difference between the cultivars in their gene expression. Tigris and Euphrates cultivars and 2H genotype gave the highest gene expression at 15ds/m and increased at 25ds/m as compared to 0.0ds/m. At the second level 15ds/m the gene expression of the two cultivars and genotype was 0.8682, 0.8190 and 0.8688 respectively, and at the third level 25ds/m was 1.656, 1.3176 and 1.2665 respectively, while at the first level 0.0ds/m was less than the other 15, 25 ds/m as compared to local cultivar that the gene salt tolerant (TaGSK1) have no gene expression at the same salt levels. This result indicated that the local cultivar does not have salt tolerant gene TaGSK1. From these results we can revealed that TaGSK1 gene was found in the Tigris and Euphrates cultivars and 2H genotype, and this gene can be considered from high salt tolerant gene, because it gave the highest gene expression at the highest salt level 25ds/m. Therefore this gene help the plant to tolerate salt stress and grow very well. The results also showed that the two salt tolerant cultivars (Tigris and Euphrates) and the salt tolerant genotype (2H) proximately have the same salt tolerance degree, so they have proximately the same gene expression as compared to salt sensitive local cultivar which have no salt tolerant gene TaGSK1. This gene is good indicator for salt tolerance at high salinity levels 15, 25 ds/m in these cultivars and genotype.

التحري عن الطفرات في جيني CNTNAP2 وIL1RAPL1 في مرضى التوحد == Mutation Screening of CNTNAP2 And IL1RAPL1 Genes In Autistic Patients

Author name: بشير كاظم خرميط
Supervisor name: عبد الكريم عبد الرزاق القزاز
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: ان اضطرابات طيف التوحد هي مجموعة من الظروف التي تتميز بضعف في التواصل الاجتماعي ونمطية في السلوك. يختلف الاشخاص المتوحدين اختلافا كبيرا في التطور المعرفي والتي يمكن ان تتراوح من فوق المتوسط الى العجز في التفكير. رغم ان اضطرابات طيف التوحد هي تورث بشكل كبي | The autism spectrum disorders (ASDs) are a group of conditions characterized by impairments in reciprocal social interaction and communication, and the presence of restricted and repetitive behaviors. Individuals with an ASD vary greatly in cognitive development, which can range from above average to intellectual disability (ID). While ASDs are known to be highly heritable (~90%), the underlying genetic determinants are still largely unknown. The research studies correlate between Contactin - associated Protein - Like 2 (CNTNAP2), Interleukin - 1 Receptor Accessory Protein - Like1 (IL1RAPL1) genes and ASDs. This study includes forty autistic patients and forty non autistic children as control groups (twenty unaffected sibling and twenty unrelated children). The age of autistic and non autistic children was ranged from 3 to 10 years. Blood samples of autistic patients were collected from Rahman specialist centre for the care and service autistic children in Baghdad. DNA was extracted from blood samples for molecular detection of CNTNAP2 and IL1RAPL1 mutations associated with ASDs by the use Polymerase Chain Reaction (PCR) technique and sequence analysis. PCR reaction was performed to amplify exons (14, 17 and 20) of CNTNAP2 gene that encode to CASPR2, a member of the neurexin family which functions in the nervous system as cell adhesion molecules and receptors. The PCR results revealed that identical bands related to the CNTNAP2 exons were present in all samples. Therefore, five samples (four from autistic patients and one from control sibling) were selected for genotype analysis of CNTNAP2 exons (14, 17 and 20) by direct sequencing. Genotype analysis revealed that there were no any variants in CNTNAP2 exons, but it shows that four different mutations were identified in non coding region (introns) of the CNTNAP2 gene. These mutations were seen only in autistic patients but absent in control sample. Three of these mutations are single nucleotide polymorphisms (SNPs) (rs3779031 A/G in 2118282 position, rs3779032 A/C in 2118436 position and T/G in 2117905 position). The other mutations were deletion in one nucleotide (Del A/ - in 2117901 position). SNP rs3779032 A/C are located at intron 21 while other mutations are located at intron 19. The current study showed that two common SNPs (rs3779031 and rs3779032) in CNTNAP2 were strongly associated with ASDs, where the frequencies of these SNPs were relatively high. SNP rs3779031 identified in two autistic patients while rs3779032 identified in three autistic patients from four unrelated families with ASDs. PCR reaction also was performed to amplify exons (3, 4, 5, 6, 7, 8 and 9) of IL1RAPL1, a gene implicated in calcium - regulated vesicle release and dendrite differentiation. The PCR results show a large intrgenic deletion (Deletion of exons 3 and 4) in six autistic patients, two of these patients were twin. This deletion may be incomplete penetrance due to phenotypic heterogeneity of these patients. This study provides evidence of the role of genetic factors in the etiology of ASD and the important CNTNAP2 and IL1RAPL1 genes mutation of pathogencity ASDs.

الازالة الحيوية لليورانيوم والسيزيوم من الترب الملوثة بواسطة نبات الشعير == Phytoextraction of Uranium And Cesium From Contaminated Soil By Hordeum Vulgare Plants

Author name: سيف صبار كامل
Supervisor name: ناظم حسن حيدر
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: Phytoremediation is defined as the use of green plants to remove pollutants from the environment or to render them harmless. Phytoremediation process can be classified based on the contaminant fate as; Phytoextraction, Phytotransformation, Phytostabilization, Phytodegradation, Rhizofiltration. In this work, the phytoextraction process was employed. A piece of land in the Botanical Garden at the University of Baghdad with an area of 25 m2 was prepared to plant the barley plant. Then, seeds of barley of type "Samir", which is an Iraqi kind that suitable for cultivation in Iraq, have been seeded. For U and Cs experiments, soil was mixed with a limited quantity of each U isotope for three different concentrations; 152 Bq/kg, 95 Bq/kg and 81 Bq/kg and for Cs106.5 Bq/kg, 79 Bq/kg and51 Bq/kg. For NPK and Urea experiments, different concentrations were tested to establish the requirements of these experiments. The LB4100 - W counting system includes the most complete data analysis software package was used to measure and calculate the alpha and beta concentrations and subsequently the overall activity concentration of any studied sample. Samples were prepared by following the Indian Standard method. For U, the experiment achieved by dividing it into four groups that differ in the spent time of agriculture in contaminated and clean (reference) soils. The results illustrated that the phytoextracted of U with planted period in contaminated soil, which were 31, 50, 63, 34 days, were 36.22, 54.84, 76.24, and 66.30 Bq/kgm, respectively. However, the 4th group differs comparing with other groups in the spent time of cleaned soil, which was 73 days. For Cs experiments, the work grouped similar to U experiment. The results of Cs phytoextraction showed that the absorbed Cs were 54.34, 100.69 and 109.07 for spent times in contaminated soil; 23, 43 and 57 respectively. Furthermore, barley plant has significant ability to phytoextract U and good ability to phytoextract Cs for all the three different concentrations. Besides, the results illustrated that the increase in the planted time in contaminated soils led to increase the quantity of phytoextracted isotopes. The results of adding K fertilizer showed a decrease in the ability of barley to absorb U, while the addition of urea enhanced the ability of barley. Finally, the following conclusion can be drawn from the present study that : barley is a good tool to phytoextract Cs rather than U and the use of urea fertilizer is suitable for enhanced the phytoextraction process.

الكشف الجزيئي عن التغيرات في جين MSX1 المسؤول عن حالة فقدان الاسنان باستخدام سلسة تفاعل البلمرة في عينة من المرضى العراقيين == Molecular Detection of Msx1 Gene Changes Responsible For Causing Hypodontia Using Polymerase Chain Reaction (PCR) In Sample of Iraqi Patients

Author name: اماني احسان الصقر
Supervisor name: اسماعيل حسين عزيز | اكرم فيصل الحويزي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: مصطلح الهايبودونشيا يعني نقص الاسنان الخلقي ويعد من اكثر التشوهات الفموية حدوثا لدى الانسان. مائة وخمسة وسبعون من مرضى نقص الاسنان الخلقي سجلوا في هذه الدراسة لديهم على الاقل سن واحد مفقود ولايزيد عدد الاسنان المفقودة عن ستة، قصدوعلاج الاسنان التقويمي في | Hypodontia means congenitally missing teeth, and considers as one of the most common oral alteration in human. One hundred and seventy five of hypodontia patients were matriculated in this study, having at least one missing tooth and no more than 6 missing teeth, seeking orthodontic treatment, who attended Al - Huwaizi Dental Center at AL - Harethia District - Baghdad, and control group consists of twenty five apparently healthy subjects (15 females, and 10 males). The age of both groups ranged from14 to 65 year. Buccal swabs were collected (for molecular study) from 25 of those patients who attended the clinic in a period between the beginnings of October 2013 to the end of April 2014, and from control group. Information were taken from the subjects under study according to a questionnaire that includes, name, gender, age, family and medical history, and the place of residence. Hypodontia was diagnosed according to the history of patients, clinical examination, orthopantomogram (OPG), and dental casts. The result of demographic study of hypodontia patients revealed that hypodontia was found in 129 of females which were more than that in males(46) with significant difference (p < 0.05). The ratio of females to males was 2.8 : 1. The missing teeth in hypodontia patients were found in right, left or both sides. The number of hypodontia patients, who had missing teeth in the right side were 37, in the left side were 48, while in both sides reached to 90 with non - significant differences and the results clarified that the missing teeth in anterior region found in 81 patients were higher than that in posterior region (63) while the least in both regions which recorded in 31 patient. It was found that hypodontia was more common in the maxilla (73) (upper jaw) than that in mandible (65) (lower jaw), whereas 37 suffering from missing teeth in both jaws, with non - significant differences. Present results showed that the maxillary lateral incisor (LI) was the most frequently missing tooth (124), the second most missing tooth was mandibular second premolar (PM2) (101) followed by lower central incisor (CI) (33), the maxillary second premolar(PM2) (27) whereas the lowest frequently missing teeth were canine (C), and the first premolar (PM1). The molecular part of present study used polymerease chain reaction (PCR) technique for amplification of DNA samples extracted from buccal swabs of twenty five hypodontia patients and control group.Four pairs of primers X1.1F, X1.3R; X1.4F, X1.4R; X2.1F, X2.3R, and X2.3F, X2.4R of the MSX1 gene, obtained from Bioneer Company (Korea), were used to amplify overlapping regions of the 2 exons of the MXS1 gene. The first pair of primers was used to amplify fragment with product size of 421 bp., while the second, third, and forth pairs of primers were used to amplify fragments with product size of 152 bp., 493 bp., and 264 bp., respectively. The outcome of MSX1 gene amplification showed that four patients with the first pair of primers and nineteen patient with the third pair of primers gave negative result (no bands) which differed from the result of the other patients and control. The disappearance of bands may be attributed to MSX1 microdeletion in those patients.The sequencing of MSX1 gene for the PCR product of second, third and fourth pairs of primer showed no genetic mutation, while the PCR product of the first pair of primers reveled nine missense and two silent mutations.It was concluded that hypodontia occurre frequently in Iraqi population and its occurrence in females was higher than that in males, and the gene MSX1 is responsible for many teeth missing in hypodontia patients.

تقويم صفة تحمل بعض اصناف البطاطا المكثرة نسيجيا للجفاف على المستوى الجزيئي == Evaluation of Some Potato Varieties Grown In Vitro To Drought Tolerance At The Molecular Level

Author name: هوازن حليم صالح
Supervisor name: علي عبد الامير مهدي الصالحي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: Arabic
University location: Baghdad
First pages:
Abstract: درست استجابة اربعة اصناف من البطاطا Solanum tuberosum L. وهي (Lusa, Ambo, Arizona, Riviera)، استجابتها للزراعة النسيجية، وتقييم صفة تحمل الجفاف، كما اجريت التحاليل الجزيئية لتحديد البصمة الوراثية لها.اوضحت نتائج تجربة اختبار استجابة الزراعة النسيجية، ان ا | Four varieties of Potato (Solanum tuberosum L. Lusa, Ambo, Arizona and Riviera) were studied for their response to tissue culture and evaluation of the treatment of drought tolerance and molecular analysis. Results showed that the varieties differ in their response to growth, where as Arizona was the best variety in shoot tip culture and the survival percent (100%) and gave the highest average of branch length and number of leaves which were 9.810 cm, 8.100 leaf/ plant, while Riviera showed lower rate of survival and growth (40%). Riviera was excluded from the experiments. Cytokines and auxins were used in the following concentrations Kinetin (kin) with 2, 4 mg/L and Benzel adenine (BA) with interacted with Indoleacetic acid (IAA) at 1 or 2 mg/l or Napthalene acetic acid (NAA) at 0.5 or 1.0 mg/L in multiplication experiment. Arizona variety exhibited the highest length shoot, number of nods, number of leaves, root length, dry weight of roots, fresh and dry weights of shoots when using kinetin 2mg/L; interacted with NAA at 0.5 mg/L. while it showed a significant increase in the average of branches number, stem height, fresh and dry weights of shoot as well as the number of roots after treatment with using BA at 1 mg/L in combination with 0.5 mg/L (IAA). Drought tolerance of the studied varieties was studied by using Poly Ethlene Glycol (PEG) at the concentrations 20, 40, 60, 70, or 80 g/l. It showed that proline content was at its highest in Ambo variety reached 22.811 ? /g at the concentration 80 gm/L PEG. Arizona gave the highest mean in number of branches, number of nods, number of leaves, number of roots, under water stress conditions when the medium was supplemental with 20 mg/L PEG. Molecular analysis the studied varieties was conducted by using 6 primers. Results showed that the inheritance approximation between them, 0.314, 0.297, 0.273 for Lusa, Ambo and Arizona respectively. These results indicate that Rivera variety is genetically different from the others.

الكشف الجزيئي عن بكتريا Oxalobacter formigenes باستخدام جينات 16SrRNA، oxc في البراز من اشخاص اصحاء == Molecular Identification of Oxalobacter Formigenes Bacteria Based On 16S rRNA, Oxc Genes In Stool Samples Healthy Subjects

Author name: طارق طلال خلف
Supervisor name: زهرة محمود الخفاجي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: Arabic
University location: Baghdad
First pages:
Abstract: بكتريا Oxalobacter formigenes من مكونات المكنون البشري الميكروبي، تتغذى على الاوكزالات السامة وبذا تخلص الجسم منها. البكتريا سالبة لصبغة كرام، ولا تكون الابواغ ولاهوائية المعيشة، ويمكن عزلها من الغائط البشري، ولدراسة البكتريا من النواحي الجزيئية، تم جمع ع | Oxalobacter formigenes is one of the human microbiome, uses the toxic oxalate for their growth (Oxalotroph), and helps the body to get rid of excess oxalate. The bacterium is Gram negative, non - spore former, obligate anaerobe, and can be isolated from human stool.To carry out molecular studies on this bacterium, 50 samples of human stool were collected from subjects with wide range of ages (3 - 63 years) and different genders, subjects were healthy and have no medications and especially antibiotics for at least for 3 months. Genomic DNA was extracted with special kit, to amplify certain housekeeping genes, these are 16S rRNA (3' end) segment having the anti - Shine - Dalgarno sequence (ASD), and the oxc gene responsible for production of oxalyl - CoA decarboxylase (middle region responsible for active site at where the Thiamin pyrophosphate binds, the enzyme co - factor). It was possible to amplify the 16S rRNA gene in 46 sample out of 50 (92%), and 7 samples out of 50 (14%) for oxc gene. Analysis and alignments of 16S rRNA sequences put the Iraqi samples in the O. formigenes i.e., the same species depending on Distance Score of alignments, it has been found that ASD of bacteria is conserved and represented by the sequence " 5' CGCGGUGAACGUUCCCGGG3' " in the studied samples and in strains found in the International Databases. Segment of oxc (~ 600 bp) translated into proteins (190 - 194 amino acids) were aligned with oxc protein of O. formigenes (M77128) reference strain, it has been found that this segment similar to the TPP binding site and characterized by its high hydrophobicity

التحري عن طفرة JAK2V617F والمستويات المصلية لانزيمي الفوسفتيز القلوي واللاكتيت ديهايدروجنيز في مرضى ابيضاض الدم النخاعي المزمن == Detection of Jak2V617F Mutation And Serum Levels of Alkaline Phosphatase And Lactate Dehydrogenase In Chronic Myelogenous Leukemia

Author name: استبرق اكرم بيرام الحسيني
Supervisor name: عصام فاضل علوان الجمیلي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: يعد مرض ابيضاض الدم النخاعي المزمن (CML) من الاورام النخاعية التكاثرية، وينشا نتيجة تكون جين Bcr - Abl (الذي يسمى كروموسوم فيلادلفيا) داخل الخلايا الجذعية المكونة للدم. يؤدي هذا الشذوذ الجيني الى تفعيل متواصل لانزيم التايروسين كاينيز وبالتالي نمو وتكاثر غ | Chronic myelogenous leukemia (CML) is a myeloproliferative neoplasm arises from Bcr - Abl gene translocation (called Ph chromosome) in hematopoietic stem cells (HSCs). This genetic abnormality results in constitutive activation of tyrosine kinase and subsequent uncontrol growth and multiplication of granulocytes. The cornerstone in treatment of CML are tyrosine kinase inhibitors, of which imatinib is the most effectively used. JAK2V617F mutation is an acquired single nucleotide polymorphism (SNP) occurs in JAK2 gene and is associated with many hematological malignancy other than CML. It was thought that the two genetic abnormalities (Bcr - Abl and JAK2V617F) occur mutually; however, growing body of evidences suggested the reverse. This study aimed to investigate the prevalence of JAK2V617F mutation associated with serum levels of alkaline phophatase (ALP) and lactate dehydrogenase (LDH) in Ph+ CML Iraqi patients treated with imatinib. A total of 43 Ph+ CML patients (25 males and 18 females, age range 16 - 80 years) who attend Iraqi National Center of Hematology for Research and Treatment/Baghdad were enrolled in this study. Each patient has been received at least six month therapy with imatinib. A consent form involving age, gender, height, weight, smoking status, residency and first family relative history of leukemia was obtained from each patient. Besides, blood samples were collected, from which the granulocytes were separated and then DNA was extracted using a ready kit. Two assays were used for detection of JAK2V617F mutation; real time polymerase chain reaction (qPCR) using specific primers and probe, and allele specific PCR (AS - PCR) using specific primers. Total white blood corpuscles (WBC) as well as serum levels of ALP and LDH were measured. qPCR assay revealed 5 patients out of 43 (11.62%) were heterozygous for the muatant allele of JAK2V617F mutation (genotype GT). The concentration of this allele ranged from 0.01% to 0.12%. None of blood sample gave positive result for AS - PCR assay. From the all risk factors, only gender had significant association with the incidence of JAK2V617F mutation. Average total WBC count, and serum levels of ALP and LDH were higher in JAK2V617F - positive patients (9042±1512.55, 146.05±8.028 IU/L and 204±10.85 IU/L respectively) than that of JAK2V617F - negative patients (6039±1772.239, 64.45±40.15 IU/L and 178.33±13.693 IU/L respectively) with significant differences. These results indicate that JAK2V617F mutation can occur simultaneously with Ph chromosome in CML patients, and qPCR is a highly sensitive method for the detection of this mutation. Furthermore, serum activity of ALP can be used as an indicator for the presence of JAK2V617F mutation in CML patients.

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

Author name: جودت نوري غائب
Supervisor name: عبد الحسين مويت الفيصل
General topic: Biology
Specific topic: Biotechnologies
Degree: Doctorate
Language: English
University location: Baghdad
First pages:
Abstract: 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.

تاثير بعض المستخلصات النباتية في معايير بيولوجية مختلفة للفئران == Effect of Some Plant Extracts On Different Biological Parameters On Mice

Author name: عقيل حيدر عطا الله
Supervisor name: مؤيد صبري شوكت
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: اجريت الدراسة الحالية للتحري عن المركبات النشطة الموجودة في المستخلص الخام المائي والميثانولي لاوراق الاس والنعناع والريحان ودراسة تاثيرها على فعالية انزيم الاسيتل كولين استريز ومستوى الدهون ومستوى السكر ومستوى انزيمات الكبد(ALP وSGPT وSGOT) ومستوى الانتر | The present study was conducted to investigate the active constituents found in aqueous and methanolic crude leaf extracts of Myrtus communis, Mentha piperita and Ocimum basilicum, and studies it effect on the acetylcholinesterase activity, levels of lipids, level of glucose, level of liver enzymes (ALP, SGPT and SGOT) and level of Interleukins (IL - 2, IL - 4, IL - 6 and IL - 10) in laboratory mice (in vivo). The results of the phytochemical analysis of the crude aqueous and methanolic extracts of Myrtus communis, Mentha piperita and Ocimum basilicum contain active compounds : Phenols, Flavonoids and Tannins and missing of Steroids and Coumarines in all extract but Saponins and Alkaloids found in alcoholic extract only, while terpens were present in Mentha piperita and Ocimum basilicum and absent in Myrtus communis. The results of administrating animals with different extracts showed no significant difference on blood Acetylcholinesterase (AchE) compared with ethanol liquid diet, while the alcoholic and aqueous extracts of M. communis, M. piperita and O. basilicum in the serum of decreased Acetylcholinesterase level significantly(p?0.05), liver and brain [(1.25 ?pH/30 min, 1.23 ?pH/30 min, 1.28 ?pH/30 min, 1.20 ?pH/30 min, 1.26 ?pH/30 min, 1.28 ?pH/30 min), (0.35 ?pH/30 min, 0.34 ?pH/30 min, 0.34?pH/30 min, 0.36?pH/30 min, 0.42?pH/30 min, 0.39?pH/30 min), (0.32?pH/30 min, 0.37?pH/30 min, 0.39?pH/30 min, 0.36?pH/30 min, 0.34?pH/30 min, 0.37?pH/30 min)] respectively compared with animals fed on ethanol liquid diet [(1.37 ?pH/30 min), (0.47 ?pH/30 min), (0.45 ?pH/30 min)] respectively. The methanolic and aqueous extracts of M. communis, M. piperita and O. basilicum reported a significant decrease in level of the total cholesterol and Low Density Lipoprotein (LDL) [(181mg/dl, 186mg/dl, 175mg/dl, 172mg/dl, 181mg/dl, 184mg/dl), (118mg/dl, 121mg/dl, 114mg/dl, 109mg/dl, 118mg/dl and 120mg/dl)] respectively, when compared with animals fed on ethanol liquid diet [(195 mg/dl), (132mg/dl) ] respectively while no effect was reported on High Density lipoprotein. The level of triglyceride reduced after the treatment with ethanol liquid diet, and then the level increased after the treatment with M. communis and O. basilicum extracts. The methanolic and aqueous extracts of M. communis, M. piperita and O. basilicum have led to a reduction in the level of glucose in the serum which increased after the treatment with ethanol liquid diet. Methanolic and aqueous extracts decreased the liver enzymes (ALP, SGPT and SGOT) significantly to the normal level (18, 17, 21, 18, 20 and 21) KAU, (19, 19, 16, 13, 17, 17) IU/L and (21, 24, 17, 15, 17 and 19) IU/L respectively after the increase by the treatment with ethanol liquid diet (24) KAU, (26) IU/L, (28) IU/L as compared to control (20) KAU, (12.2) IU/L, (13.5) IU/L, respectively. The level of Interleukin - 2 and Interleukin - 4 in the serum significantly increased in the treatment of alcoholic and aqueous extract of M. communis, M. piperita and O. basilicum [(18 pg /ml, 17 pg /ml, 20 pg /ml, 18 pg /ml, 20 pg /ml, 20 pg /ml), (100 pg/ml, 110 pg/ml, 119 pg/ml, 108 pg/ml, 90 pg/ml, 92pg/ml)] in comparison with ethanol liquid diet treatment [(14 pg /ml), (77 pg/ml)] respectively. While the level of Interleukin - 6 and Interleukin - 10 increased significantly in the serum, when animals were fed with ethanol liquid diet [(259pg/ml) and (501pg/ml)] respectively, and then decreased significantly after the treatment of methanolic, aqueous extract of M. communis, M. piperita and O. basilicum reported [(198 pg/ml, 202pg/ml, 202pg/ml, 201pg/ml, 214pg/ml, 217pg/ml), (370 pg/ml, 385pg/ml, 200pg/ml, 280pg/ml, 350pg/ml and 350pg/ml)] respectively.

التحري الجزيئي عن بعض التغيرات في الدنا المايتوكونديري للنطف لمرضى يعانون من وهن حركة النطف == Molecular Screening of Some Changes In Sperm Mitochondrial DNA In Asthenozoospermic Patients

Author name: عدي عدنان مهدي
Supervisor name: اسماعيل عبد الرضا عبد الحسن
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: من خلال البحوث والدراسات التي اجريت على اسباب وهن حركة النطف لدى الرجال الا انه هناك عدد من المسببات لم تحدد والى الوقت الحاضر. الا انه بالرغم من ذلك بعض الدراسات تشيربان العامل الوراثي يلعب دور في ذلك والمتمثل بعضيات المايتوكوندريا والحاوية على الدنا الم | Through researches and studies done on the causes of the asthenozoospermia in men, although there was no specific reason so far responsible for the infertility in people, but some of the studies were indicated that the genetic factors plays a role in the sperm dysmotility through the mitochondria that contained mitochondrial DNA responsible for providing the energy required for the sperm motility by production of ATP, which serve as the fuel that is consumed by sperm during motility. This study is conducted to determine the changes that could occur in the sperm mtDNA which included (common deletion, mtDNA copy number per cell and mutations in the ATPase8 and ATPase6 genes).This study was consisted of two parts as follow : The first part was consisted of two steps : The first step conducted on 71 samples from subjects suffering from the weakness of the movement of sperm and 12 samples from subjects who have a normal movement of sperm, total samples were 83 samples were divided patient samples into 5 groups depending on the percentage of the sperm motility under the classification adopted by the World Health Organization as well as a control group, that it has become 6 groups, these samples were subjected for investigation about the mitochondrial DNA deletions and mitochondrial DNA copy number by using real time PCR.On the other hand was taking 66 sample dispersed sample 56 patients and 10 healthy samples were subjected to the tests of ATPase6 and ATPase8 genes sequences for detection about mtDNA mutation.The second step was included the use of discontinuous gradient centrifugation (percoll) , which are 40 % and 80 % which represent both progressive and non progressive motile sperms, bringing the total samples 166 samples that collected and fractionated in a hospital, Kamal Al - Samarrai to treat infertility and IVF in Baghdad and examined for further molecular investigations for the period from February 2012 to October 2013.The second part was included the molecular tests that carried out after DNA extraction from the cells and this part include three steps : The first step was to investigate the common deletion in mtDNA where the results showed that the total number of deleted samples in the group (0 - 5) in the nonprogressive sperm was 81.25 %, which ranged between 6.8 - 74.7% while the ratio between the other groups 6 - 10, 11 - 15 , 16 - 20, 21 - 25 where were 75, 35.72, 28.5 , 36.3, and 12% for control group respectively. The second step was to investigate the sperm mtDNA copy number. The same groups where the results showed that the content of DNA in subjects with impaired movement of sperm was high compared with the control group where scored in some samples the highest level of gene expression 246.9 copies per cell in the group (0 - 5%) either other groups have shown lower levels were observed in groups (6 - 10) , (11 - 15) , (16 - 20) and (21 - 25) were 203.1, 180.7, 133.5 and 128.3 copies per cell, either the highest level of DNA content of the control group was 94.7 copies per cell, there was also a significant difference (P>0.05) between the normospermic motile isolated from 80% class and poor sperm movement isolated from 40 % class. The third step was included the detection of mutations by analyzing the sequences of the MT - ATAase8 and MT - ATPase6 genes which documented 59 mutations that were 23 missense and 36 silent mutations where all of them were heteroplasmic mutation except for a single mutation of the type missense (A8860G) which was homoplasmic mutation was noticed in all mtDNA copies of patients and control subjects also in progressive and non progressive sperm cells. Thought this current study novel nucleotides changes in MT - ATAase8 and MT - ATPase6 genes among groups where 3 novel missense mutations in MT - ATAase8 gene at positions (8378, 8483 and 8558), the changes of nucleotides bases were A>G, A>C, A>C and A>C respectively, replacing Asparagine to Aspartic acid, Leucine to Proline and Proline to Alanine respectively. Also two novel missense mutations were observed in MT - ATPase6 gene at positions nt (8822 and 9055) where the changes nucleotides were (C >T and G>A) that replaced amino acids (Serine to Phenylalanine and Alanine to Threonine) respectively. Silent novel heteroplasmic change as a transition substitutions in ATPase8 gene at position nt (8371) by replacing C>T without any changing in amino acid of protein. Results of this present study showed novel heteroplasmic silent mutations in ATPase6 gene at positions nt (8899, 9048 and 9060) nucleotides changes (C>T, T>C and C>A) respectively, without changing in amino acid of protein which were observed in infertile group

تشخيص بكتريا Neisseria gonorrhoeae بالطرق التقليدية والجزيئية في المرضى الذكور ودراسة مدى تاثيرها في حدوث الحذوف في موقع AZF == Conventional And Molecular Diagnosis of Neisseria Gonorrhoeae In Male Patients And Study Its Suspected Effects In Microdeletions In Azf Locus

Author name: غزوان علي مسلم الرماحي
Supervisor name: عبد الرضا عبد الحسن اللامي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: This study includes 82 clinical samples from male patients suspected to have gonorrhea (urethral discharge with dysuria). Two types of samples were collected from each patient, urethra swab and blood samples from patients who attended Al - Yarmouk teaching hospital in Baghdad city, Clinics and private laboratories during a period from December 2012 to April 2013. All of them were married and have children's, and 20 samples were taken from apparently healthy subjects.Cases caused by gonococci were diagnosed by finding the Neisseria gonorrhoeae bacteria in the samples using microscopic examination and culturing on rich media chocolate agare for growth bacteria and selective media modified Thayer martin agar (MTM) contact antibiotic to prevent growth of all the types of bacteria except N. gonorrhoeae.Microscopic examination using specific kit for gram stain, showed gram negative diplococcus, like bean shape. Culturing on rich media revealed that 82 (100%) samples positive but re culturing on Modified Thayer martin media revealed that 76 (92.68%) out of 82 positive. Then biochemical test had the same results.Results of the molecular diagnosis of the bacteria causing gonorrhea using specific primers that were specific for the Orf1 gene, revealed that 80.26% of samples (61 out of 76) were positive. Results of the microdeletion in Y chromosome AZFc region revealed that no microdeletion were occurred in SY - 254 STS and BPY - 2 gene.This study, provided high specificity and sensitivity for the diagnosis of gonorrhea using PCR technique which is cheaper and faster than traditional methods. Also, PCR - based method for detection of N.gonorrhoeae can be readily used in hospitals and laboratories.

دراسة بعض تاثيرات اللقاح المحضر من العزلة المحلية لبكتيريا Klebsiella pneumoniae == Study of Some Effects of Prepared Vaccine From Local Strain of The Klebsiella Pneumoniae

Author name: ياسر عبد الجبار عبود السوداني
Supervisor name: عصام فاضل علوان الجمیلي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: English
University location: Baghdad
First pages:
Abstract: تم جمع خمسين عينة سريرية من قشع مرضى مصابين بذات الرئـــة. وذلك للمدة من تشرين الثاني 2013 ولغاية ايار 2013 من مستشفى ابن البلدي ومستشفى بغداد وذلك لعزل وتشخيص بكتريا Klebsiella pneumoniae التي تعد احدى العوامل المهمة المسببة لاصابات الرئة. واخضعت عينات | Fifty clinical samples collected from sputum of patients who suffered from pneumoniae in Ibn - Balady hospital and the hospital in Baghdad city during the period from November 2012 to May 2013 for the isolation and identification of Klebsiella pneumoniae, one of the important causative agents of infection occurs in the lungs. Sputum samples were subjects to the standard laboratory procedures including identification by biochemical test and VIETK system. The results showed 15 isolates were revealed as Klebsiella Spp, only 10 isolates represented K.pneumoniae, The isolates were examined to produce extracellular toxic complex (ETC) it was found that the isolate named K2 was the higher production. Two method for purification the extracellular toxic complex (ETC) were used, first Aqueous two phase systems, In this method polymer - salt aqueous two phase system was evaluated in crude extract of K. pneumoniae at varying concentration of Dextran T - 150 with 20% with polyvinyl pyrrolidone to final rate (1 : 1) (wt : wt) with 0.2M sodium sulphate. The results showed the best concentration dilution sample given as (4.25 : 0.75) with protein concentration (97.173 mg/ml) which contained ETC in the lower layer and the mice died within 4 hours, while the second method performed by using two step column chromatography, ion exchange DEAE - Cellulose and gel filtration (Sepharose - 4B). In the first step sample given lethal activity by injection to the mice after six hours with protein concentration (55mg/ml), More purification by the second step animal died after 3hours with contain protein (27.75mg/ml). Furthermore, the results of the extracellular toxic complex characterization proved that molecular weight was 39810 Dalton determined through Gel - filtration chromatography using Sepharose 6B gel. The LD50 value of purified toxin was calculated, and the result was (6.52 mg/ml) of toxin.This quantity was found effective to cause killing of 50% of the total toxin treated animals. The biological effect of purified toxin of K. pneumoniae K2 have been examined in vivo by injection of dose (0.5 ml) of purified ETC toxin that contain (10.875 mg/ml ) protein. The final part of the study involved the histopathological changes were noted, abundant mononuclear infiltrate of inflammatory cells with necrosis of lung parenchyma. The second group of mice injected with (0.05 ml of ETC) that contain protein (1.085mg/ml) represented as sub lethal dose Histopathological changes were noted showing near of the normal appearance of alveoli and alveolar space, with presence of congestion of blood vessels. The third group of mice inject with (0.5 ml from Tris - base buffer only) represented control showed normal alveoli and alveolar space with presence of bronchial. In the immunological test the sample ETC examined with ELISA and given IgG titer (189.68+50.70 ng/ml) compared with control (46.78+12.45). This titer of IgG tested with Double immune diffusion assay and gave precipitation line with antigen compared with control.

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.

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.
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