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تاثير تعريض بذور واعضاء بادرات القرع الجبلي Cucurbita maxima والكالس الناشئ منها للاشعة فوق البنفسجية في محتوى الزيت وتشخيص مركب Cucurbitacin == Exposure Effect of Seeds and Seedlings Explants of Cucurbita maxima and Their Calli to UV in Oil Content with Diagnosis of Cucurbitacin Compound

اسم المؤلف: اسلام ياسر عبد الله الحمداني
اسم المشرف: عبد الله نجم عبد الله النعيمي
الموضوع العام: علوم الحياة
السنة: 2007
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة الموصل
اللغة: العربية
مكان الجامعة: الموصل
الصفحات الاولى:
المستخلص: تعرفت الدراسة الحالية على استجابة الاوراق والسيقان والاوراق الفلقية والسيقان تحت الفلقية لنبات القرع الجبلي Cucurbita maxima لاستحداث الكالس على الوسط الغذائي MS الصلب المدعم بتراكيز مختلفة من الاوكسينات (2,4 - D, IAA,NAA) والسايتوكاينينات (Kin,BA) وقد ابدت قطع الاوراق والسيقان استجابة عالية في الوسط المنتخب MSI(MS+1.0 ملغم/ لترBA+0.5 ملغم/ لترNAA)، اذ بلغت نسبة استحداث الكالس فيهما 100% ويليهما السيقان تحت الفلقية 88% ثم الاوراق الفلقية 77% التي اعطى كالسها اعلى وزن طري على هذا الوسط 5.2غم ثم السيقان 4.3 غم وعليه اعتمد هذا الوسط في هذه الدراسة لادامته الجيدة لكالس جميع الاجزاء النباتية المستخدمة فضلا عن نسبة الاستحداث العالية، كما ان الوسط الثاني MS) + 3.0 ملغم/ لتر Kin+ 0.5 ملغم/ لتر 2,4 - D) حقق نسبة استحداث عالية كذلك للاجزاء النباتية نفسها لكنه استبعد لانه لم يحافظ على حيوية الكالس عند الادامة لمدة طويلة.كما نجحت هذه الدراسة في زيادة نسبة استحداث الكالس في كل من الاوراق الفلقية والسيقان تحت الفلقية الى 100% باستخدام الوسط MSI المضاف اليه نترات البوتاسيوم بما يعادل ثلاثة اضعاف وسط MS وكالس السيقان تحت الفلقية الى 100% باستخدام الوسط MSI المضاف اليه نترات الامونيوم بما يعادل ثلاثة اضعاف وسط MS.استخدمت اربع جرع اشعاعية 10، 20، 40، 60 دقيقة للمطفر الفيزيائي (الاشعة فوق البنفسجية) في هذه الدراسة ولوحظ من خلال النتائج تاثير هذا المطفر على عمليات الانبات والصفات المظهرية وشكل خلايا كالس النبات اذ شجعت اشعة UV على تبكير عملية انبات البذور عند الجرعتين 10، 20 دقيقة كما ان الجرعتين العاليتين 40، 60 دقيقة زادتا من معدل طول النبات وطول الجذر في حين قللت الجرعتان الواطئتان 10، 20 دقيقة ذلك.كذلك شجعت غالبية الجرع الاشعاعية القطع النباتية المختلفة على استحداث الكالس خلال مدة زمنية قصيرة وحصول زيادة في الوزن الطري للكالس المعرض لاشعة UV وخاصة كالس السيقان تحت الفلقية عند الجرعتين 20 ، 40 دقيقة اذ بلغت نسبة الزيادة 2.4 و3.3 على التوالي وكذلك زادت كمية الزيوت في العينات التي عرضت للاشعة وخاصة عند الجرعتين 10 ، 20 دقيقة في قطع الاوراق والسيقان تحت الفلقية الناتجة من البذور المشععة وكالسهما وكذلك عند الجرعة 20 دقيقة في الكالس المشعع.تمكنت الدراسة من تشخيص الاحماض الدهنية Palmitic acid وStearic acidو Oleic acid وLinoleic acidبالاضافة الى تشخيص ثلاثة احماض دهنية جديدة وهي Myristic acid وArachidic acidو Lauric acid لاول مرة في هذا النبات، فضلا عن التشخيص النوعي للمركب Cucurbitacin والتعرف على ايزومراته Cuc E وCuc B وCuc I وCuc L باستخدام كروماتوغرافيا الطبقة الرقيقة (TLC). | The present study found out the response of leaves, stems, cotyledons and hypocotyls of Cucurbita maxima to produce callus on solid MS medium enriched with different concentrations of auxins such as 2,4 - D, IAA and NAA, and cytokinins such as Kin and BA. The explants of leaves and stems showed high response in the selected medium MSI (MS+1.0 mg/L BA + 0.5 mg/L NAA) where callus production from leaves and stems reached 100%, in hypocotyls 88% and cotyledons 77% whose callus had the highest fresh weight in this medium 5.2gm and stems 4.3gm therefore, the study adopted this medium for its good sustenance of the callus of explants in addition to the high rate of production. The medium MS + 3.0 mg/ L Kin + 0.5 mg/L 2,4 - D achieved a high production rate for the same explants. However it was excluded since it did not keep the vitality of callus when maintained for a long time.The study succeeded in increasing the callus production rate in cotyledons and hypocotyls to 100% by using MSI medium added to it potassium nitrate which equals three times the MS medium)] and the callus production rate in hypocotyls to 100% by using MSI medium added to it ammonium nitrate which equals three times the MS medium)]. Four radiational doses 10, 20, 40 and 60 min of ultraviolet were used by the physical mutant. It was noted from the results that the physical mutant affected the germination processes and morphology of the plant and cells shape of plant callus. The UV activated the early germination of seeds at the two doses 10 and 20 min. The high doses 40 and 60 min raised the average length of plant and roots whereas 10 and 20 min decreased it.Most of UV doses activated the explants to produce callus early. There was also an increase in the fresh weight of callus exposed to UV especially the hypocotyls callus of the at the doses 20 and 40 min which reached 2.4 and 3.3 gm respectively. The oil rate in the samples exposed to UV showed an increase at the doses 10 and 20 min in the leaves and in the hypocotyls which grew from radiated seeds and their callus and also at 20 min in radiated callus.The study singled out fatty acids like palmitic acid, stearic acid, oleic acid, Linoleic acid, and also singled out new three fatty acids like myristic acid, arachidic acid and lauric acid first time in this plant besides the qualitative diagnosis of cucurbitacin and defining it’s isomers Cuc E, Cuc B, Cuc I and Cuc L, by using thin layer chromatography

دراسة السمية الوراثية لحبوب منع الحمل على الانسان والفئران البيض == Study the genotoxicity for oral contraceptive pills on human and white mice

اسم المؤلف: اسراء ايوب علوان منصور القره غولي
اسم المشرف: عبد الامير ناصر الركابي | سعد محمد الندا
الموضوع العام: علوم الحياة
السنة: 2009
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: الهدف من الدراسة الحالية هو تسليط الضوء على التاثيرات الوراثية الخلوية لحبوب منع الحمل الفموية المركبة على عينات دم ماخوذة من نساء اعتمدن هذا العقار كمانع حمل وفي الحيوانات المختبرية (الفار الابيض) بوصفه نظاما اختباريا ، وتم الاعتماد على بعض التحليلات الوراثية الخلوية شملت الانحرافات الكروموسومية في خلايا نقي العظم والطحال ، الانوية الصغيرة ومعامل الانقسام بالاضافة الى تشوهات الهيكل العظمي للاجنة في الفار الابيض، والانحرافات الكروموسومية ، الانوية الصغيرة ومعامل الانقسام في عينات دم الانسان . درست التاثيرات الوراثية لمجموعة من النساء بفئتين عمرية مختلفة بين )25 - 34) و(35 - 45) سنة والتي قسمت الى اربع مجاميع ، اظهرت النتائج عدم وجود فرق معنوي باحتمالية (P<0.05) في النسبة المئوية للانحرافات الكروموسومية الكلية لمجموعة من النساء اللواتي ياخذن حبوب منع الحمل المركبة (COCPs) ذوات الفئة العمرية )25 - 34) سنة ، بينما اظهرت وجود فرقا معنويا في النسبة المئوية للانحرافات الكروموسومية للنساء ذوات الفئة العمرية (35 - 45) سنة . كذلك لوحظ وجود فرق معنوي باحتمالية (P<0.05) في اعداد الانوية الصغيرة (MN) لمجموعة من نساء ياخذن (COCPs) في كلا الفئتين العمريتين. في التجربة الاخرى درست التحليلات الوراثية الخلوية (الانحرافات الكروموسومية) للمانع (COCPs) على الفئران، التي قسمت الى خمس مجاميع شملت مجموعة السيطرة السالبة التي تمثل الحيوانات المجرعة بدارئ (PBS) ومجموعة السيطرة الموجبة المعاملة بعقار السايكلوفوسفومايد (CP) وثلاث مجاميع معاملة بالمانع (COCPs) بالتراكيز(2.44), (1.25) (0.60) , ملغم / كغم ، اشارت النتائج ان التركيزين (2.44), (1.25) ملغم / كغم سجلا فروقا معنوية باحتمالية (P<0.05) في نسبة الانحرافات الكروموسومية بينما لم تسجل المجموعة ذات التركيز (0.60) ملغم / كغم فرقا معنويا في نسبة الانحرافات الكروموسومية لخلايا نقي العظم والطحال . واظهرت نتائج التحليل الاحصائي وجود فروقا معنوية باحتمالية (P<0.05) للجرع المختلفة للمانع (COCPs) على اعداد الانوية الصغيرة اذ كانت معدلات اعداد الانوية الصغيرة هي (1.26) , (2.50) , (3.53) للجرع ذات التراكيز (2.44) ،(1.25) ، (0.60) ملغم /كغم على التوالي. وكذلك اظهرت النتائج ان هناك فروقا معنويا باحتمالية (P<0.05) في معدل اوزان اجنة فئران المجاميع المدروسة ، فقد اظهرت مجموعة السيطرة السالبة اعلى معدل وزن للاجنة بلغ (1.55) غم، في حين سجلت معاملة السيطرة الموجبة اقل معدل وزن بلغ (0.56) غم. بينما كانت معدلات اوزان الاجنة للمجاميع المجرعة بالمانع ((COCPs ذات التراكيز (0.60) , (1.25) , (2.44) ملغم /كغم هي (1.34) , (1.32) (1.40),غم على التوالي . اما بالنسبة لحساب عدد هلاكات الاجنة فلم يوجد اي تاثير ملحوظ لحبوب منع الحمل المركبة (COCPs) على هلاكات الاجنة في حين ظهرت الهلاكات بنسبة عالية جدا في السيطرة الموجبة ، لكن عند دراسة تاثير حبوب منع الحمل المركبة على الهيكل العظمي للاجنة لم نلحظ اي تشوهات بالهياكل العظمية لجميع اجنة الفئران للمجاميع الثلاث المعاملة بحبوب منع الحمل المركبة . | Our work aimed to study the cytogenetic effects of Combined oral contraceptive pills (COCPs) on blood samples obtained from women utilized this contraceptive and lab animals (mice) treated with (COCPs). Several cytogenetic analyses (Chromosome aberrations of bone marrow and spleen , Micronuclei , Mitotic index and skeleton abnormal of embryos) in mice and (CA , MN , MI) in blood samples of human were used in this study . Studied the genetic effect for women in two age groups rang between (25 - 34) , (35 - 45) years in four groups. Result show non significant difference (P<0.05) in percentage of total chromosome aberration in women taking (COCPs) of age group rang between (25 - 34) years. While show significant differences (P<0.05) in percentage of (CA) in women of age group rang between (35 - 45) years. Also it was noticed significant differences (P<0.05) in Micronuclei (MN) of women taking oral contraceptive pills of two age groups. In other experiment studied the cytogenetic analysis for (COCPs) on mice that were divided into five groups as follows : Negative control group that animals orally treated with PBS buffer, Positive control group was treated with (CP) and three groups treated with (COCPs) with concentration (2.44) , (1.25) , (0.60) mg/kg and the result show was concentration (2.44) , (1.25) mg/kg recorded significant differences (P<0.05) while group with concentration (0.60) mg/kg non significant difference. also it was noticed Significant differences (P<0.05) in three groups treated with (COCPs) in Micronuclei rate and were the rates (3.53%) , (2.50%) , (1.26%) of concentration (2.44) , (1.25) , (0.60) mg/kg. Also result show statistically significant differences (P<0.05) in body weight of mice embryo of studied groups, Negative control group recorded rate highly in embryo body weight was (1.55) gm. While recorded Positive control group rate less in weight was (0.56) gm while were rates in embryo body weight of orally treated groups with (COCPs) with concentration (2.44) , (1.25) , (0.60) mg/kg were (1.40) , (1.32) , (1.34) gm . but non show any influences to (COCPs) in embryo number .while show died embryos in positive control group. also the results show non any abnormality in skeleton of embryos for three groups that treated with oral contraceptive pills.

دراسة فعالية بعض مستخلصات الطحلب الاخضر Enteromorpha intestinalis (L.,1820 على الاكياس العذرية للمشوكة الحبيبية Echinococcus granulosus (Batsch,1786 وكمضادات لخط الخلايا السرطانية نوع HeLa == Study of the activity of some of the green alga Enteromorpha intestinalis (L.,1820) extracts on Hydatid cysts of Echinococcus granulosus (Batsch,1786) and as anticancer cells type HeLa

اسم المؤلف: غزوان طالب نوري الجابر
اسم المشرف: صبيح هليل المياح | احمد محسن عذبي
الموضوع العام: علوم الحياة
السنة: 2016
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
الجامعة: جامعة البصرة
اللغة: العربية
مكان الجامعة: البصرة
الصفحات الاولى:
المستخلص: The present study was conducted in order to evaluate the activity of green alga Enteromorpha intestinalis extracts (methanol and hexane extracts) on growth and development of hydatid cysts of Echinococcus granulosus in vitro and in vivo by using mice type Mus musculus Balb\c that in comparing with albendazole drug. Anti - cancer activity was also evaluated in vitro for both algal extracts by using HeLa cells. Chemical compositions of algal extracts were analyzed by using Gas chromatography - mass spectroscopy (Gc - mass)technique . The hydatid cysts were collected from livers and lungs of infected sheep which slaughtered in Basrah abattoir. While, The algae specimens were collected from Abu Sokhir marshland area /north of Basra. The results revealed that E.intestinalis chemical compounds are carbohydrate , alkaloids, phenols , saponins , triterpenes and sterols compounds. In vitro study showed that the 1 mg / mouse weight of albendazole drug was able to reduce the percentage of protoscolices activity to zero after the third day of treatment. While, the dose 6 98 mg / kg of methanol extract inhibited all protoscolices after five days of treatment, followed by the dose 804 mg / kg of hexan extract that inhibited all protoscolices after six days of treatment.In vivo study revealed that the methanol extract (698 mg / kg) treatment was the best in reducing the weights of organs significantly in comparison with the positive control group it was 2.192 , 0.192 , 0.434 and 0.282 g of liver, spleen, lung and kidney respectively. The methanol extract (698 mg / kg) treatment lowered the average number of hydatid cysts (2.6) more than other extracts treatments , then followed by hexane extract (804 mg / kg) treatment it was 2.8 cyst .This study has identified that the liver enzymes Aspartate transaminase (ALT) and Alanine transaminase (AST) values in the serum of positive control group were higher significantly than negative control group the value of the enzyme ALT was 79 IU / l and the value of the enzyme AST was 118.4 IU / l.All methanolic extracts groups showed a preference in enzyme's values compared with other treatments groups , the value of ALT enzyme of 698 , 688 and 678 mg / kg of methanol extract groups were 46.8 , 45.8 and 44.6 IU / l respectively, while all hexane extracts groups showed a preference in AST enzyme values compared to the other treatments groups, the value of AST enzyme of 804 , 794 and 784 mg / kg of hexane extract groups were 73.6 , 70.2 and 70.4 IU / l respectively. Found a significant increase in the number of white blood cells WBCs in a positive control group ( 4.6 × 013μ), compared with a negative control group and the other groups, While it has been found significant decrease in the Hb (5..8deciliter) and Packed cell volume (PCV) (31..%) values in positive control group as compared with control negative group. The study found that the number of white blood cells WBCs n the methanolic extract group (678 mg / kg) was 4.06 × 103 microliter was as same as in the negative control group, In contrast, hexane extract group (804 mg/ kg) has did not significant difference with negative control group in the Hb and PCV were 11.5 g / dl and 36.8% respectively .Histological study confirmed that there are the histological changes in positive control group included congestion , bleeding , degeneration , infiltration of inflammatory cells and hyperplasia. Whereas the histological changes in group treated with albendazole were bleeding, infiltration of inflammatory cells , atrophy of the glomerulus and fibrosis in some areas . In algal extracts groups the histological changes were infiltration of inflammatory cells, bleeding and congestion.The Gc mass results revealed that the ethanol extract contains loliolide , ethyl stearate , palmitic Acid , ethyl palmitate , phytol , ethyl oleate , ethylhexyladipate and squalene compounds. While, hexane extract has N, Ndimethyltetradecylamine , diisobutyl phthalate , palmitic Acid , ethyl palmitate and ethylhexyladipate compounds.Anti - cancer study showed the algal extracts have an inhibition activity against cancer cells, The half maximal inhibitory concentration (IC50) value of the methanol extract was 79.08 μ/ ml, While, the IC50 for hexane extract was 156.3 μ/ ml.

السيطرة الاحيائية واللااحيائية على نمو الفطر Aspergillus flavus والتعبير الجيني للجينين aflDو aflR وانتاج الافلاتوكسين B1 == Biotic and Abiotic Control on Aspergillus flavus Growth, aflD and aflR Expression and Aflatoxin B1 Production

اسم المؤلف: لبيد عبد الله نجم السعد
اسم المشرف: عدنان عيسى البدران | سامي عبد الرضا الجميلي
الموضوع العام: علوم الحياة
السنة: 2015
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
الجامعة: جامعة البصرة
اللغة: الانكليزية
مكان الجامعة: البصرة
الصفحات الاولى:
المستخلص: Fifteen isolates of Aspergillus flavus (AFL1 - AFL15) were isolated from multiple sources included soil, compost, seeds, fruits, feed and air. The isolates were identified morphologically then confirmed by molecular techniques. Only thirteen isolates were confirmed positively (AFL1 - AFL5 and AFL8 - AFL15) while two of them were not. A phylogenetic analysis was made which revealed that the isolates grouped according to their source of isolation. The aflR profile and phenotypic AFB1 production confirmed that all thirteen isolates posses the ability to produce AFB1 with significant differences (P ≤ 0.05) among them. Growth rate profile was performed under 30ºC which showed significant differences (P ≤ 0.05) among isolates. According to the above tests, A.flavus AFL14 was selected to be the experimental isolate for the rest of experiments in this study. Comparing to A.flavus NRRL3357 type strain, the impact of temperature, water activity aw and isolate factors was examined to determine the limits and optimum ecological conditions of growth of A.flavus AFL14. The results displayed that the best growth rate was (7.217 mm/day) at 0.98 aw where the lowest was (4.069 mm/day) at 0.9 aw with no growth at 0.85 aw while the impact of temperature demonstrated by outweigh of growth rate at 35ºC (6.201 mm/day) followed by 30ºC (5.272 mm/day) which exceeded 25ºC and 40ºC (4.604 and 4.051 mm/day), respectively.Bacillus subtilis (isolates : BSS1, BSS2, BSS3, BSS4 and BSW) and Pseudomonas fluorescens (isolates : PFMst and PFDL) were elected to be used as biocontrol agents. All the isolates were passed the biochemical identification tests moreover, BSS4 and BSW isolates of B.subtilis and PFMst and PFDL isolates of P.fluorescens were confirmed molecularly which used later in all biocontrol experiments in this study. The ability ofbiocontrol agents to inhibit fungal growth was investigated and the resultsshowed significant inhibition impact represented by significant outweighof B. subtilis BSS4 (99%) on the rest of B. subtilis isolates followed by P.fluorescense PFDL and PFMst (92.29 and 86.19) %, respectively, with nosignificant differences (P ≤ 0.05) among them. The extra - cellular abilityof biocontrol agents to degrade AFB1 showed a high degradation abilityIpreformed by B. subtilis BSS4, BSW isolates and P. fluorescence PFMst,PFDL isolates (100, 100, 97.805 and 97.396%), respectively. Thedegradation residues administrated to rats to determine their effect onbiosystems, the blood parameters showed a significant reduction ofWBC, HB, RBC, and P.C.V while there was a significant increase inUrea, Glutamic pyruvate transaminase (GPT) Glutamic oxaloacetictransaminase (GOT) and MCV in AF+DMSO treatment comparing tothe negative and positive control. Water activity and biocontrol factorswere examined to control aflD and aflR expression and phenotypic AFB1production. When the experiment held using Nutrient Agar medium, asignificant inhibition of aflD and aflR expression (P=0.000 and 0.003),respectively performed by BSW at 0.94 aw comparing to 0.98 aw whileno significant inhibition was observed in the rest of the treatments. Allbiocontrol agents revealed high significant reduction of aflD and aflRexpression (P≤0.001) in each water activity level separately whichconfirmed by HPLC results. The phenotypic results showed that 0.94 awexceeded 0.98 aw in AFB1 reduction (2011 and 4280 ng/gm),respectively which agreed with gene expression results. When MaizeMeal Agar used as a medium, aflD expression presented a highsignificant increase at 0.98 aw relating to the NM level of each individualtreatment (P= 0.000 - 0.03) except PFDL+AFL14 which showed nosignificant aflD expression between both levels. aflR expression revealeda high significant reduction (P= 0. 000) caused by 0.98 PFDL+AFL14and 0.98 BSW+AFL14, respectively, while no significant differenceswere observed in the rest of the treatments or control. The HPLC resultsdisplayed a high significant reduction of AFB1 at 0.98 aw (8447 ng/gm)compared to (219000 ng/gm) at NM aw. The biocontrol agents caused ahigh significant reduction for both aflD and aflR expression (P=0. 000 - 0.043) under each individual water activity level while, HPLC resultsshowed non significant low reduction to the AFB1 performed by BSS4and BSW (25160 and 44790 ng/gm), respectively, followed by asignificant increase in AFB1 amount caused by PFMst and PFDL(267600 and 184100 ng/gm), respectively

عزل وتشخيص احد قلويدات الايض الثانوي لبعض الطحالب ودراسة فعاليته الحيوية == Isolation and identification one alkaloid of secondary metabolites from some algae and study of biological activity

اسم المؤلف: انفال نوري عباس اللفتة
اسم المشرف: احمد محسن عذبي | اقبال جاسم الاسدي
الموضوع العام: علوم الحياة
السنة: 2015
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
الجامعة: جامعة البصرة
اللغة: العربية
مكان الجامعة: البصرة
الصفحات الاولى:
المستخلص: The current study includes an isolation, Identification and purification of three species of algae, two of them belonging to cyanobacteria they are Oscillatoria brevis and Nostoc carneum . The third one was Enteromorpha intestinalis which belong to green algae which was from different location in Basrah .There are two extract were prepared from the algal species Alcohol , and alkaloid extracts , Alcoholic extract prepared to know what was compound it had and alkaloid extracts to test bioactivity of algae Cytotoxcity also was carried out on human red blood cells, the results revealed that alkaloid extract from E.intestinalis was nontoxic, whereas bioactive compounds isolated from O.brevis , N.carneum. Showed hemolytic action .The bioactivity of alkaloid extract was examined to elucidate their on ability to inhibit the growth of gram Positive and negative bacteria. .Biological activity of alkaloid extracts of three algae isolated were determined by using the Minimal Inhibitory Concentration (MIC) againsttwo bacterial isolates. And also test the effect of filtrate pure algal culture on growth of plant pathogenic fungi ,also apparently that the algal alkaloid extract and isolated compounds exhibits antioxidant.The antitumor activity of the algal alkaloid extract from E.intestinalis against Rhabdo myosarcoma cell line was examined. The results showed that crude alkaloid extract possessed an antitumor bioactivity at low concentration 0.78mg / ml .In this study also was carried out to investigate the possibility of preventing the hypercholesterolemia by using alkaloid extract of E.intestinalis .it also aimed to study the effect of use this extract in reducing the Plasma Total Cholesterol (TC) , Triglyceride (TG), Low - Density Lipoprotein (LDL) , Very Low - Density Lipoprotein (VLDL) and no effect on High - Density Lipoprotein (HDL).The identification of the compound was made depending on the active groups test and spectroscopic analyses including : Infrared (IR) and Gas Chromatography / Mass spectrum (GC - MS). The results of such analyses showed that alkaloid extract from E.intestinalisa has three compound these are Hexadecanamide , Methenamine and Benzenedicarboxylic acid, diisooctyl est and alkaloid extract from N.carneum showed that has three compound 1,2 - Benzenedicarboxylic acid diisooctyles,ا9 - Octadecenamide, (Z) - )ااااا and Didecyl phthalate .Hexadecanamide purification from alkaloid extract of E.intestinalis depended on physical and chemical properties The identification of the compound was made depending on the active groups test and spectroscopic analyses including : Infrared (IR); Gas Chromatography / Mass spectrum (GC - MS) and this component showed bioactivity on gram. Positive and negative bacteria and it has also antioxidant activity

التحلل والمعالجة الاحيائية للمركبات الهيدروكاربونية النفطية بوساطة سلالات Streptomyces وNocardiopsis المعزولة من الترب الملوثة بالنفط في محافظة البصرة - العراق == Biodegradation and bioremediation of petroleum hydrocarbons by Streptomyces and Nocardiopsis strains isolated from oil contaminated soil of Basrah Governorate - Iraq

اسم المؤلف: احمد عبد برغال الاسدي
اسم المشرف: كوثر هواز مهدي | نادية عبد الامير المظفر
الموضوع العام: علوم الحياة
السنة: 2015
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
اللغة: العربية
مكان الجامعة: البصرة
الصفحات الاولى:
المستخلص: سبعة سلالات من البكتريا الخيطية تعود الى جنسي Streptomyces وNocardiopsis عزلت من ترب شركة مصافي الجنوب الملوثة بالنفط الخام في البصرة جنوب العراق، وقد تم اختبار قدرتها على النمو بوجود النفط الخام باستعمال وسط ملحي سائل مزود بالنفط الخام كمصدر وحيد للكربون. شملت السلالات على S. variabilis 7525 وS. variabilis SW75 وS. cellulosae XFB - T وS. parvus S86 وS. bacillaris S4BW2 وS. flavoviridis VITHM - 1 وN. synnemataformans ITD - 3 بنسبة تشابه من 99٪ الى 100٪. في هذه الدراسة تم العثور على سلالات جديدة من البكتريا الخيطية المعزولة من التربة الملوثة شخصت باستعمال تحديد تتابعات جين الحامض النووي الريبوزي 16S rRNA وقد اظهرت السلالات قدرات مختلفة على تحلل النفط الخام في الوسط الملحي السائل من 50.2٪ الى 72.4٪. كما تم تقييم تاثير الظروف الزرعية من درجة الحرارة والدالة الحامضية وتركيز النفط الخام والعناصر النزرة في نمو سلالات البكتريا الخيطية وتحلل النفط الخام، وقد لوحظ ان اعلى نمو للخلايا واعلى تحلل للنفط الخام كان 80٪ في ظروف زراعة مثالية ( درجة حرارة 30 ᵒم ودالة حامضية 7 بعد 10 ايام من الحضن وزادت هذه النسبة الى 85٪ من قبل السلالة S. cellulosae XFB - T بعد اضافة العناصر النزرة.في حين اظهرت نتائج اختبار التحلل الحيوي للنفط الخام في التربة من قبل سلالات البكتريا الخيطية منفردة بعد 60 يوما من الحضن تحت ظروف مختلفة ، اظهرت ان اعلى قيمة للتحلل كانت 87 ٪ من قبل النوع S. bacillaris عند درجة حرارة 30 ᵒم وحجم لقاح مضاف 10مل/ 25غم تربة ومستوى رطوبة 40٪.المعالجة الاحيائية هي ستراتيجية فعالة لتنظيف المواقع الملوثة بالهيدروكربونات. ونتيجة تاثر الكائنات الحية الدقيقة من تلك التربة يحدث لها اضطراب وبالتالي يتم انتقاء الاحياء المجهرية القادرة على استهلاك الهيدروكربونات لاستخدامها في هذه التقنية الفعالة في التعامل مع الملوثات الهيدروكربونية. الهيدروكربونات التي تتفاعل مع نسجة التربة والكائنات الحية الدقيقة هي التي تحدد مصير الملوثات وطبيعتها الكيميائية والقدرات الهادمة للميكروبات. الاثار المحتملة لنشارة الخشب، وخليط من روث الابقار والغنم وتعديل المكملات الغذائية لتحفيز الكائنات الدقيقة الاصلية وزيادة المعالجة البيولوجية للهيدروكربونات، هذه العملية تم اجراءها باستعمال طريقة المعالجة خارج الموقع بطريقتي الكومة والمفاعل الحيوي، بخلط التربة الملوثة مع نشارة الخشب والروث الحيواني ودعم التربة بالمغذيات والعناصر النزره مع توفير الرطوبة الملائمة والظروف المواتية لنمو الكائنات الحية الدقيقة كما قدمت التهوية الصناعية للتربة في الكومة والمفاعل بوساطة شبكة من الانابيب المثقبة لمحاكاة المعالجة الحيوية التي استمرت مدة 90 يوما.خلال تلك الفترة، رصدت الهيدروكربونات النفطية الكلية (TPHs) وازالة الالكانات الاعتيادية والتغيرات في المجتمعات البكتيرية. وقد اظهرت النتائج ان (TPHs) قد انخفضت من 52غم/كغم الى 10.6 غم / كغم بمعدل تحلل بلغ 79.6٪ ومعدل تحفيز كان 71.4٪ في تجربة الكومة ومن 52غم/كغم الى 13.5 غم / كغم بمعدل تحلل بلغ 74٪ ومعدل تحفيز كان 63.5٪ في تجربة المفاعل الحيوي. وخلال مدة 45 يوما من الاختبار بلغت اعداد الكائنات الحية الدقيقة من البكتيريا المضافة والمستوطنة التي تحلل المواد الهيدروكربونية اعلى مستوى لها 2 × 710 خلية/ غم و1.1 × 710 خلية/ غم في الكومة والمفاعل الحيوي على التوالي.بناء على هذه المعطيات، نستنتج ان المعالجة الحيوية خارج الموقع افضل ستراتيجيه غير مكلفة وفعالة وصديقة للبيئة وبالتالي قد توفر خيارا قابلا للتطبيق لمعالجة التربة من الملوثات الهيدروكربونات النفطية. | Seven actinomycetes strains from genera Streptomyces and Nocardiopsis isolated from hydrocarbon contaminated soil in Basra Governorate, Iraq were selected for their capacity to grow in the presence of crude oil. Their growth rates and biodegradation ability were investigated in mineral salt media supplemented with light Remella crude oil. These strains consist of Streptomyces variabilis 7525, Streptomyces variabilis 5W75, Streptomyces cellulosae XFB - T, Streptomyces parvus S86, Streptomyces bacillaris S4BW2, Streptomyces flavoviridis VITHM - 1 and Nocardiopsis synnemataformans ITD - 3, that the sequence identity range from 99% to 100%. Our study find a new actinomycetes strains isolated from contaminated soil. these strains appears different capacity on the degradation of crude oil in mineral salt media , the highest was found to vary from 50.2% to 72.4%.The effect of the cultivation factors (temperature, pH, and concentration of crude oil and trace elements ) on growth of the actinomycetes strains and crude oil degradation was evaluated. The highest cell growth and the amount of crude oil degraded 80% were observed in optimized cultivation conditions (30oC and initial pH 7) after 10 days, this ratio reached to 85% by addition of trace elements solution at same condition by S. cellulosae XFB - T.The biodegradation test lasted 60 days in soil by actinomycetes strains at different factors shown the highest of the crude oil was removed in experiment that inoculums with 10ml of each bacterial strain, value 87% by Streptomyces bacillaris S4BW2 at 30oC. Bioremediation is an efficient strategy for cleaning up sites contaminated by hydrocarbons. Microorganism of soil is affected by hydrocarbon disturbance thus selective enrichment of hydrocarbon utilizers occurs. The technology is very effective in dealing with petroleum hydrocarbon contamination. Hydrocarbons react with the soil matrix and microorganisms determining the fate of the contaminants relative to their chemical nature and microbial degradative capabilities. The potential effects of sawdust, and mixture of cow and sheep dung as amendment nutrient supplements to biostimulate autochthonous microflora and augmentation for hydrocarbon bioremediation were investigated in test biopile and bioreactor. The soil was ground and fluffed by admixture of 1.5% sawdust, then supplemented with the necessary minerals and watered to provide conditions favoring microorganism growth industrial aeration was provided in pile by system of a abundant perforated drainage - pip network to simulate bioremediation treatments through a 90 - day period. During this period, we monitored total petroleum hydrocarbons (TPHs) and n - alkanes degradation and changes in bacterial communities. The (TPHs) had been reduced from 52 to 10.6 g/kg (79.6 %), the rate of Biostimulants Efficiency (BE) was (71.4%) in biopile and from 52 to 13.5 g/kg (63.5 %) in bioreactor . In soil, the dominant microorganism population comprised Gram - positive bacteria from actinomycete group and autochthonous microorganisms which decompose hydrocarbons reached highest level 2 x 107 and to 1.1 x 107 CFU/g at 45 days in biopile and bioreactor respectively . Based on these data, the study conclude that is ex situ ( Biopile and bioreactor ) experiment the best strategy, inexpensive, efficient, and environmentally friendly and may thus offer a viable choice for petroleum hydrocarbons - contaminated soil remediation.

انتاج وتوصيف لنقي المنتجات الايضية لعزلة Streptomyces ودراسة تاثيراتها المضاد حيوية والسرطانية == Production and Characterization of Purified Metabolites from Streptomyces Isolate and Study

اسم المؤلف: ليندا حميد تركي عطية الغزالي
اسم المشرف: رباب عمران راضي الجيلاوي
الموضوع العام: علوم الحياة
السنة: 2017
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
اللغة: الانكليزية
مكان الجامعة: بابل
الصفحات الاولى:
المستخلص: شملت الدراسة الحالية جمع 100 عينة من الترب بواقع 50 عينة من ترب صحراوية جمعت من المناطق التي تبعد مسافة 3 كيلومترات من بحيرة الرزازة و50 عينة من ترب زراعية جمعت من مناطق مختلفة من محافظه كربلاء المقدسة. حيث عزلت 26 عزلة من البكتريا الخيطية من الترب الصحراوية و36 عزلة من البكتريا الخيطية من الترب الزراعية ودرست الفعالية التضادية لهذه العزلات ضد اربعة انواع من البكتريا المرضية ( بكتريا الموجبة لصبغة كرام Staphylococcus aureus and Streptococcus agalactiae وبكتريا السالبة لصبغة كرام (Escherichia coli and Pseudomonas aeru¬ginosa وبطريقة التخطيط العمودي, واظهرت النتائج ان جميع actinomycetes المعزولة من الترب الزراعية ليست لها اي فعالية مضادة للبكتريا المرضية بينما كانت لبكتريا actinomycetes المعزولة من الترب الصحراوية فعالية مضادة للبكترياالمرضية وبمديات مختلفة تبعا لنوع البكتريا المرضية المدروسة, وقد وجد ان لثمانية من عزلات actinomycetes الصحراوية فعالية مضادة للبكتريا المرضية السالبة والموجبة لصبغة كرام وتبعا لذلك المدى الواسع في التثبيط اختبرت فعاليتها المضادة بطريقة الانتشار بالحفر, واظهرت النتائج ان العزله LHR 9 actinomycetesلها فعالية مضادة للبكتريا المرضية اعلى من العزلات الاخرى المختبرة وباستخدام طريقة الانتشار بالحفر, لذلك اختيرت لاكمال محاور الدراسة المتقدمة. درست الصفات المظهرية والبايوكيمياوية والزرعية للبكتريا actinomycetes LHR 9 بعد زرعها على اوساط زرعية مختلفة من وسط استخلاص Streptomyces الدولي واعتمادا على الفحص المجهري باستخدام المجهر الضوئي المركب وتحت قوى تكبير 1000و400مره لدراسة شكل المايسليوم الهوائي والارضي, وقد تبين ان للعزلة مايسليوم هوائي متفرع له مظهر مفصص وغير مجزا وباستخدام المجهر الاليكتروني الماسح تبين ان المايسليوم يحمل ابواغا ملساء وغير متحركة, وبالاعتماد على مجمل الصفات الزرعية والفسيولوجية المدروسة تبين ان البكترياLHR 9 actinomycetes المعزولة تعود لجنس Streptomyces. درست عدد من الظروف الزرعية والغذائية المثلى لتحسين انتاج مواد الايض الثانوي من البكتريا في وسط التخمر وقد كان وسط فول الصويا الحاوي على الدكستروز كمصدر كاربوني والبيبتون كمصدر نتروجيني وK2HPO4 كمصدر فوسفاتي وبظروف تخمر : بدرجه حراره 35م0 , وحموضه 7 , وقوه اهتزاز 200دوره \ دقيقه ولمده 7 ايام كفتره حضن هو وسط الانتاج الامثل . استخدمت مذيبات عضوية مختلفة القطبيةn) - بروبانول وكلوروفورم وخلات الاثيل ووالاسيتون والميثانول والايثانول (في استخلاص مواد الايض الثانوي المنتجة من بكتريا Streptomyces sp. LHR 9 المضادة للبكترياالمرضية وقد كان المذيب العضوي خلات الاثيل افضل المذيبات العضوية في الاستخلاص اذ اعطى مستخلصه البكتيري اعلى قيم في التثبيط البكتيري . نقيت مواد الايض الثانوي بعد استخلاصها بخلات الاثيل بعمود الفصل : جيل السيليكا ذو الابعاد (1.6× 14) سم وباستخدام الميثانول والكلوروفورم بنسبه 8 : 2 حجم\حجم كمحلول الغسل لسم3 واحد من المستخلص البكتيري , وقد جمع 50 جزء من المحلول الناتج بعد الغسل واختبرت فعاليتهاالتثبيطية لنمو البكتريا المرضية , وقد اعتمدت الاجزاء الفعالة لقياس التركيز المثبط الادنى باستخدام طريقة الانتشار من القرص, وقد كانت التراكيز (4.5 , 5 , 2.5 ,3)مايكروغرام/مل هي التراكيز المثبطه الدنيا لبكتريا Escherichia coli , Pseudomonas aeruginosa, Staphylococcus aurues , Streptococcus agalactiae على التوالي. قيمت السمية الخلوية لمواد الايض الثانوي والمنقاة جزئيا لتحديد مدى فعاليتها المضاده للسرطان وبالاعتماد على فحص MTT (3 - (4, 5 - dimethylthiazol - 2 - yl) - 2 - 5 - diphenyl tetrazolium bromide) وباستخدام خطوط خلايا الثدي السرطانية وخطوط خلايا سليمة, وقد اظهرت النتائج بان التركيز 400مايكوغرام\ مل هو التركيز الامثل لتثبيط نمو الخلايا السرطانية بنسبه 60% مع المحافظة على 80% من الخلايا السليمة. اعتمدت الكروماتوغرافيا السائلة Liquid Chromatography - Mass Spectroscopy (LC - MS) في تنقية المركب الفعال وتوصيفة اذ اظهرت نتائج المخطط اللوني ان هنالك خمسة قمم وبالمقارنة مع الاطياف الكتلية للمركبات حسب المعهد الامريكي للمعايير والتقنية National Institute of Standards and Technology ان للمركب صيغة كيمياوية C18H32O2 ووزن جزيئي 282دلتن . درست الفعالية المضادة للاكسدة والمضادة للاورام لمستخلص Streptomyces sp. LHR 9 (المنقى جزئيا) في اناث الجرذان المختبرية المحفزة للاصابة بسرطان الثدي كيمياويا باستخدام مادة 7,12 dimethyl Benz (A) anthracene (DMBA) واظهرت النتائج ان لمستخلص LHR 9 Streptomyces sp. (المنقى جزئيا) القدره على المحافظة على تركيز الانزيمات المضادة للاكسدة في مصل الحيوانات المعاملة بمادة DMBA وبقيم مقاربة لما هي موجودة في مصل الحيوانات السليمة وقد تمثلت اثار ذلك بصغر وزن وحجم الورم فيى منطقة الثدي والتغيرات النسيجية الملحوظة, وعلى مستوى جزيئ ايضا اذ لوحظ وباستخدام تقنية Polymerase Chain Reaction - Single Strand Conformation Polymorphism(PCR - SSCP) التغيرات الوراثية الحاصلة للجينين ( Bcl - 2 وCaspase - 3) بفعل مادة DMBA ودور مستخلص Streptomyces sp. LHR 9 (المنقى جزئيا) في الحفاظ على الشكل الطبيعي للجينين اذ تبين ان هناك83.66% من الحيوانات السليمة و50% من الحيوانات المعاملة بالمستخلص البكتيري تمتلك جين Caspase - 3 بشكله الطبيعي بينما 66.66% من الحيوانات التي عوملت بماده DMBA فقط تمتلك جين Caspase - 3 من النوع الطافر وكذلك بالنسبة للجين Bcl - 2 اذ تبين ان هناك 78.66% من الحيوانات السليمة و60% من الحيوانات المعالجة بالمستخلص البكتيري تمتلك جين Bcl - 2من النوع الطبيعي بينما لوحظ وجود نفس الجين ولكن من النوع الطافر بنسبة 66.67% في الحيوانات المصابة بسرطان الثدي بتاثير مادة DMBA. | The present study included the collection of 100 soil samples from desert regions away from about 3 km from Al - Razzaza lake and agricultural regions at Karbala Province, Iraq. From these samples 26 and 36 actinomycetes isolates were obtained from desert and agricultural regions respectively . All actinomycetes isolates were tested for their antibacterial activity using primary screening against pathogenic bacteria by perpendicular streaking method .The results were showed that all the actinomycetes isolates were isolated from agricultural regions hadn't any antibacterial activity while the 26 isolates isolated from desert regions were had antibacterial activity in different spectra . Eight actinomycetes isolates from all the 26 desert regions isolates were selected for secondary screening due to their antibacterial activities with wide spectrum against pathogenic bacteria namely Gram positive (Staphylococ¬cus aureus and Streptococcus agalactiae) and Gram negative (Escherichia coli and Pseudomonas aeru¬ginosa) by the agar diffusion methods and the results revealed the selection of actinomycetes LHR 9 isolate for farther studies because it gave us the highest inhibition against the tested pathogenic bacteria growth. The active actinomycetes LHR 9 isolate was identified using several morphological and biochemical methods. The morphological characterization was involved the study of aerial and substrate mycelium on many International Streptomyces Project agar medium and using light microscopy at 1000X and 400X to study the mycelium morphology, as well as using the scanning electron microscope (SEM) to examine the spore surface. The results exhibited that the isolate had the aerial mycelium which formed unfragmented, branched, looped and rectus - flexibilis hyphae type bearing non - motile spores with smooth surface. In addition to results of biochemical tests we could revealed that the isolate was belong to Streptomyces genus . The active isolate Streptomyces sp. LHR 9 was selected for optimization critical cultural and nutritional parameters for increasing the antibiotic production titer in fermentation medium (Soybean broth medium), the results showed the dextrose as carbon source , peptone as nitrogenous source and K2HPO4 as a phosphate source with fermentation cultural condition included ; temperature of 35 °C, pH 7 with agitation rate 200 rpm for 7 days as incubation period gave high values in active secondary metabolites production. The antibacterial compound from fermentation medium was extracted using liquid - liquid extraction method with various organic solvents n - propanol, chloroform, ethyl acetate, acetone, methanol and ethanol) 1 : 1 (v/v) . the results was revealed the ethyl acetate the most efficient in extraction . The Streptomyces sp. LHR 9 crude extract was purified by using silica gel column (1.6 X 14) chromatography. Chloroform and methanol (4 : 1) (v/v), was used as an eluting solvent. The active fractions ( had the antibacterial activity ) were selected for determination the minimum inhibitory concentration and evaluation the cytotoxic activity using MTT (3 - (4, 5 - dimethylthiazol - 2 - yl) - 2 - 5 - diphenyl tetrazolium bromide) assay. The results of the MIC of partially purified Streptomyces sp. LHR 9 extract were 4.5 µg/ml for Escherichia coli, 5 µg/ml for Pseudomonas aeruginosa, 2.5µg/ml for Staphylococcus aurues and 3µg/ml for Streptococcus agalactiae . In vitro screening for cytotoxic activity was done by MTT assay for evaluation the anticancer activity of the partially purified antibacterial compounds produced by Streptomyces sp. LHR 9. The cytotoxic activity of the partially purified Streptomyces sp. LHR 9 extract was determined against established MCF - 7 cell line ( breast cancer cell lines ) and WRL cell as a representative of normal breast cell line, depending on their viability ratio. The viability of MCF - 7 cell line was decreased dramatically with the increasing in extract concentration in the same time the viability of WRL cell was decreased in slowly manner. The using of multiple purification methods Liquid Chromatography - Mass Spectroscopy (LC - MS) along with a sensitive measuring method and possibly isolation of these compounds should allow us to evaluate the entire potential of secondary metabolites production, the results from LC - MS chromatogram of the partially purified Streptomyces sp. LHR 9 extract showed five peaks, on comparison of the mass spectra of the constituents with the National Institute of Standards and Technology( NIST) library the compound was had the Formula : C18H34O2 and molecular weight : 282 D. The partially purified Streptomyces sp. LHR 9 extract was evaluated for its in vivo antioxidant and antitumor activity depending on the ability of 7,12 dimethyl Benz (A)anthracene (DMBA) to inducing breast tumor in female rat model, the results revealed the partially purified Streptomyces sp. LHR 9 extract had the ability to keeping the enzymatic antioxidant in approximately its natural concentrations (as the same concentrations in serum of negative group) in serum of tested rats which were induced breast cancer by DMBA and treated with partially purified Streptomyces sp. LHR 9 extract, so these results were more studied on histological plane and the results of that were achieved by decreasing in weight and size of the breast tumor of tested rats and other histological change in breast tissue. In addition to that partially purified Streptomyces sp. LHR 9 extract effect was more studied on molecular plane depending on the Bcl - 2 and Caspase - 3 genes mutation using Polymerase Chain Reaction - Single Strand Conformation Polymorphism ( PCR - SSCP) analysis. The results were two conformations polymorphism in both Caspase - 3 gene ( about 83.66 % of negative group and 50% of treated group had three bands (wild type) where's positive control had 66.66% four bands (mutant type)) and Bcl2 gene (the wild type had one band such as negative group was 78.66% and test group 60%, compared with positive group (60.67%) have two bands(mutant type)). so these results suggest the active apoptotic property of Caspase - 3 and Bcl - 2 and the role of partially purified Streptomyces sp. LHR 9 extract effect in keeping them in their wild type.

دراسة فسلجية وجزيئية للتغاير الجيني لمستقبلات فيتامين د في النساء المصابات بمرض السكري من النوع الثاني == Physiological and Molecular Study of Vitamin D Receptor Gene Polymorphism in Type II Diabetic Women

اسم المؤلف: نورا ماجد حميد ابو خمرة
اسم المشرف: حيدر كامل زيدان السعدي | محمد عبد الله جبر
الموضوع العام: علوم الحياة
السنة: 2017
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
اللغة: العربية
مكان الجامعة: بابل
الصفحات الاولى:
المستخلص: هدفت هذه الدراسة الى تقييم دور تعدد اشكال مستقبلات فيتامين (د) وارتباطها بتطور مرض السكري وتاثيرها في الاناث قبل وبعد سن الياس. جينات مستقبلات فيتامين (د)، تتضمن (Apa - I, Taq - I, Fok - I وCdx2) و(جين الفاEstrogen receptor - ) التي حددت بواسطة تفاعل سلسلة البوليميريز - تباين اطوال قطع التقييد وتقنية تفاعل سلسلة البوليميريز المتعدد المتخصص في الاليل, في حين ان الهرمونات والتي تتضمن (الاستراديول والانسولين) والمؤشرات الفسيولوجية (فيتامين د، الكولسترول، البروتين الدهني عالي الكثافة، الدهون الثلاثية، البروتين الدهني منخفض الكثافة، البروتين الدهني منخفض الكثافة جدا، الهيموجلوبين A1C، الجلوكوز الصيامي في الدم، مقاومة الانسولين، الحساسية للانسولين ، ضغط الدم الانقباضي وضغط الدم الانبساطي). اجريت هذه الدراسة في الفترة من اذار 2016 الى تشرين الاول 2016، حيث تم جمع العينات من مستشفى الصدر التعليمي في محافظة النجف. وقد بلغ عدد العينات (80) مريضا مصابين بداء السكري من النوع الثاني بمتوسط عمر (36 - 65 سنة ، وقسمت العينات الى مرحلة ما قبل انقطاع الطمث وبعد انقطاع الطمث, كذلك قسمت حسب مدة المرض وحسب التاريخ العائلي، ومؤشر كتلة الجسم، ومضاعفات المرض (ضغط الدم المرتفع). كما تضمنت الدراسة 40 شخصا اصحاء على ما يبدو بمتوسط عمر (36 - 65 سنة)، حيث ان مجموعة السيطرة تقارن مع مجموعة المرضى. قسمت الدراسة الحالية الى جزئين رئيسيين : الاجزاء الفسيولوجية والجزيئية. واظهرت النتائج ان المرضى الذين يعانون من مرض السكري من النوع الثاني، كان لديهم معنويا (p ≤ 0.05) مستوى اعلى من الكوليسترول ، الدهون الثلاثية، البروتين الدهني منخفض الكثافة، البروتين الدهني منخفض الكثافة جدا، الهيموغلوبين A1C، الانسولين، مقاومة الانسولين ونسبة السكر الصيامي في الدم مقارنة بمجموعة السيطرة، في حين ان معدلات الحساسية للانسولين تنخفض معنويا (P≥0.05) في مرضى السكري من النوع الثاني عند مقارنتهم بمجموعة السيطرة. كان معدل الاستراديول معنويا ( ≤ 0.05) اعلى في الاناث قبل انقطاع الطمث في حين انخفض في الاناث بعد سن الياس لكل من مجموعة السيطرة ومرضى السكري من النوع الثاني. واظهر الضغط الانقباضي والانبساطي فرقا معنويا بين المرضى الذين يعانون من السكري النوع 2 ومجموعة السيطرة. وفقا للمقارنة بين النساء قبل انقطاع الطمث وبعد سن الياس في كل من مرضى السكري ومجموعة السيطرة، اظهرت النتائج ارتفاع معنوي (p ≤ 0.05) في معدلات مستوى هرمون الاستراديول، ومعدلات مستوى هرمون الانسولين ومقاومة الانسولين في كلتا المجموعتين. من ناحية اخرى، اظهرت المقارنة بين المرضى قبل انقطاع الطمث الذين يعانون من السكري ومرحلة ما قبل انقطاع الطمث ارتفاع معنوي (p ≤ 0.05) في معدل مستويات الكوليسترول، البروتين الدهني منخفض الكثافة، هيموغلوبين A1c معدل مستويات (HBA1c)، الجلوكوز الصيامي في الدم , ومستويات هرمون الاستراديول، ومعدلات مستوى هرمون الانسولين، ومقاومة الانسولين، وحساسية الانسولين وضغط الدم الانبساطي. وفقا لمقارنة مرضى السكري بعد انقطاع الطمث ومجموعة السيطرة بعد سن الياس، كانت هناك اختلافات معنوية (p ≤ 0.05) في معدل مستويات الكوليسترول، البروتين الدهني منخفض الكثافة، معدل هيموغلوبين A1c، الجلوكوز الصيامي في الدم , هرمون الاستراديول ومعدلات مستوى هرمون الانسولين، ومقاومة الانسولين وحساسيةالانسولين. اظهرت نتائج تحليل الارتباط وجود ارتباط معنوي (p ≤ 0.05) بين فيتامين (د) مع الدهون الثلاثية والبروتين الدهني منخفض الكثافة جدا في النوع الثاني من مرضى السكري. في مرضى السكري قبل انقطاع الطمث، اظهرت نتائج تحليل الارتباط وجود علاقة عكسية معنويه بين فيتامين (د) مع كل من الدهون الثلاثية والبروتين الدهني منخفض الكثافة جدا في حين تم العثور على علاقة ايجابية معنوية بين فيتامين (د) والبروتين الدهني عالي الكثافة في مرحلة ما بعد انقطاع الطمث في مرضى السكري. في مجموعة السيطرة بعد انقطاع الطمث تم العثور على علاقة عكسية معنويه بين فيتامين (د) ومستوى السكر الصيامي في الدم. اظهر تحليل الارتباط وجود علاقة معنوية عكسية بين معدلات مستوى الكالسيوم وضغط الدم الانقباضي في مجموعة السيطرة، وبينما في مرضى السكري قبل انقطاع الطمث، اظهرت نتائج تحليل الارتباط وجود علاقة ارتباط معنوية بين الكالسيوم ومؤشر كتلة الجسم، بينما وجدت علاقة معنوية عكسية بين الكالسيوم وكلا من البروتين الدهني عالي الكثافة والعمر بالاضافة الى ارتباط ايجابي معنوي مع الاستراديول في المرضى بعد سن الياس. من ناحية اخرى، في مجموعة السيطرة قبل انقطاع الطمث، تم العثور على ارتباط عكسي معنوي بين الكالسيوم وضغط الدم الانقباضي، في حين وجود علاقة طردية معنوية مع هرمون الانسولين ومقاومة الانسولين. في حين لوحظ وجود علاقة عكسية معنوية في مجموعة السيطرة بعد سن الياس بين الكالسيوم ومستوى السكر الصيامي في الدم ومؤشر كتلة الجسم. هنالك ارتباط معنوي كبير بين هرمون الانسولين مع البروتين الدهني عالي الكثافة والعمر وحساسية الانسولين، في حين كان هناك علاقة طردية معنوية بين الانسولين مع مؤشر كتلة الجسم ومقاومة الانسولين في النوع الثاني من مرضى السكري. في مجموعة السيطرة كانت هناك علاقة ارتباط معنوية طردية بين الانسولين ومقاومة الانسولين، ارتباط معنوي عكسية مع حساسية الانسولين. في كل من مرضى السكري قبل انقطاع الطمث وبعد انقطاع الطمث ومجموعة السيطرة كانت هناك علاقة سلبية كبيرة جدا بين الانسولين وحساسية الانسولين، في حين وجود علاقة طردية معنوية مع مقاومة الانسولين. في مرضى السكري قبل انقطاع الطمث، كانت هناك علاقة ايجابية كبيرة بين الانسولين والدهون الثلاثية , البروتين الدهني منخفض الكثافة جدا ومؤشر كتلة الجسم. في مرحلة ما بعد انقطاع الطمث مرضى السكري، كانت هناك علاقة سلبية كبيرة بين الانسولين والهيموغلوبين .A1c ايضا هنالك علاقة طردية معنوية كبيرة بين الهيموغلوبين A1c والسكر الصيامي في الدم في مجموعة السكري. في مجموعة السيطرة كان هناك ارتباط طردي كبير بين الهيموغلوبين A1c والسكر الصيامي في الدم ,وعلاقة عكسية معنوية مع الكولسترول وحساسية الانسولين. وفقا لمجموعة السيطرة قبل انقطاع الطمث، كان هناك ارتباط ايجابي كبير بين الهيموغلوبين A1c والجلوكوز الصيامي في الدم، في حين وجود علاقة عكسية كبيرة مع الكولسترول، الدهون الثلاثية، البروتين الدهني منخفض الكثافة جدا وحساسية الانسولين. من ناحية اخرى، في مجموعات مرضى السكري قبل انقطاع الطمث وبعد انقطاع الطمث وجدت علاقة طردية معنوية بين الهيموغلوبين A1c والسكر الصيامي في الدم , وارتباط ايجابي كبير مع الدهون الثلاثية والبروتين الدهني منخفض الكثافة جدا في مجموعة ما قبل انقطاع الطمث فقط. اظهرت نتائج التحليل الوراثي لتباين النوكليوتيدات المفردة، للجينات المستقبلة لفيتامين د باستخدام تقنية تفاعل سلسلة البوليميريز - تباين اطوال قطع التقييد ان هناك فرقا غير معنوي في الاشكال الجينية (Apa - I, Fok - I, Taq I - ) و(Estrogen receptor - alpha ) بين المرضى ومجموعة السيطرة باستخدام نفس التقنية اعلاه. بينما وجد فرق معنوي بين المرضى ومجموعة السيطرة وفقا للتغاير الجيني في الجين (CdxII)وفقا الى الجين (Apa - I)، كان النمط الوراثي المتماثل (AA) اكثر تواترا في المرضى الذين يعانون من داء السكري من النوع 2 (41٪) من مجموعة السيطرة ، في حين ان النمط الوراثي المتغاير (CA) اكثر تكرارا في مجموعة السيطرة من المرضى. نتائج الجين (Taq - I)، كان النمط الوراثي المتماثل (TT) اكثر تواترا في مجموعة السيطرة من المرضى الذين يعانون من السكري من النوع 2 ، في حين ان النمط الوراثي المتغاير (TC) كان اكثر تكرارا في المرضى من مجموعة السيطرة. وفقا الى الجين (Fok - I)، كان النمط المتماثل (CC) اكثر تواترا في مجموعة السيطرة من المرضى الذين يعانون من داء السكري من النوع 2 ، في حين ان النمط المتغاير (CT) اكثر تكرارا في السيطرة من المرضى ، والنمط المتماثل (TT) كان اكثر تواترا في المرضى من مجموعة السيطرة بينما الجين (ER_alpha)، كان النمط المتماثل (AA) اكثر تواترا في المرضى الذين يعانون من داء السكري من النوع 2 من مجموعة السيطرة ، في حين ان النمط المتغاير (AG) اكثر تواترا في السيطرة من المرضى ، والنمط المتماثل (GG) كان اكثر تكرارا في المرضى من مجموعة السيطرة | This study aimed to evaluate vitamin D receptor gene polymorphism role and its correlation with the development of diabetes disease and its effect in pre and post menopausal females. Vitamin D receptor gene including ( APA - I, FOk - I, Taq - I and CdxII) and (Estrogen receptor alpha gene) determined by Polymerase Chain Reaction (PCR) , restriction fragment length polymorphism (PCR - RFLP) and Allele specific multiple (ASM - PCR), while the hormones including (Estradiol and Insulin) and the physiological parameters ( Vitamin D, Cholesterol, High density lipoprotein, Triglyceride, Low density lipoprotein, very low density lipoprotein, Hemoglobin A1c, Fasting Blood Glucose, insulin resistance, insulin sensitivity, systolic and diastolic blood pressure). This case - control study was done in a period of March 2016 to October 2016 ,where the samples collected from Al - Sadr Teaching Hospital in Najaf Province. The number of samples was (80) females with type 2 diabetes with an average age (36 - 65 year), all of them were with type 2 diabetes are divided to pre menopause and post menopause, also divided according to duration of disease and according to age groups, family history, body mass index and complications with disease (hypertensive blood pressure). Also, the study included 40 apparently healthy people with an average age ( 36 - 65 year) , as control matched with disease group. The present study divided into two main parts : physiological and molecular parts . The results showed that patients with type 2 diabetes , had significantly( p ≤ 0.05 ) higher cholesterol, Triglyceride, Low density lipoprotein, very low density lipoprotein, Hemoglobin A1c, insulin, insulin resistance and Fasting blood glucose levels rate than control group, while the levels rate of insulin sensitivity is decreased significantly (p ≤ 0.05) in type 2 diabetic patients when compared with control group . The Estradiol levels rate was significantly (p ≤ 0.05) higher in premenopausal females while it decreased in postmenopausal females of both control and type 2 diabetic patients. Systolic and diastolic pressure showed significant difference between patients with type 2 diabetic and control. According to the comparison of premenopausal and postmenopausal women in both diabetic patients and in control group, the results showed significant ( p ≤ 0.05) elevation in Estradiol hormone level rates, Insulin hormone level rates and insulin resistance in both groups. On the other , the comparison of premenopausal patients with diabetic and premenopausal control groups the statistical analysis showed significant elevation (p ≤ 0.05) in cholesterol levels rate, Low density lipoprotein (LDL), HemoglobinA1c levels rate (HBA1c), fasting blood glucose (FBG), Estradiol hormone level rates, Insulin hormone level rates, insulin resistance ,insulin sensitivity and diastolic blood pressure. The results of correlation analysis in pre menopausal diabetic patients showed there are significant negative correlation between vitamin D with both Triglyceride (TGC) and very low density lipoprotein (VLDL) while significant positive correlation has been found between vitamin D and high density lipoprotein (HDL) in post menopausal diabetic patients. In post menopausal control group a significant negative correlation has been found between vitamin D and fasting blood glucose (FBG) level.In pre menopausal diabetic patients, the results of correlation analysis showed there were significant positive correlation between calcium and BMI, while significant negative correlation has been found between calcium and both high density lipoprotein (HDL) and age in addition to significant positive correlation with Estradiol in post menopausal diabetic patients.On the other hand, In pre menopausal control group, a significant negative correlation has been found between calcium and systolic blood pressure, while a significant positive correlation with insulin hormone and insulin resistance. While, in post menopausal control group a significant negative correlation has been found between calcium and fasting blood glucose (FBG) level and body mass index. In both premenopausal and postmenopausal diabetic patients and control group there were highly significant negative correlation between Insulin and insulin sensitivity, while highly significant positive correlation with insulin resistance. In pre menopausal diabetic patients, there were significant positive correlation between insulin and Triglyceride (TGC), very low density lipoprotein (VLDL) and BMI. In post menopausal diabetic patients, there were significant negative correlation between insulin and HBA1c.On the other hand, in premenopausal and postmenopausal diabetic patients groups there were highly significant positive correlation between hemoglobinA1c (HBA1c) and fasting blood glucose (FBG), and significant positive correlation with TGC and VLDL in premenopausal group only. The genetic analysis of the Single Nucleotide Polymorphisms , for vitamin D receptor gene by using RFLP - PCR technique showed that there non - significant difference in the genotype polymorphisms of (APA - I, FOk - I and Taq - I) and (Estrogen receptor - alpha gene) between patients and control by using the same technique above, While there were significant differences according to (Cdx - II gene) by using (ASM - PCR) technique. According to (Apa - I gene), the homozygote pattern (AA) were more frequent in patients with type 2 diabetes than control group, While the heterozygote (CA) pattern more frequent in control than patients.The results of (Taq - I gene), the homozygote pattern (TT) were more frequent in control than patients with type 2 diabetes group, While the heterozygote (TC) pattern more frequent in patients than control. According to (Fok - I gene), the homozygote pattern (CC) were more frequent in control group than patients with type 2 diabetes, While the heterozygote (CT) pattern more frequent in control than patients, and the homozygote (TT) pattern was more frequent in patients than control group.While the (ER_alpha gene), the homozygote pattern (AA) were more frequent in patients with type 2 diabetes than control group, While the heterozygote (AG) pattern more frequent in control than patients, and the homozygote (GG) pattern was more frequent in patients than control group

التباين الوراثي في جينات TNF - ?, VEGF - A and IL - 10 المصاحبة لداء الصدفية == Genetic Polymorphism in TNF - ?, VEGF - A and IL - 10 genes associated with Psoriaasis

اسم المؤلف: خالد عبد الكاظم هادي الطائي
اسم المشرف: علي حمود السعدي | وسام علي امين عوض
الموضوع العام: علوم الحياة
السنة: 2017
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
اللغة: الانكليزية
مكان الجامعة: بابل
الصفحات الاولى:
المستخلص: هدفت هذه الدراسة الى تقصي بعض التغايرات الوراثية لكل من (TNF - α, VEFGF and IL10) لدى مرضى داء الصدفية وعلاقتها بشدة الاصابة باستخدام تقنيات تفاعل البلمرة المتسلسل وتباين اطوال قطع التقيد . بينما قدر مستوى الحركيات المناعية باستخدام تقنية الايلايزا.اجريت هذه الدراسة في الفترة من نيسان 2016 الى اذار 2017، حيث جمعت العينات مرضى داء الصدفية المراجعين لمستشفى مدينة مرجان الطبية، محافظة بابل / العراق. شملت الدراسة 64 عينة لمرضى داء الصدفية 40) ذكر و24 انثى) يبلغ متوسط اعمارهم (9 - 70 سنة)، كما شملت الدراسة 38 شخصا من الاصحاء (21 ذكر و17 انثى) يبلغ متوسط اعمارهم 15 - 65 سنة كمجموعة السيطرة جارت مجاميع المرضى.اظهرت النتائج زيادة معنوية في مستوى كل من TNF - α وIL10 في المصل لدى مرضى داء الصدفية مقارنة بمجموعة السيطرة (P<0.01) . كما اظهرت الدراسة ارتباط معنوي بين مستوى TNF - α وشدة الاصابة ( - r=0.49, p=0.0001) بينما كان معامل الانحدار عكسي غير معنوي بين مستوى IL10 في المصل وشدة الاصابة ( - r= 0.29, P=0.0109) , وكان الانحدار عكسي معنوي بين مستوى IL10 و( - r= - 0.29) (p=0.0028) TNF - α. كما اظهرت النتائج ارتفاعا معنويا (P<0.01) في مستوى VEGF في المصل لدى مرضى داء الصدفية مقارنة بمجموعة السيطرة بينما كان الانحدارغير معنويا بين مستوى VEGF وشدة الاصابة( - r= - 0.05) .اظهرت النتائج وجود ثلاث انماط وراثية للتغايرات في موقع الجين TNF - α - 308 حيث كانت النسب AA 5 (7.81%) ,AG 24 (37.50% وGG 35 (54.69% في مجموعة السيطرة كانت 10 (26.31%) , 16 (42.11% و12 (31.58% اما توزيع الانماط الوراثية الناتجة من تحليل جين VEGF+405 كان النمط الوراثي CG هو السائد 24 (37.05) لدى مرضى الصدفية والنمط الوراثي CC هو الاقل ظهورا 1 (1.56%).. اضافة الى ذلك اظهر نتائج تحديد تتابع النيكليوتيدات للموقع الوراثي IL10 - 1082 نمطين وراثين حيث كان النمط الوراثي GG هو الاكثر تواجدا لدى مرضى داء الصدفية ( 47.5%) والنمط GA بنسبة (28.5%) مقارنة بمجموعة السيطرة التي جاءت جميعها بالنمط الوراثي GG. | Psoriasis is an immune mediated skin disease with a wide - world prevalence involves both genders and different age, it is a multifactorial disease influenced by genetic and environmental factors. The current study aims to evaluate the association of (Interleukin 10, Tumor Necrosis Factor - α and Vascular Endothelial Growth Factor) genes polymorphism in psoriasis. Single Nucleotide Polymorphism (SNPs) determined by Polymerase Chain Reaction (PCR), restriction fragment length polymorphism (PCR - RFLP) and DNA sequencing techniques, while there serum levels determined by ELISA. This case - control study is done in a period of March 2015 to December 2017, where the samples collected from Marjan Medical City in Babylon Province. The number of samples was (64) patients psoriasis (40 males and 24 females ) with an average age ( 9 - 70 year), (50) of them were with type 1 psoriasis and (14) type 2. In addition, the study included 38 apparently healthy people who were (21 males and 17 females) with an average age (15 - 65 year), as control matched with disease group. The present study revealed a significant increasing (P<0.01) of Tumor Necrosis Factor - α (TNF - α) and Interleukin 10 (IL10) in patients compare with controls. In addition, there is a significantly regression between severity of disease and TNF - α serum level (r =0.49, p=0.0001) and non - significant invers regression with IL10 serum level (r= 0.29, P=0.0109). In addition, the results established an A significant negative correlation which has been found between IL10 and TNF - α (r= - 0.29) (p=0.0028). Physiological, the study shows significant increasing in Vascular Endothelial Growth Factor (VEGF) level in patient compare with control (P<0.01). There is no correlation between Vascular Endothelial Growth Factor (VEGF) serum level and severity or onest of disease (r= - 0.05).Genetically, analysis of the Single Nucleotide Polymorphisms PCR - RFLP genotyping for TNF - α - 308 G/A (rs1800629) SNP, Genotype distribution in patient were GG 35 (54.69%), AG 24 (37.50%) and AA 5 (7.81%) while in control were AG16 (42.11%), GG 12 (31.58%) and AA10 (26.31%).The genetic analysis of the Single Nucleotide Polymorphisms of VEGF +405 SNP, genotype distribution CC homozygous was predominant in patient 39(60.94%) and GG homozygous was 24 (37.05) While the GCwas the least 1 (1.56%). Comparing with the control group was CG 18(47.37%), CC 13(34.21%) and GG 7(18.42%).On the other hand , the statistical analysis for the role of genotyping for TNF - α - 308 SNP in the Serum levels of TNF - α were different between TNF - 308 AA (126.90 ± 14.96) patients and TNF - α GA(106.55 ± 16.42) and GG (57.94 ± 7.72) genotypes (p < 0.01). Analysis for the role of genotyping for VEGF +405 SNP non - significant difference between homozygous CC (408.30 ± 0.00), homozygous was GG (326.00 ± 34.81) and heterozygous CG (376.41 ± 58.63).This result promotes that the both of TNF - α - 308 SNP polymorphism and VEGF +405 SNP strongly implicated in the pathogenicity, severity and onest of psoriasis.PCR - sequencing for IL10 - 1082 A/G (rs1800896) locus shows a significant difference of percentage distributions of alleles and genotypes for IL10 - 1082 (rs1800896) locus between patient and control, GG homozygous was predominant in patient (71.5%) while the AG heterozygous was (28.5 %). Comparing with control group GG (100%).

تقدير الاسبارجين في مرضى سرطان الدم بوساطة الاسبارجينيز المقيد والمنقى من Portulaca oleracea L == Estimation of Asparagine in Serum Leukemia Patients by Immobilized L - asparaginase Purified from Portulaca oleracea L

اسم المؤلف: هادي ساجد عبد العباس عبد الله
اسم المشرف: محمد عبد الله جبر جاسم
الموضوع العام: علوم الحياة
السنة: 2017
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: بابل
الصفحات الاولى:
المستخلص: جمعت عينات النباتات قيد الدراسة من منطقة الرارنجية \مدينة الحلة\العراق. وصنفت النباتات من قبل د.نداء عدنان في قسم علوم الحياة \كلية العلوم \جامعة بابل بالاعتماد على الصفات المظهرية للنباتات . تم التحري عن فعالية انزيم الاسباراجينيز والمحتوى البروتيني في المستخلص الخام لهذه النباتات , تبين ان اعلى فعالية نوعية لانزيم الاسباراجينيز في نبات البربينة Portulaca oleracea L., ثم قدرت الفعالية الانزيمية في اجزاء النبات والتي شملت (الاواق, السيقان,الجذور, نبات كامل) حيث ظهرت اعلى فعالية في النبات الكامل مقارنة بالاجزاء الاخرى ,بالاعتماد على هذه النتائج استخدم نبات البربينة Portulaca oleracea L. كمصدر لانتاج وتنقية وتوصيف والاستخدام الطبي لانزيم الاسباراجينيز.درست الظروف المثلى لفعالية انزيم الاسباراجينيز من النبات الكامل وقد اشارت النتائج الى ان اعلى فعالية للانزيم قد تحققت بحضن الانزيم الخام مع الاسبراجين بتركيز 200mM بنسبة 1 : 3 في محلول دارئ فوسفات البوتاسيوم المنظم بتركيز 0.05 مولار بدالة حامضية 8 وبدرجة حرارة 37 م مع الانخفاض التدريجي في الفعالية النوعية للانزيم الخام بعد خزنه لفترات زمنية محددة .تم تنقية انزيم الاسباراجينيز الخام المستخلص من نبات البربينة بثلاث خطوات تضمنت الاولى الترسيب بكبريتات الامونيوم بنسبة 90% ثم التنقية بكروماتوغرافيا الترشيح الهلامي باستحدام السيفاديكس G - 100 وبعدها التنقية بكروماتوغرافيا الترشيح الهلامي باستخدام السيفاديكس G - 150 وقد ظهرت ثلاثة اشكال للانزيم حيث بلغت الفعالية النوعية للانزيم النقي 7440 وحدة\ملغم وعدد مرات التنقية 4.08 وبحصيلة انزيمية بلغت 22.7%.تم توصيف الاسباراجينيز النقي ,اذ بلغ الوزن الجزيئي52±1 كيلو دالتون باستخدام كروماتوغرافيا الترشيح الهلامي باستخدام السيفاديكس G - 100 بوجود البروتينات القياسية والتي هي Trypsin (23 KD), Pepsin (34.5 KD), Egg Albumin (43KD) , and Bovine serum albumin (67 KD) وكان الرقم الهيدروجيني الامثل لفعالية وثبات الانزيم هو 8 في حين كانت درجة الحرارة المثلى لفعالية وثبات الانزيم هي 37 م وعند دراسة الثوابت الحركية للانزيم , ظهر ان معدلات قيم ثابت ميكالس (Km) والسرعة القصوى (Vmax) وعدد التحول(Kcat ) كانت 250 ملي مولار و100 ملي مولار \دقيقة و350 1\ثانية على التوالي. قيد الانزيم باستعمال الاكار,الاكاروز والجيلاتين وتم توصيف الانزيم المقيد ومقارنته مع الانزيم الحر. اظهرت نتائج الدراسة كفاءة التقييد باستعمال مادة الاكاروز والاكار والجلاتين حيث بلغت ( (70,72,78 % على التوالي وكما تبين من النتائج ان اعلى فعالية للانزيم الحر والمقيد بالاكاروز كانت عند الدالة الحامضية 8 اما الانزيم المقيد بالاكار والجيلاتين فكانت اعلى فعالية عند الدالة الحامضية 9. بالمقابل كانت درجة الحرارة 37 م هي المثلى للفعالية للانزيم المقيد بالاكار والجيلاتين والحر و40 م للانزيم المقيد بالاكاروز. واستخدم الانزيم المقيد في تقدير تركيز الاسبارجين في مصل الدم , اظهرت النتاج ارتفاع واضح في تركيز الاسباراجين لمرضى اللوكيميا (سرطان الدم) حيث بلغت〖10〗^( - 2) - 〖10〗^( - 3) مولار مقارنة مع الاشخاص الطبيعيين حيث بلغت〖10〗^( - 4) - 〖10〗^( - 7) مولار | Plants samples were collected during 2016 (August to September) from Al - Raranjia Hamlet field / Al - Hilah city / Iraq. These samples were classified by Dr. Nidaa Adnan Muhammed ,Biology Department/ Collage of Science / University of Babylon , according to their morphological characteristics. Activity of asparaginase was detected in whole plant, leaves, steems and roots extracts. Results showed that Portulaca oleracea L. was the best one from group of tested plants for research due high yield of L - asparaginase enzyme . Maximum asparaginase activity was detected in whole plant extracts which was 43.55 U/ml in comparison with 37.74,33.16 and 36.8 U/ml in extracts of leaves, steems and roots respectively. According to these results whole plant of Portulaca oleracea L. were used as a source for asparaginase production, purification , characterization, and medical application . Activity of crude asparaginase extracted from whole plant were under optimal conditions studied. Results revealed the maximum activity of asparaginase was achieved when the enzyme was incubated with asparagine 200mM at 37°C in the presence of potassium phosphate buffer 0.05 M solution/plant sample (g) (3 : 1 ratio) at pH8 , with gradually increase in activity with increase of the substrate and gradually activity decrease after particular storage period. Crude asparaginase extract was purified in three steps ( Ammonium sulfate precipitation (90%), gel filtration chromatography technique by Sephadex G - 100 and gel filtration chromatography method by Sephadex G - 150. There are three isoform of enzyme while Specific activity of the purified asparaginase was 7440 U/mg and 4.08 Purification folds, 14.5 % yield. Characterization of the purified asparaginase showed that the molecular weight of the enzyme was 52±1 Kilo Dalton by gel filtration chromatography using Sephadex G - 100 in the presence of four standard proteins which they are Trypsin (23 KD), Pepsin (34.5 KD), Egg Albumin (43KD) , and Bovine serum albumin (67 KD). pH 8 was the optimum pH for enzyme activity and stability, while 37°C was the optimum temperature for both enzyme activity and stability, with Km of 250 mM toward L - asparagine as substrate and the maximum velocity (Vmax) and Kcat of 100 mM/min, 350 sec^( - 1) respectively . The activity of immobilized enzyme was determined and compared with free enzyme ,where immobilizing Efficiency of agar , agarose and gelatin techniques was ( 72 %,78 % and 70 % ) respectively, all immobilizing technique proved that the activity of the enzyme was increased significantly compared with free enzyme, where agar and gelatin methods showed more stability of enzyme at alkaline pH (pH 9) , whereas only agarose method get the enzyme more heat stability to enzyme than other immobilizing methods. There are significant elevation level of asparagine concentration in leukemic human serum samples than normal human serum samples , where reached to 〖10〗^( - 4) - 〖10〗^( - 7) M of asparagine in normal human serum samples while it was 〖10〗^( - 2) - 〖10〗^( - 3) M of asparagine in leukemic serum blood samples.

تقييم دور الجين في دم نساء عراقيات مصابات بسرطان الثدي == Evaluation The Rule of Mammoglobin A Gene In Blood of Iraqi Women With Breast Cancer

اسم المؤلف: نانسي فيصل رشيد
اسم المشرف: امنة نعمة الثويني
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: Mammaglobin A is a member of the superfamily of secretoglobins. Its expression is highly specific for mammary tissue and has been shown to be overexpressed in breast tumor tissue, indicating that mammaglobin A might confer a growth advantage to mammaglobin A - expressing tumor cells. The present study was undertaken to develop the mammaglobin A as a serum biomarker for the early detection of breast cancer, and breast cancer patients follow up after treatments. This study had taken ten months, it involved 84 patients with age ranged 15 - 74 years who had diagnosed with either stage I to III breast cancer or benign lesions of breast at certain Iraqi hospitals and breast cancer centers, and a total number of 10 apparently healthy individuals, with comparable age range of patients as control group.Ten ml of venous blood was collected from studied groups in order to be used for measuring estrogen and progesterone levels, Reverse transcription polymerase chain reaction (RT - PCR) test for detection of their serum m - RNA mammaglobin A existence, and serum mammaglobin A levels using Enzyme - Linked ImmunoSorbent Assay (ELISA) technique. Demographic study revealed that most cases enrolled in this study were women within their reproductive age 25 - 44 years old. Distribution of patients according to age group, marital status, and lactation status showed no statistically significant association (P>0.05) with any of these variables, in all comparison.The hormonal study states that elevated levels of estrogen were observed in 52 (62%) patients, also decrease in progesterone levels were observed in 50 (59%) patients out of the 84 cases studied. This indicate a highly significant SUMMARYassociation (p < 0.01) between breast cancer risk, elevated estrogen levels, and decrease in progesterone levels. Mammaglobin A protein was detectable by ELISA in 100% of breast cancer patients (65 case), and not in any benign tumor patients (19 case) nor healthycontrol women (10 females) which indicate the highly specificity of ELISA test (P<0.01) for measuring mammaglobin A levels in patients serum. With high concentration of protein in sera of women with late stage of breastcancer compared with a low protein concentration in sera of women with early stage of breast cancer. By applying a statistical analysis on the result it was found that serum mammaglobin A ELISA test is highly significant (P<0.01) for, differentiation between patients with metastatic breast cancer and patients with early diagnosed breast cancer. In this work, SCGB2A2 gene which coded for mammaglobin A was detected in 64 out of 65 breast cancer cases, but not in the benign or healthy individuals indicating its high specificity as a marker gene (SCGB2A2) for cells derived from mammary glands. A statistical analysis was made for comparison between the three groups, E - Malignant Tumors \ Early Stage, L - Malignant Tumors \ Late Stage, and B - benign, (E, L, and B) which show that the results of molecular study of mammaglobin A detected by RT - PCR is statistically high significance (P<0.001) when used for the differentiation between E group and B group, also between L group and B group. The results showed that mammaglobin A can become an important tool for detecting primary and metastatic breast cancer, and predicting disease outcome.

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

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

التحري عن التشوهات الكروموسومية وطفرات المورث FLT3/ITD في مرضى ابيضاض الدم النخاعي الحاد == Screening For Chromosomal Aberrations And Gene Mutations FLT3/ITD In Patients With Acute Myeloid Leukemia

اسم المؤلف: سمارة كاظم محمد
اسم المشرف: عبد الحسين مويت الفيصل
الموضوع العام: علوم الحياة
السنة: 2015
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: صممت الدراسة الحالية من اجل تسليط الضوء على تاثيرات مرض ابيضاض الدم ((AMLفي بعض معايير الوراثة الخلوية والجزيئية الحاصلة في كريات الدم البيضاء في ثلاثة حالات للمرضى اشتملت على مرحلة ما قبل العلاج, في اثناء العلاج وبعد العلاج الكيميائي. كما شملت الدراسة ا | The present study was designed to shed light on the cytogenetic and molecular effects caused by acute myeloid leukemia (AML) pathogenesis in three stages before, during and after treatment with chemotherapy (in vitro) in lymphocytes. It was also aimed to investigate chromosomal aberrations, micronucleus formation and FLT3/ITD point mutations in CN - AML region 100 - 300 bp compared with healthy control. The study was comprised of forty - seven AML Iraqi patients their ages ranged between 2.5 - 81 years. It included twenty females and twenty seven males compared with twenty - six healthy control. The following results were obtained : AML is most common leukemia in adults and uncommon in children. It was found that 46.8% patients suffer from AML, who were under 15 years old, while 19.15% patients their age ranged between 16 - 30 years; 19.15% of patients their age were more than 45 years and 14.9% of patients their age ranged between 31 - 45 years of the total AML patients. AML is more common in males than females. The percentages of females and males were 42.5% and 57.5% respectively representing 1.35 : 1.00 male : female ratio. Two AML patients 5.3% have diploid cells when examined under light microscope. The highest mitotic index was 7.498±1.7 occurred in patients before taking treatment while recorded 6.784±4.5 during treatment and 7.000±2.5 after treatment. Nuclear anomalies such as nucleoplasmic bridges and nuclear buds were observed in AML patients, Micronucleus mean values recorded 0.033±0.018, 0.020±0.00015 and 0.036±0.01for AML patients before, during and after treatment respectively when compared with the control which recorded 0.002±0.00002. Nuclear division index (NDI) means before, during and after treatment of AML patients were 1.658±0.2, 1.000±0.022 and 1.424±0.19 respectively. Significant differences occurred among the three groups when compared with the control group (1.282±0.09). Extracted DNA from 30 AML patients was amplified by PCR to obtain FLT3/ITD gene from exon 11 to intron 12 and ten of them sent for sequencing. Two patients showed larger bands (470 and 460) bp when compared with wild type (330) bp. Among six patients, three displayed point mutations of deletion and substitution while others were normal since no mutations were detected. The percentages of mutation types were substitution 57.78% and deletion 22.2%. The effect of mutations was missense mutation (55.54%), deletion (22.22%) and nonsense (22.22%). It has been concluded from the current work that AML is more common in adult males, patients suffer from AML exhibited high MI, NDI, MN and point mutations are present in such patients including deletion and substitution causing missense, nonsense and deletion.

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

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

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

اسم المؤلف: مرتضى عادل الشامي
اسم المشرف: محفوظة عباس عمران | احمد مجيد الشمري
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

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

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

اسم المؤلف: لمى ياسين موسى
اسم المشرف: محمد ابراهيم نادر
الموضوع العام: علوم الحياة
السنة: 2015
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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.

التحري عن فيروس التهاب الكبد نمط B باستخدام تقنيات ELISA وReal Time PCR في دم ولعاب المرضى في محافظة الانبار == Detection of Hepatitis B Virus By Using Elisa And Real Time PCR Techniques in Patients' Blood And Saliva in Al - Anbar Governorate

اسم المؤلف: ثائر عبد الله حسن الدليمي
اسم المشرف: امين سليمان بدوي الجنابي
الموضوع العام: علوم الحياة
السنة: 2012
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: صلاح الدين
الصفحات الاولى:
المستخلص: اجريت هذه الدراسة في مختبرات قسم علوم الحياة في كلية التربية وفي مختبرات مستشفى الرمادي التعليمي العام ومستشفى الرمادي للنسائية والاطفال ومختبرات النظائر المشعة في بغداد للفترة من 1/11/2011 ولغاية 1/5/2012 والتي تضمنت التحري عن فيروس التهاب الكبد نمط B في | The present study has been carried out at the laboratories of the department of biology at the college of Education and the laboratories of the public - teaching Al - Ramadi hospital and Al - Ramadi hospital for women and children and laboratories of Al - Nathaer Al - Mushe'ah in Baghdad for the period from 1/11/2011 to 1/5/2012. The study consisted of the investigation about Hepatitis B virus in the saliva as compared with serum by using the ELISA and Real Time PCR techniques as well as the detection about antigens and antibody to this virus in serum. It also seeks about the liver functions enzymes and evaluating the extent of its importance for diagnosing liver disease resulted that virus. However, the obtained results showed that HBsAg antigen was found in 99.4% and (1.101± 0.335) mean ±SD in serum as compared with 48.6% in saliva with a (0.330±0.278) mean ±SD, that gave a noticeable increase when comparing it with the control group which was (0.0039± 0.0024) and (0.0017±0.0014 ) for serum and saliva respectively. The value of T test for serum and control group was (16.25) , while the value of T test for saliva and control group was (4.18). The proportion of correlation between saliva and serum in ELISA test was equal to (0.361) with high significance of test while the value of T test for them was (28.902) with high significance of test. The results have also stated that the age groups involved in the experiment contributed as being high proportion of infection in the serum, but in the saliva, these proportions were different and it has been found that the high proportion of infection was 53.3% in the group (10 - 20) yrs., While the lowest proportion was 38.4% in the (31 - 40) yrs. Moreover, it has been concluded that the number of males exceeded the number of females, but the number of females was the most infected with HBsAg in the experiment group. However, the total number of males was 91, It was found that 90 males have positive test to the HBsAg in serum with a proportion equals 98.9% whiles 42 males have a positive test to the HBsAg in saliva with a proportion equals 46.1%. on contrary, the number of female was 84 female and all of them had a positive test to the HBsAg in serum with a proportion 100% and 43 females have a positive test to the HBsAg in the saliva with 51.1 % as compared with control group which included 15 males and 10 female. All of them were negative to the HBsAg test in serum and saliva... The levels of HBV DNA were determined in the samples of positive saliva to ELISA test by using Real Time PCR. 95% from tested serum samples were positive to the viral load While in saliva 50% from the samples was HBV DNA positive. The results also showed that soluble antigen HBeAg has been detected in 24 patients with 13.7% from a total 175 patient as compared with the control group that showed a negative result to this antigen. As for the antibodies, it has been found that anti - HBc IgM was found in only 5 patients with 2.5% while anti - HBc IgG was found in 170 patients with 97.2% from the total number of patients. The control group revealed negative result for these two antibodies. The anti - HBeAg was in 150 patients with 85.7% from the total number of patients. There were no positive patients to this antibody in control group. The anti - HBs wasn’t found in any patient but it was found in the half of control group. This emphasizes to immunize against HBV in those members. The results also exposed that there was a noticeable increase in liver enzymes that it was found that ALT the elevated levels of were found in 76% of patients. As compared with the control group which has normal values 100%. As for AST enzyme was elevated in 60.6%, As compared with the control group which was normal 100%.It has been observed that ALP enzyme was elevated with a 38.4% in the experiment patients as compared with the control group which was normal 100%. Finally, the total values of TSB was elevated in 35.5%, As compared with control group which was normal 100% with (0.52±0.19). Nevertheless, it has been noticed that it cannot be judged about the person's blood on the basis that it is healthy in case of absence of surface antigen unless it is certain that his/her blood is free from HBV DNA by using the technique of Real Time PCR - since many patients who heal from past infection or their infection in a time of passive and amounts of HBV DNA can't be diagnosed by using the ELISA technique

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

اسم المؤلف: انتصار طه لفتة
اسم المشرف: واثق عباس الدراغي
الموضوع العام: علوم الحياة
السنة: 2015
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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.

مستوى هرمون الليبتين في مصل النساء المصابات بمتلازمة تكيس المبايض قبل وبعد العلاج بالميتفورمين == Serum Leptin Level In Women With Polycystic Ovary Syndrome Before And After Treatment With Metformin

اسم المؤلف: بتول حسين كاظم
اسم المشرف: وليد حميد يوسف سندال النصيري
الموضوع العام: علوم الحياة
السنة: 2012
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دبلوم عالي
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: Polycystic ovary syndrome is the most common endocrinopathy in women, affecting 5 - 10% of women of reproductive age.The principal features of PCOS are anovulation, resulting in irregular menstruation, amenorrhea, ovulation - related infertility, and polycystic ovaries; excessive amounts or effects of androgenic hormones. Leptin, a key hormone in energy homeostasis and neuroendocrine function, has a permissive role in the pathogenesis of reproductive dysfunction. Aim of the study To assess the role of serum leptin in women with PCOS and to evaluate leptin levels in PCOS women before and after treatment with metformin. Materials and Methods This prospective experimental study included 60 women of reproductive age (18 - 38years) were allocated to four groups : 15 obese women with PCOS (BMI >30 kg/m2), 15obese controls, 15 non - obese women with PCOS (BMI 18 - 30 kg/m2), and 15 non - obese controls. Serum leptin and insulin levels were measured and compared between case and control subjects also comparison done before and after treatment with metformin. Results There was a significant increase in leptin in non - obese PCOS group (8.2±2.73) compared to non - obese control (5.64±1.43), (P value=0.0032), insulin level was significantly higher in PCOS group (15.87±6.65) than control (5.47±1.68), (P value<0.001). There was significant decrease in BMI, leptin and insulin levels after 12 weeks of metformin treatment in obese and non - obese PCOS subjects. Conclusions Leptin level increased remarkably with increasing body weight. It is higher in non - obese PCOS women in comparison with non - obese healthy women. Treatment with metformin for 12 weeks resulted in significant reduction in leptin levels in both obese and non - obese PCOS subjects.

مقارنة نسب الحمل للنساء اللواتي يعانين من توقف الطمث لاسباب ثانويه بعد استعمال ادوية المنشطه للحويصلات المبيضيه باستخدام التمنيه داخل الرحم والطريقه الطبيعيه للحمل == Comparison of Pregnancy Rate Between Intrauterine Insemination And Spontaneous Pregnancy After Ovulation Induction In Women With Secondary Amenorrhea

اسم المؤلف: سوسن كاظم عبد الامير
اسم المشرف: محمد باقر محمد رشاد فخر الدين
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: انقطاع الطمث لاسباب ثانوية هو انعدام الدوره الشهريه لمدة ثلاثة الى ستة اشهر للنساء اللواتي لهن دوره شهريه منتظمة.واستنادا لمنظمة الصحة العالمية ان انقطاع الطمث يتضمن مجموعة (WHOI) و(WHOII) والتي تكون (30%) من حالات العقم.وتعاني اكثر النساء ممن يكونون من م | Secondary amenorrhea refers to the lack of cycle for three months in a row or for more than six months in a woman who has had regular menstrual periods before, and its associated with conditions classified as World Health Organization (WHO) group 1 and WHO group 2 and has been estimated to represent almost (30%) of all causes of infertility. In clinical practice, WHO group 2 subjects present much more frequently than WHO group 1 subjects and they appear to be much harder to treat successfully. Among women classified as WHO group 2, those diagnosed with polycystic ovary syndrome (PCOS) constitute the largest group (60 - 85% of cases). These women account for most cases with oligomenorrhea (90%) and represent about (30%) of those with amenorrhea.ObjectiveThe aim of this study was to find out the pregnancy rate following intrauterine insemination (IUI) in women complaining of secondary amenorrhea after induction of ovulation and compare it with those achieve pregnancy by natural way.Material and MethodsSeventy five females with secondary amenorrhea were involved in this study, there ages were ranged between (?30, 30 - 39, ?40) years old with infertility duration ranged between (?2, 3 - 4, 5 - 6, 7 - 8, ?9) years. History and medical examination were done for all women, with tubal patency assessment by HSG or laparoscopy.By cycle day two, hormonal assay (S.FSH, S.LH, S.E2, S.Prolactin and S.Testosterone) were done for all females in addition to ultrasonography to exclude any ovarian pathology.Ovulation induction program was applied for all females which include either clomiphene citrate (CC) alone, recombinant Follicular Stimulating Hormone (r - FSH) alone or combined CC and r - FSH. Vaginal ultrasonography was done for all females on day 12 or13 to assess ovarian response (size and number of mature follicles) and endometrial thickness. Meantime all male partners undergo seminal analysis to assess sperm parameters (sperm concentration, sperm motility and morphology).Thirty eight of females participated for natural pregnancy and thirty seven of females participate for IUI. Washing swim up technique was used for sperm activation for all male partner involved in IUI group.Follow up of all these females for pregnancy test to assess the pregnancy rate for both groups (natural and IUI).ResultsThe pregnancy rate achieved by natural way for a women with secondary amenorrhea is (23.7%), and (18.92%) by IUI way.In this study, it was found that female age, infertility duration, hormonal balance and ovulation induction program affect the pregnancy outcome where the young age group and short infertility duration correlate positively with pregnancy rate. Furthermore, early diagnosis of underlying causes of secondary amenorrhea to achieve hormonal balance by suitable ovulation induction program, have an effect on pregnancy rate, were combined CC and r - FSH produce a significant result among females achieve pregnancy by natural way (13.5%) compared with other ways. While ovulation induction by r - FSH drugs produce a significant results (13.5%) among females achieve pregnancy by IUI way.ConclusionFrom results of this study, it was concluded that young female age, short infertility duration, early diagnosis of underlying causes of secondary amenorrhea and the use of r - FSH drugs for ovulation induction produce a high significant pregnancy rate using IUI program.

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

اسم المؤلف: بنان محمود سليمان
اسم المشرف: ناظم حسن حيدر
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: رؤى اياد يوسف
اسم المشرف: غازي منعم عزيز | حسن فياض
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: ايمان نعمان اسماعيل
اسم المشرف: نورية عبد الحسين علي | مجيد ارشيد سباح
الموضوع العام: علوم الحياة
السنة: 2013
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: بشير كاظم خرميط
اسم المشرف: عبد الكريم عبد الرزاق القزاز
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: سيف صبار كامل
اسم المشرف: ناظم حسن حيدر
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: اماني احسان الصقر
اسم المشرف: اسماعيل حسين عزيز | اكرم فيصل الحويزي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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.

دراسة انتاج صبغة البرودجيوسين من بكتريا Serratia marcescens المعزولة من مصادر سريرية وبيئية وتاثيرها في بعض الميكروبات المرضية == Study of the Prodigiosin productivity from Serratia marcescens isolated from environmental and clinical sources and the effect on some pathogenic microbes

اسم المؤلف: فرزدق ناظم حربي البديري
اسم المشرف: مكرم ضياء جعفر شكارة | صالح عبد الرضا الصالح البكري
الموضوع العام: علوم الحياة
السنة: 2009
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: The cotemporary world witnesses currently a rapid change due to the challenges imposed by the information technology in all scientific aspects.Hence, the scientific knowledge has become a prerequisite for each individual in this world. This led to a new view on education, in that it is no longer to be a process of providing information but it is a technique for a better learning.However, the main goal of education has shifted to focus on how to make the student to think appropriately and how to utilize the skills of the scientific thinking in order to motivate their thinking and understanding of knowledge as well as discovering the facts and relationships concerning the scientific phenomena aiming to form the generalities. The problem of the current research is mainly concerned with specifying the classical methods adopted in the Iraqi schools which do not match the contemporary scientific development. This research aims at utilizing the strategy of the strategy of forming generalizations according to Helda Taba Inductive Model as a teaching technique in order to identify its effect on : 1 - achievement in Biology.2 - the skills of the scientific thinking of the Fifth - Year female pupils.For sake of achieving the above two aims, two null hypotheses have been formed as follows : 1 - There are no statistically significant differences at level of significance of 0.05 between the mean scres of the female pupils (the experimental gronp), who have been taught according to the strategy of forming generalizations the strategy of forming generalizations, and the mean scores of those female pupils who have been twght classically in the achievnent test. 2 - there are no statistically siguificant differences at level of significance between the experiemental group, who have been taught according to the strategy of forming generalizations, and the mean scores of those female pupils who have been taught classically in the test of the scienhific thinting skills. In order to verify the above two hypotheses, the researcher has conducted an experiment which lasted for one whole academic semester.This experiment inluded : - The lesson plans. - The scientific thinking skills test (30 items). - An achievement test (60 items).The discrimination and difficulty power as well as the alternative action of the two test have been computed.Moreover, validity and reliability of the two tests have been secured. The reliability of the scientific thinking test has been found out to be (0.75) and the reliability of the achievcment test has been found out to be (0.92). This was done by using koder Richardson formula - 20. The researcher has prepared an experimental design, comprising two groups, the first one of which is called the experimental group which is to be taught according to the strategy of forming generalizations.While the second group is called the control one which is to be taught classically. The sample of the study covers (46) female pupils at the fifth - year secondany school (scienbific branch).The whole sample have been randomly distributed into two groups, the experimental one is of (24)pupils and the control one is of (22)pupils. The equalization between the testees in the variables of age, intelligence, exinformation, previous achivement in Biology and the scientific thinking skills has been conducted. The experiment group was conducted at the second academic semester (7weeks).The researcher himself had taught the two groups.The results obtained has been statistically analyzed by using two - indepenbent samples t - test.The results gained showed a better performance of the experimental group more than that of the control one in the scientific thinking and the achievement tests. In the light of the above results, the researcher reconmends the use of the strategy of forming generalizations in accordance to Helda Taba Inductive Model.Also, the researcher recommends to conduct other studies and research on other subjccts other than Biology on different school stages to find out the impact of the strategy of forming generalizations on thinking skills.

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

اسم المؤلف: هوازن حليم صالح
اسم المشرف: علي عبد الامير مهدي الصالحي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: طارق طلال خلف
اسم المشرف: زهرة محمود الخفاجي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: العربية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: استبرق اكرم بيرام الحسيني
اسم المشرف: عصام فاضل علوان الجمیلي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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.

انتاج انزيم الفوسفاتيز القاعدي من العزلة المحلية لبكتريا Bacillus sp. وتنقيته جزئيا

اسم المؤلف: جعفر انور قدوري الخاجي
اسم المشرف: رباب عمران راضي الجيلاوي
الموضوع العام: علوم الحياة
السنة: 2015
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: بابل
الصفحات الاولى:
المستخلص: Fifteen local isolates of Bacillus bacteria and previously isolated were screening a from laboratory of advanced biotechnology and Genetic Engineering in Babylon University, College of Science to know their ability on production of extracellular alkaline phosphatase after its growth in liquid production in PH8 medium for 4 days and temperature 37°C. and destinguished 11 isolates by their different abilities on produc of enzyme and Bacillus sp.I bacteria was their best and perfected it selection to complete the study and described by depending on microscopic and cultured and biochemical characteristics studying of effect some of environmental condition for production of alkaline phosphatase enzyme by submerged cultural method were appeared that the optimum PH for production was 8, and optimum temperature 40°C, after incubation time 4days and explain that the production by static incubator was better from shaker incubator.Alkaline phosphatase of the isolate Bacillus sp.I was purified partial by fractionating with ammonium sulphate and the best ratio for saturation 95%. And on purification of enzyme by ion exchange chromatography with batch wise by using DEAE - cellulose and enzymatic recovery was 37.1%, and fold 3.64 once, and enzyme passed after concentrated on gel filtration column (Sepharcyl S - 200) one peak was appeared for enzyme and enzymatic recovery was 23 % and fold 36.80 once.Results of electrophoresis by presense of denaturation condition appeared that the molecular weight of alkaline phosphatase produced from gel filteration by using Sepharcyl S - 200 gain approximately 29.5 KDa and 28.8 KDa and this enzyme composed of homodimer.The characterization results for the enzyme indicated that the optimal pH for activity of enzyme was (9), while the optimal pH for the enzyme stability was (8 - 9), and found that the optimal temperature for activity of anzyme was 30 ? C, and on studying optimal temperature for stability of enzyme indicated that better stability for this enzyme was on temperature 0 ?C while the activity lost completely on temperature 60°C. and on studying effect some chemical compounds in activity of enzyme found that the enzyme needs to cofactor and it is zinc andmagnessium ion to increase catalytic activity and needs to calessium ion to increase stability of enzyme. At that time potassium and sodium ions appeared a negatively effect on activity, and heavy metals ions Fe and Hg appeared an inhibited effect to activity and so that the enzyme was inhibited by presence EDTA and and potassium ferro cyanide indicating that the enzyme from metal enzymes.Determination of constant kinitics of the enzyme was completed of Michaelis - Menten constant (Km) of para - nitrophenyl phosphate and this gained 500 micromolar and maximum velocity of this enzyme (Vmax), and this gained 2500 unit/ml.

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

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

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

اسم المؤلف: عقيل حيدر عطا الله
اسم المشرف: مؤيد صبري شوكت
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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.

التاثيرات المعدلة - مناعيا للمعززات الحيويه ضد بكتريا Salmonella enterica Serovar Typhimurium المعزولة من حالات الاسهال لدى الاطفال في ذكور الفار الابيض == Immunomodulatory Effects of Probiotics On Salmonella Enterica Serovar Typhimurium Isolated From Diarrheal Children In Albino Male Mice

اسم المؤلف: صفا خليل ابراهيم
اسم المشرف: عبد الواحد باقر الشيباني | علي حسين ادحيه
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: This study aimed to determine the frequency of Salmonella Typhimurium and its multi - drug resistance (MDR) status as a diarrheal causative pathogen in children younger than five years old, as well as to investigate the role of four probiotic microorganisms(L.acidophillus, L.casei, S.cerevisiae, S.boulardi) in controlling such infection.For this purpose, 128 stool samples of patients (76 males and 52 females) was included in this study. They were children suffering from diarrhea and fever who admitted to the “Central Pediatric Hospital” in Baghdad during the period from April to September, 2012. Their ages ranged from six days up to five years.The immunological effects of probiotics and MDR S. Typhimurium isolate in the experimentally infected and probiotic - treated mice were investigated via assessing the level of ten cytokines (IFN - ?, IL - 1?, IL - 4, IL - 10, IL - 12, IL - 17A, IL - 21, GM - CSF, RANTES and IP - 10) in the lavage of small intestine. Accordingly, five groups of mice were used in the in vivo part of this study; Mice in group I received probiotic for 7 successive days, challenged with S. Typhimurium on day 8, and dissected on days 14 and 21. Group II wassimilarly treated, but the probiotic was continued for 14 days. Group III was given the probiotic only, and group IV was challenged with S. Typhimurium, while group V was the control.Results obtained in this study could be summarized as follows : 1. Out of 128 stool samples, S. Typhimurium was isolated and identified in 9 samples only (7.03%). All isolates were totally resistant to nalidixic acid, with the exception of one isolate which showed intermediate sensitivity.Furthermore, only one of these isolates (symbolized B) was found to be resistant to three antibiotics (ampicillin, amoxicillin and nalidixic acid).Therefore, this isolate was considered as an MDR isolate and selected for further experiments in the study.2. When the susceptibility of MDR S. Typhimurium isolate B was further assessed in vitro by using four probiotics (Saccharomyces cerevisiae, S. boulardi, Lactobacillus acidophilus and L. casei), results showed that S.cerevisiae and L. acidophilus were the most efficient by recording the highest inhibition zones (12.6 and 16.3 mm, respectively), therefore, they were further investigated for their anti - S. Typhimurium effects in vitro and in vivo. 3. By using the unconcentrated and (one - fold, two - fold, three - fold) concentrated filtrates of these two probiotics, it was found that the three - fold filtrates were most efficient in their antibacterial activity by recording the highest inhibition zones (25.0 mm for S. cerevisiae and 31.0 mm for L. acidophilus.4. Significant increases in the values of liver index were observed in mice of group I treated with L. acidophilus at 21 days (10.73%) compared to the corresponding group treated with S. cerevisiae (7.41%) or other four groups. For spleen, index value in L. acidophilus groups was higher than the corresponding groups in S. cerevisiae. Mice treated with a probiotic alone or in a combination with the pathogen showed significant increases in the spleen index values of all groups compared to the untreated mice (group V).5. Both probiotics were effective in reducing S. Typhimurium colony forming units per plate (cfu/plate) in the liver and spleen. In liver, mice in group IV showed a count of 224.4 cfu/plate, which was significantly higher than any count in groups of L. acidophilus and S. cerevisiae. Group II mice recorded better results than group I mice, while the lowest counts (21.6 and 27.8 cfu/plate for L. acidophilus and S. cerevisiae, respectively), were observed at day 21.6. The ten investigated cytokines showed different levels in the small intestine wash; such differences were subjected to the group of mice underinvestigation and type of probiotics used. In addition, variations in IFN - ?/IL - 4, IFN - ?/IL - 10, IL - 4/IL - 10 and IL - 17A/IL - 10 were also observed.

العلاقة بين متلازمة تكيس المبايض وهرمون المضاد لمولر وبرامج تحريض الاباضة عند اجراء عملية التمنية داخل الرحم

اسم المؤلف: عبير ناظم جاسم
اسم المشرف: محمد باقر محمد رشاد فخر الدين | صباح مهدي حسين
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دبلوم عالي
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: تعد متلازمة المبيض متعدد الا كياس هي من اكثر امراض الغدد الصماء شيوعا عند النساء حيث يصاب من النساء في سن الانجاب مايقارب (5 - 10%). الاعراض الرئيسيه لمرض متلازمة المبيض متعدد الاكياس هو اضطراب الدوره الشهريه مع فشل لاباضه وظهور اعراض زيادة الهرمون الذكر

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

اسم المؤلف: عدي عدنان مهدي
اسم المشرف: اسماعيل عبد الرضا عبد الحسن
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: غزوان علي مسلم الرماحي
اسم المشرف: عبد الرضا عبد الحسن اللامي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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 وراثية وبكتريولوجية على بكتريا تثبيت النيتروجين التكافلية Ensifer meliloti == Bacteriological And Genetic Study On Symbiotic Nitrogen Fixing Bacteria Ensifer Meliloti

اسم المؤلف: محمد زهير ناجي المرعب
اسم المشرف: حسن فاضل ناجي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: بابل
الصفحات الاولى:
المستخلص: عزلت عشر عزلات من بكتريا Ensifer meliloti من العقد الجذرية لنبات الجت Medicago sativa L. بعد ان جمعت من مناطق مختلفة من محافظة بابل خلال المدة من شهر تشرين الثاني/2012 لغاية شهر شباط/2013. شخصت العزلات جزيئيا بتقنية تفاعل البلمرة السلسلي فضلا عن طرق ال | Ten isolates of Ensifer meliloti were isolated from root nodules of Medicago sativa L. collected from different areas of Babylon province during a period from November - 2012 to February - 2013. The isolates were molecular identified using polymerase chain reaction technique, in addition to traditional methods (morphological and biochemical). All isolates were harboured nodbox and mucR genes and produced cyclic ? - (1, 2) glucans, cellulose fibers and lipopolysaccharide.The ability of isolates to grow at different degrees of pH, temperatures and different concentrations of sodium chloride were tested. It was found that the optimum pH for growing of Em1, Em3, Em4, Em6 and Em8 isolates was pH7, while for the isolates Em2, Em5, Em7, Em9 and Em10 was pH8. The optimum temperature for growing of Em1, Em3, Em5, Em7, Em8 and Em9 isolates was 30?C, while the optimal growth of Em2, Em 4 and Em6 isolates was 35?C, Em10 isolate showed significantly extremism to grow at temperture reached to 45?C. The optimal percentages of NaCl for growing of Em1, Em3, Em4, Em5, Em6, Em8 and Em9 isolates were 2%, for Em7 isolate was 3% and for Em2 and Em10 isolates were 4%. The effect of different degrees of pH on symbiotic properties was studied. Em1 and Em8 isolates that grew at pH6.5 ; all isolates that grew at pH 7, pH 7.5 and pH 8 were able to infect the root hairs of alfalfa plants. The isolates that grew at pH6.5 (except Em1 and Em8 isolates) and all isolates that grew at pH8.5 were failed to infect the root hairs and did not formed nodules and resembled to the control plants (without inoculum).Different concentrations of NaCl on symbiosis was studied, it was found that all isolates that grew at 2% and Em2 and Em10 isolates that grew at 3% formed nodule, while Em1, Em3, Em4, Em5, Em6, Em7, Em8 and Em9 isolates that grew at 3% and all isolates that grew at 4%, 5% and 6% did not elicited nodules on the roots of alfalfa plant. Light microscopic observations of root hairs inoculated with Ensifer meliloti isolates showed the root hair deformations like curling, waviness, swelling and formation of shepherd`s crook and infection thread. These consequent changes lead to emerge the mature nodules having five distinct zones viz., distal meristematic, infection, inter, nitrogen - fixing and proximal senescence zones.The gel electrophoresis results of plasmid DNA of isolates showed that these isolates harboured two megaplasmids. When the isolates exposed to extreme factors like pH, temperture and NaCl, it was found that the isolates lost these plasmids and its ability to elicite the nodules and failed to fix the nitrogen. This study showed that the nitrogen fixing bacteria had essential role to support the plant growth and increase their productivity, and recommended to use the isolate Em10 as example for biofertilizers and the best alternative to the chemical fertilizers

دراسة العلاقة بين فقدان الجينين الـ GSTM1 وGSTT1 والاصابة بمرض الربو القصبي في محافظة ذي قار == Study The Relationship Between The Null Genotype of GSTM1, GSTT1 Genes And Bronchial Asthma In Thi - Qar Province

اسم المؤلف: نـور عمران عبد الكريم الغـزي
اسم المشرف: حسن ريسان الركابي | عدنان عيسى البدران
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: ذي قار
الصفحات الاولى:
المستخلص: تم تصميم هذه الدراسة للتحقق من دور جينات الـ Glutathione - S - Transferase (Mu - 1) gene و(Theta) - 1 Glutathione - S - Transferase ومدى مساهمتها بخطر الاصابة بمرض الربو القصبي في محافظة ذي قار. حيث شملت (105) شخص مصاب بالربو باعمار من (20 - 69) سنة كان | This study has been designed to investigate the roles of the gens GSTMI and GSTTI the extent to which they contribute in asthma at Thi - Qar governorate. The study has been conducted on (105) patients with asthma whose ages from (20) to (69) years. (20) of them are males and (85) are females, and (110) are healthy with no infection of asthma or any sickness of respiratory system as a control group of the same ages, (60) of these are males and (50) are females. Blood samples have been collected from both groups in ampoules with EDTA and stored under temperature ( - 20) until they are used in getting the DNA from the groups. The genes GSTTI, GSTMI, and Albumin have been enlarged as internal control employing Polymerase chain Reaction (PCR). Some dangerous aspects of this disease have been investigated, the results showed that the disease in the urban areas (85. 71 %) higher than it rural areas, and Asthma among females is (80. 95%) higher than males. The results also indicated that asthma in ages (50 - 59) is more than other categories; it is (34.28%). The majority of patients are smokers with (63.80%). The study revealed that (73.33 %) of the patients have a family history with the disease while (26.66 %) have not. Statistical analysis showed that the loss of the gene GSTM1 is about three times in the patients more than the case in the control group. (OR=2.90) , while the loss of the gene GSTT1 is five fold more than it compared to the control group (OR= 5.55). The results indicated a significant effect of the loss of the two genes in having asthma in the urban and rural areas, it is six times and a half larger in the urban areas compared to the control group of the same area. Asthma probability is ten times more in the case of the loss of both genes in the rural areas compared to the control group in the same area. And no significant difference has been discovered when gene patterns of patients in both areas. The results also indicated that the probability of having asthma in male patients is two times and a half as many as it in the females when the gene GSTM1 is lost compared to the control group. While the same probability is eleven times in the case of the loss of both genes in the females as many as it the control group. The loss of Missing the gene GSTT1 is three times in the male patients and five times in the females compared to the two control groups. The results indicated significant differences when gene patterns of males and females are compared in the case of the loss of both genes. The statistical analysis indicated that the loss of both genes in the smoker patients contributes 17 fold in the probability of having the disease compared to the control group. While the probability increases four times approximately in the nonsmokers when the gene GSTM1 is lost compared to the control group (OR=3.77; 95% C1=1.69 - 8.407). No difference has been noticed in the gene patterns of the smoker and nonsmoker patients. The results indicated that the loss of both genes has been in all the age categories. No significant differences have been noticed among the gene patterns between the patients of less and those who are more than 50 years. Asthma probability has been found 7.77 times more in the case of the loss of both genes in the patients who have a family disease history compared with the control group. The asthma probability is twice when the gene GSTM1 is lost in those patients who do not have a family history disease compared to the control group. Upon comparing the gene patterns of patients according to the family disease history, there has been an effect of the loss of both genes and having a family disease history on the probability of asthma, it is almost six times when both genes are lost. The results indicated that the probability increases three times in the loss of the gene GSTT1 in the educated patients compared to control group. The probability also increases seven times when both genes are lost in the uneducated patients compared to the control group. When comparing the gene patterns of patients according to levels of education, significant differences between the educated an uneducated have been approximately twice in the loss of the gene GSTT1.

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

اسم المؤلف: ياسر عبد الجبار عبود السوداني
اسم المشرف: عصام فاضل علوان الجمیلي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: ياسمين ابراهيم فرحان
اسم المشرف: امنة نعمة الثويني
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: زينب صالح هادي الزبيدي
اسم المشرف: محمد ابراهيم نادر
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: هاشم محمد زهراو الصبيحاوي
اسم المشرف: عصام فاضل علوان الجمیلي | فارس عبد الكريم
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
الجامعة: جامعة بغداد
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: هالة كمال محسن القزاز
اسم المشرف: نورية عبد الحسين علي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: ضياء خليل اسماعيل
اسم المشرف: نورية عبد الحسين علي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: 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

اسم المؤلف: مياسة مثنى خالد
اسم المشرف: اسماعيل عبد الرضا عبد الحسن
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
الجامعة: جامعة بغداد
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: اجريت الدراسه الحاليه في معهد الهندسه الوراثيه والتقنيات الاحيائيه - جامعة بغداد خلال الفتره من كانون الثاني ولغاية حزيران - 2013 للكشف عن علاقة بعض الطرز الجين كالبين - 10 بحدوث مرض السكري من النوع الثاني في العراق. تم استخلاص الدنا من الدم الكلي باستخد | The present study was carried out in Genetic Engineering and biotechnology Institute - Baghdad University during a period from January to June, 2013, for detecting the association of some calpain - 10 gene polymorphisms with the incidence of type 2 diabetes mellitus in Iraq. Genomic DNA was isolated by using Geneaid DNA extraction kit from the whole blood; conventional PCR (SNP - 44 and Del/Ins - 19) and PCR - RFLP (SNP - 43 and SNP - 63) were used to detect the calpain10 variants by using specific primers and restriction enzymes. The study population consisted of 50 subjects with type 2 diabetes and 50 with normal fasting blood glucose (80 - 110 mg/dl). The type 2 diabetic subjects were recruited from the National Center for Diabetes treatment and Research. The non - diabetic control subjects were recruited from the same area as the comprising blood donors, healthy volunteers, or hospital /university staff members. Previous studies have detected a role for Calpain - 10 (CAPN10) polymorphisms in susceptibility to Type 2 diabetes mellitus (T2DM) in many populations. This study aimed to evaluate possible associations between these polymorphisms in the CAPN10 gene (SNP - 44, SNP - 43, Del/Ins - 19, and SNP - 63) and T2DM incidence in Iraqi population. Enrichment of allele 1(2R) in Del/Ins - 19 and 2R/2R genotype were found in T2DM patients. While the alleles and genotypes distribution of SNP - 44, SNP - 43 and SNP - 63 were not significantly different between patient groups and non - diabetic control subjects. The genotype AA in SNP - 43 and genotype TT in SNP - 63 were not found neither in T2DM nor in control subjects. of the eight haplotypes detected, enrichment of both haplotype 112 defined by variants of SNP - 43, Del/Ins - 19, and SNP - 63 and haplotype 2112 defined by variants of SNP - 44, SNP - 43, Del/Ins - 19, and SNP - 63 were seen in patients. The distribution of the other haplotypes was comparable between patients and control subjects. The calpain10 haplotype combinations were also obtained, and the haplotype combinations 111/111 and 111/112; which are created by variants of SNP - 43, del/ins - 19 and SNP - 63 and; haplotype combinations 1111/2111, 1111/2112 and 1121 / 2222; created by SNP - 44, SNP - 43, del/ins - 19 and SNP - 63; were associated with increasing the risk of T2DM.

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

اسم المؤلف: شیماء عصام عبد الوهاب البرزنجي
اسم المشرف: عصام فاضل علوان الجمیلي
الموضوع العام: علوم الحياة
السنة: 2014
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: دكتوراه
الجامعة: جامعة بغداد
اللغة: الانكليزية
مكان الجامعة: بغداد
الصفحات الاولى:
المستخلص: In this study, natural lignan dimer was isolated from nutmeg seeds (Myristica fragrans) using organic solvent, partially purified using liquid/liquid partiation, purified using anion exchanger and chemically characterized using Benedict’s Reagent, Fehling’s Reagent and Molish’s Reagent. Then, by the aid of UPLC - PDA - IT - TOF - MS System, the molecular weight (626.221 Dalton) and the molecular formula (C39H45O7) of this dimer were determined. After that, the free radical scavenging activities were studied using stable free radical compound 1, 1 - Diphenyl - 2 - Picryl - hydrazil (DPPH). Results showed that 100, 10, 1 and 0.1 ?g/ml of purified lignan had 76.7 %, 65%, 28% and 8% scavenging activity respectively, while the same concentrations of partial purified lignan had 44.3%, 18.5%, 11% and 0% scavenging activity respectively.MTT(3 - (dimethylthiazol - 2 - yl) - 2, 5 - diphenyl tetrazolium bromide) assay was conducted to determined the IC50 for both purified and partial purified lignan using 4 different cell lines A549 (human lung adenocarcinoma epithelial cells), MCF7 (breast cancer cells), PC3(human prostate cancer cells), and HepG2(liver hepatocellular cancer cells), and to determine which cells type were be affected more by this natural lignan dimmer.The IC50 values for purified lignan were 85.17, 51.16, 108.4 and 60.21 ?g/ml while the IC50 values for partial purified one were 170.1, 84.14, 154.4 and 151.3 ?g/ml using A549, MCF - 7, PC - 3 and HePG2, respectively.The high content screening analysis (HCSA) and Cellomics Thermo Scientific maltiparametric Kits were used for the evaluation of cell - lignan interaction; 100, 50 and 25 ?g/ml of purified lignan caused 87.22, 69 and 53.36% reduction in MCF - 7cell count respectively and the same concentrations caused 98.1, 97.5 and 98.55% nuclear morphology changes. Results also revealed that these concentrations caused 7.7, 7.0 and 5.83% increase in MCF - 7 cells permeability respectively and they also caused 12.22, 11.15 and 0.2%decrease in mitochondrial membrane potential respectively, while these concentrations caused 11.12, 10.1 and 10% increase in Cytochrome C releasing from mitochondria to cytoplasim respectively.Reactive oxygen species (ROS) induction in MCF - 7 cells in the presence of 200, 100 and 50 ?g/ml of purified lignan caused 20, 11.48 and 9.61% ROS reduction respectively.MCF - 7 cell cycle was studied in the presence of 100, 50 and 25 ?g/ml of purified lignan, and results revealed that this compound blocked cell cycle at Sphase and the percentages of S - phase cells reduction were 74.33, 52.4 and 67%, respectively. This reduction was dose dependent while the same concentrations had no effect on MCF - 7 mitotic cells. Cell cycle arrest was detected immunofluorescently using BrdU antibodies (S - phase cell staining) and phosphor - Histone H3 antibodies (M - phase cells staining

عزل وتشخيص البكتريا العنقودية السالبة لانزيم الخثرة من حالات مرضية مختلفة ودراسة مقاومتها للمضادات الحيوية == Isolation And Identification of Coagulase Negative Staphylococci From Different Clinical Cases And Study of Their Antibiotic Resistance

اسم المؤلف: مجاهد خلف علي احمد الجميلي
اسم المشرف: عباس علي هادي الدجيلي | وعد محمود رؤوف
الموضوع العام: علوم الحياة
السنة: 2008
الموضوع الدقيق: التقنيات الاحيائية
الدرجة: ماجستير
اللغة: العربية
مكان الجامعة: صلاح الدين
الصفحات الاولى:
المستخلص: جمعت 265 عينة من مستشفى تكريت التعليمي العام وشملت هذه العينات عينات سريرية لمرضى راقدين في المستشفى ومرضى مراجعين لها. اخذت العينات من المرضى المشتبه باصابتهم بخمج السبيل البولي وخمج الجروح والحروق وخمج الاذن الوسطى ولكلا الجنسين وباعمار مختلفة. انجزت هذ | A number of 265 samples had been collected from Tikrit Teaching Hospital (T.T.H.).These samples were taken from inpatients and outpatients suspected to have Urinary Tract Infection, Wound & Burn Infections and Mid - Ear infection for males and females in varied ages. The practical part had been fulfilled during the period from the beginning of December 2006 till the beginning of March 2007.Isolates diagnosed were identified depending on morphological and cultural characteristics. The results revealed 60 Staphylococci isolates, which shows 27 isolates (45%) were S. epidermidis, 13 isolates (21.66%) S. saprophyticus, 10 isolates (16.67%) S. xylosus, 3 isolates (5%) S. lentus, two isolates (3.33%) S. heamolyticus, and one isolate (1.67%) for each of S. simulans and S. hominis Ten (34.48%) of patients are located in the age group (15 - 30) years. while concerning the classification according to sex, females were (65.5%). Otitis media and Wounds and Burns infections, their ratio was high (61.1%, 53.8%) consequently both in males in comparison with females. Concerning the relationship between the infection and the residence and the level of education, the results revealed that isolation of Coagulase Negative Staphylococci (CoNS) were high in rural areas and in uneducated persons regarding the various infections than those live in urbans and educated ones. A of main tests were carried out in order to achieve the purpose of this study according to the planned goals, some of which are the virulence factors of Coagulase negative Staphylococci. These include DNase, Urease, Lipase, Lecithinase, and Alkaline phosphatase.The results revealed the ability of 67.5% of isolates to produce Alkaline phosphatase, and 46.6% of isolates to produce Urease, also 27.5% of isolates were able to produce Lecithinase , and the ability of 19.16% to produce Lipase, while 22.5% of isolates were capable of producing DNase. In addition to these tests, the test for producing ? - lactamase was done, 85% of isolates were positive to this test because ? - lactamase is one of the significant enzyme which are produced by bacteria since it improves the resistance of bacteria for antibiotics. Thirteen different antibiotics used for the Antibiotics susceptibility tests which show that most effective antibiotic is Nitrofurantoin (95%), followed by Amikacin (93.3%), Novobiocin(90%), Chloramphenicol(88.3%), Rifampicin (78.5%), Vancomycin (75%), Ciprofloxacin (73.3%), Clindamycin (70%), Gentamicin (58%), Penicillin G (50%), Trimethoprim (50%), Tetracycline (40%) and Cefoxitin (18.3%).
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