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

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

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

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

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

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

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

Author name: فرزدق ناظم حربي البديري
Supervisor name: مكرم ضياء جعفر شكارة | صالح عبد الرضا الصالح البكري
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: Arabic
University location: Baghdad
First pages:
Abstract: 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

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

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

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

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

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

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

Author name: جعفر انور قدوري الخاجي
Supervisor name: رباب عمران راضي الجيلاوي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: Arabic
University location: Babylon
First pages:
Abstract: 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.

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

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

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

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

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

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

دراسة وراثية وبكتريولوجية على بكتريا تثبيت النيتروجين التكافلية Ensifer meliloti == Bacteriological And Genetic Study On Symbiotic Nitrogen Fixing Bacteria Ensifer Meliloti

Author name: محمد زهير ناجي المرعب
Supervisor name: حسن فاضل ناجي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: Arabic
University location: Babylon
First pages:
Abstract: 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

Author name: نـور عمران عبد الكريم الغـزي
Supervisor name: حسن ريسان الركابي | عدنان عيسى البدران
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: Arabic
University location: Dhi Qar
First pages:
Abstract: 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

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

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

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

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

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

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

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

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

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

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

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

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

Author name: مجاهد خلف علي احمد الجميلي
Supervisor name: عباس علي هادي الدجيلي | وعد محمود رؤوف
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
Language: Arabic
University location: Salahaddin
First pages:
Abstract: 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%).

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

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

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

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

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

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

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

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

التفكك الاحيائي لبروتينات اللعاب بواسطة المسبحيات الفموية-ودراسة الطوافر المضعفة == Proteolytic Degradation Of Salivary Proteins By Oral Streptococcus And Selection For Attenuated Mutants

Author name: علي جبار رشك الساعدي
Supervisor name: صبحي جواد حمزة | عصام فاضل الجميلي
General topic: Biology
Specific topic: Biotechnologies
Degree: Master
University: University of Baghdad
Language: English
University location: Baghdad
First pages:
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