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
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
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
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
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
التاثيرات المضادة للاكسدة والسمية الخلوية لمركب اللكنان المنقى من بذور نبات جوزة الطيب == Antioxidant And Cytotoxic Effects of Lignan Purified From Myristica Fragrans Seeds
Author name:
شیماء عصام عبد الوهاب البرزنجي
Supervisor name:
عصام فاضل علوان الجمیلي
Abstract:
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
