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تاثير الانزيمين الكلوكواميليز والكلوكوز اوكسيديز في تثبيط بكتريا Streptococcus mutans المسببة لتسوس الاسنان The Effect of Glucoamylase And Glucose Oxidase On Inhibition of Streptococcus Mutans Causes The Dental Caries

اسم المؤلف: بيداء عبود حسن الجنابي
اسم المشرف: زهرة محسن علي محمد عبد الله جبر
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2014
مكان الجامعة: النجف
المستخلص: تضمنت الدراسة استخلاص الكلوكواميليز والكلوكوز اوكسيديز من الفطرين Aspergillus niger وPenicillum notatum على التوالي وتنقية وتوصيف الانزيمين لغرض زيادة تركيز بيروكسيد الهيدروجين المحفز لانزيم اللاكتوبيروكسيد الموجود باللعاب على تكوين ايونات الهايبوثايوسيان | This study aims at producing glucoamylase and glucose oxidase from the Aspergillus niger and Penicillium notatum respectively , purifying and characterizing the enzymes to increase H2O2 concentration which induce lactoperoxidase in saliva to the formation of hypothiocyanite ion to inhibite Streptococcus mutans which causes the dental caries. For glucoamylase and glucose oxidase production, the results shown the the highest enzymes production were occurred using the synthetic medium, it gave high titer of glucoamylase and glucose oxidase activity (0.212, 0.605 U/ml) respectively, the optimum incubation period for glucoamylase was occurred after 96 h (0.125 U/ml), while the maximum activity of glucose oxidase was occurred after 72 h (0.662 U/ml), the optimum temperature for enzymes production was occurred in 30°C, (0.257, 0.690 U/ml) respectively, the maximum production of glucoamylase and glucose oxidase were occurred in the pH = 6 (0.174, 0.391 U/ml) respectively, the optimum nitrogen source for enzymes production was yeast extract and potassium nitrate (0.157, 0.571 U/ml) respectively, the optimum carbon source for glucoamylase and glucose oxidase production was starch and glucose (0.167, 0.459 U/ml) respectively. For glucoamylase and glucose oxidase purification, the results found the ammonium sulfate (80 %) was selected as the best ratio for precipitate of glucoamylase , the specific activity reached to (3.626 U/mg) , with a purification fold (1.241) and yield (0.38) % , while the (60 %) was selected as the best ratio for precipitate of glucose oxidase, the specific activity reached to (10.304 U/mg) , with a purification fold (1.219) and yield (0.34) % , two peaks of glucoamylase were appeared in the gel filtration the specific activity of the first form (4.195 U/mg) with purification fold (1.436), while the second (31.214 U/mg) with purification fold (10.689), while one peak of glucose oxidase was appeared in the gel filtration, the specific activity was reached (62.382 U/mg) with purification fold (7.385). For characterization of enzymes, the results reported the highest activity of glucoamylase (Aand B) occurred in pH = 8, 6.5 (0.253U/ml, 0.511 U/ml) respectively , while the maximum activity of glucose oxidase (1.289 U/ml) in pH = 5.5, the optimum temperature of glucoamylase (Aand B) activity (0.243U/ml, 0.703 U/ml) in 40, 30 °C respectively , while the optimum temperature of glucose oxidase activity (1.424 U/ml) in 45 °C, the optimum pH for glucoamylase (B) stability ranging between (5.5 - 6.5), while the optimum pH for glucose oxidase stability ranging between (5 - 6) , the optimum temperature for glucoamylase (B) stability ranging between (10 - 35) °C for 30 min, while the optimum temperature for glucose oxidase stability ranging between (25 - 35) °C for 15min, the molecular weights of glucoamylase (Aand B) approximately 52 & 66 kDa respectively using the electrophoresis technique, while approximately 68 kDa for glucoamylase (B) using the gel filtration technique, the molecular weight of glucose oxidase approximately 78 and 87 kDa electrophoresis electrophoresis and gel filtration respectively, the values of Km and Vmax of glucoamylase and glucose oxidase were (2.4 mM , 9.6 mM/min and 19.6 mM, 7.5 mM/min) respectively. The result of this study showed the Streptococcus mutans growth was killed in the using the first and the second concentration of glucoamylase and glucose oxidase (0.09, 0.009 and 0.3, 0.03 µl) respectively , while the growth was appered in third concentration of enzymes (0.0009, 0.003 µl) respectively. The NaF, ZnF2 , C 12H 7 Cl 3 FNaO2 , NaCl , NaHCO3 and KSCN with (0.5 and 1M ) leads to increase the enzymes activity , while Na3PO4 , SrCl2 , Fe2O3 and Ca(HCO3) 2 caused decreased the activity of enzymes when treated with (0.5 and1 M) from these solutions, the glucoamylase activity also decreased when it treated with (0.5 and 1M) from KNO3 solution but this solution increased the activity of glucose oxidase when it treated with (0.5 and 1 M) from this solution
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الكشف عن الجينات المقاومة للكاربابنيم لبكتريا Acinetobacter baumannii المعزولة من عينات سريرية في محافظة بابل Detection of Carbapenem - Resistant Genes of Acinetobacter Baumannii Isolated From Clinical Samples In Babylon Province

اسم المؤلف: رعـد عبد العباس حمزة الهرموش
اسم المشرف: ايمان محمد جار الله
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بابل
المستخلص: للفترة من اذار 2014 الى اذار 2015 تم جمع 1300 عينة سريرية مختلفة (عينات ادرار ومسحات جروح وحروق وعينات دم ومسحات من الفم والاذن والعيون بالاضافة لعينات البراز والقشع) من اثنين من مستشفيات محافظة بابل (مستشفى الحلة التعليمي ومستشفى بابل التعليمي للنسائية و| During the period between March 2014 and March 2015, a total of 1300 clinical specimens (urine, wounds, burns, blood, throat, ear, eye, stool, and sputum) were collected from two hospitals in Babylon province; Al - Hillah Teaching Hospital, and Babylon Hospital for Maternity and Pediatric. All specimens were cultured and 10 Acinetobacter baumannii isolates were obtained from clinical specimens in a percentage of (0.76%) distributed in blood 1 (0.6%), urine 2 (1%), wound infection 1 (0.75%), burn 5 (0.85%) and throat 1 (2%). This study was conducted to determine the occurrence of carbapenem resistant A. baumannii isolates obtained from Hillah hospitals. Isolates were identified according to API20E system and further confirmed using PCR technique. Antibiotics susceptibility was assayed by using disks diffusion method according to CLSI recommendations. All isolates revealed positive results with rapid iodometric test indicting the ability of beta lactamase production. Phenotypic detection of carbapenemase production was performed using the imipenem - EDTA disk and modified Hodeg's test (MHT). Then isolates were subjected to monoplex PCR targeting blaOXA - 51, blaOXA - 23, blaOXA - 24, blaOXA - 58, blaIMP, blaSIM blaNDM - 1, blaNDM - 2 and blaVIM genes, as well as, efflux pumps genes. One of the identified isolates (10%) was found to be imipenem, meropenem and ertapenem resistant, this isolate gave positive result with the imipenem - EDTA disk and (60%) of isolates were positive to MHT. PCR experiments showed ; all isolates were harbored blaOXA - 51 gene, four (40%) isolates were harbored blaOXA - 23 gene, three (30%) isolates were harbored blaOXA - 24 gene, four (40%) isolates were harbored blaOXA - 58 gene, five (50%) isolates were harbored blaIMP gene, six (60%) isolates were harbored blaSIM gene, two (20%) isolates were harbored blaNDM - 1 gene, four (40%) isolates were harbored blaNDM - 2 gene, while none of these isolates harbored blaVIM gene. The present findings suggest that emergence of (OXA - 23, OXA - 24, OXA - 58, IMP, SIM, NDM - 1, NDM - 2) carbapenemase producing A. baumannii clinical isolates in Hillah City hospitals. Also there are indicative appearance of efflux pump genes ; (50%) for Ade - A and Ade - B each other, (40%) for Ade - C, (80%) for all of Ade - R and Ade - S, (100%) for all of Ade - I and Ade - J, and (50%) for Ade - Y. All isolates of A. baumannii appear as MDR, while only one appear to be as PDR. The trans - mobility of resistant genes was examined by trans - conjugation experiment ; the results indicated that only blaIMP and blaOXA - 23 genes were transferred horizontally in the current study. In attempting to investigate any blaNDM gene variation the results showed some different than NCBI - Blast A. baumannii blaNDM - 1 and blaNDM - 2 it may as a unique genotyping.These results revealed that carbapenemase producing A. baumannii were detected in both phenotypic and genotypic methods (PCR). This underlies the importance of their accurate identifications and reporting to prevent the emergence of complete resistance to the most potent drugs against A. baumannii in Babylon province.
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توصيف العاثيات الحالة للمكورات العنقودية الجلدية (S. epidermidis) Characterization of Lytic Phage Against Staphylococcus Epidermidis

اسم المؤلف: ريام سليم هندي الزيادي
اسم المشرف: غانم عبود جابر المولى
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: ماجستير
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بغداد
المستخلص: لحصول على عزلات Staph. epidermidisتم جمع (160 عينة) من حالات سريرية مختلفة كالتهاب المجاري البولية, التهابات جلدية, البلعوم, الانف, والاذن. تم الحصول على 51 (875¸31?) عزلة للمكورات العنقودية الجلدية Staph. epidermidis خلال فترة امتدت من ايلول 2014 الى كان | In order to isolate Staph. epidermidis, 160 clinical specimens were collected from (urinary tract infection, skin infection, pharynx, nose, and ear). Only 51(31.875 %) Staph. epidermidis isolates were obtained during a period from September 2014 to January 2015, depending on some biochemical tests and VITEK2 system. The Staph. epidermidis was given (gram stain, catalase , urase) positive, (coagulase, manital fermentation, oxidase, motility) negative, non haemolytic to human blood with some exception, most of the strain were able to produce biofilm, and (100%), (64.70%), (74.50%) resistant to penicillin G, ampicillin, and amoxicillin respectively. The Staphylococcus species identified during study were 44 (27.5%) isolates of Staph. aureus, 5 (3.125 %) Staph. haemolyticus, 4 (2.5 %) Staph. saprophyticus, 8 (4%) isolates other Staphylococcus spp. such as Staph. hominis ssp. hominis, Staph. capatis, Staph. xylose, Staph. simulanus, Staph. lentus, and 48 (30 %) from unknown genera. To isolate bacterioghages from sewage water, several sewage water samples were assayed using plaque assay of double agar overlay as a source of Staph. epidermidis phages. The bacteriophages were described depending on plaques size and shapes, phage 1 was the most predominant and frequent in the bacterial lawn and able to infect other S. species such as S. aureus. It has been selected to study it is titer, latent, rise period, and burst size were calculated. The effect of temperature, pH , and NaCl ions on it is original titer were studied.The results revealed a gradual decreasing in the phage titer with increasing dilution number. Latent period extended to (30 minutes) , while rise period was started with (40 minutes) extending to (60 minutes) , burst size was 2.346. Each temperature at several incubation periods , pH , and NaCl ions was significantly varied depending on phage titer. The optimum temperature was 40 ° C, while the 80 ° C was represented the inhibitor temperature. L.S.D. at level (0.05) for interaction was 39.552. The pH 6.5 - 7.5 represented the optimal pH for the best phage activity while the phage titer beginning to declining in above and below this range of optimal pH, L.S.D. at level 0.05 was 17.898 , the optimum NaCl ions concentrations were (0.1 M and 0.25 M), while the titer was significantly decreased with increasing the NaCl ion concentration in the culture solution, the L.S.D. at level 0.05 was (10.696). In conclusion of this study found that Phage1 was considered as predominant phage because of their ability to infect other Staphylococci species such as Staph. aureus.
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النمو، الفعالية الانزيمية والتعبير الجيني للانزيمات المحللة للبروتين (Protease) في الفطر الجلدي Trichophyton rubrum Growth, Enzyme Activity And Gene Expression of Proteases In A Dermatophyte Trichophyton Rubrum

اسم المؤلف: سارة كريم كاظم الزبيدي
اسم المشرف: جواد كاظم الجنابي عدنان حمد الحمداني
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بابل
المستخلص: صممت الدراسة الحالية لتوصيف الانماط الجزيئية والمظهرية للنمو والفعالية الانزيمية للانزيم المحلل للبروتين والتنوع الوراثي وعلاقته بالتعبير الجيني بين عزلات الفطر الجلدي.Trichophyto rubrum ولهذا الغرض جمعت 150 عينة سريرية (قشطات جلدية (80), اجزاء شعر (60) | The present study was designed to characterize the morphological and the molecular patterns of growth, enzymic activity of proteases, genetic diversity and their correlation with gene expression among strains of Trichophyton rubrum. For these purpose, A total of 150 clinical specimens (skin scrapings (80), hair fragments (60) and nail clippings (10)) were collected from 100 patients (70 males&80 females) whom clinically diagnosed with dermatophytosis after attendingto the dermatology and venereal diseases centre of Mergan Teaching hospital and private clinic in Babylon city from Feb. 2014 to May 2014. T. rubrum were isolated and identified using cultural, biochemical and physiological tests. Isolates were then subjected for confirmation by PCR, genotyping using RFLP - PCR, sequencing and registration of these sequences in GeneBank for obtaining accession numbers then phylogeny. The growth conditions, were tested, in addition to, the genetic expression of proteases (exocellular protease and endocellular aminopeptidase) were determined using Real - Time PCR. Out of 150 specimens, 24 (16%) isolates were dermatophytic fungi and only 5 (20.8%) of them belong to T. rubrum. The influence of cultural conditions in growth of T. rubrum showed that the 30 ?C, pH 6, SDA medium and 7 days incubation were the optimal conditions for its growth. Proteolytic activity of isolates of T. rubrum against casein as a substrate revealed an ability to produce protease in solid and liquid media after 14 days of incubation. This activity was varies according to the type of isolate where the isolate No.1 (isolated from skin) gave a high proteolytic activity (5.6cm) in solid media and (80.1U/ml) in liquid media in comparison with other tested isolates (No.2 - No.8). on the other hands, the 9 days of incubation, 30 ?C, pH 7 and 0.5% substrate concentration were the optimal condition for proteolytic activity of these isolates. The molecular tests confirmed that all tested isolates belong to T. rubrum with amplicon size (601bp) after amplification of ITS1 primers using PCR technique. While the RFLP - PCR technique showed the presence of two genotypes (I%II) belong to T. rubrum with subgenotypes (Ia - Id) and (IIa - IId) respectively. The relative quantification of proteolytic activity (exocellular protease and endocellular aminopeptidase) produced by T. rubrum genotypes were expressed by using Real - Time PCR after amplification of the target gene of ptotease and aminopeptidase in comparison with housekeeping gene (? - actin) as a reference gene. The results showed the up - regulation of gene encoded to exocellular protease than the down - regulation of endocellular aminopeptidase produced by T. rubrum in the presence of casein as a substrate. The internal transcribed spacer 1, partial sequence, 5.8S ribosomal RNA gene and internal transcribed spacer 2, complete sequence and 28S ribosomal RNA gene, partial sequence of local isolates of T. rubrum were used for sequencing, registration in Genebank - NCBI and phylogeny. Five accession numbers were recorded and available to NCBI, EMBL in Europe and the DNA Bank of Japan. These accession numbers were : KP979787, KP979788, KP979789, KP979790, KP979791. Phylogenetic relation between local strains and world strain showed a high identicasl with T. rubrum (GQ376105.1). This is the first study in Iraq which employed sequencing, registration of sequences in Genebank - NCBI and carrying out phylogeny of local and world strains of T. rubrum.
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الكشف عن بكتريا الكوكسيلا بيرنتي في حالات الاجهاض في الانسان والمجترات الصغيرة في محافظة ذي قار Detection The Role of Coxiella Burnetii In Abortion of Human And Small Ruminants In The Thi - Qar Province

اسم المؤلف: عباس دخيل مطر جبر الجوراني
اسم المشرف: عبد الله كاظم هندي محمد عبد الله جبر
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بابل
المستخلص: الحمى المجهولة من الامراض الواسعة الانتشار في العالم تحدث بسبب جرثومة الكوكسيلا بيرنتي.هذه الجرثومة تسبب عدة امراض الاجهاض هو الاكثر حدوثا. المخاطر البيولوجية وانشار هذه البكتريا على النساء المجهضة والحيوانات في العراق غير معروفة. لذلك هدفت هذه الدراسة ال | Query fever is a worldwide distributed disease caused by Coxiella burnetii bacteria causes several disease main of this disease is abortion, the biological hazard and prevalence of this bacterium on the aborted woman and small ruminants are not known in Iraq. Therefore this study was aimed to detection and isolation of Coxiella burnetii as a causative agent of abortion in woman and female of small ruminants.352 samples were collected includes human samples and animals samples. A total of 120 blood aborted women, 7 breast milk samples, 20 placental samples and 50 blood samples from normal women as control groups, these samples collected from Bent Al - Huda hospital in Thi - Qar province. Animals samples, a total of 80 aborted animal blood samples, 15 milk samples, 10 placental samples and 50 blood samples, these samples from normal animals as controls group, these samples collected from veterinary hospital in Thi - Qar province. The results of this study showed a high incidence of abortion occurs in first trimester of gestation (41.666%) and high incidence in rural regions (64.16%) and also age group from 21 to 30 years old (50%).The methods used for detection of Coxiella burnetii includes serology tests, polymerase chain reaction (PCR) technique for confirmation serological test and isolation on embryonated chicken eggs and detection by PCR technique. Out of the 120 women serum samples analyzed by enzyme ELISA, the results of percentage of anti - Coxiella burnetii IgM and IgG in human samples were 36 (30 %). The percentage of anti - C. coxiella IgM in human samples were 10 (8.333%), while the percentage of anti - C. Coxiella burnetii IgG in human 26(%21.667%).. In animals total of (80) serum samples, the percentage of anti - C. coxiella IgM and IgG in animals' serum samples were 31(38.75%) The percentage of anti - Coxiella burnetii IgM in animals 10 (12.5%), while, the percentage of anti - C. coxiella IgG in animals are 21(26.25 %). PCR technique used for identification of Coxiella burnetii in human and animals samples by targeting three genes including outer membrane protein (com1 and com2), 16S rRNA and transposase insertion element (IS1111) genes. In human blood samples the com1 and com2 genes detected in 23of 120 (19.166%) samples and in breast milk samples 1 of 7 (14.28%). and not detected in placental samples. The 16S rRNA gene was detected in 16 of 120 blood samples (13.33%) and in breast milk samples and placental samples were not detected. The IS1111 gene was detected in 9 of 120 blood samples (7.5%) in human blood samples, also were not detected in milk and placental samples.. In animals blood samples the com1 and com2 genes were detected in 19 of 80 blood samples (23.75%) and were not detected in milk and placental samples. The 16S rRNA gene detected in the same percentage of com1 and com2 genes 19 of 80(23.75%) and in the milk samples and placental samples not detected. The IS1111 gene detected in 10 of 80 (12.5%) in animal blood samples, also were not detected in placental samples. An isolation of Coxiella burnetii which identified by ELISA and PCR via inoculated Coxiella burnetii buffy coat samples in embryonated chicken eggs from 6 to7 days old and then completed an incubated to 10 - 15 days, harvested yolk sac contains then detecting Coxiella burnetii by PCR. The results of PCR after inoculation of (43) samples were positive in PCR (100%). Statistical analysis revealed no significant different between ELISA test, PCR and isolation results in human and animals samples. These mean that we can use the ELISA in the primary diagnosis. The correlation coefficient was highly significantly between human and animal samples at 0.01%.
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تقييم قابلية بعض انزيمات السليليز الفطري على تخمير تخمير بقايا النباتات لانتاج الايثانول Evaluation of Capability of Some Fungal Cellulase In Fermentation of Plant Residues For Ethanol Production

اسم المؤلف: عذراء حرجان محسن الدحيدحاوي
اسم المشرف: فاطمة عبد الحسين التميمي محسن هاشم رسن
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2014
مكان الجامعة: النجف
المستخلص: هدفت الدراسة الحالية الى تقييم قابلية بعض انزيمات السليليز الفطري على تخمير بقايا النباتات على انتاج الايثانول من بعض الفطريات المحلية المعزولة من 50 نموذج من التربة والذي تضمن Aspergillus oryzae , A. niger, A. terreus , A.flavus , A.fumigatus, A.parasiti | The present study was conducted to proceed a Bioethanol production using some of the waste fermentation plant by cellulosic hydrolysis enzyme produced by some local fungi isolated from 50 samples from soil which were including Aspergillus oryzae, A.niger, A.terreus , A.flavus , A.fumigatus, A.parasiticus, A.nidulans, Penicilliuum chrysogenum, Trichoderma longi, Rhizopus stolonifer, Cladosporium spp, Mucor indicus, Trichothecium spp. The prominent isolates were A. oryzae, A.niger, A. terreus which were chosen for further studies and screening to producing cellulase enzymes the fungal isolates revealed variation values of clear zone It’s (5.1, 5, 5, 4.8) cm to A. oryzae, A.niger, A. terreus , A.flavus respectively. The results revealed that the suitable carbon source to the cellulase activity was induced in different raw plant substrate, the highest was produced when using Corn cobs, Rice husk and Reed reached to (1.72, 2, 1.26)IU/ ml respectively when using A. oryzae and (1.24, 1.17, 1.89) IU/ ml when using A.niger, and (1.31, 1.19, 1.93)IU/ ml when using A. terreus. but using the cellulose powder and CMC, Avical sole carbon source total cellulase given least activity compare with raw plant substrate.on the other hand, using Mandels - Weber medium activity of enzyme production by A. oryzae, A.niger, A. terreus recorded (6.51, 4.14, 5.61) U/ ml respectively when using Corn cobs and (5.79, 4.03, 5.24) U/ ml and (5.55, 3.97, 5.19) U/ ml when using Rice husk and Reed by different fungi. The results refers that , the best chemical pretreatment results were appear when using NaOH at 100Co in 30min on Corn cobs, the total cellulase activity were (1.63, 1, 1.21) U / ml when using fungal isolated to degradation after treatment, while using hot water at100Co without NaOH in 15 min to pretreatment Rice husk activity were (2.16, 2.07, 2.12) U / ml but Reed are appear high activity when pretreatment with NaOH at 100C0 in 15min activity It’s (2.28, 1.65, 2.09) U / ml. The results revealed that, The optimized conditions of the enzymes were different in this study that give activities of Fpase, CMCase and ? - glucosidases to fungus A. oryzae compared to the rest of the fungi using Corn cobs as carbon source, it reached to(2.21, 2.30, 46.72) IU / ml respectively when pH6, and temperature tested for these enzymes recorded (1.64, 1.61, 30.86) IU\ ml respectively at a temperature of 30 C° and when using the concentration of substrate gave activity (2.4o, 2.48, 42.34) IU / ml of 6 % from Corn cobs and recorded when using nitrogen source concentration at 6 % (1.92, 1.89) IU\ ml in KNO3 and (NH4)2SO4 to Fpase and CMCase and the effectiveness of ? - glucosidase at the same nitrogen source (43.54, 50) IU / ml. on the other hand, showed enzymes Fpase, CMCase and ? - glucosidase high activities to ? - glucosidase, CMCase to fungus A. oryzae compared to the rest of the fungi using Reed as carbon source reached (2.31, 2.21, 46.15) IU/ml respectively at pH 6, and at different temperatures were (1.66, 30.94) IU/ml to Fpase and ? - glucosidase at 30 C°, but CMCase gave the highest efficacy (1.60) IU \ml at 25 C° and decreased effectiveness to (2.60, 2.57, 49.69) IU / ml at 6 % of the carbon source recorded effectiveness of the enzyme Fpase (1.19, 1.09) IU / ml when using (NH4)2PO4 and (NH4)2SO4 at 6 % as a nitrogen source of from either enzymatically CMCase and ? - glucosidase was effective at same nitrogen source (1.29, 2) and (35.19, 40.41) IU / ml respectively.The results showed when you use a Rice husk recorded higher effective enzymatic Fpase and CMCase and ? - glucosidases of fungus A. oryzae was (2.32, 39.27) IU /ml for Fpase and ? - glucosidases at pH 6 but CMCase gave the highest efficacy at 5 pH as recorded (2.04) IU / ml at temperatures 30 C°, reached to (1.64, 1.58, 29.13) IU / ml respectively for Fpase and CMCase and ? - glucosidases at a temperature of 30 C°, and when using 6% of the substrate was effective (2.47, 2.19, 44.86) IU / ml, activity recorded when using (NH4)2PO4 and (NH4)2SO4 at 6 % as a nitrogen source of (1.1) IU / ml to FPase on either enzymatically CMCase and ? - glucosidase was effective when use same a nitrogen source (1.18, 1.83) and (31.92, 36.91) IU/ml, respectively. In addition, The results refers when precipitation by ammonium sulfate specific activity were (38.69, 1.25, 5767.5) IU of the enzyme with 0.012 mg protein and then, Elution protein of crude enzyme solution of A.oryzae from DEAE - cellulose column was shown to the protein was separated in (33) fractions.It was found that only the fractions (F - 8) contained cellulase activity, while fraction (F - 30) eluted in column containing buffer and different concentrations of NaCl then using gel filtration and gave three peak of enzymes alone and Characterize enzymes purified FPase activity was found at 5.5 pH which (0.91) U and temperature was having activity at 25 and 30 C°, were (0.14, 0.15) U and CMCase found that the enzyme exhibited maximum activity at 25 C° was (0.33) U but 30 C° for ? - glucosidase was (38.22) U and the molecular weight of the protein was found to be about (38, 52 and 49) kD for (FPase, CMCase, ? - glucosidase) respectively. Finally, estimated ethanol density (0.80 - 0.91) g /ml and ethanol concentration (60 - 97) % but using chromic acid test of purified ethanol the complex enzyme with Reed, complex enzyme with Corn cobs recoded high value of ethanol which were (0.98, 1.72, 1.87, 1.89 ) % and (1.09, 1.83, 1.92, 1.99) % respectively but complex enzyme with Rice husk were given (1.01, 1.12, 1.48, 1.79) % in 4, 5, 6, and 7 days respectively on the other hand, when using the A.oryzae and A.terreus with Reed gave high ethanol value from other fungi with substrate in this study, and observed Rice husk in all results was recorded less absorbance at 350 nm and determination of ethanol concentration by gas chromatography mass used ethanol concentration 99 % as standard, also we showed the ethanol when using enzyme pure with substrate in which were (93.7, 97.1, 88) % ethanol to enzyme with Corn cobs and Reed and Rice husk respectively
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دراسة بعض المؤشرات المناعية لدى بعض المرضى المصابين بفيروس المليساء المعدية في محافظة ديالى A Study of Some Immunological Parameters In Some Patients With Molluscum Contagiosum In Diyala Province

اسم المؤلف: رغد ياسين اعويد
اسم المشرف: عباس عبود فرحان الدليمي
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: ماجستير
اللغة: الانكليزية
السنة: 2014
مكان الجامعة: ديالى
المستخلص: المليساء المعدية مرض يسببه فيروس (MCV) الذي ينتمي لعائلة POXVIRUS.المليساء المعدية حيث انه مرض غير قاتل وشائع في جميع انحاء العالم, قد يشترك مع اورام المليساء التهابات قليله, المليساء المعدية يستمر لشهور او سنوات..اجريت الدراسة الحالية للفترة من 1 تشرين ا | The molluscum contagiosum (MC) virus (MCV) is a dermatotropic poxvirus, and the causative agent of MC. MC is nonlethal, common andworldwide. Additionally, little inflammation is associated with MCpapules, and MC can persist for months to years. The present study was conducted for the period from1 November 2013 to 30April, of 2014 in outpatient clinic of Baquba Teaching Hospital in city. The study aimsat assessing the immune status of patients infected with molluscumcontagiosum through the measurement ofthelevel of immunoglobulin(IgG, IgM)and the level of complementcomponents especially (C3, C4) by radial immune diffusion assay and measurement of the level of interleukin 18 R1 by Elisa assay which immune responses are key for the eventual resolution of MC. 75 patients were diagnosed with clinical lesions of MCV on different areas of the body, age of patients ranged from(2 - 50 years) including 40(53.3%) males and 35(46.7%) females.The control were 26.6±15.4 years (range from 2 - 50 years), 8 (53.3%) of them were males and 7(46.7%) females.After theexamination by single radial immune diffusion and Elisa kit.The results showed the patientswere equal or less than 16 years, 24 (32%) from 17 - 30 years, 19 (25.3%) from 31 - 45 years and 12 (16%) above 45 years and 40(53.3%) males and 35(46.7%) females, no static significant difference showed between the MCV infection and either the sex or age.The results revealed the level of IL 18R1in patient the mean±SD were 677.15±874.22 ng/ml while in control were 178.46±31.79 ng/ml. There was significant statistical difference between both groups as patients with MC had high level of IL18R1 than control.Themean±SD of IgMinpatients were 1946.6±825.6 mg/dlwhile in control were 140.1±68.7mg/dl This result was highly significant which indicates that patients with MC had higher level of IgM than control In contrast, patients had lower levels of IgG than control, The mean±SD of IgG in patientwere 221.9±96.7mg/dl while in control were 1229.9±299.7mg/dl. This result was highly significant. Also the patients had lower level of C3 and C4 than control. The mean±SD of C3 in patients were 109.6±64.8mg/dl while in control were 120.8±22.1mg/dl. The mean±SD of C4 in patients were 27.8±12.7mg/dl while in control were 38.7±9.8mg/dl. These results were statistically significant. Eleven 9 (14.7%) of patients had positive CRP while 12 (80%) of control had positive CRP which was statistically highly significant. Forty seven (62.7%) patients were from rural area while 28 (37.7%) from urban. There is no statistical difference between both groups as. Family history of MC and Recurrence of MC in patients were nostatically significant difference to infection by MCV.
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دراسة امكانية الاستفادة من تركيز بروتين المصل الفعال نوع سي بروتين التفاعلي CRP في التشخيص المبكر للاصابات البكتيرية لحديثي الولادة وتحديد توقف اعطاء العلاج من خلاله The Study of Usefulness of C - Reactive Protein Concentration In The Early Diagnosis And Determining The Duration of Antibiotic Therapy of Suspected Neonatal Bacterial Infection

اسم المؤلف: قاسم ابراهيم حسين
اسم المشرف: ايمان ناطق ناجي البياتي
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: ماجستير
اللغة: الانكليزية
السنة: 2014
مكان الجامعة: بغداد
المستخلص: ان التشخيص السريع لاصابات تجرثم الدم في الاطفال حديثي الولادة من الامور المهمة وذلك لخطورة الاصابة المؤدية الى الموت.الغرض من دراستنا هو امكانية استخدام فحص سي - البروتين الفعال في التشخيص المبكرلهذا المرض البكتيري وكذلك استخدامه في تحديد نقطة التوقف عن ا | Rapid diagnosis of bacterial infections of newborn infants is very important itcause of morbidity and mortality. Aim of the studyThe aim of the studyis to examin the possibility of using C - reactive protein in the early diagnosis of neonatal sepsis, as well as its use in determining thestop point of the antibiotic treatment.Patent, Materials and methods This study was conducted at the Hospital of Ibn - Albaladi for children and women in Rusafathe Healthoffice / Baghdad.Iraq from February 2013 to February 2014.The number of patients with bacterial infection was 102 (62%) of the total number of patients (165) who were admitted to the neonatal care unit aged less than 28 days and their weights was more than 1500 g,. The number of males was 92 (56%) and the number of females was 73 (44%), the number of patient withearly onset sepsis EOS (less than three days) was 43 (42%) and with late onset sepsis LOS for (more than three days) was 59 (58%), 43 (57%) ofEOSwas normal deliveries infected and cesarean deliveries infected 33 (43%), while51 (57%) of LOS was normal deliveries infected and cesarean deliveries in LOS was 38 (43%). Blood samples were collected and divided into three portion : bloodculture, CRP andhematological profile.1 - Blood culture.Blood was collection from neonatal sepsis patientinto special blood culture bottle and incubate in bact /Alert 3 D device when growth of bacteria then subcultured on blood agar, MaCconkey agar, and incubatedusingvitek /2 to diagnose bacterial infection anddeterminetheir susceptibility to antibiotic treatment.ResultThe Compact results indicated that 63 (38%)of patient were negative and 102 (62%) were positive out of them 23 patients (22.6%)were infected with gram negative bacteria including {Klebsiella pneumonia 30%, Acinetobacterlowffi 17%, Enterobacter cloaca 13%, } while 79 (77.4%) were infected with gram positive bacteria including { staph Homins and staph hemolyticus each 28% then staph epidermidis 13% and staph aureus 10%, while strep spp was 6% }. 2 - c - reactive protein.It was done by twomethod for the same samples in order to characterize the accuracy of the diagnosis. The first method was agglutination test (method 1) the cut off value for positive result ? 6 µg % 79 (48%)were negative, while 86 (52%) werepositive, while the second method was immune test (method 2) µµµusing Nycocard device werethe cut off value for positive result ? 5 µg % the results 102 (62%) were positive while 63 (38% negative.3 - hematological profile.The hematological profile of our patient included PCV, WBC, neutrophil, lymphocyte, monocyte, count were measured using Emerald automated Hematology device. The results of these tests werenot compatible with the results of the blood culture, but normal result were compatible with negative blood cultures. The results adopted of blood culture confirm the presence of bacterial infection and then compared with theresults CRP test and blood profilefor the same samples.Negative result of blood culture and CRP test were 54 (33%) while positive results of blood culture and CRP test were recorded and improved after taking treatment for (, 3, 5, 7,) days for (40, 31, 5)patients respectively.ConclusionWeconcluded C - reactive protein test was useful in the diagnosis of bacterial blood infections and determine the stop - point otreatment. Gram negative bacterial isolates were sensitive impinim, livofloxacin and Amikacin while they were resistant to Ampicillin, Ampicillin /Sollbectam and Gentamycin. on the other hand gram positive bacteria were sensitive to Amikacin, Ampicillin/sulbactam Ciprphloxacin, and Tigecyclin and were resistant to Erythromicin, and Oxacillinand Benzylpencillin.
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دراسة كفاءة بعض المضادات الحيوية ذات المناشئ المختلفة على بعض انواع البكتريا المعزولة من حالات مرضية مختلفة The Efficacy Study of Some Antibiotics From Different Origins On Some Pathogenic Bacteria Isolated From Different Cases

اسم المؤلف: ليلى عاصي خزعل
اسم المشرف: نجدت بهجت مهدي
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: ماجستير
اللغة: الانكليزية
السنة: 2014
مكان الجامعة: كركوك
المستخلص: شملت الدراسة جمع 278عينة سريرية تضمنت نماذج من اخماج المجاري البولية ومسحات الجروح والحروق وعينات الدم ومسحات الاذن الوسطى من مستشفى كركوك العام ومستشفى طوز العام وللمدة من ( 2013 - 4 - 15) ولغاية ( (2013 - 10 - 15. اظهرت نتائج الزرع البكتريولوجي الاولي | The study included the collection (278) clinical samples included the same of urinary tract infections and swabs of wounds and burns, and blood samples and swabs from ear from Kirkuk General Hospital and Tuz General hospital in period from (2013 - 4 - 15) until (2013 - 10 - 15). The results of the first Bacteriological implant using the blood Agar and Macconkey agar emergence of planting positive in (148) of the total sample (278) sample rate (53.33%). Diagnosed colonies developing depending on planting characters and biomicroscopy and tests biochemical as possible diagnosis (90) the isolation of clinical divided to (30) isolation of each of the bacteria (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa) divided to (28) isolates of Urine and 21 isolated from wounds and 25 isolates of burns and 11 isolated from blood and 5 isolates from the ear of the total isolates, confirmed diagnosis using the API Staph System and API 20E system. The study included examination of sensitivity using discs toward the 15 antibiotic for three types. The isolates showed to different in resistant rate, and was Anti Chloramphenicol is the most effect on the bacteria S.aureus where all isolates were sensitive to him 100%, and was an anti Imipinem, Nitrofuranation the most effect on bacteria E.coli where the percentage of sensitivity to two anti - (90%), either Anti Amikacin was the most impact on the isolates of Ps. aeruginosa where all isolates were sensitive to him ratio (100%) I tested the sensitivity of bacterial species toward (5) antibiotic (Amoxicillin Augmantine, Cefotaxime, Ciprofloxacin, Tetracyclin) concentrations of certain several different origins for each antibiotic using the method of casting dishes, use an anti Amoxicillin concentration (25) mg / ml and origins different (Iraqi, Indian, Turkey, United Arab Emirates) The results show that the origin United Arab Emirates is the most efficient than the rest of other. Use the adversaries (Cefotaxime, Augmantine) concentration (10) mg / ml and three - origins (Turkish, Indian, United Arab Emirates), oukd show that Origin United Arab Emirates is the most efficient of the rest of the origins Other. also used anti Tetracyclin concentration (10) mg / ml for three origin (Iraqi, Indian, Chinese). show that Origin Iraqi is the most efficient of the rest of the origins Other.and used anti Ciprofloxacin concentration (5) mg / ml for four origin (Iraqi, Indian, Turkish, United Arab Emirates) and show that the United Arab Emirates and Turkish origin is more efficient than the rest of other origins.Bacterial isolates showed multiple drug resistance to antibiotics where all isolates Ps. aeruginosa with resistance to multiple antibiotics by (100%) and showed isolates of S.aureus resistant multi - rate (93.3 %) and showed isolates of E.coli (96.6%) as well characterized bacterial isolates in multiple resistance to antibiotics Alpittalaktam particularly where the rates of multiple resistance (93.3 %) of the isolates of E.coli and (100%) isolates Ps.aeruginosa and (93.33%) of the isolates by S.aureus.the rapid Iodine standard method was used for screening for susceptibility of bacterial isolates to produce enzymes Beta - Lactamase have shown (73) the isolation of a positive result from the total (90) isolation rate (81.1%), were distributed among the isolates produced (27) isolated from bacteria by S.aureus (90 %) and (25) isolates of E.coli bacteria by (83.3%) and (21) isolated from bacteria Ps. aeruginosa (70%).to detect broad - spectrum Beta - Lactamase enzymes is used discs method adjacent and the results showed that there are (24) isolation of the total (73) is positive for examination by (32.87%) distributed on (12) isolated from bacteria S.aureus by (44.44%) and (5) isolates of E.coli bacteria (20%) and (7) of the bacterial isolates Ps. aeruginosa by (33.33%).Tested susceptibility bacterial isolates to produce mineral Beta - Lactamase enzymes Metallo ? - Lactamase using IMP - EDTA Combination disc and three isolates can from the production of the enzyme (4.10%) by two isolates of bacteria Ps.aeruginosa by (9.52%) and one isolate of the bacterium E.coli (4%).
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التحري عن جين الاوتولايسين في المكورات العنقوديه البشروية المقاومة لمضاد الفانكومايسين Autolysin Gene Detection In Vancomycin Resistant Staphylococcus Epidermidis

اسم المؤلف: مريم خميس عبد ربة بريس
الجامعة: جامعة بغداد
اسم المشرف: مي طالب فليح
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: ماجستير
اللغة: الانكليزية
السنة: 2014
مكان الجامعة: بغداد
المستخلص: من مجموع مئة عينة سريرية جمعت من مصادر مختلفة شملت الحروق والدم والجروح ومسحات انفية, تمكنت 90 عزلة من النمو على اكار المانيتول الملحي , ومن بينهم 40(44.4 %) عزلة عائدة لبكتريا المكورات العنقودية المنتجة للانزيم المخثر للبلازما و50 (55.5%) عائدة لبكتريا ا | Out of one hundred clinical samples were taken from different sources which include burns, blood cultures, wounds and nasal swabs infections ; 90 isolates developed growth on mannitol salt agar. Among these, 40 (44.4%) were Coagulase positive (Staphylococcus aureus) isolates, 50 (55.5%) belong to coagulase negative staphylococci in which Staphylococcus epidermidis isolates were 30(60%). The pattern of antibiotic susceptibility of Staphylococcus epidermidis isolates to 12 antibotics (Amoxiclav, Ceftazidim, ciprofloxacin, clindamycin, Erythromycin, Gentamycin, Imipenime, Penicillin G, Tetracycline, Rifampin , Methicillin, and Vancomycin) were determined using disc diffusion method. The results revealed that resistance to Penicillin G10 and Amoxiclav (Amoxicillin - clavulanic acid) were 100%, Methicillin were 93%, Erythromycin were 90%, Gentamycin and Clindamycin were 70%, Tetracycline and ceftazidim were 75%, Ciproflaxacin were 60%, Rifampin were30%. 95% of S.epidermidis isolates were sensitive to Imipenim and 5% of them were intermediate resistant, while these isolates showed 90% sensitivity to vancomycin. 19 isolates were multidrug resistance. Minimum inhibitory concentration of S.epidermidis isolates to vancomycin, was determined. The results revealed that (12) S.epidermidis isolates (40 %) were vancomycin resistant, the MIC of them were between 256 ?g\ml and 32 ?g\ml, (4) S.epidermidis isolates (13.3%) were intermediate resistance, the MIC to 3 of them were 16 ?g\ml and the last was 8 ?g\ml. Some virulence factors of VRSE and VSSE were detected including the hemolysin, protease, lipase and urease. The S. epidermidis isolates were produce hemolysin, protases, lipase and urease, were 100%, 100%, 25%, 100% respectively in VRSE, while in VSSE 100%, 100%, 60%, 100% respectively. The isolates were subjected to polymerase chain reaction (PCR) technique in monoplex pattern to amplify resistant incoding gene : the vanA, vanB and autolysine gene aae gene. The results by this study showed that 12 (40%) S. epidermidis isolates gave the implicone size (1030 base pair) of the vanA gene. However the results of MIC and PCR were similar but no any isolates gave product for presence of vanB gene. All S.epidermidis were able to produce implicone size(858bp) of aae gene. The effect of vancomycin resistant S. epidermidis on cell autolysis activity was detected by whole cell autolytic assay.The results revealed that there was significant difference among three isolates, the VSSE isolate (S.epidermidis 22) have the highest autolytic activity in the presence of antibiotic, followed by the VRSE isolate (S. epidermidis 1) and the VISE isolate (S. epidermidis14) which was the lowest autolytic activity with the presence of antibiotic. The result of transmission electron microscope (TEM) showed that the VRSE isolates (S.epidermidis 1) have thicker cell wall followed by VISE (S.epidermidis 14) isolates.However, the VSSE (S.epidermidis 22) didn't showed any cell wall thickening.
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التحري عن التعبير الجيني لل FOXP3 وTGF - ?1 باستخدام الطرائق الجزيئية والمناعية في سرطان الرئة اللاصغير الخلية Detection of FOXP3 Gene Expression And TGF - ?1 Using Molecular And Immunological Methods In Non - Small Cell Lung Carcinoma

اسم المؤلف: سهاد فيصل حاتم المقدادي
اسم المشرف: امنة نصيف جاسم بان عباس عبد المجيد
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2014
مكان الجامعة: بغداد
المستخلص: Studies have linked FOXP3 and TGF - ? expression to the outcome of certain cancers. FOXP3 is a marker known to be expression in T - regulatory cells while TGF - ? is a secreted protein usually detected in the extra cellular matrix.The present study aimed at focusing on the identification of immune markers namely FOXP3 and TGF - ? with their expression patterns in lung cancer patients as a useful tool to predict disease progression.Also it is aimed to design molecular evaluation of m RNA expression of both FOXP3 and TGF - ? in peripheral blood mononuclear cells and bronchial (brush) cells of patients with lung cancer and benign lesions, using qRT PCR; determining the T - reg level in the peripheral blood employing the High Rsolution Melting (HRM) as a novel method to detect Treg - specific demethylated region (TSDR); molecular DNA analysis of somatic mutation of exons 3, 6, 7 of FOXP3 in patients with lung cancer tissue and benign lesions and immunohistochemical (IHC) estimation of FOXP3 and TGF - ?1 in T - reg and cancer cells in formalin fixed paraffin embedded(FFPE)lung cancer tissue and benign lesions.Blood samples were collected from 30 patients with newly diagnosed, non small cell lung carcinoma and 30 patients with benign lesions.Patients were recruited at The Specialized Surgery Hospital and Oncology Teaching Hospital/Baghdad.Samples from 16 apparently healthy donors were used as control during the period from June 2012 to June 2013. The samples preservation with TRIzol reagent were subjected to molecular study including RNA and DNA extraction; reverse transcription; RT - PCR; HRM assay and DNA sequencing were done in the Molecular Oncology Unit/Guys and ST Thomas? s hospital/ King College/London/UK.The expression level of FOXP3 was high in 16(61.5%)in lung cancer cases.A significant difference was noticed between cancer cases from one side and benign lesions or healthy control on the other side p<0.05.Mean of FOXP3 expression(fold change)was significantly high(2.64±0.09)in cancer cases than in benign cases(1.32±0.04)and healthy control(1.38±0.06)with p<0.05.A significant association between high expression level and >60 age and squamous cell carcinoma in cancer cases P<0.05.The expression level of TGF - ? was high in 16(61.5%)in lung cancer cases.A significant difference was noticed between cancer cases from one side and benign lesions or healthy control on the other side p<0.05.Mean of TGF - ? expression (fold change) was significantly high (6.27±0.56) in cancer cases more than healthy control (2.87±0.09) with p<0.05.The association was significant between TGF - ? expression level (high and low) and age>60in cancer and benign groups (p<0.05), while no significant association with gender and cancer types were noticed.For FOXP3 mRNA expression in bronchial brush cells, the result showed no significant difference between the mean fold change of malignant(3.57 ± 0.06) and benign(4.02 ± 0.06) patients. The low expression was predominated both in cancer and benign cases. No significant differences were found between FOXP3 expression (high and low) and age; gender ; cancer types..According to FOXP3 T - reg specific demethylated region detection, results showed that the mean percentage of FOXP3demethylation in lung cancer patients (4.32 ± 0.04) was significantly higher than in benign lesions (3.22 ± 0.02) patients andhealthy controls(3.33±0.04). A positive correlation coefficient with high significant, was found in the group of cancer samples (R² = 0.6653;r = 0.69;P : 0.0017)on correlating percentage of Treg and demethylation of FOXP3 from one side with its m RNA expression on the other side.In benign lesion group was(R² = 0.5334;r= 0.59;p= 0.0027), While in the control group a positive correlation but a weak significance was found (R²= 0.2383;r=0.28;P=0.0437).FOXP3 gene sequencing revealed high frequency of missense mutations c.715 GTA>CTA : V 239 L in 17 (94.44%) in malignant sample and non cancerous cases7 (87.5%)without statistical differences. Missence mutations were also detected in exone 3 in 3(16.67%) cancer cases and in 1(12.5 %) benign lesion.No missense mutations could be detected in exon 6. Intronic mutations and silent mutation were variable in three exons without statistical differences. Many cases of adenocarcinoma have shown multiple mutations either of missense or Intronic types. Missense mutations of exon 7 were correlated significantly with an age of 60?years. Exon 3 mutations were significantly associated with adenocarcinoma. Positive FOXP3 Immunohistochemistry (IHC) staining in tumor cells was associated with high missense mutations frequency 10(55.55%) in exon7, while in exone 3 was 2(11.11%). Negative FOXP3 IHC staining in the tumor cells was associated with seven missense mutations in exon7 and one (5.55%) in exon 3, in addition 4(23.53%) cases of the exon 7 missense mutations were associated with negative FOXP3 expression in lymphocytes.The result showed that FOXP3 by using IHC staining was positive in 21(70%) of nuclei of cancer cells, and 22(73.3%)in Treg infiltrates.The positive cancer cells and Treg infiltrates associated significantly with age>60 (p<0.05).No significant association was found withgender, cancer type, while there is association with moderate differentiation compared to poorly differentiation (p<0.05). High frequence of FOXP3 expression score 3 and high intensity were appeared in nuclear cancer cells compared to benign lesions cells, while Treg infiltrates with score 1 and high intensity was high frequency in malignant and benign.The result showed that TGF - ?1 by using IHC staining was positive in 25(83.3%) in the cancer cells and 21(70%) in stromal cells. No significant difference was noted between positive expression in malignant and benign lesions p>0.05.No significant association was noticed between positive cells expression and age, gender, cancer type and differentiation p>0.05. High frequency of TGF - ?1 expression score 3 and high intensity in malignant cells and benign. Also stromal cell expression score 3 and high intensity were predominated in malignant and benign lesions. The high and moderate intensity expression was more frequent in matrix surrounding cancer cells compared to non cancerous.Total agreement and kappa coefficient between FOXP3 and TGF - ?1were poor in malignant and benign epithelial cells and stroma, while the perfect agreement was between expression of TGF - ?1in stromal cells
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تعدد الاشكال الوراثي للحركيات الخلوية وHLA - DQB1 في مرضى السل الرئوي Genetic Polymorphisms of Cytokines And HLA - DQB1 In Pulmonary Tuberculosis Patients

اسم المؤلف: خلود كريم حسن
الجامعة: جامعة بغداد
اسم المشرف: علي حسين ادحية
الموضوع العام: علوم الحياة
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2014
مكان الجامعة: بغداد
المستخلص: The present study aimed to understand the correlation between serum level of nine cytokines (IL - 1?, IL - 1RA, IL - 2, IL - 4, IL - 6, IL - 10, IL - 12, IFN - ? and TNF - ?) and their genetic polymorphisms at 16 gene positions defined by sequence specific primer - polymerase chain reaction (SSP - PCR) in pulmonary tuberculosis (PTB) patients, and in addition HLA - DQB1 gene polymorphism was also defined by SSP - PCR to determine their role in susceptibility or resistance to M. tuberculosis. Finally, serum level of cortisol was also determined in the patients.Ninety four Iraqi Arabs PTB patients (70 males and 24 females) were enrolled in the study. They were referred to the Institute of Chest and Respiratory Diseases in Baghdad for diagnosis and treatment during the period May - October 2012. A control sample of 80 apparently healthy individuals was also included and matched patients for gender (60 males and 20 females) and ethnicity. The results are summarized in the following : 1. A significant increased serum level of IL - 1? (24.16 ± 8.82 vs. 3.20 ± 1.18 pg/ml), IL - 1RA (41.31 ± 6.64 vs. 16.85 ± 5.50 pg/ml), IL - 2 (17.63 ± 3.53 vs. 7.80 ± 1.10 pg/ml), IL - 4 (9.56 ± 2.60 vs. 3.81 ± 1.70 pg/ml), IL - 10 (34.49 ± 4.60 vs. 7.61 ± 1.70 pg/ml), IL - 12 (25.16 ± 5.85 vs. 7.70 ± 1.12 pg/ml) and TNF - ? (22.52 ± 4.41 vs. 4.97 ± 1.15 pg/ml) was recorded in PTB patients compared to controls. Also, Cortisol serum level was significantly increased in patients (215.47 ± 1.33 vs. 38.63 ± 1.74 ng/ml).2. Cytokine gene polymorphism analysis revealed that neither genotypes nor alleles of IL1A - 889, IL2 - 330, IL2+166, IL4 - 590, IL4 - 33, IL6+565, IL10 - 819, IL10 - 592, IL12B - 1188 and TNF - 238 genes showed a significant variation between PTB patients and controls. In contrast, the frequency of TT genotype of IL1RN gene at position mspal 11100 showed a significant (P = 0.004) increase in PTB patients compared to controls (65.9 vs. 43.7%). For IL4 - 1098, the frequency of TT genotype was also significantly (P = 0.048) increased inpatients (82.9 vs. 70.0%). At position - 174 of IL6 gene, a significant (P = 0.002) increased frequency of GG genotype was observed in patients (55.3 vs. 31.2%). For IL10 gene, only GG genotype at position IL10 - 1082 was observed with a significant (P = 0.045) increased frequency in patients (18.1 vs. 7.5%). At position - 308 of TNF gene, a significant (P = 6.9 x 10 - 5) decreased frequency of GG genotype was observed in patients (60.6 vs. 87.5%), while GA genotype was significantly (P = 1.3 x 10 - 4) increased (38.2 vs. 12.5%). Finally, the frequency of AA genotype of IFNG gene at position +874 demonstrated a significant (P = 0.006) increase in PTB patients (55.3%) compared to controls (33.7%).3. To determine the impact of cytokine genotypes on cytokines serum level, PTB patients and controls were distributed according to their serum level in the three genotypes of each cytokine. It was found that CC genotype of IL1RNmspal 11100 in patients was observed with the highest IL - 1RA level (52.16 ± 5.81 pg/ml) compared to TT (41.39 ± 3.23 pg/ml) or TC (38.10 ± 4.54 pg/ml) genotype. The TT genotype of IL2 at position - 330 also showed the highest level of IL - 2 (22.16 ± 4.31 pg/ml) compared to TG (17.59 ± 3.40 pg/ml) or GG (13.68 ± 3.53 pg/ml) genotype in patients. The IL4 - 1098 TT genotype showed the highest level of IL - 4 in patients (10.38 ± 2.21 pg/ml) compared to TG (6.09 ± 1.20 pg/ml) or GG (3.93 ± 0.80 pg/ml) genotype. For IL10 gene, the GG genotype of IL10 - 1082 recorded the highest level of IL - 10 (40.67 ± 2.96 pg/ml), which was significantly different from AA genotype (26.66 ± 5.65 pg/ml). At position - 308 of TNF gene, serum level of TNF - ? in GG genotype of patients demonstrated a significant increased mean compared to genotype GA (24.76 ± 1.30 vs. 19.15 ± 1.12 pg/ml). At position - 238, TNF GG genotype showed a significant increase level of TNF - ? (23.02 ± 2.91 pg/ml) in patients compared to AA genotype (17.18 ± 1.53 pg/ml) of patients. Finally, IFNG+874 AA genotype was observed with the highest IFN - ? level in patients (11.07 ± 1.12 pg/ml) compared to AT (7.97 ± 1.81 pg/ml)or TT (6.10 ± 2.20 pg/ml) genotype. In contrast, no such differences were observed in controls.4. Out of the five encountered HLA - DQB1 alleles, DQB1*03 showed a significant (P = 0.005) increased frequency in PTB patients compared to controls (71.3 vs. 50.0%). It was also observed that heterozygosity at such gene locus was significantly (P = 0.03) more frequent in patients than in controls (93.6 vs. 82.5%), while homozygosity was observed with a less percentage frequency in patients compared to controls (6.4 vs. 17.5%) and the difference was also significant (P = 0.03).Accordingly, it is possible to conclude that the cytokine profile was deviated in PTB patients, and such deviation was correlated with the genotypes of some cytokines, which might also together with HLA - DQB1polymorphism confer the individual an immunogenetic predisposition to develop M. tuberculosis infection.
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دراسة الملف المصلي وتقييم بعض الخصائص المناعية لدى مرضى الحزام الناري Serum Profile Study And Evaluation Of Some Immune Features Among Patients With Shingles

اسم المؤلف: زينب عبد النبي طليفح النصراوي
اسم المشرف: حبيب صاحب نهر محمد كاظم طاهر الحطاب
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2017
مكان الجامعة: بابل
المستخلص: اجريت الدراسة الحالية للفترة من 1شباط 2015 لغاية 3شباط 2016 وجمعت خلالها عينات المرضى المصابين بالحزام الناري من قسم الامراض الجلدية في العيادات الاستشارية في مستشفى مرجان التعليمي للتحري عن بعض الجوانب المناعية في المرضى, وكان العدد الكلي للمرضى 50 مريضا | This study was conducted during the period from 1st February 2015 to 3rd February 2016, and the samples were collected from dermatology department of the consultant clinic in Marjan Teaching Hospital to investigate certain immunological markers of patients with shingles. The total number of patients were whose their ages ranges were 14 - 80 years. Age group (40 - 61) years have the highest percentage of infection reaching 46%.The immunofluorescence test for Varicella Zoster virus - specific IgG showed 48 out 50 are positive(96%). The higher percentage of infection with shingles was in female (54%), while male patients the percentage was(46%).The results showed that patients with blood group (O+) have 50% of infection rate.When CD4 was investigated by ELISA test , the results showed that the mean± SD of serum level of CD4 in shingles patients and control group is (6.70± 0.97 and 9.36±2.02) ng/ml respectively. The concentration of CD4 was lower in patients with shingles than healthy control group, while concentration of CD8 and CD56were higher than healthy control group ( the mean± SD of serum level of CD8 in shingles patients and control group was 21.42±5.43 and19.11±3.29 ng /ml respectively, whereas the mean ± SD of CD56 concentration in shingles and control group were 107.58± 40.39 and 67.59 ± 36.31 ng/ml, respectively).The results also showed that the concentrations of IFN ? and TNF? were lower in patients with shingles compared with non - infected persons , as the following : the mean± SD of serum level of IFN ? in shingles patients and control group was 184.31±21.95 and 218.03±26.21 pg /ml respectively, and the mean ± SD of TNF? concentration in shingles and control group were 51.55± 5.14 and 62.35 ± 6.74 pg/ml, respectively. For IL 10, the concentration was higher in patients with shingles than that in the healthy control group; the mean± SD of serum level of IL 10 in shingles patients and control group was12.42±5.59 and 4.47±0.90 pg /ml respectively. This study was concluded following : • Gender represents a risk factor for the occurrence of shingles mainly at ages over 40 years old (higher in females than males).• Varicella Zoster virus specific IgG is a sensitive and specific test that can be used for the laboratory diagnosis of shingles.• Varicella Zoster virus reactivation has a suggested link with decreased level of soluble CD4 molecules and, for lesser extend with increased soluble CD8 and CD56 molecules in patients serum. • Blood group O+ might be consider as a risk factor for reactivation of VZV infection as the majority of patients were within this blood group.• A diminished role of the inflammatory TH1 cells in reactivated shingles patients is more likely as evident by the reduced concentration of IFN? and TNF? profiles. The Treg cells (CD4+ CD25+) seems to play a role in such reduction of TH1 proinflammatory cytokines as evident by the significant elevation of IL10 in shingles patients.
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عزل وتحديد هوية فيروس الحصبة في علاقته ببعض الجوانب المناعية من الاشخاص النازحين الملقحين في محافظة بابل Isolation And Identification Of Measles Virus In Relation To Some Immunological Aspects From The Vaccinated Displaced Individuals In Babylon Governorate

اسم المؤلف: ليث احمد عمران كيف الكيف
اسم المشرف: محمد عبد كاظم حسن السعدي يونس عبد الرضا كحيوش الخفاجي
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: ماجستير
اللغة: الانكليزية
السنة: 2016
مكان الجامعة: بغداد
المستخلص: اجريت الدراسة لتقييم مناعة المجتمع لكلا النازحين والمقيمين الملقحين بلقاح فيروس الحصبة ضد خمج الحصبة في محافظة بابل. لذا تضمنت هذه الدراسة معايرة الحالة المناعية في امصال النازحين والمقيمين محافظة بابل مع عزل وتشخيص فيروس الحصبة من الحالات المشتبه بها. ت | The study was conducted to evaluate the herd immunity for both the displaced and residents vaccinated with measles virus vaccine against measles infection in Babylon governorate. Therefore this study includes the evaluation of the immune status in sera of Babylon governorate displaced and residents with specific aspect for isolation and identification of measles virus from suspected cases. The immune response was evaluated by means of ELISA test (enzyme linked immunosorbet assay) for titration of IgM and IgG immunoglobulin level, in addition to evaluation of perforin level in the sera of 90 measles vaccinated individuals included in this study (50 displaced and 40 residents) from different areas in Babylon governorate, that were collected during the period extent from January up to April of 2016. Data about individuals were fixed according to information of formula including age, sex, geographical location, place displace of displaced and the vaccination date. Successful virus isolation from suspected cases on two types of cells culture included Vero cells line and chicken embryo fibroblast was carried. The titer of the isolated virus on Vero cell line reached (105.8 TCID50 / 0.1ml),which was higher than the titer of the virus isolated on CEF which reach (105.4 TCID50 / 0.1ml) after the third passage of the virus. The isolated virus was confirmed by indirect immunofluorescence technique.The age of subjects included in this study ranged from <1 year up to 24 years old. The results revealed that 15 (30%) have positive IgM index in sera of displaced, whereas residents sera sample shown only 6 (15%) positivity. Howover, all 90 (100%) subjects of both groups showed positive IgG index. The results of the present study showed that there were no significant difference of perforin level in sera of refugees and residents at p<0.05.Assaying of immune response against measles virus by measuring IgM immunoglobulin in sera of vaccinated displaced and resident with measles vaccine reflected a highly significant difference between two groups which reached 0.43 mIU/ml in sera of displaced in comparison with residents sera which reached 0.34 mIU/ml. The result showed also high significant differences in IgM between females and males. The females sera IgM levels of displaced and residents were higher than its level in males sera which reached 0.47 mIU/ml and 0.35 mIU/ml, respectively, in comparison with male sera level which reached 0.40 mIU/ml and 0.32 mIU/ml, respectively. According to age group, the highest IgM value was recorded in displaced and residents sera in age group 10 - 14years (0.50 mIU/ml) and 5 - 9years (0.36 mIU/ml) respec - tively. Whereas the lowest IgM serum value was detected in age group <1 year (0.34 mIU/ml) and 15 - 19years (0.31 mIU/ml) for displaced and residents subjects. According to geographical location, the highest IgM level was detected in sera of displaced from Tal Afar city (0.44 mIU/ml) followed by displaced from Ramadi (0.41 mIU/ml) and the lowest IgM value was detected in sera of displaced from Mosul city (0.33 mIU/ml). Whereas the highest IgM value in sera of residents in the subjects of Al - Mahaweel city (0.37 mIU/ml) followed by subjects of Hilla and Al - Qasem city (0.29 mIU/ml and 0.28 mIU/ml) respectively. For IgG serum immunoglobulin the results revealed that there was no difference in significant levels in both group population. But according to age groups significant difference between displaced and residents was detected at p <0.05 in which the 1 - 4 years old showed the highest IgG serum level in both group (1.98 mIU/ml and 2.18 mIU/ml) respectively. While the lowest IgG sera value was recorded in displaced age group of < 1 year old (0.60 mIU/ml) compared with residents age group of 20 - 24 years old (1.65 mIU/ml).It could be concluded that the results showed that the displaced do not have any effect on the residents of the province of Babylon for measles as a result of the good immunization by Babil Health directorate following to the Ministry of Health.
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التعدد الشكلي للمستقبل الثاني لعامل الورم التنخري والجين المحفز للـ CD4 والانترلوكين 37 في تقييم التهاب المفاصل الرثوي Tumor Necrosis Factor Receptor II, Cd4 Enhancer Gene Polymorphisms And IL - 37 In Assessment Of Rheumatoid Arthritis

اسم المؤلف: وسناء جمعة محمد
اسم المشرف: محمد شمخي جبر باسم شهاب احمد
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2016
مكان الجامعة: بغداد
المستخلص: التهاب المفاصل الرثوي (Rheumatoid Arthritis) مرض جهازي التهابي مزمن يصيب المفاصل وتحديدا الاغشية الزلالية والتراكيب المفصلية، والذي يحدث للعديد من الناس وبنسبة تقارب 0.5 الى 1% من السكان في العالم. لوحظ خلال الدراسات الوبائية لمرض التهاب المفاصل الرثوي | Rheumatoid arthritis (RA) one of the most common systemic autoimmune diseases characterized by chronic joint inflammation and subsequent joint destruction. To date, it well known that RA is characteristic of the expansion of the synovium and infiltration of the inflammatory cells coupled with destruction of adjacent articular cartilage and bone. This is strongly dependent on CD4 T cell. CD4 Cells stimulate monocytes, macrophages, synovial fibroblasts, and other cells to produce cytokines such as tumor necrosis factor - ? (TNF - ?, interleukin - 1 (IL - 1), IL - 6, IL - 15, IL - 17 and metalloproteinases that produce tissue damage. TNF is a major inflammatory cytokine contributing to the pathogenesis of RA, which provides rational for development of anti - TNF biological agents in the treatment of RA.Recently have shown that IL - 37 is a key cytokine in regulating inflammatory response, mainly by inhibiting the expression, production, and function of proinflammatory cytokines. Objectives This study planned to evaluate the association of TNFRII and CD4 enhancer genes polymorphisms in development and severity of RA in Iraqi patients, evaluate IL - 37 in patients with RA and investigate the correlation between IL - 37 levels with disease activity and relation of inflammatory parameters (ESR, CRP, ACPA, and RF) with TNFRII, CD4 enhancer genes polymorphisms.Patients and methods This study was performed during the period from May to September 2015. The patients were attending the out patients' Clinic in Medical City/Baghdad Teaching hospital/rheumatology unit and the laboratory ELISA tests were done in nursing home hospital laboratory, polymerase chain reaction (RFLP) analysis was performed in the specialist private molecular laboratory (ASCO Lab) in Al - Harthia / Baghdad. Fifty patients and 50 apparently healthy control individuals, Patients received disease modifying anti rheumatic drugs (DMARDs) and newly diagnosed patients.The diagnosis of RA patients has performed under the supervision of rheumatic disease consultant physician at the consultation clinic of Baghdad teaching hospital of the medical city. Blood samples had taken from RA patients to measure (ESR), white blood cells count and hemoglobin. Also, analyze the serum levels of ACPA, RF screen CRP and estimate the levels of IL - 37 in patients and healthy individuals using ELISA test kits. TNFRII and CD4 enhancer genes polymorphisms genotyping had achieved by polymerase chain reaction restriction fragment length (PCR - RFLP).Results The results showed the estimation of RF by screening test revealed that its mean level was high among RA patients (168.87+31.62) in comparison with apparently healthy control (4.96+0.71) with highly significant difference (p<0.01). There was a higher positivity of Anti CCP in the patients sera (114.85+21.06) in comparison with apparently healthy control (1.71+0.13) (P<0.01). while the estimation of CRP showed that its level was higher among sera of RA patients (38.39+4.31) in comparison with healthy control group (16.49+2.51) (P<0.01). Furthermore, the ESR level in RA patients were higher than apparently healthy individuals (52.96+3.68) and (10.44+0.74) respectively with significance (P<0.01).The frequencies of the MM, MR, RR genotypes of TNFRII gene polymorphisms were 60%, 32%, 8% in RA patients and 52%, 42%, 6% in controls. There were no significant differences in the genotypes frequencies polymorphisms of the TNFRII 196 MR polymorphism between apparently healthy control and RA. The frequencies of AA, AG, GG genotypes of CD4 10845 A/G in patients was 36%, 42%, 22% and 8%, 44%, 48% in control group. There were significant differences in the genotypes frequencies polymorphisms of the CD4 10845A/G polymorphisms between apparently healthy control and RA patients (p<0.01). AA genotype were significantly more likely to develop RA (OR=1.355). There was significant increase in disease activity and severity in patients carries AA genotype (p<0.01).IL - 37 levels were elevated markedly in RA patients (101.31+10.41) compared with apparently healthy control (43.90+0.91) (p<0.01). More importantly, IL - 37 showed a significant correlation with disease activity (CRP) in RA patients (p<0.05). Also, IL - 37 show non - significant relationship with (MM, MR, RR) exon6 TNFRII in RA patients compared with controls, and non - significant difference between level of IL - 37 with CD4 10845 A/G genotypes compared with healthy control.Conclusion All findings suggested that TNFRII - 196R genotypes not associated with RA diagnosis. In addition, genetic polymorphisms at the CD4 enhancer gene are one of important factors that associated with susceptibility and severity of RA and can serve as a genetic marker for the risk of development of RA.
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دورا محتملا لفيروس ابشتاين بار في عملية تسرطن الغدة الدرقية الحليمية A Possible Role Of Epstein - Barrvirus In Carcinogenesis Of Papillary Thyroid Carcinoma

اسم المؤلف: هند علي خميس
اسم المشرف: عامر رشيد النجار سعد حسن محمد علي
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2016
مكان الجامعة: بغداد
المستخلص: This study was designed as a retrospective research. A total number of (90) formalin - fixed, paraffin embedded tissues were studied. Malignant and normal thyroid tumors tissue blocks were collected from the archives of histopathology laboratories of different general hospital including ALYarmouk Teaching Hospital (Baghdad), Baghdad Medical City Teaching Hospital as well as many private laboratories in Baghdad, during the period from May 2013 to May 2014. These cases include ? Thirty tissue specimens from patients with papillary thyroid cancer. ? Thirty tissue specimens with benign thyroid lesions as control thyroid tissues group. ? Thirty (30) thyroid specimens were obtained from the normal thyroid tissues from the same those totally - thyroidectimized patients for thyroid papillary cancer (PTC) and have served as the second control group for this study. This study was therefore a paired case - control design. The sections were stained by hemotaxyline and eosin staining so as to confirm the diagnosis and assure that the intervening sections ( that were specified for the in situ hybridization and Immunohistochemistry study have containing the questioned tissues from papillary thyroid cancers and benign thyroid tissues (control group).After histopathological sectioning of these thyroid gland tumors and staining by Hematoxylin and Eosin, a final definitive diagnosis was done by histopathologist consultant. The practical part of this study was designed in four pathways : 1. Molecular detection of Epstein - Barr virus in those tissue blocks were performed by using ultra - sensitive version of in situ hybridization (ISH) for detection of EBV - EBERs. 2. To perform immunohistochemical study to demonstrate EBV - latent gene (LMP1, EBNA - 2) in the papillary thyroid cancer compared to control group. 3. To perform immunohistochemical screening study to demonstrate the over expression state of p53 - tumor suppressor gene in those tissues with papillary thyroid cancer compared to control group. 4. To perform immunohistochemical screening study to detected the CD8 and CD56 in tissues with papillary thyroid cancers compared to control group. The obtained results are summarized as follows : 1. The mean age of patients with papillary thyroid cancer and simple nodular goiter was (39.87&41.13) years respectively, with standard deviation ±(11.773,9.243) years. 2. In this study, the percentage of the females with papillary thyroid cancer was higher (86.7%; 26 cases) than the percentage of their PTC - male counter parts (13.3%; 4 cases). The patients whom sub totally thyroidectimized for simple noduler goiters , the percentage of females was also higher (73.3%; 22 cases) than the percentage of their male counter parts with simple noduler goiters ( 26.7%; 8cases). Female / male ratios of the patients with PTC and simple noduler goiters were 6.5 and 2.75 respectively. 3. The percentage of EBV results in the total group of PTC was (50%) while the percentage in the total group of normal thyroid tissue was (26.7%) and lastly in the total simple nodular goiter group was (10.0%). 4. The positivity rate of EBV EBERs - ISH technique in the total group of PTC was (30%), where as its percentage in NTT tissues was (16.7%) and in SNG tissues was (3.3%). 5. The percentage of positive - EBNA - 2 IHC technique in the total group of PTC was (20%), where as its percentage in NTT tissues was (10.0%) and in SNG tissues was (6.7%). 6. The percentage of positive - EBV - LMP - 1 - IHC technique in the total group of PTC was (43.3%), where as its percentage in NTT tissues was (20.0%) and in SNG tissues was (6.7%). 7. Mutated P53 - protein was detected in 28 cases (70%) of the studied cases. Twelve cases (30.0 %) showed negative IHC reactions. 8. The highest total percentage of CD8 - IHC reactions (21 cases; 70%) was found in those with papillary thyroid cancer followed by normal thyroid tissues cases ( 5 cases; 16.7%) and then ( 2 cases : 6.7%) in simple nodular goiter. 9. It was found that the highest percentage of CD56 - IHC reactions was in those with normal thyroid tissues (25 cases; 83.3%) followed by an equal percentages of CD56 - IHC reactions in either thyroid papillary or simple nodular goiter (2 positive cases; 6.7%, each). 10. The importance of CD56 marker has been found to play a role or it may be better to be used as a negative - diagnostic biomarker for papillary thyroid cancer in differentiating it from other malignancies as well as benign lesions of the thyroid gland, individually as well as in combination with other markers for clinical evaluation of those patients. 11. The evident high mutated p53 - over expression, as reflected by abnormal gene product, among papillary thyroid cancer patients indicates for a pivotal role of such genetic mutation in their carcinogenesis as well as could be useful in the clinical evaluation of patients with papillary thyroid cancer, too. 12. The high coexistence of p53 - mutation with EBV in PTC could point for participation of EBV - transformation genes in the p53 activation in high proportion of PTC. 13.The high percentage of CD8 lymphocytes in EBV - positive PTC as compared to their EBV - negative counterparts could reflect the participation of specific cellular immunity against both, the papillary thyroid cancers and this EBV infection during the initiation and progression of EBV - associated papillary thyroid cancers.
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تشخيص مصلي وجزيئي لفيروس الحلا البشري النوع السادس المصاحب لبعض سرطانات الدم Serological And Molecular Detection Of Human Herpesvirus Type 6 Associated With Certain Hematologic Malignancies

اسم المؤلف: هديل محمد فياض
اسم المشرف: علاء فاضل علوان داود سلمان داود
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بغداد
المستخلص: Human herpesvirus 6 (HHV - 6) is largely ubiquitous double stranded DNA viruses within the betaherpesvirinae subfamily and of the genus Roseolovirus.HHV - 6A and HHV - 6B infects very high percentage of population around the world, primarily during childhood through respiratory droplets. HHV - 6 has been found out in many types of cancers : lymphomas, leukemia, cervical cancer, and brain tumors.This cross - sectional case control study was carried out in Baghdad Teaching Hospital and National Center of Hematology - AL - Mustansiriyah University - Baghdad, for the period from 1 September 2013 to 1 April 2015.The aims of the study were to explore the seropositivity rate of HHV - 6 antibodies among Iraqi patients with different hematological malignancies using different laboratory assays, beside, the molecular determination of plasma viral DNA load by quantitative polymerase chain reaction (qPCR).The study included 109 patients with different hematological malignancies; 24 patients with acute lymphoblastic leukemia (ALL), 30 patients with acute myeloid leukemia (AML), 11 patients with Hodgkin lymphoma (HL), 39 patients with non hodgkin lymphoma (NHL) and 5 patients with chronic lymphocytic leukemia(CLL). All patients were newly diagnosed and enrolled before receiving chemotherapy. The age range was between 14 - 80 years. Fifty three (48.6%) and 56 (51.4%) patients were males and females, respectively. The diagnosis of their malignancies was based on either bone marrow aspirate biopsy and lymph node biopsy as well as cytochemical investigations. Additionally, 59 apparently healthy blood donors were enrolled as control group. The age range was between 18 - 59 years, twenty eight (47.4%) and 31 (52.6%) were males and females, respectively.The serum anti - HHV - 6 IgG was detected by indirect immunofluorescent technique (IFAT) as well as by enzyme linked immunosorbant assay (ELISA).While, the serum anti - HHV - 6 IgM was detected by ELISA only. The detection and quantification of plasma viral DNAemia was carried out by quantitative polymerase chain reaction. Serological as well as molecular assays were done in Central Public Health Laboratory in Baghdad. The highest IFAT positivity rate was among patients with CLL (100%), followed by patients with AML (83.3%), and the least positivity rate was among patients with NHL (64.1%) compared to that of healthy control. Generally, the total anti - HHV - 6 IgG by IFAT was insignificantly higher among patients compared to healthy controls (74.3% vs 61.0%, p=0.074). The anti - HHV - 6 IgG positivity rates by ELISA were insignificantly higher in all groups of haematological malignancies except in patients with AML in whom it was significantly higher compared to healthy controls (96.7% vs 72.9%, p= 0.007 ). A part from AML, the highest anti - HHV - 6 IgG by ELISA was among patients with ALL (87.5%) followed by patients with NHL (84.6%), and the least was among patients with CLL (80.0%) compared to healthy control. Generally, the total anti - HHV - 6 IgG positivity rate detected by ELISA was significantly higher compared to IFAT in patients (p= 0.013) as well as in healthy control (p= 0.0005).The results of anti - HHV - 6 IgM positivity rate as detected by ELISA was significantly higher in patients with AML (36.7%, p=0.0004), and patients with HL (27.7%, p= 0.038) compared to healthy controls (6.8%). Whereas, other groups of hematological malignancies showed insignificantly higher positivity rate versus control group. However, the highest positivity rate was among patients with AML (36.7%), and the least positivity rate was among patients with CLL (0%). Generally, the total ELISA anti - HHV - 6 IgM positivity rate was significantly higher in patients versus healthy controls (22.0% vs 6.8%, p= 0.011). The results of PCR showed that 5 out of 109 patients had detectable HHV - 6 DNA in the plasma. The highest detection rate was among patients with HL (27.3%), followed by patients with ALL (4.2%), and then patients with AML (3.3%), while the virus was not detected in other disease categories as well as in healthy controls. The plasma viral load (mean ± SD) among the three Hodgkin lymphoma patients was 1.4± 0.3*102 particle/milliliter.The study concluded that the seropositivity rate of HHV - 6 infection is highly prevalent among Iraqi patients with hematological malignancies compared to healthy population. However, primary or reactivation infection as determined by anti - HHV - 6 IgM in patients as well as healthy individuals is relatively low. Furthermore, the HHV - 6 DNA detection rate as well as plasma viral load among malignant patients before receiving immunosuppressive treatment were low.
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دراسة Toll like receptors(TLR2& ( 4 ومؤشرات حيوية مختارة في المرضى المصابين بسرطان المثانة Study Of Toll Like Receptors (TLR - 2 And 4) And Selected Biomarkers In Bladder Cancer Patients

اسم المؤلف: هدى سعدون البياتي
اسم المشرف: ميسون علي سليم ناهي يوسف ياسين عصام سلمان العزاوي
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بغداد
المستخلص: Worldwide, cancers of the urinary bladder are complex and involve genetic abnormalities and may be due to different environmental chemical carcinogens, as well as chronic infection like Schistosomiasis, all these will allow normal transitional cells to become cancerous which are typically transitional cell carcinoma(TCC).This study was conducted at AL - Yarmouk and Baghdad Teaching Hospitals in Baghdad from June 2013 to April 2014, of which urine and bladder tissue were taken from 57 bladder cancer patients and 80 controls to assess Interleukin - 8 (IL - 8), Matrix MetalloPeptidase - 9 (MMP - 9), Bladder Cancer - Associated Protein (BLCA - 4),Toll - Like Receptors (TLR - 2and TL - R4) level among them and to evaluate their roles in cancer development.One hundred thirty seven Iraqi individual were divided to 28(20.43%) newly diagnosed and 29(21.16%) recurrent bladder cancer patients (relapse), 40(29.19%) apparently healthy volunteers and 40(29.19%) autopsies (apparently normal urothelium), of whom urinesample and tissue biopsy were collected, clinical diagnosis of patients carried out by cystoscopic and histopathological examination, the mean ±SD age of bladder cancer patients were (63± 9.3) with (M : F ratio7.1 : 1).Predominant cancer type was Transitional cell carcinoma (TCC). According to risk factors of bladder cancer, out of 57 bladder cancer patients, 34(59.64%), 29(50.87%), 9(15.78%), 7(12.28%), 6(10.52%) were smoker, had UTI, history of Schistosomiasis, family history ofcancer and stones respectively. In this study 30 patients had non - muscle invasive bladder cancer, most of them (21) had newly diagnosed tumors, and only 9 had recurrent disease, as well as, 27 patients with muscle - invasive bladder tumor, just 7 had newly diagnosed tumors and 20 with recurrent disease.According to the result of ELISA technique, the mean±SD urine level IL - 8(pg/ml) for recurrent bladder cancer patients was 359.6±170.1pg/ml, for newly diagnosed was 265.95±139.09pg/ml and for healthy volunteers was 62.04±37.66pg/ml. The mean ±SD concentration of urine MMP - 9(pg/ml) was 7368.3±1730.19pg/ml for recurrent, 6176.8±2366.9 pg/ml for newly diagnosed and 1131.53±1554.80 pg/ml for healthy volunteers. Mean±SD Level of urine BLCA - 4 (ng/ml) for recurrent was 1.45± 0.26 ng/ml, for newly diagnosed was 1.30± 0.22 ng/ml and for healthy volunteers was 1.02±0.067 ng/ml. Urine level of these markers was significantly higher in recurrence group when compared with newly diagnosed and these levels decreased in normal volunteers which was statistically significance (P<0.001 for IL - 8, MMP - 9and BLCA - 4),also there was a significant increase in urine IL - 8 level according to grade(P=0.0002) and muscle invasion(P= 0.0009) as well as BLCA - 4 showed this significant increase in high grade(P=0.0001), and muscle invasion(P=0.0001),but no significant difference was found in the level of urine MMP - 9 with grade(P=0.23) and muscle invasion(P =0.069)In comparison, IL - 8 and MMP - 9 IHC staining of bladder tissue, showed low expression in cancerous tissue with no significant difference between cancer and normal urothelium(P=0.140 for IL - 8, P=0.265 for MMP - 9 ), and unrelated to grade, muscle invasion and recurrence.Significant increase in TLR - 2 and TLR - 4 expression in bladder cancer tissue than in normal urothelium (P=0.0001 for each), and according to grade and muscle invasion, TLR - 2 showed significant over expression in high grade than low grade (40.7%vs36.6%) with P=0.018, muscle invasion than non - muscle invasion (44.4%vs33.3%) with P value of 0.02, while TLR - 4 was none significantly correlated with grade andmuscle invasion. As well as, there was no correlation between these receptors with tumor recurrence.In regard to risk factors, Smoking, schistosomiasis and family history showed correlation with study marker in different manner as, IL - 8 was significantly highly expressed in smoking associated bladder cancer group, as well as patients with family history of bladder cancer showed high expression IL - 8, for Schistosoma associated TCC patients and UTI they demonstrated a statistically significant high expression of TLR - 2 p= (0.0001).From this study we conclude that urine IL - 8, MMP - 9and BLCA - 4 measured by ELISA showed specificity in diagnosis of bladder cancer, besides that, urine IL - 8, MMP - 9 and BLCA - 4 have a role in discrimination between newly diagnosed vs recurrent, with a significant association between urine IL - 8 with BLCA - 4 (P= 0.0001) and IL - 8 with MMP - 9 (P= 0.005) in patients with recurrent bladder cancer. Urine IL - 8 and BLCA - 4 concentration were statistically with significant increase in high grade than low grade and muscle invasion than non - muscle invasion, On the other hand, bladder cancer cells over express TLR - 2 and TLR - 4,while TLR - 2 showed correlation with high grade and muscle invasion,TLR4 did not show such correlation. Lastly, IL - 8 and MMP - 9 staining showed low expression in tumor tissue of different grades, stages and groups.
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دراسة بكتريولوجية على التفاقم الحاد لمرضى الانسداد الرئوي المزمن Bacteriological Study On Acute Exacerbation Of Chronic Obstructive Pulmonary Disease (COPD) Patients

اسم المؤلف: هبة اسماعيل علي العزي
اسم المشرف: عروبة خالد عباس عبد الحميد القصير
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: ماجستير
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بغداد
المستخلص: لا يزال مرض الانسداد الرئوي المزمن مشكلة رئيسية في مجال الصحة العامة. ويرتبط التفاقم الحاد مع فقدان سريع في وظيفة الرئة وخلل في نوعية الحياة وهي من الاسباب الرئيسية للامراضية والوفيات في مرضى الانسداد الرئوي المزمن. تعتبر العدوى البكتيريا هي من الاسباب | Chronic obstructive pulmonary disease remains a main problem in public health. The acute exacerbations are related with rapid loss in lung function and defect in the quality of life and are the main causes of morbidity and mortality in COPD. This study was carried out to isolate and identify the bacteria that cause acute exacerbation in chronic obstructive pulmonary disease and sensitivity of isolated bacteria for most of the antibiotics currently in use.Eighty sputum specimens from patients were included in this study and their age was range forty and above , during the period from January 2015 to June 2015 from three places Al - Yarmouk Teaching Hospital, Medical City Hospital and Medical Al khademeain Emamaain City.Bacteria were identified in 52 (65%) while the patient is not specified any reasoned in 28 patients (35%) patients. The diagnosis of bacteria through culture and microscopic examinations and biochemical tests necessary for diagnosis and then the diagnosis has been confirmed of isolation by the Vitek method for each bacterium, the percentages were as follows : Streptococcus pneumoniae was the most frequently isolated bacteria, which recovered from 21 (26.25%) patients. Followed by Klebsiella pneumoniae, Escherichia coli, Pseudomonas spp., which isolated from (7.5%, 7.5%, and 5%) of patients respectively, while (3.75%) for each of Acinetobacter baumannii & Staphylococcus aureus. The proportion of Stenotrophomonas maltophilia was (2.5%) while the proportion (1.25%) for each of Enterobacter cloacae, Leclercia adecarboxylata, Moraxella catarrhalis & Raoultella ornithinolytica.Through the search three cases were infected with Mycobacterium tuberculosis diagnosed by direct Acid Fast Stain test and their percentage was (3.75%).Sensitivity of bacterial isolates to antibiotics Carbapenems and Doxycycline and Chloramphenicol were the most effective against tested bacteria in vitro while Erythromycin, Tetracycline, and Amoxicillin were the least sensitive
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دراسة جزيئية ومناعية لدى عينة من مرضى التدرن الرئوي Molecular And Immunological Study In A Sample Of Pulmonary Tuberculosis Patients

اسم المؤلف: مي يحيى عبد الحسين
اسم المشرف: عروبة خالد عباس احمد اسمر منخي
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: ماجستير
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بغداد
المستخلص: يعتبر مرض السل الرئوي tuberculosis الذي تسببه بكترياMycobacterium tuberculosis من الامراض المعدية والشائعة واحد الاسباب الرئيسية لحالات الوفاة في العالم, ولاهميته اجريت هذه الدراسة لتسليط الضوء اكثر حول التشخيص المناعي للمرض وعلاقته بالتشخيص الجزيئي.تضم | Tuberculosis (TB) is caused by Mycobacterium tuberculosis; which consider as one of the most common, infectious diseases and major causes of morbidity and mortality worldwide. A prospective study was conducted to diagnose disease by immunological methods and its association with molecular diagnosis.The study included immunological diagnosis by Quantiferon - TB Gold In Tube Test, estimation the serum levels of IFN - ?, TNF - ? and IL - 10 in TB patients and control by a sandwich ELISA test using commercially available kits, in addition to molecular diagnosis by using Gene Xpert MTB/RIF assay, conventional bacteriological tests such as smear microscopy by Ziehl - Neelsen stain for sputum samples. Furthermore, estimation of ESR and WBCs.Fifty TB patients attending the National Reference Laboratory of Tuberculosis /Baghdad from the period between April to July 2014 were recruited in this study. For the purpose of comparison, 50 control samples (respiratory disease and healthy) matched by age and gender were also included.All TB patients' samples were positive by smear microscopy. The results revealed that there were significance differences between AFB scoring and G - Xpert results (p Forty five samples of TB patients with significance difference (p?0.01) were positively detected by Quantiferon - TB Gold Test.The mean serum level of QFT among TB patient and control (3.54 IU/ml vs. 0.866 IU/ml and 0.556 IU/ml), IL - 10 (53.02 Pg/ml vs. 22.24 pg/ml and 7.51 pg/ml) and TNF - ? (74.34 Pg/ml vs. 49.12 Pg/ml and 27.81 pg/ ml) were significantly higher in TB patients as compared to controls (P?0.05).In conclusion, high levels of IFN - ?, TNF - ? and IL - 10 in TB patients serum indicate an important role of theses cytokines in the pathogenesis of tuberculosis, so they could be considered as a good biomarkers for diagnosis and considered as a target for future therapy. The present study revealed significant difference in the WBCs and ESR values between TB patients and control (P?0.05)
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دور الايبشتاين بار فايرس كعامل محتمل لتقدم سرطان الدم اللمفاوي المزمن Molecular Characterization Of Metallo ? - Lactamase (MBL) Genes

اسم المؤلف: لمى عامر ياسر
اسم المشرف: عامر رشيد النجار داود سلمان داود
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2016
مكان الجامعة: بغداد
المستخلص: Epstein Barr virus (EBV) is ubiquitous virus which infects the majority of the human population and is the causative agent of infectious monocleosis and a variety of B - cell tumors including Hodgkin lymphoma, Burkitt lymphoma, human immunodeficiency virus - associated lymphomas and post - transplant lymphoproliferative disorders , this virus expressed many protein and it is associated with a variety of B - cell tumors. Recently many studies have suggested a causal relationship between EBV and chronic lymphocytic leukemia (CLL). A prospective study of thirty samples of formalin - fixed, paraffin - embedded tissue of bone marrow aspirates samples and blood from newly diagnosis BCLL of 30 patients and our control included : 1) 20 bone marrow of patients who they have hematological disease other than leukemia as control. 2) Blood serum from healthy controls to study the level of IL - 10 and IL - 8. The present study conducted at Baghdad Teaching hospitals and the National Center of Hematology from September 2013 to Jun 2014. In this study we investigated the association between EBV and CLL. The detection of EBV encoded RNAs (EBER1,EBER2) by in situ hybridization and also latent membrane protein - 1 (LMP - 1) and EBV nuclear antigen - 2(EBNA2) by immunohistochemistry and detection for the level of Interleukin n (8 and 10) in the serum of CLL patients by ELISA. In situ hybridization study revealed that all the controls were negative for EBERs and 46.7% patients were found to be EBERs positive. There was a correlation between positive EBERs and tumors stage and also EBERs and IL10 and with LMP - 1 and EBNA - 2. Immuno - histochemical method was used to demonstrate the rate of (LMP - 1 and EBNA - 2) in CLL patients where a positive results of EBNA2 and LMP1were found in (43.3 % and 56.6s %), respectively in CLL patient group. All controls were negative with an exception of two patients were LMP1 positive. Enzyme linked immunosorbent assay test (ELISA) was used to detect the level of IL - 8 and IL - 10 in serum of newly diagnosed of CLL patients. Interleukin 8 was significantly higher in CLL cases group (33.1 pg/ml) compared to control group.On the other hand It was found that high increase in IL - 10 level ,in CLL patients when compared with controls. Conclusion : It can be concluded that there is a significant relationship between EBV and Chronic lymphocytic leukemia, mainly EBERs, also the association of significant increase in IL - 10 and IL - 8 with CLL.
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مؤشرات جزيئية حول مقاومة الزوائف الزنجارية المرتبطة بخمج المجاري البولية للمضادات الحيوية Molecular Markers Of Antibiotics Resistance Of Pseudomonas Aeruginosa Associated With Urinary Tract Infections

اسم المؤلف: كريم عليوي حمادي سليمان
اسم المشرف: عامر رشيد النجار
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بغداد
المستخلص: الزوائف الزنجارية : وهي بكتريا سالبة لصيغة الكرام والمنتشرة بشكل واسع في البيئة مثل التربة والمياه. وهذه البكتريا تتسبب في كثير من الالتهابات مثل التهاب المجاري البولية والتهاب الحروق وهي بكتريا انتهازية. وتشكل تهديد لحياة المصابين بالمجاري البولية وخاص | Pseudomonas aeruginosa is a Gram |( - ve) bacteria, widely presents in the environment such as soil, water, its incubation period (1 - 3) days, generation time 1 - 2 hrs. This bacteria causes many infections such as (UTI), and burn infections. It is an opportunistic pathogen, life threating for immune compromised patients such as diabetic patients. Pseudomonas aeruginosa is a nosocomial pathogen, and it is ranking the second among Gram ( - ve) hospitals acquiring pathogen. Its antibiotic resistance increased due to acquiring many antibiotics resistance genes. The spread of these genes among bacteria is via plasmids. Plasmid profiling of Ps. aeruginosa isolates by gel electrophoresis is a best technique for detection of plasmid content. Through this study, which was carried out in two main teaching hospitals, the diabetic center and private labs in Baghdad, during the period from January to October 2014. A total of (620) urine specimens were collected during (9) months. Urine specimens from inpatients were (420 / 620, 67.75%) and from out patients were (200 / 620, 32.25%). Urine specimens from Dialysis unit were (280 / 620, 45.1%) and from burn units were (140 / 620, 22.5%). The male represented (n = 334, 52.38%) while female were (n = 286, 46.20%). The positive cultures of urine samples obtained from hospitalized patients were (72 / 420, 17.1%) while for urine samples obtained from out patients (n = 40 / 200, 20%) were positive culture. A total of (112) positive culture out of (620) collected with (18.06%) percentage. A total of (30) bacterial isolates were tested by using disc diffusion method for susceptibility test forward to (15) antibiotics. Ps. aeruginosa isolates showed highest resistance to gentamicin (87.5%) while the lowest resistant toward meropenem, and imipenem with resistance percentage (16.7% and 8.0%) respectively.The occurrence of multi - drug resistance in (30) selected isolates were grouped as follows to (MDR) for those isolates which showed resistance to at least (3) antibiotics which was found to be (5 / 30, 6.6%). While (XDR) referred to those isolates which showed resistance to (4 - 5) antibiotics with (19 / 30, 63.3%). In related to (PDR) which referred to isolates that showed resistance to more than 5 with (6 / 30, 20%). The extracted plasmid DNA were resolved by gel electrophoresis for detection of plasmid content for (20) isolates including (10) isolates from inpatients, and (10) isolates from out patients. The selection of these isolates were grouped according to their antibiotic resistance pattern and the number of plasmids that they carried. The results showed that out of (20) there were (12 / 20, 60%) had no plasmid. While the remaining (8 / 20, 40%) had plasmids with different sizes and numbers. By using PCR technique the most common kinds of genes such as bla CTX - M, bla OXA, which encoded extended spectrum ? - lactamase (ESBLS) and those for metallo ? - lactamase such as bla IMP, were investigated with specific primers. Five isolates were selected depending on the numbers of plasmids which they carried and their resistance to antibiotics.
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دراسة مؤشرات مناعية وبايوكيميائية مختارة لمرضى السكري النوع الثاني المصحوب وغير المصحوب باعتلال الكلى ذات الادرار قليل الزلال Study Of Selected Immunological And Biochemical Markers In Type 2 Diabetes With And Without Microalbuminuric Nephropathy

اسم المؤلف: علي ناصر محمد علي
اسم المشرف: عبد العظيم ياسين عبود البراك حازم عبد الرزاق عبد الوهاب
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بغداد
المستخلص: Diabetic Nephropathy (DN) is a common complication of diabetic diseases occuring all over the world including Iraq. This type of microvascular complication of diabetes mellitus represents the most common reason of end stage renal disease (ESRD) in the world which considers the main reason for hemodialysis, kidney transplantation and death in developed countries. For this reason the assessment of some biomarkers as an early predicator before onset of microalbuminuria stage of diabetic nephropathy and the correlations between these biomarkers with microalbuminuria were carried out.This study was achieved at AL - Yarmouk Teaching Hospital and the Diabetes National Center /AL - Mustansiriyah University/Baghdad from January 2013 to September 2014. The study involved ninety individuals, twenty volunteers selected from the local community; apparently healthy (group I), 40 type 2 diabetics with normal urinary albumin to creatinine ratio (UACR) (<30 mg/g) (group II), and 30 type 2 diabetic patients with abnormal UACR (? 30 - 300 mg/g) (group III). Mean of diabetes duration (D.D.) in group II is about 4 years and in group III is about 8 years. The mean age for group I, II and III were 53, 55 and 58 years respectively. The concentration was measured of each microalbuminuria by turbidmeteric method, estimated glomerular filtration rate (eGFR) by using chronic kidney diseases - epidemiology equation, glycated heamoglobin (HbA1c) by using high performance liquid chromatography (HPLC), both fasting blood sugar (FBS) and creatinine (serum and urine) by using colorimeteric method, body weight by using body mass index (BMI) and all of IL - 18, IL - 12 IL - 4, IFN - ? and urinary vitamin binding protein (VDBP) by using ELISA method in three groups.The present study showed that there is inverse correlation between UACR and eGFR. The cause of this inverse correlation is that the decline in renal function of diabetics can be predicted accurately by using both UACR to show the increase in microalbuminuria while eGFR show a measure of the decrease in the ability of kidney for filtration.The difference of FBS mean between group I and II and group I and III was significant (P= 0.000 for both). The P - value between II and III groups was significant (P= 0.024). The difference of HbA1c mean between group I and II, group I and III and group II and III was significant (P=0. 000 for all). These results were expected as high HbA1c level is in consequence of high FBS. The difference of BMI mean between group I and II, group I and III and group II and III was not significant (P= 0.870, 0.885 and 0.968 respectively). Because the BMI levels were approximately constant in three groups. The difference of D.D. between group II and III was significant (P= 0.000).There was no significant correlation in group II between urinary albumin to creatinine ratio with each of diabetic duration, glycated haemoglobin, fasting blood sugar and body mass index (r=0.219 with P=0.244, r=0.039 with P=0.840, r=0.080 with P=0.673 and r= - 0.126 with P= 0.506 respectively); but there was positively significant correlation between urinary albumin to creatinine ratio with each diabetic duration, HbA1c and fasting blood sugar in group III (r=0.298 with P=0.043, r=0.869 with P=0.000 and r=0.518 with P=0.003 respectively) which may be due to the chronic nature of the disease in this group that require these risk factors to initiation; however, the correlation was non significant between UACR and BMI (r= - 0.127 with P=0.228) in group III.The difference of S.Cr. among I and II groups, I and III groups and II and III groups were not significant (P= 0.998, P= 0.331 and P= 0.145 respectively).The correlation was not significant in group II between serum creatinine with each of diabetic duration, glycated haemoglobin, fasting blood sugar and body mass index (r= - 0.140 with P=0.462, r= - 0.124 with P=0.515, r=0.168 with P=0.374 and r=0.007 with P= 0.969 respectively). Also, there was no significant correlation in group III between S. Cr. with each D.D., HbA1c, FBS and BMI (r=0.187 with P=0.077, r=0.109 with P=0.220 and r=0.175 with P=0.124 respectively). The reason of these results backs to that S. Cr. level is not increased as the podocytes remain intact in the diabetics with and without microalbuminuria.The difference of eGFR between group I and II was not significant (P= 0.303). The difference between group I and III and group II and III was significant (P= 0.001 and 0.010 respectively). The correlation was not significant in group II between eGFR with each of diabetic duration, glycated haemoglobin, fasting blood sugar and body mass index (r=0.121 with P=0.524, r= - 0.180 with P=0.341, r= - 0.310 with P=0.096 and r=0.021 with P= 0.911 respectively). Also, there was no a significant correlation in group III between eGFR with each diabetic duration, HbA1c, FBS and BMI (r= - 0.179 with P=0.07, r= - 0.188 with P=0.061, r= - 0.123 with P=0.388 and r= - 0.112 with P=0.557 respectively). The reason of these results back to that eGFR level is at the normal range in group II and slightly beneath the normal range in group III in consequence of the podocytes remain intact in the diabetics with and without microalbuminuria.The difference of urinary VDBP level between group I and II, group I and III and group II and III was significant (P= 0.000 for all). The correlation between urinary albumin to creatinine ratio levels and VDBP level (r=0.963) with P - value 0.000 which was positively significant in group III. The correlationbetween eGFR and VDBP was - 0.524 with P - value 0.003 which is inversely significant in group III. The reason of this inverse correlation may be similar to the reason of inverse correlation between eGFR and UACR, as the cubilin - megalin receptors are common receptors for albumin and vitamin D binding protein. Otherwise, the correlation between urinary VDBP and eGFR in the group I was a weak negative with non significant P - value (r= - 0.188, P=0. 428). The reason of this non significant correlation is that the cubilin - megalin receptors are not damaged by inflammatory process to elevate VDBP in urine; also the podocytes in glomeruli are intact from the damage by inflammatory process, so the eGFR was at the normal range in the control group.The correlation was a positive between HbA1c and VDBP levels in group II and group III (r=0.579, P=0. 001 and r=0.686, P=0.000 respectively). This positive correlation was explained on the basis that deterioration of sugar level control lead to increase the proinflammatory cytokines that damage cubilin - megalin receptors then VDBP increase in urine.The difference of serum IL - 18 level between group I and II, group I and III and group II and III was significant (P=0.000 for all).The correlation between urinary albumin to creatinine ratio and IL - 18 was 0.983 with P - value 0.000 which was positively significant in group III. This may back to damage the cubilin - megalin receptors by IL - 18 action that lead to increase of UACR in urine. In other words, the correlation between serum creatinine and serum IL - 18 in group III was not significant (r=0.041 with P= 0.830). This may back to that podocytes remain intact in this early stage of diabetic nephropathy. There is a significant positive correlation between serum IL - 18 levels and HbA1c levels in group II and group III (r=0.641, P=0.000 and r=0.721, P=0.000 respectively). These two positive correlations support the suggestion of choosing serum IL - 18 as an excellent biomarker for avoidance an early stage of the disease.The difference of IFN - ? levels between group I and group II, between group I and group III and between group II and group III were not significant as follows : (P=0.640, P=0.292 and P=0.522 respectively). The correlation between UACR and IFN - ? (r=0.047) with P - value 0.830 which was not significant in group III which means that IFN - ? is not a good biomarker for prediction of the microalbuminuria as an early stage of DN.The difference of IL - 12 level between group I and group II, between group I and group III and between group II and group III were not significant (0.884, 0.431 and 0.439) respectively. The correlation between UACR and IL - 12 (r=0.190) with P - value 0.314 which was not significant in group III.The difference of IL - 4 levels between group I and group II, between group I and group III and between group II and group III were not significant (0.943, 0.704 and 0.648 respectively). The correlation between UACR and IL - 4 (r=0.169) with P - value 0.371 which was not significant in group III.These results of IFN - ?, IL - 12 and IL - 4 might lead to conclude that both cytokines can’t be selected as a biomarker for an early detection of DN. Finally from the all presented data it can be concluded that IL - 18 and VDBP are considered more sensitive and more efficient than a classic diagnostic method (UACR and eGFR) for avoidance and detection the early stage of DN.
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مميزات التحويرات المناعية للريسفراترول المستخلص من قشور العنب الاحمر على التهاب الكبد المحدث باستخدام ديكالاكتوزامين والذيفان المعوي نوع ب للمكورات العنقودية Immunmodulatory Properties Of Resveratrol Extracted From Skin Of Red Grape On Acute Liver Injury Induced By D - Galactoseamine And Staphylococcus Enterotoxin B

اسم المؤلف: صباح زيارة كاظم المالكي
اسم المشرف: عبد العظيم ياسين عبود البراك متزي نكاركاتي
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2015
مكان الجامعة: بغداد
المستخلص: تعتبر اصابة الكبد الحاد احد الامراض ذو الاعراض السريرية التي تتصف بارتشاح الخلايا اللمفية الى الكبد وتجمع السوائل وارتفاع انزايمات الكبد مثل ناقل امين الاسبارتات Aspartate aminotransferase(AST) في مصل الدم. هنالك الكثير من الاسباب التي تؤدي الى حدوث ضرر | Acute Liver Injury is a vital clinical syndrome characterized by infiltration of lymphocytes in the liver, fluid accumulation, and elevation of liver enzymes such as Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the serum. There are many causes of liver injury, but in this particular study Staphylococcal Enterotoxin B (SEB) was used to induce acute liver injury in mice, it is previously known that (SEB) it act as superantigen that bind with T cell receptor variable region beta chain (VB8), and MHC II of Antigen Presenting Cells (APCs), which leads to activation of T - cells, and huge secretion of pro - inflammatory cytokines, all of these events cause an acute liver injury lead to liver failure, and death. In the last few decades the importance of some natural products appeared, in which these botanicals have an anti - inflammatory properties. Resveratrol is one of these botanicals with an anti - inflammatory effects, which was used as treatment in this study.This study was particularly intended to study the influence of Resveratrol on acute liver injury induced by Staphylococcus Enterotoxin B, lead to liver failure which includes estimation of aspartate aminotransferase levels (AST), histopathological analysis, immune cell counts of liver and spleen, in addition, study whether resveratrol has the ability to cause immunological changes on different immune cells that lead to suppress acute liver injury by using flowcytometry technique,these cells are inflammatory cells carrying CD3+ T cells, CD4+ T cells, CD8+ T cells, NK1.1+ T cells, CD44+ T cells, and suppressive cells such as Myeloid Derived Suppressor Cells (MDSCs), T regulatory cell (FOXP3), as anit - inflammatory markers. Furthermore evaluate the pro - inflammatory, and anti - inflammatory cytokines by using Bio - plex. Also to determine the molecular mechanism in which resveratrol can lead to induce changes in microRNAs (miRNAs) by microarray analysis. Finally validation (miRNAs) resulting from microarray analysis and genes of interest associated with these miRNAs by Real time polymearase chain reaction (RT - PCR).After sixteen hours blood samples were collected to determine AST levels in differents groups using Nanodrop, (Fisher - USA), after sixty hours animals sacrificing organs (liver, spleen) were collected afterward liver samples in 10% formalin were sent for histopathological study, liver immune cells were isolated for counts and molecular study (microarray analysis and quantitive (RT - PCR) total RNA was isolated from liver immune cells for microarray analysis then preparation of complementary Deoxyribonucleic acid (cDNA) by Reverse transcriptase PCR. Validation of the microarray analysis data was achieved by Real time PCR. Spleen immune cells were collected afterward for flowcytometric analysis of CD3+, CD4+, CD8+, NK1.1+, CD44+ T cells, MDSCs, and T regulatory cells using cell surface staining and intra cellular staining.This study showed that Resveratrol oppose the effect of SEB induced acute liver injury through decrease AST concentrations (P< 0.0001). Also Resveratrol 50mg/Kg body weight led to cause reduction in the size of the spleen, compared to SEB+ Vehicle group. Histopathology study reported that resveratrol led to decrease of hepatic necrosis up to 10% in comparison to vehicle group with 60% of hepatic necrosis and decrease in the inflammatory response represented by immune cells as in liver immune cell counts which showed a significant decrease (P< 0.014) after 50mg/ Kg body weight of Resveratrol treatment. Spleen immune cell counts result reported a significant decrease (P<0.013) in the total cells when Resveratrol was used as treatment compared to disease group. Regarding to Flowcytometric analysis, it was found that Resveratrol significantly decrease the absolute count of CD3+, CD4+, CD8+, NK1.1+, CD44+, Foxp3+ T cells in C57BL/6 mice (P< 0.0019, P< 0.001, P< 0.009, P< 0.002, P< 0.03, and P< 0.03 respectively). While MDSCs result reported significant increase with P< 0.01 in absolute cell counts after Resveratrol treatment. Depending on the Bio - plex data showed significant decrease in serum concentrations of inflammatory cytokines IL - 1?, IL - 1?, IL - 3, IL - 5, IL - 12p(40), IL - 12p(70), IL - 13, IFN?, MIP - 1?, MIP - 1?, KC and Rantes cytokines with significant P values (0.009, 0.004, 0.034, 0.004, 0.0009, 0.01, 0.0007, 0.009, 0.0037, 0.0001, 0.0035, 0.006 respectively), while anti - inflammatory cytokines IL - 6, IL - 10 and G - CSF c showed a significant increase in the serum concentration after Resveratrol treatment with a P values of, 0.012, 0.02 and 0.001 respectively. Microarray analysis results recognized 451 miRNAs with P< 0.05, according to fold change expression, 78 miRNAs have fold change greater than - 1.5 fold of expression in Resveratrol treatment group (downregulated), while 87 miRNAs have fold change greater than 1.5 fold of expression (upregulated). According to the ingenuity tools predicted target gene for some of these miRNAs by using miRNA.org database, the database showed that miR - 130a - 3p with - 2.28 fold expression have direct relation of binding with CSF1 gene, Real time PCR results confirmed the results obtained from microarray analysis, the result of miR - 130a - 3p showed a significant decrease of relative fold expression with P< 0.013, and Colony Stimulating Factor - 1 CSF1 (M - CSF) relative expression increased significantly after resveratrol treatment with P< 0.0001. From all of previous data, it can be concluded that Resveratrol can counteract acute liver injury induced by SEB, by decrease AST concentration. Resveratrol acts as an anti - inflammatory compound due to decrease of immune cell numbers, decrease of inflammatory markers, and increase of anti - inflammatory markers. miR - 130a - 3p with inflammatory properties downregulated after resveratrol treatment. Finally Resveratrol treatment increased relative fold expression of CSF1(M - CSF); gene which play a role in the MDSCs proliferation
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دور الانترلوكين RA - 1 والسكليروستين والخلية التائية المستجيبة CD4 في مصل مرضى التهاب المفاصل الرثوي Role Of Serum IL - 1RA, Sclerostin And Effector T - Cell (CD4) In Rheumatoid Arthritis Patients

اسم المؤلف: سنن ثائر عبد الوهاب
اسم المشرف: حيدر صباح كاظم علاء الدين مظفر زبير
الموضوع العام: الطب
الموضوع الدقيق: الاحياء المجهرية
الدرجة: دكتوراه
اللغة: الانكليزية
السنة: 2016
مكان الجامعة: بغداد
المستخلص: التهاب المفاصل الرثوي هو احد امراض المناعة الذاتية الاكثر شيوعا والذي يتميز بالتهاب المفاصل المزمن والذي يؤدي فيما بعد الى تدمير المفاصل.لا يزال مرض التهاب المفاصل الرثوي يعرف على انه سمة من التوسع في الغشاء الزليلي وارتشاح الخلايا الالتهابية اضافة الى | Rheumatoid arthritis (RA) is one of the most common systemic autoimmune diseases characterized by chronic joint inflammation and subsequent joint destruction. It is well known that RA is characterized by the expansion of the synovium and infiltration of the inflammatory cells coupled with destruction of adjacent articular cartilage and bone. This is strongly dependent on CD4 T cell. CD4 Cells stimulate monocytes, macrophages, synovial fibroblasts, and other cells to produce cytokines such as tumor necrosis factor - ? (TNF - ?), interleukin - 1 (IL - 1), IL - 6, IL - 15, IL - 17 and metalloproteinases that produce tissue damage. TNF is a major inflammatory cytokine contributing to the pathogenesis of RA, which provides rational for development of anti - TNF biological agents in the treatment of RA.Wnt pathway (a complex protein network) important in control of the bone formation through the regulation of osteoblast activity, and sclerostin is an important in the regulator of the Wnt pathway by blocking Wnt binding to its receptor and thereby inhibiting bone formation. Blockage of Wnt antagonists such as sclerostin will trigger repair or even healing of bone erosion.Recently have shown that IL - 1Ra has been given therapeutically in several experimental models of arthritis with weak effect in RA. This study planned to evaluate the frequencies of CD4+CD25+ Treg cells in Iraqi patients treated with RA under treatment with biological therapy (Etanercept). The level of serum sclerostin, serum level of IL - 1RA, and investigate the correlation between serum sclerostin and Treg expression. Correlate the level of serum sclerostin and Treg expression with disease activity by (CDAI or DAS28).This study was performed during the period from January 2016 to May 2016.The patients were attending the out patients' Clinic in Medical City/Baghdad Teaching hospital/Rheumatology Unit and the laboratory Enzyme Linked Immunosorbent Assay tests were done in Bio - technology center at the University of Al Nahrain, flow cytometery analysis was performed in the private laboratory (Al Rawabi Laboratory) at Yarmouk/Baghdad. Thirty patients and 30 apparently healthy control individuals were included in this study, Patients under treatment of etanercept.The diagnosis of RA patients has performed under the rheumatologist consultant at the consultation clinic of Baghdad Teaching Hospital of the medical city. Blood samples had taken from RA patients to measure Erythrocyte Sedimentation Rate, Complete Blood Picture, Rheumatoid Factor and High - Density Lipoprotein. Also, analyze the serum levels of Anti - Citrullinated Protein Antibodies and estimate the levels of sclerostin and IL - 1RAin patients and healthy individuals using Enzyme Linked Immunosorbent Assay test kits. CD4+CD25+ Treg cells had achieved by flow cytometery analysis.The results showed the reduction in Treg cells in patients (0.00264 ± 0.002) % under biological therapy than control group (0.026 ± 0.012) % with significant difference (p< 0.05). Also body mass index showed significant difference between patients (32.10 ± 5.81) kg/m2 and control group (28.84 ± 4.67) kg/m2 (p< 0.05). The results showed that sclerostin level higher in healthy (1.072 ± 0.59) ng/ml than patients (0.801 ± 0.28) ng/ml which showed significant difference (p<0.05). Furthermore IL - 1RA serum level was higher in healthy (0.979 ± 0.34) ng/L than patients (0.633 ± 0.35) ng/L with significant difference (p<0.05). Anti - Cyclic Citrullinated Peptide results showed non - significant difference between patients and healthy control. Treg cells is reduction in the patients with RA. Sclerostin and IL - 1RA levels are low in patients treated with etanercept.
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