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Article
mRNA in situ hybridization analysis of VEGF in chronic lymphocytic leukemia

Authors: Haythem A. MJ Al-Rubaie --- Ban A. Abdul Majeed بان عباس عبد المجيد --- Abdulkareem M. Jaafar
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2011 Volume: 53 Issue: 2 Pages: 186-189
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Several factors render chronic lymphocytic leukemia (CLL) an interesting subject for study by researchers. These include marked progress in understanding the molecular biology of normal and neoplastic lymphocytes and recent advances in molecular genetics techniques. Among molecular markers, vascular endothelial growth factor (VEGF), have been widely studied.Objective: The aim of the study is to evaluate the role of VEGF in the pathogenesis of CLL and its role in disease progression.
Patients, materials and methods: A retrospective cross-sectional study was done on 60 patients with chronic lymphocytic leukemia (45 males & 15 females) compared with 20 controls (anemic patients), all recruited at the Medical City Hospital/ Teaching Laboratories/ Baghdad from January 2004 to December 2007. The bone marrow biopsy of each was re-examined histologically. In situ hybridization was performed utilizing biotin labeled VEGF cDNA probes.
Results: The frequency of VEGF positivity was 95% (57 of 60 cases). A statistically significant inverse correlation was found between increasing VEGF scores and lower packed cell volume level and lower platelet count. A statistically significant association was found between advanced modified Rai stage and patients with high score. In addition, there was a statistically significant positive correlation between increasing scores of VEGF and advancing clinical stage.
Conclusions: The score of VEGF is high among patients with advanced clinical stage of the disease.


Article
Detection of CagA in H. Pylori Gastric Illness Using in Situ Hybridization

Authors: Nidhal A. Mohammed --- Ahmed S. Abood
Journal: Diyala Journal For Pure Science مجلة ديالى للعلوم الصرفة ISSN: 83732222 25189255 Year: 2010 Volume: 6 Issue: 3 Pages: 85-90
Publisher: Diyala University جامعة ديالى

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Abstract

Background: H. pylori had been recognized as the causative agent of several gastric diseases and possessed wide variety of virulence factors, one of most important factors is CagA. Objective: This study investigated the expression of CagA cytotoxin on the gastric epithelial cells in patients with Helicobacter pylori gastric illness by using In Situ Hybridization technique. Methods: Paraffin embedded tissue had been made form biopsies taken from 30 patients undergo O.G.D. and selected according to exclusion criteria. In addition 10 apparently healthy volunteers were included as a control group. Results: The CagA cytotoxin was detected in 21(70.0%) patients out of 30 and 9 (30.0%) gave negative results, on other hand control group showed no positive results. Conclusions: The CagA positive H. pylori strains cause the majority of H. pylori related gastric illness.


Article
The possible role of EBV in carcinogenesis of colorectal carcinoma

Author: Manal A. habib منال حبيب
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2010 Volume: 52 Issue: 2 Pages: 174-176
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Except for the tight correlation to nasopharyngeal carcinoma, accumulating evidences show that Epstein-Barr virus (EBV) is correlated to other carcinomas. This study was to investigate the association of EBV with colorectal tumors.Materials & methods: Forty paraffin embedded blocks of colorectal tumors (thirty were adenocarcinoma and ten were benign tumors) were all examined for the presence of EBV DNA with the application of In Situ hybridization.Results: In Situ EBV DNA signals was detected in 6 out of 30 (20%) of colorectal carcinoma with no observed signals in the sections from benign group.Conclusion: Our results showed that infection of EBV exists in human colorectal adenocarcinoma, which indicates that EBV may be involved in the carcinogenesis process.Key words: EBV, In Situ Hybridization, colorectal tumors.


Article
Molecular Localization of Human Papilloma Virus genotypes (16, 18, and 6/11) in Patients with Colorectal Cancer by DNA- Insitu Hybridization
الموضعه الجزيئيه للفيروس الحليمي البشري ذو الانماط الجينية (16 و18 6 و11) في مرضى سرطان القولون والمستقيم باستخدام تقنيه الدنا التهجين الموضعي

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Abstract

Human papilloma viruses (HPV) have been detected in several types of cancers. Over the last few years, a possible correlation between HPV infection and colorectal cancer has been suggested. The aim of this study was to investigate the association between human papilloma virus (HPV) infection and colorectal cancer. To determine the relationship between HPV and colorectal adenocarcinoma, a retrospective study was done. This study was carried out on 50 patients with hisopathologically confirmed primary colorectal cancer. Two samples were collected from each patient: one sample from the tumor site and the other one from adjacent normally appearing colorectal tissues, as well as ten (20) colorectal tissues from control individuals with no cancer. In situ hybridization (ISH) was used to detect HPV DNA (HPV 16 and 18 DNA ISH and 6/11 DNA CISH) in colorectal tissues. HPV 16 was detected in 16(32%) of tumor samples, and in 7 (14%) of adjacent normal tissues, and in additionally two cases of apparently healthy group gave positive results for it. HPV 18 was detected in 11 (22%) of tumor samples and in 6 (12%) of adjacent normal tissue. HPV 6/11 was detected in 24 (48%) of tumor samples and in 7 (14%) of adjacent normal tissue.Our results suggest that colorectal HPV infection is common in patients with adenocarcinoma colorectal, albeit at a low DNA copy number, with HPV16 being the most prevalent type. HPV infection may play a role in colorectal carcinogenesis.

تم الكشف عن فيروسات الورم الحليمي البشري في عدة أنواع من السرطانات. على مدى السنوات القليلة الماضية ، وقد اقترح وجود علاقة محتملة بين عدوى فيروس الورم الحليمي البشري وسرطان القولون والمستقيم.هدفت الدراسة الحالية لتحري عن العلاقة بين الاصابة بفيروس الحليمي البشري وسرطان القولون والمستقيم. لتَحديد العلاقةِ بين فيروس الحليمي البشري وسرطان القولون والمستقيم , صممت هذه الدراسة كبحث ذو أثر رجعي، اذ اشتملت على (50) مريضِ مَع اجراء الفحص النسيجي لتاكيد تشخيص سرطانَ القولون والمستقيم. تم جمع عينتان مِنْ كُلّ مريض: عيّنة واحدة مِنْ موقعِ الورمَ والآخرَ مِنْ الانسجة المجاورةِ التي تبدو سليمة، بالإضافة إلى (20) عينه من أنسجة القولون والمستقيم مِنْ أفرادِ غير مصابين ظاهريا كمجموعة سيطرة لهذه الدراسة. استخدمت تقنية التهجين الموضعي للكشف عن الدنا للفيروس الحليمي البشري ذو النوع الجيني 16 و18و 6/11 في انسجة سرطان القولون والمستقيم. ظهرت النتائج بان الفيروس نمط 16 وجد في 16حالة وبنسبة (32 %) من عيّنات الورمِ بينما في الأنسجةِ الطبيعيةِ المجاورة كانت 7 حالة وبنسبة (14 %)، وحالتلان فقط من مجموعه السيطرة اعطيت نتائج ايجابية. اكتشف الفيروس نمط 18 في 11 حالة وبنسبة (22 %) مِنْ الورمِ وفي 6 حالة وبنسبة (12 %) مِنْ النسيجِ الطبيعيِ المجاورِ. الفيروس الحليمي البشري نمط 6/11 إكتشفَ في 24 حالة وبنسبة (48 %) مِنْ انسجةالورمِ ووجد في 7 حالة وبنسبة(14 %) مِنْ النسيجِ الطبيعيِ المجاورِ.تَقترحُ نَتائِجَنا بأنّ الاصابة بفيروس الحليمي البشري شائعة في مرضى سرطان القولون والمستقيم، ولو أنْه عدد نسخ الدنا منخفض، الفيروس الحليمي البشري نمط 16 النوعَ الأكثر شيوعا». الاصابة بفيروس الحليمي البشري قد تلعب دور في تكون سرطان القولون والمستقيم.


Article
Detection Human Papilloma Virus genotype (16/18) in Iraqi Women Patients with endometrial carci- noma by using Chromogen - Insitu Hybridization (CISH) Technique
الكشف عن وجود فايروس الورم الحليمي البشري نوع 16/18 بمرضى النساء العراقيات المصابات بسرطان بطانة الرحم بواسطة تقنية التهجين الموضعي المولد للون

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Abstract

Aim: To investigate the possible association between human papillomavirus (HPV) and endometrial carcinoma. Does HPV play any role in the initiation or prognosis of endometrial adenocarcinomas among Iraqi women patients.Method: To determine the relationship between HPV and endometrial carcinoma , a retrospective study was done. The study was carried out on 30 patients with hisopathologically con rmed primary endometrial adenocarcinoma ,samples were col- lected from each patients , as well as twenty (20) endometrial tissues from control individuals with no cancer. Chromogen In situ hybridization (CISH) was used to detect HPV DNA genotype 16 /18 in endometrial tissues.Results: HPV 16/18 was detected in 19/30 (63.3%) tumor sample while negative cases were 11/30 (36.7%) of tumor sample and HPV 16/18 was detected in 5/20(25.0%) of endometrial control group while negative in 15/20 (75.0%) cases of endo- metrial control group.Conclusion: Our results revealed that high risk of HPV(16 / 18 ) being the most frequent type in Iraqi women patients with endometrial adenocarcinoma. Association between HPV infection and endometrial adenocarcinoma was observed and this research considered the rst study which evaluated score ,intensity and pattern of replication system of HPV by using CISH technique which re ected role of HPV in endometrial carcinogensis.

الهدف من الدراسة : لتحري عن الع قة المحتملة بين فايروس الورم الحليمي البشري وسرطان بطانة الرحم . وهل ان فايروس الورم الحليمي البشري يعمل على اي دور في نشوء او تشخيص سرطان بطانة الرحم الغدي بين النساء العراقيات المصابات بسرطان بطانة الرحم . طريقة العمل : لغرض تحديد الع قة بين فايروس الورم الحليمي البشري وسرطان بطانة الرحم صممت هذه الدراسة ذو اثر رجعي . وهذه الدراسة شملت 30مريضة اوكدت اصابتها بسرطان بطانة الرحم الغدي نسيجيا . العينات تم جمعها من كل المرضى با ضافة الى 20 عينة من نسيج الرحم من اشخاص غير مصابين بالسرطان كمجموعة سيطرة. تقنية التهجين الموضعي المولد للون استخدمت لتشخيص دنا فايروس الورم الحليمي البشري نوع 16/18 في نسيج الرحم. النتائج : فايروس الورم الحليمي البشري تم كشفه في 19من اصل 30)%63.3( عينة مصابة بالسرطان بينما الحا ت السالبة كانت 11 من اصل 30 )%36.7( عينة مصابة بالسرطان بينما فايروس الورم الحليمي البشري نوع 16/18 تم كشفه في 5 من اصل 20 )%20( عينة غير مصابة بالسرطان من مجموعة السيطرة بينما كان غير موجود في 15 من اصل 20 )%75( عينة غير مصابة بالسرطان من مجموعة السيطرة لبطانة الرحم . ا ستنتاجات: نتائج هذه الدراسة تشير الى ان فايروس الورم الحليمي البشري نوع 16/18 العالي الخطورة هو من ا نواع ا كثر انتشارا بالنسبة للنساء العراقيات المصابات بسرطان بطانة الرحم الغدي و قد لوحظت الع قة بين ا صابة بفايروس الورم الحليمي البشري وسرطان بطانة الرحم الغدي وهذا البحث يعتبر اول دراسة تشير الى كمية وكثافة ونمط التكاثر للفايروس باستخدام تقنية التهجين الموضعي المولد للون التي تعكس دور فايروس الورم الحليمي البشري بسرطنة بطانة الرحم .


Article
Molecular Detection and Genotyping of Human Papilloma Virus Infections in Iraqi Patients with Esophageal Carcinoma.

Authors: Tural Y. Bakir * MSc --- Saad H. Mohammed Ali ** PhD --- Intisar M. Jumaa * PhD
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2009 Volume: 51 Issue: 2 Pages: 204-210
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Molecular DNA hybridization has confirmed more than 120 different human papilloma virus (HPV) genotypes. A small group of them have high- risk oncogenic potential. Many studies have described an association of such high risk-HPV genotypes with a variety of esophageal benign tumors as well as malignant squamous cell carcinomas.Patients and Methods: A total number of 90 tissue specimens were collected from 50 patients with esophageal squamous cell (SCC), adenocarcinoma (AC) and carcinoma in situ (CIS); 20 patients with squamous acanthosis (SA); and 20 individuals with apparently-healthy esophageal tissues (AHET). The molecular detection methods for HPV detection and genotyping were performed by in situ hybridization using cocktailed- and specific high- risk HPV DNA probes, respectively.
Results: The overall percentage of HPV in the total group of esophageal carcinoma was 20%.The percentage of HPV DNA in the subgroup of SCC and AC was 26.7% and 13.3%, respectively,. However, neither HPV DNA was detected in CIS subgroup nor in both control groups (SA and AHET).The overall genotyping results showed that HPV 18 constituted the majority of the detected high-risk oncogenic HPV genotypes, followed by HPV 16 then HPV 31/33.<.Conclusions: Despite the low prevalence of HPV infection and rarity of invasive esophageal carcinoma in the general Iraqi population, the detection of high percentage of such high oncogenic risk- HPV genotypes in these carcinomas indicating for a relevant importance in esophageal carcinogenesis.


Article
The In Situ Hybridization Expression of Fas and Fas Ligand (FasL) in Trophoblastic tissue of Aborted Women Compared with Normal Pregnancy.

Authors: Dhamraa W. Ahmad *** MSc --- Dalya B. Hana ** MSc --- Amal H. Salman * PhD
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2009 Volume: 51 Issue: 2 Pages: 211-215
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Estimation of the in situ expression of Fas and Fas ligand (FasL) in aborted women compared with normal human pregnancy .Patients and Methods: A technique utilizing in situ hybridization(ISH) was performed to detect and determine the in situ expression of Fas and Fas ligand (FasL) mRNA using paraffin embedded sections of curettage samples obtained from 42 women, who were divided into two groups: 30 women with first trimester abortion and 12 women with induced abortion as control.Results: The levels of the in situ expression of both Fas and Fas ligand (FasL) mRNA were found to be highly significant increased in group 1 as compared with group 2 (p<0.01), with a significant positive correlation between Fas; and abortion in group I , FasL & abortion in group I , and between these two parameters(Fas & FasL) (p<0.01) in group1 .Conclusions: The increasing expression of Fas and FasL trophoblasts might influence pathogenesis of first trimester abortion.


Article
mRNA in situ hybridization analysis of p-53 cancer suppression gene and Bcl-2 oncogene in chronic lymphocytic leukemia

Authors: Saad A. Mustafa --- Haythem A. Al-Rubaie --- Abdulkareem M. Jaafar
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2010 Volume: 52 Issue: 2 Pages: 177-181
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Several factors render chronic lymphocytic leukemia (CLL) an interesting subject for study by researchers. These include marked progress in understanding the molecular biology of normal and neoplastic lymphocytes and recent advances in molecular genetics techniques. Among molecular markers, p53 cancer suppressor gene and the antiapoptotic gene Bcl-2 have been widely studied. Patients and methods: A retrospective cross-sectional study done on 60 patients with chronic lymphocytic leukemia compared with 20 controls (anemic patients), all recruited at the Medical City Teaching Hospital laboratories from January 2004 to December 2007. The bone marrow biopsy of each was re-examined histologically. In situ hybridization was performed utilizing biotin labeled p53 and Bcl-2 cDNA probes.
Results: The frequency of p53 positive signals in the study group was 28.3% (17of 60 cases). A significantly larger number of patients, with high score for p53 signal, were associated with high-risk clinical stage than patients with low score (p = 0.005). There was a significant direct positive correlation between increasing scores of p53-positive chronic lymphocytic leukemia cells and advancing clinical stage of the disease (p =0.002).
The frequency of Bcl-2 positivity was 50% (30 of 60 cases). No significant correlations found between Bcl-2 scores and clinical stage of the disease.
Conclusion: Although p53 alteration may occur early in the course of the disease, as shown by the p53 positivity in a proportion of patients in low and intermediate-risk stage of the disease, the highest frequency p53-positive cells, has been observed in high-risk stage of the disease. Therefore, p-53 score is an important prognostic variable in patients with chronic lymphocytic leukemia. However, Bcl-2, as assessed by ISH, is not regarded as an important prognostic marker.


Article
Detection and Genotyping of Human Papilloma Virus-Associated

Author: Saad H. Mohammed Ali* MBChB , MSc , PhD (Medical and Molecular Virology). د. سعد محمد
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2009 Volume: 51 Issue: 3 Pages: 276-282
Publisher: Baghdad University جامعة بغداد

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Abstract

Summary:Background: Molecular DNA hybridization has confirmed more than 120 different human papilloma virus (HPV) genotypes. A small group of them have high- risk oncogenic potential. Many studies have described an association of such high risk-HPV genotypes with a variety of oral lesions including squamous cell carcinoma, leukoplakia, and lichen planus. .Materials and Methods: A total number of 42 tissue specimens, representing 27 patients with oral lichen planus and 15 apparently-healthy oral tissues, were included in this study. The molecular methods for HPV detection and genotyping were performed by in situ hybridization(ISH) using cocktailed- and specific high- risk HPV DNA probes, respectively.Results:The overall percentage of HPV in the total group of patients with oral lichen planus (OLP) was 33.3 %. Negative HPV DNA- ISH reactions were detected in all tissues of the control group. The overall genotyping results revealed HPV 16- DNA in all HPV- positive oral lichen planus tissues , while none of these OLP tissues showed ISH reactions for HPV 18-DNA or HPV 31/33-DNA.Conclusions: The significant incidence of such high oncogenic HPV genotype in those patients with oral precancerous lesions could have a relevant importance along its pathogenesis and the multi-steps oral carcinogenesis , HPV-16-associated oral lichen planus that has mostly previliged the site of cheek mucosa represents a herald indicator for spread of such sexually important transmitted infection among Iraqi general population.Key Words: Oral lichen planus; Human Papilloma Virus; In Situ Hybridization


Article
VEGF EXPRESSION IN TROPHOBLASTIC TISSUE OF WOMEN WITH SPONTANEOUS MISCARRIAGE AND INFECTED BY TOXOPLASMA GODNII USING INSITU HYBRIDIZATION
التعبير الموقعي لعامل النموVEGF في النسيج المغذي للجنين (التروفوبلاست) لنساء تعرضن للإجهاض الفجائي نتيجة الإصابة بداء المقوسات

Author: Nidhal A. AL-Mohaimen
Journal: Iraqi Journal of Biotechnology المجلة العراقية للتقانات الحياتية ISSN: 18154794 Year: 2012 Volume: 11 Issue: 2 Pages: 494-502
Publisher: Baghdad University جامعة بغداد

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Abstract

A total of fifty women, aged between (16-42) years, were involved in this study. Enzyme linked immunosorbent assay (ELISA) test was used for the detection of specific IgM (using serum samples) and insitu hybridization method was used for the detection of VEGF m RNA in trophoplastic tissue. Samples were classified into three groups: Group A-patients with spontaneous miscarriage and Toxoplasma godnii positive (n= 20 women), with a mean age of(23.8±1.631);Group B-patients with spontaneous miscarriage and Toxoplasma godnii negative (n=20 women), with a mean age of (25.5± 1.60); Group C- control group, women with induced abortion for medical causes (n=10 women), with a mean age of (26.4± 1.628). The highest percentage of Vascular Endothelial Growth Factor mRNA expression (65.30± 3.587)% was within the induced abortion group, while the lowest expression was within (A) and (B) group (35.45± 2.184)% and (37.65± 2.964)% respectively. These results indicated that VEGF is essential factor in the process of blood vessel formation in the successful pregnancy, and its low level in miscarriage groups indicated its role in the etiology of miscarriage irrespective of toxo status.

تضمنت الدراسة الحالية خمسون إمرأة يعانين من الاجهاض التلقائي وعشرة نساء عندهن إجهاض مُحث لأسباب مرضية. قسمت الدراسة المرضى الى مجاميع على أساس نتائج الفحوص الى:المجموعة الاولى(A) والتي شملت 20 إمرأة تعاني من الاجهاض التلقائي وهي موجبة لمُستضدات طفيلي داء المقوسات. المجموعة الثانية(B) وشملت 20 إمرأة تعاني من الإجهاض التلقائي وهي سالبة لمُستضدات طفيلي داء المقوسات. المجموعة الثالثة(C) (مجموعة السيطرة) وشملت عشرة نساء عندهن إجهاض مُحث لأسباب مرضية. تم جمع الدم الوريدي من النساء لغرض إستخراج المصل وفحص نسبة المضاد المناعي نوع IgM بواسطة فحص ELISA. وفي الوقت نفسه تم جمع نسيج "التروفوبلاست " أثناء عملية الكشط (الكرتاج), وقد أُستعمل التعبير الموقعي للحامض الرايبوزى الناقل (ISH) لعامل النمو(VEGF mRNA). بينت النتائج إن التعبير الموقعي لـVEGF mRNA ظهر بأعلى نسبة في المجموعة(C) (65.30±3.587)% , أما النسب الأقل فهي في المجموعة(B)(37.65±2.964)% و(A),(35.45±2.184)% على التوالي وكان هناك فرق معنوي عالي بين المجموعة (A) والمجموعة (C) (P<0.001). ومن هذه النتائج يظهر أن عامل النموVEGF له دور مهم في عملية الإجهاض التلقائي.

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