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Article
Digital Image Analysis versus Visual Scoring for Bcl-2 and P53 Protein Expression in Glioma

Author: Hayder F. Ghazi
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2010 Volume: 52 Issue: 2 Pages: 200-204
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Traditionally, evaluation of the results of immunohistochemistry was done by visual quantification. Materials and methods: for reliable evaluation, more time-efficient and user friendly method we used simple computer program with image analysis options as independent parameters for reading positive results. To test the validity of visually scored results, we compare and correlate the results of Digital image analysis (DIA) variables with the visual scores of 280 pictures taken from entire stained glioma tumor sections for Bcl-2 and P53 oncoproteins in different glioma tumor grades.
Results: In this study, rates expression of both oncoproteins was evaluated visually in glioma tumor samples (Bcl-2=72.41% and P53=82.76%), no statistical significant differences were observed according to pathologic grades. Similarly, these results were also explained by data obtained by DIA variables that has been closely correlated with visual scores. More importantly, the DIA variables explained little discrepancy in the visual scores of both oncoproteins. Conclusions: the quantitative DIA measurements of the immunohistochemically stained sections made the results more objective and supported pathomorphological diagnosis.


Article
Characterization of lymphocyte subsets as a tool of assessment of histological disease activity pattern in ulcerative colitis

Author: Hayder F. Ghazi
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2010 Volume: 52 Issue: 3 Pages: 304-308
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: In Ulcerative colitis (UC), the presence of a persistently elevated number of T cells in the inflamed area with respect to the activity of the disease suggesting bad prognosis. This study was done to evaluate cellular immune response in different disease activity patterns and its possible implication in evaluation of disease activity.Materials and methods: This study included Forty seven archived paraffin-embedded samples of ulcerative colitis; these samples diagnosed and graded for disease activity. Then dual immunofluorescence staining was used for phenotyping of lymphocytic infiltrate (CD3- CD19) and (CD4-CD8). A total and differential T cell as well as plasma cell count was recorded in these UC tissue samples. Non parametric Kruskal-Wallis test was used to compare the median cell counts among different study groups.Results: There is higher lymphocytic infiltrate for all types of cells when UC samples compared with control samples with highly statistical significant difference, T cells represents the major constituents of colonic mucosal infiltrate (86.89%) and about 75.79% were CD4 positive T cells. T-cell subsets and plasma cell have high statistical significant difference (p≤0.001) according to histological grade. Furthermore, with the histological grade the highest association found with T lymphocytes (r=0.944) followed by T helper (r=0.821), T cytotoxic (r=0.653) and B lymphocyte (r=0.237).Conclusions: Qualitative and semi quantitative characterization of lymphocytes subsets was useful in the assessment of different histological grades of UC disease activity.


Article
RELATIONSHIP OF PERIPHERAL BLOOD LYMPHOCYTES IMMUNE ALTERATION PHENOTYPE TO DISEASE ACTIVITY IN RHEUMATOID ARTHRITIS PATIENTS.
علاقة التغير المظهري المناعي لخلايا الدم اللمفية المحيطية مع مقياس فعالية المرض في المرضى المصابين بالتهاب المفاصل الرثوي

Authors: Abdul-Razak H. Ahmad عبد الرزاق حردان احمد --- Hayder F. Ghazi حيدر فيصل غازي
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2010 Volume: 8 Issue: 2 Pages: 10-17
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Background: Rheumatoid arthritis (RA) is a systemic autoimmune disorder causing synovitis of diarthroidal joints in which activated T-lymphocytes has a prominent position in RA pathology and may be important in prediction of disease outcome. Objective: To evaluate cellular expression of certain activation marker on PBLs and their relevance to disease activity pattern of RA patients.Patients and methods: this study included forty six RA patients, seven patients with Osteo-arthritis (OA) and 10 apparently healthy individuals. The collection of our baseline data based on routine laboratory and clinical assessment of disease activity Score (DAS). Blood sample was taken from each subject in all groups, at the time of attendance. Lymphocytes were separated; slides were prepared fixed on charged slides, foiled, and kept at -20ºC until assayed. CD3 and CD54 expression was detected using Immunocytochemistry staining, while CD71was detected using direct immunoflouresence staining.Results: The results of CD3 and CD54 revealed a statistically higher percentage of expression in rheumatoid arthritis patients when compared with that of the apparently healthy and OA control groups. The CD71 showed statistically significant higher expression in minimum disease activity group without any correlation with clinical and laboratory disease activity indices. Conclusions: we provide further evidence of a T-cell differentiation defect in RA, which could explain some of the well-characterized immunologic features of the disease but it is not related to the disease activity state.Keywords: Rheumatoid arthritis, disease activity, lymphocytes activation, immunocytochemistry, immunofluorescence and CD marker.

خلفية الدراسة: إلتهاب المفاصل الرثوي هو احد امراض ذاتية المناعة يسبب التهاب الغشاء الزلالي للمفاصل ثنائية التركيب, حيث تلعب الخلايا اللمفية المنشطة دوراً مهماً في امراضية التهاب المفاصل الرثوي ولهذا يعتقد بان لها دور مهم في تنبؤ مآل المرض.هدف الدراسة: تقدير التعبير الخلوي لبعض دوال التنشيط في خلايا الدم اللمفية المحيطية وصلتها بنمط نشاط مرض إلتهاب المفاصل الرثوي.المواد وطرق العمل: صممت هذه الدراسه للتحري عن نسب اظهار دوال التغير المظهري المناعي في خلايا الدم اللمفية المحيطية في 46 مريض مصاب بالتهاب المفاصل الرثوي, 7 مرضى مصابين بالتهاب المفاصل العظمي و 10 اشخاص اصحاء ظاهرياً. جمعت المعلومات الاساسيه اعتماداً على التقييم المختبري والسريري لفعالية المرض. اخذت عينات الدم من كل الاشخاص ولجميع المجاميع في وقت استشارة الطبيب المعالج, فصلت الخلايا اللمفية وعدت منها مسحات على شرائح زجاجية مشحونه, حفظت بدرجة حراره -20ºم لحين وقت التحري. قيست نسبة اظهار المعلمات المناعية 3 و 54 قيست باستخدام طريقة التصبيغ المناعي الخلوي الكيميائي بينما المحدد المناعي 71 فقد قيس نسبة اظهاره باستخدام التصبيغ المناعي الخلوي المتآلق المباشر.النتائج: ان نتائج المحدد المناعي 3 و 54 قد اظهرت نسبة اظهار عالية في مرضى التهاب المفاصل الرثوي وبمعنوية احصائيه عالية مقارنة بمجموعتي السيطره الصحية والمصابة بالتهاب المفاصل العظمي. اما المحدد المناعي 71 فقد لوحظ فرق احصائي معنوي وبنسبه اظهار اعلى في المجموعة متدنية شدة المرض. عدم وجود اي ارتباط معياري مع دوال المرض المختبرية والسريرية.الاستنتاج: من نتائج هذه الدراسة, قدمنا ادلة اضافية للخلل التمايزي الحاصل في الخلايا التائيه في مرضى التهاب المفاصل الرثوي والتي من الممكن ان توضح مظاهر مناعية واضحة لهذا المرض والتي ليس لها علاقة بفعالية المرضمفتاح الكلمات: التهاب المفاصل الرثوي، فعالية المرض، تغير المناعي المظهري، تصبيغ مناعي كيميائي، تصبيغ مناعي متالق و محددات مناعية.

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