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EVALUATION OF SERUM AND URINARY FIBRONECTIN AS A DIAGNOSTIC MARKER OF BLADDER CANCER

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Abstract

Background:Accurate and sensitive detection of bladder cancer is important to diagnose this deadly disease at an early stage, estimate prognosis, prediction the response to treatment and for monitoring the recurrence. In past few years, laboratory diagnosis and surveillance of urinary bladder cancer have improved significantly. Although, urine cytology remains the gold standard test, many new urinary biochemical markers have been identified.Objectives:To evaluate the value of fibronectin in serum and urine to detect bladder cancer in different grades and stages.Methods:Thirty five patients diagnosed as bladder cancer with mean age 61.94±11.66 years and thirty five aged-matched healthy volunteers as control group were included in this study. Serum and urinary fibronectin were measured by ELISA technique.Results:The mean±SEM serum and urine levels of fibronectin in patients with bladder cancer (33.11±1.90 µg/ml; 33.08±1.12 ng/ml respectively) were significantly higher than the levels in control group (8.57±1.10 µg/ml; 7.58±1.00 ng/ml, respectively). When using a serum fibronectin concentration of 25.65 µg/ml as a cutoff value for the diagnosis of bladder cancer, sensitivity was 71.4%, specificity 100%, the positive predictive value was 100% and the negative predictive value 77.78%, and the sensitivity and specificity of urine fibronectin were (94.3%, 97.1% respectively); when using a urine fibronectin concentration of 20.00 ng/ml as a cutoff value for the diagnosis of bladder cancer. The positive predictive value was 97.05%, and the negative predictive value was 94.44%.Conclusion:The measurement of fibronectin level in serum and urine is useful in discriminating bladder cancer patients from normal subjects.Key words:Serum and urinary Fibronectin, Bladder cancer.


Article
DNA Methylation Patterns of Interferon Gamma Gene Promoter and Serum Level in Pulmonary Tuberculosis: Their Role in Prognosis
طرز مثيلة الدنا لمشغل جين الانترفيرون كاما ومستواه في المصل لمرضى التدرن الرئوي– دورهما في الاستعدادية للإصابة

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Abstract

Tuberculosis (TB) still remains an important medical problem due to high levels of morbidity and mortality worldwide. A series of innate immune mechanisms that create a cytokine network control the pathogenesis of tuberculosis and this response has the capacity to modify the host genomic DNA structure through epigenetic mechanisms such as DNA methylation which could constantly alter the local gene expression pattern that can modulate the metabolism of the tissues and the immune-response. Interferon-gamma (IFN-γ) is an important pro-inflammatory cytokine regulator of the innate immune response to TB. This study aims to determine DNA methylation patterns of INF-γ gene promoter and measure serum IFN- γ level in newly diagnosed TB patients, relapse TB patients, and healthy control, in order to study the possibility of using these as a biomarker for the prognosis of TB stages in patients. The current case-control study included 66 patients with TB and 33 healthy control subjects. DNA was extracted from peripheral blood(PB) of included subjects and modified using sodium bisulfate specific kit. DNA methylation patterns of IFN-γ gene promoter was determine by using methylation specific polymerase chain reaction(MS-PCR).Serum IFN-γ level was determined using enzyme linked immune-sorbent assay(ELISA). Results showed that percentages of DNA methylation patterns in normal controls, newly diagnostic TB patients and relapse TB patients were (63.3%, 18.2% and 21.2% respectively). Also, higher significant differences (P≤0.0001) of un-methylated IFN-γ gene promoter patterns in newly diagnostic TB patients than relapse TB patients comparison with healthy controls. The percentage of un-methylated DNA patterns in healthy controls, newly diagnostic TB patients and relapse TB patients were (9.9%, 39.4% and 51.5%, respectively). The mean of serum IFN-γ levels (pg/ml) for normal controls, newly diagnostic TB patients and relapse TB patients were (59.3± 13.8,75.8±24.3 and 69.6±18.7,respectively).In conclusion, there is a relative association between methylation of IFN-γ gene promoter and predisposing to TB progression.

يعد مرض التدرن الرئوي مشكلة صحية عالمية لما يسببه من إمراضيه ووفيات. تنشا سلسلة من الاستجابات المناعية الفطرية نتيجة الاصابة بالتدرن الرئوي والتي تتمثل بشبكة من الحركيات الخلوية للسيطرة على إمراضية التدرن الرئوي وهذه الاستجابة سوف تحور تركيبة دنا الخلايا المصابة من خلال اليات التخليق المتعاقب مثل مثيلة الدنا والذي من الممكن ان يؤدي الى تحوير في طرز التعبير الجيني وبالتالي يغير في العمليات الايضية النسيجية والاستجابة المناعية. يعد الانترفيرون كاما احد الحركيات الخلوية للاستجابة الالتهابية الأولية للتدرن الرئوي . تهدف الدراسة الحالية الى تحديد طرز مثيلة الدنا لمشغل جين الانترفيرون كاما وقياس مستوى الانترفيرون كاما في مصل الدم في مرضى التدرن الرئوي المشخصة اصابتهم حديثا و مرضى التدرن الرئوي المتكرر وكذلك السيطرة من الاشخاص الاصحاء وذلك لدراسة امكانية استخدامها كمعلمات بيولوجية في دراسة الاستعدادية للإصابة بالمرض. تم جمع 66 عينه دم من المرضى الذين يعانون من مرض التدرن الرئوي بواقع 33 عينة من مصابين تم تشخيصهم حديثا و33 عينة من مرضى التدرن الرئوي المتكرر للفترة من ( 1-4 -2016 ولغاية 1- 4- 2017) و33 عينه دم من الاشخاص الاصحاء .تم الحصول على العينات المرضية من مركز التدرن في بغداد ومستشفى واسط ومستشفى الإمامين الكاظمين(ع).تم قياس مستوي الانترفيرون كاما في مصل 66 مريضًا وكذلك مصل 33 من الاصحاء بطريقة التفاعل المناعي المقترن بالأنزيم. استخلص الدنا من عينات الدم المحيطي وتم تحوير تركيبته باستخدام عدة متخصصة من الصوديوم ثنائي الكبريت. تم الكشف عن طراز المثيلة في دنا مشغل جين الانترفيرون كاما للاشخاص قيد الدراسة بواسطة طرق تفاعل البلمرة المتسلسلة المختص بالمثيلة. اشارة النتائج الاحصائية الى وجود علاقة معنوية بين مثيلة مشغل جين الانترفيرون كاما للأشخاص الاصحاء و مرضى التدرن الرئوي حديثي التشخيص ومرضى التدرن الرئوي المتكرر وكانت النسب المئوية للدنا الممثيل ( 63.3% ،18.2% و 21.2%، على التوالي)، كذلك وجود علاقة احصائية معنوية للاختلافات في طرز الدنا الغير ممثيل بين المجاميع قيد الدراسة وكانت النسب المئوية للدنا الغير ممثيل (9.9% ،39.4% و51.5% ، على التوالي). اشارت النتائج الى وجود علاقة احصائية معنوية للاختلافات في متوسط مستوى الانترفيرون كاما (بيكوغرام/ مل) في مصل الأشخاص الاصحاء و مرضى التدرن الرئوي حديثي التشخيص ومرضى التدرن الرئوي المتكرر.يستنتج مما سبق وجود ترافق نسبي بين مثيلة مشغل جين الانترفيرون كاما والاستعدادية لتطور الاصابة بالتدرن الرئوي.


Article
ASSOCIATION BETWEEN ASN142ASP GENETIC POLYMORPHISM OF GSTO2 AND SUSCEPTIBILITY TO BLADDER CANCER

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Abstract

Background:The glutathione-S-transferases (GSTs) comprise a class of enzymes that detoxify carcinogenic compounds by conjugating glutathione to facilitate their removal. Polymorphisms in glutathione S-transferase Omega 1,2 (GSTO1, GSTO2), and GSTP1 genes have been related to risk for bladder cancer.Objective:To assess a comprehensive picture of the relationship between smoking and GSTO2 gene Asn142Asp variant (rs156697) with bladder cancerMethods:A case control study was conducted at Chemistry and Biochemistry Department, College of Medicine and DNA Research and Training Center, Al-Nahrain University from February 2014 to September 2014. Forty one bladder cancer patients and 41 age matched apparently healthy controls were participated in this study. Genotyping of the GSTO2 Asn142Asp polymorphism was evaluated using a polymerase chain reaction fragment length polymorphism (PCR-RFLP) method. The odds ratio (OR) and 95% confidence interval (CI) were calculated as a measure of the combined effect of cigarette smoking and the GSTO2 Asn142Asp polymorphism on bladder cancer risk.Result:It was found that subject with the GSTO2 Asp/Asp genotype have significantly increased bladder cancer risk (OR 4.92; 95% CI =1.32 - 18.30). A statistically highly significant increased the bladder cancer risk was also found in ever smoker of the GSTO2 (Asn/Asn) (OR =11.8; 95% CI=2.43 - 57.84) and (Asn/Asp +Asp/Asp) (OR =12.8; 95% CI=3.23 - 51.41) compared with never smoker Ala/Ala genotype.Conclusion:The study suggests that smokers having GSTO2 Asn/142Asp polymorphism could play an important role as risk factor for the development with bladder cancer.Keywords:Bladder cancer, single nucleotide polymorphism, glutathione S-transferase, GSTO2, Asn142Asp, smoking, rs156697.

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