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Article
ROLE OF DNA INTEGRITY OF SPERMATOZOA IN MALE INFERTILITY

Author: Ali F Al-Hashimi علي فؤاد الهاشمي
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2012 Volume: 10 Issue: 1 Pages: 4-11
Publisher: Al-Nahrain University جامعة النهرين

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Abstract

Background Classical semen analysis gives an approximate evaluation of the functional competence of spermatozoa, but not the quality of sperm DNA. With the advent of assisted reproductive technologies (ART), the concern over using damaged DNA has become apparent.Objective To clarify the role of DNA integrity and maturity of ejaculated spermatozoa in male infertility.MethodsA randomly selected group of 50 nonazoospermic infertile patients with a history of infertility of at least 1 year duration were included in this study. Whereas control group consisted of semen samples obtained from healthy volunteers of proven fertility (n = 27). Two main assays were studied in ejaculated spermatozoa: The green Acridine Orange (AO) fluorescence test that measures DNA integrity and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labeling (TUNEL)- based ApopTag® technology that assesses DNA fragmentation as a late apoptotic marker. Results were compared with the standard sperm characteristics (concentration, motility and morphology) between infertile patients as well as some patients' subgroups against control donors.Results Significant low levels of green AO fluorescent spermatozoa were observed in all patients and patient's subgroups with asthenoteratozoospermia and oligoasthenoteratozoospermia (P = 0.00001). High percentages of TUNEL-positive spermatozoa were significantly noticed, against control donors, in all patients as well as asthenoteratozoospermic and oligoasthenoteratozoo-spermic infertile patients. Interestingly, normozoospermic patients had a significantly low percentage of green AO fluorescent spermatozoa and high levels of TUNEL-positive spermatozoa versus control donors (P = 0.0005, P = 0.0069, respectively).Conclusion From this study, it can be concluded that male infertility is associated with high rates of DNA damage in the spermatozoa, and that sperm DNA damage analysis could reveal a buried deformity of sperm nuclear DNA in infertile men classified as idiopathic, having apparently normal standard sperm parameters.Keywords DNA denaturation, DNA fragmentation, acridine orange, TUNEL.


Article
The Effect of Coumarin Derivatives(compounds) on the Vibrio cholerae Isolates from Different Clinical Iraqi Sources
تأثير مركبات الكومارين على عزلات من بكتريا Vibrio cholerae المعزولة من مصادر سريرية عراقية مختلفة

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Abstract

From a large number of bacterial samples collected from different hospital in Iraq in central health laboratory ,only ten isolates were identified primary as Vibrio. A number of morphology and biochemical test were carried out to complete this identification that showed all bacterial isolates were related to Vibrio cholerae .In this study all Vibrio isolates were investigated for Bio typing and the result showed that all (10) isolate were related to (Eltor biotypes) .Also, the susceptibility test towards eight antibiotics were carried out .Results shows that ciprofloxacin , Norfloxacin, Erythromycin, Ampicillin, ceftriaxone and Amikacin were the most effective antibiotics and their resistance percentage were 20%,20%,20, 20,30% and 30% respectively ,While Chloramphenicol and Co- trimoxazole were less effective and their resistance percentage were 90% both of them. Three (3,5,6) isolates V. cholerae were selected depending on results of antibiotics sensitivity tests as showed multiple –antibiotics resistance(100%). Then tested to study the effect of coumarin derivatives compounds (1, 2, 3 ) which showed inhibitory effect on V. cholera (3,5,6) isolates and the compound ( 3 ) showed the highest antibacterial activity of (12,15,14 mm) of inhibition zone diameter against V. cholera (3,5,6) isolates respectively. Also, these Iraqi isolates (3,5,6) used to test the effect of acridine orange (0.1%) as acuring agent , the results showed that all (3) isolates V. cholerae were sensitive to (ciprofloxacin, ceftrixone and Norfloxacin), While the rest were resistance to remained five antibiotics. The results of Agarose –gel electrophoresis of both normal V. cholerae (3,5,6) and cured isolates showed the presence of chromosomal and plasmid DNA bands in the normal case ,While only chromosomal DNA bands occur with V. cholerae (isolate 8) treated with an acridine orange at concentration of (10-2 to 10-4).

من مجموع عدد كبير من نماذج بكتيرية جمعت في مختبر الصحة المركزي من مستشفيات مختلفة في العراق للفترة من 1/11/2015- 1/4/2016 ,تم الحصول على (10) عزلات شخصت مبدئيا على انها تعود الى جنس vibrio .تم اجراء عدد من الفحوصات المظهرية والكيموحياتية لا كمال هذا التشخيص مما اظهر ان هذه العزلات تعود الى Vibrio cholerae.شملت هذه الدراسة اخضاع هذه العزلات الى التنميط الحيوي (Bio typing) وقد اظهرت النتائج ان جميع هذه العزلات العشرة كانت من نوع (Eltor).كما تم قياس حساسية هذه العزلات تجاه ثمانية (8) من المضادات الحيوية , واظهرت النتائج ان السبروفلوكساسين , نورفلوكساسين ,الارثرومايسين, الامبسلين والسيفرتوكسين والاميكاسين كانت اكثرالمضادات تاثيرا وبنسبة مقاومة تساوي 20%,20%,20%,20%,30%,30% على التوالي, بينما كانت للكلورمفنيكول والكواتراميكسول اقل تاثيرا حيث كانت نسبة المقاومة 90% لكليهما. تم اختبار ثلاث عزلات من بكتريا V. choleraeهي (6,5,3) اعتمادا على نتائج الحساسية للمضادات الحيوية والتي اظهرت مقاومة متعددة بنسبة (100%) . كما تم اخضاعها (العزلات الثلاثة) لدراسة تأثير مركبات الكومارين الثلاثة (3,2,1) (كلوريد الكوبلت, كلوريد المنغنيز, المركب الثالث هو مزيج من كليهما) على العزلات الثلاثة حيث اظهرت النتائج ان المركب (3) كان له اعلى تاثير ضد بكتيري بقطر تثبيط (14,15,12 ملم) ضد عزلات V. cholerae(6,5,3) على التوالي.كما شملت هذه الدراسة اختبار حساسية المضادات من خلال دراسة تأثير مادة الاكردين البرتقالي بتركيز(0.1%) كعامل محيد على هذه العزلات اعلاه وقد اظهرت النتائج ان جميع العزلات الثلاثة من البكتريا كانت حساسة للمضادات (سبروفلوكساسين ,سيفوتوكسيم والنورفلوكساسين) بينما بقية العزلات كانت مقاومة المضادات الخمسة الباقية.اظهرت نتائج الجانب الوراثي للترحيل الكهربائي لكل من عزلات V. cholerae(6,5,3) الطبيعية والمحيدة وجود حزمة للبلازميد في العزلات الطبيعية بينما ظهور حزمة للكروموسوم فقط لعزلات ضمات الكوليرا (العزلة 8)المعاملة مع الاكردين البرتقالي بتركيز ( 10-4 الى 10-2 ) .

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