79 EFFECTS OF ANTIOXIDANTS AND ROS ON SPERM DNA INTEGRITY IN IDIOPATHIC INFERTILE MEN

S32 78 ACCURACY OF HOFFMAN-BASED OPTIC SYSTEM FOR THE SELECTION OF MORPHOLOGICALLY NORMAL SPERMATOZOA IN ASSISTED REPRODUCTION TECHNIQUES S. Rovira1 , O. Cairo1 , A. Brassesco1 , F. Del Rio1 , M. Rodriguez1 , L. Prats1 , M. Brassesco2 . 1 CIRH, IVF Laboratory, Barcelona, Spain, 2 CIRH, Medical Director, Barcelona, Spain Introduction: When an ICSI is performed, it is expected to select the maximum number of good quality spermatozoa. The objective of this study was to assess the accuracy for the selection of “normal” spermatozoa of a standard Hoffman-based optic system, when compared with a high magnification microscope. Material and Methods: Samples from 20 patients undergoing an IVF treatment were studied. All samples were over 106 of A-grade spermatozoa/ml. 10 spermatozoa of each sample were selected at 400× using a standard inverted microscope (used normally for ICSI), and were re-assessed using a high magnification microscope (used for IMSI). Spermatozoa were assessed depending on its vacuolization pattern (Grade I to IV). Grade I and Grade II means good quality spermatozoa, while grades III and IV means low quality (abnormal vacuolization). Results: When comparing with high magnification microscope, only 27% (n = 54) of spermatozoa selected as “normal” with the 400× microscope were considered good-quality, while the remaining 73% (n = 146) presented an abnormal vacuolization pattern. Conclusions: Although the sample size was small, we observed a poor correlation rate between the two tests. The high rate of false positive results (73%) would come from the limitation of the optic system used. Even when the assessment of the samples was performed by the same operator, we used an objective evaluation method that minimises the verification bias. The evaluation of the impact of this results on embryo development as well as in implantation and miscarriage rates are still pending. 79 EFFECTS OF ANTIOXIDANTS AND ROS ON SPERM DNA INTEGRITY IN IDIOPATHIC INFERTILE MEN M.B. Shamsi1 , S. Venkatesh1 , P. Talwar2 , R. Kumar1 , R. Dada1 . 1 All India Institute of Medical Sciences, New Delhi, India, 2 ART Centre, Army research and Referral Centre, New Delhi, India Introduction: Abnormalities in the sperm genome integrity are a potential reason for infertility. Sperm DNA maintains its high degree of inertness, stability and integrity by processes as absence of transcription, replacement by protamine, presence of DNA repair genes and presence of an antioxidant mechanism. Reactive Oxygen Species (ROS) in pathological concentrations is an important factor challenging the antioxidant and DNA repair machinery of the sperm. ROS damages the lipid in the sperm membrane affecting the motility and morphology and can also induce mutations in the nuclear and mitochondrial genome. Sperms subjected to increased free radical concentrations, have decreased energy production and motility defects which reduces the chance of reaching the female gamete. The antioxidant machinery regulate the free radical concentration in physiological levels. The imbalance between the anti oxidant and free radicals generated either due to increased free radical or lower activity or decreased expression of antioxidants produces an oxidative environment in the sperm. The structural biomolecules as well as the nuclear and mitochondrial genome in this oxidative stress are prone to oxidation which leads to

WARM 2010 disruption in their functions. Oxidation of nucleotides produces DNA breaks which, if unrepaired are lethal to the accurate transmission of genetic information. The current study was designed to study the affect of ROS and antioxidant levels on the sperm DNA integrity. Materials and Methods: Semen samples from 97 idiopathic infertile patients and 52 normozoospermic healthy control were examined. Level of DNA damage was assessed by Comet assay, ROS by chemiluminescence and seminal antioxidants (Superoxide Dismutase- SOD, CatalaseCat, Gluatathione-GSH) by biochemical methods. Semen analysis was done according to WHO 1999 guidelines. Statistical analysis was done using Mann Whittney test. Results: On the basis of sperm parameters the infertile men were divided into two groups- one with the abnormal sperm pathology (Group 1) and the other with normal sperm parameters (Group 2). In group 1 the mean sperm concentration is 6.7±2.19 million/ml, progressive motility is 38.65±4.37, abnormal sperm morphology is 42.37±3.18. In group 2, mean sperm concentration is 23.03±1.34 million/ml, progressive motility is 57.39±3.71, abnormal sperm morphology is 20.59±4.29. ROS levels were significantly higher in group 1 and 2 as compared to controls (p = 0.018, p = 0.053 respectively). Seminal SOD and catalase was significantly lower in both, group 1 and 2 (p= 0.012, p = 0.037 respectively) as compared to controls. There was no significant difference in the levels of seminal glutathione in group 2 patients as compared with controls (p = 0.062) whereas group 1 patients had significantly lower glutathione (p = 0.029). The number of sperms with high DNA damage was significantly greater in both the patients groups as compared to controls (p = 0.009; p = 0.017). Conclusion: There is accumulating evidence linking sperm nuclear DNA anomalies to elevated ROS, lowered antioxidants and poor reproductive outcome. Antioxidant supplementation may help to protect sperm from oxidative stress induced damage and there by help to improve the rate of conception and successful pregnancy. Oxidative stress induced paternal DNA damage leads to pathological sperm parameters, poor quality blastocysts (during assisted conception), less than optimal pregnancy rates, and increased chances of miscarriages, which gives emotional and psychological trauma to the couples experiencing it. Therefore assessment of oxidative stress should be included in the work up of infertile men. 80 INCREASED PERCENTAGE OF SPERMATOZOA WITH FRAGMENTED DNA CORRELATES WITH SPERM HEAD ABNORMALITIES E.M. Shilnikova1 , I.D. Fedorova2 , T.V. Kuznetzova2 , A.M. Gzgzyan2 . 1 St-Petersburg State University, SaintPetersburg, Russia, 2 Ott’s Institute of Obstetrics and Gynecology, Saint-Petersburg, Russia Sperm DNA fragmentation is an important parameter of semen quality which affects ART outcomes. However, basic semen analysis does not include assessment of sperm DNA fragmentation. Are there any relationships between conventional semen parameters and the incidence of sperm DNA fragmentation? The aim of this study was to estimate correlation between sperm head morphology and the level of sperm DNA fragmentation. In semen samples from 17 men from infertile couples portion of sperm with vacuole and bulb head morphology was assessed in 300 spermatozoa per sample. The evaluation incidence of sperm DNA fragmentation was performed in 2000 sperm per