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Robotic assisted microsurgical varicocelectomy
P-426 Wednesday, October 21, 2009 ANTIOXIDANT SYSTEM IN SEMINAL PLASMA AND ITS RELATIONSHIP TO SEMEN PARAMETERS IN INFERTILE MEN. A. C. Varghese, R. Gutgutia, A. K. Bhattacharyya, S. Suresh, J. Bhattacharya, P. Dasgupta. Fertility Clinic & IVF Div., AMRI Medical Center, Kolkata, West Bengal, India; Department of Biochemistry, University of Calcutta, Kolkata, West Bengal, India; IVF & Infertility Research Center, Kolkata, West Bengal, India; Institute of Reproductive Health & Toxicology, Kolkata, West Bengal, India. OBJECTIVE: The purpose of this investigation was to determine whether there exists a relationship between the antioxidant system in seminal plasma with clinical parameters of infertile men. DESIGN: Prospective study. MATERIALS AND METHODS: Fifty semen samples of infertile men were analyzed by WHO method and categorized into Normospermic, Asthenospermic and Oligospermic subjects. The semen antioxidant parameters such as G-6-PDH, catalase, GPX, GSH, SOD and MDA were analyzed by established spectrophotometrical methods. RESULTS: G-6-PDH (P<0.001), GPX (P<0.001), GSH (P<0.001) and SOD (P<0.001) were decreased in asthenospermic subjects compared to the normozoospermics. In oligospermics, all the antioxidant parameters were found to be increased than normospermic subjects, except GSH. The concentration of MDA was increased significantly (p<0.001) in semen of asthenospermic and oligospermics compared to normospermics. TABLE 1. Antioxidant parameters and semen variables Variables
Antioxidant Parameters
G-6-PDH (IU/L)
Catalase (U/mg of protein)
GPX (U/mg of Protein)
GSH (mg/dl)
SOD (U/mg of MDA Protein) (nmoles/ml)
Normospermic (12) 24.620.73 2.071.45 72.070.67 9.013.63 0.336.57 Asthenospermic (24) 18.990.58 2.281.42 60.880.99 0.106.26 0.216.43 Oligospermic (14) 26.573.92 2.772.41 116.182.23 6.621.41 0.451.46
0.411.61 0.722.11 0.582.89
CONCLUSIONS: The decreased activity of G-6-PDH causes increased ROS generation along with decrease in the activity of GPX and SOD results in sperm cell membrane fragility by the action of ROS and it affects ion exchanges essential for maintaining normal motility. Thus, this study reveals that there exists a strong relationship between the antioxidant system in the seminal plasma of asthenospermic subjects and clinical semen parameters. These findings substantiate the use of antioxidant therapy in infertile men especially those undergoing assisted conception procedures.
P-427 Wednesday, October 21, 2009 ROBOTIC ASSISTED MICROSURGICAL VARICOCELECTOMY. S. J. Parekattil, M. S. Cohen. Urology, University of Florida, Gainesville, FL. OBJECTIVE: Our goal was to compare standard microsurgical varicocelectomy (MVV) to robotic assisted varicocelectomy (RAVx) and present our initial human results. DESIGN: A prospective randomized control trial of MVV to RAVx performed in a canine varicocele model by an experienced surgeon (SP) and a review of our initial 18 human cases with the same surgeon. MATERIALS AND METHODS: For the canine study, the surgeon performed cord dissection and ligation of 3 veins with 3-0 silk ties. 12 canine varicocelectomies were performed, randomized into 2 arms of 6: MVV vs. RAVx. Procedure duration, vessel injury and knot failures were recorded. 18 human RAVx cases from Jun’08-April’09 were reviewed (follow up 110 months: mean 4). RESULTS: For the canine study, the RAVx duration (mean 9.5min) was significantly faster than MVV (mean 12min), p 0.04. There were no vessel injuries or knot failures in either group. In the human series, mean duration for the RAVx per side was 42 mins (25-80). Indications for the procedures were: 3 azoospermia, 12 oligospermia and 3 testicular pain (who had failed all other treatment options). 3 month follow up available for 10 patients: 6 with oligospermia had significant improvement in sperm counts (two achieved pregnancy), 1 with oligospermia had decrease in count and 1 with azoospermia remains unchanged. For testicular pain patients: 2 had complete resolution of pain, 1 only 50% reduction in pain. 4th robotic arm allowed the surgeon to control one additional instrument (Potts scissors or
S208
Abstracts
Doppler probe for arterial localization) during the cases. This improved surgeon efficiency and less reliance on the microsurgical assistant. CONCLUSIONS: Robotic assisted microsurgical varicocelectomy appears to be safe. There are advantages in decreasing operative duration and improving surgeon efficiency. The preliminary human results appear promising. Further evaluation and follow up is needed.
P-428 Wednesday, October 21, 2009 DIFFERENT SPERMATOZOAL PROTEIN PHOSPHORYLATION BETWEEN HEALTH MEN AND VARICOCELE OF SPERMATIC VEIN. C.-C. Chan, G. J. Wu, H.-A. Shui. ObGyn, Taipei City Hospital, Taipei, Taiwan; ObGyn, Tri-Service General Hospital, Taipei, Taiwan; Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan. OBJECTIVE: A clinical study targeting infertile males found that the majority of them suffer from varicocele of spermatic vein. Whenever spermatic vein dilates, it will adversely affect blood circulation. A current theory speculates that the expanded vein will cause elevation of testicle temperature and reduction of hormones inside the testicles, hence impeding the normal functions of the testicles. However, the exact mechanism linking male fertility and varicocele of spermatic vein is still not fully uncovered. DESIGN: This study involves 20 healthy males, and 20 varicocle of spermatic males. MATERIALS AND METHODS: Sperms from health men and varicocele of spermatic vein were isolated according to their motilities determined after liquefaction. Sperm protein samples were subjected to 2D electrophoresis. Western blot analyses were performed after 2D electrophoresis using to study protein phorphorylation. RESULTS: The research comparing normal males and those patients suffering from varicocele of spermatic vein, it was found that the spermatozoon of latter patients has more phosphorylation in 28 proteins included COPG2, Gamma-2 COP, exoribonulease 2, oxysterol-binding protein, Neutral alphaglucosidase AB precursor, Protocadherin alpha C1 precursor, Heat shock protein 1, hypothetical protein FJ, DNA-directed RNA-polymerases III, keratin cytokeratin-3, cytokeratin-9, FBXO21 protein, Heat shock protein 1L, HSV-1 stimulating-related protein, Myotonin-protein kinase, EH-domain protein 4, T-complex protein 1 alpha, Glyceraldehyde-3-phosphate dehydrogenase, protein disulfide-isomerase A3 precursor when comparing to the normal population. CONCLUSIONS: We have identified several proteins present at different amount in varicocele of spermatic vein sperm samples. These proteins could be candidates towards the development of diagnostic markers and open up the opportunity to gain further insight into the pathogenic mechanism involved in varicocele of spermatic vein. Supported by: The work were supported by grants from the TPECH and TSGH.
TESTIS P-429 Wednesday, October 21, 2009 THE ROLE OF VASCULAR ENDOTHELIAL GROWTH FACTOR IN ISCHEMIA-REPERFUSION INJURY OF TESTIS. H. Hashimoto, T. Ishikawa, K. Yamaguchi, K. Chiba, M. Shiotani, M. Fujisawa. Reproduction Center, Hanabusa Women’s Clinic, Kobe, Hyogo, Japan; Urology, Kobe Univercity Faculty of Medicine, Kobe, Hyogo, Japan. OBJECTIVE: Testicular torsion causes ischemia-reperfusion injury of testis and leads to male factor infertility. Its injury initiates a pathophysiologic cascade. In recent studies, protective effect of VEGF has been shown in various forms of ischemia- reperfusion injury. However, no study has investigated the role of VEGF in testicular ischemia-reperfusion injury. The aim of our study was to investigate the time course expression of VEGF, VEGF-receptor (R)1, VEGF-R2, inducible and endothelial nitric oxide synthase (iNOS and eNOS) and to evaluate the effect of VEGF in ischemiareperfusion injury of testis. DESIGN: Animal study. MATERIALS AND METHODS: Using 8-week-old SD rat, surgery for making unilateral experimental torsion was performed. After 1 hour of torsion, the testis was counter-rotated back to the natural position and reinserted into the scrotum. Bilateral castrations were performed in various time courses after reperfusion. Western bolt analysis and immunohistochemical
Vol. 92., No. 3, Supplement, September 2009
Antioxidant Parameters
G-6-PDH (IU/L)
Catalase (U/mg of protein)
GPX (U/mg of Protein)
GSH (mg/dl)
SOD (U/mg of MDA Protein) (nmoles/ml)
Normospermic (12) 24.620.73 2.071.45 72.070.67 9.013.63 0.336.57 Asthenospermic (24) 18.990.58 2.281.42 60.880.99 0.106.26 0.216.43 Oligospermic (14) 26.573.92 2.772.41 116.182.23 6.621.41 0.451.46
0.411.61 0.722.11 0.582.89
CONCLUSIONS: The decreased activity of G-6-PDH causes increased ROS generation along with decrease in the activity of GPX and SOD results in sperm cell membrane fragility by the action of ROS and it affects ion exchanges essential for maintaining normal motility. Thus, this study reveals that there exists a strong relationship between the antioxidant system in the seminal plasma of asthenospermic subjects and clinical semen parameters. These findings substantiate the use of antioxidant therapy in infertile men especially those undergoing assisted conception procedures.
P-427 Wednesday, October 21, 2009 ROBOTIC ASSISTED MICROSURGICAL VARICOCELECTOMY. S. J. Parekattil, M. S. Cohen. Urology, University of Florida, Gainesville, FL. OBJECTIVE: Our goal was to compare standard microsurgical varicocelectomy (MVV) to robotic assisted varicocelectomy (RAVx) and present our initial human results. DESIGN: A prospective randomized control trial of MVV to RAVx performed in a canine varicocele model by an experienced surgeon (SP) and a review of our initial 18 human cases with the same surgeon. MATERIALS AND METHODS: For the canine study, the surgeon performed cord dissection and ligation of 3 veins with 3-0 silk ties. 12 canine varicocelectomies were performed, randomized into 2 arms of 6: MVV vs. RAVx. Procedure duration, vessel injury and knot failures were recorded. 18 human RAVx cases from Jun’08-April’09 were reviewed (follow up 110 months: mean 4). RESULTS: For the canine study, the RAVx duration (mean 9.5min) was significantly faster than MVV (mean 12min), p 0.04. There were no vessel injuries or knot failures in either group. In the human series, mean duration for the RAVx per side was 42 mins (25-80). Indications for the procedures were: 3 azoospermia, 12 oligospermia and 3 testicular pain (who had failed all other treatment options). 3 month follow up available for 10 patients: 6 with oligospermia had significant improvement in sperm counts (two achieved pregnancy), 1 with oligospermia had decrease in count and 1 with azoospermia remains unchanged. For testicular pain patients: 2 had complete resolution of pain, 1 only 50% reduction in pain. 4th robotic arm allowed the surgeon to control one additional instrument (Potts scissors or
S208
Abstracts
Doppler probe for arterial localization) during the cases. This improved surgeon efficiency and less reliance on the microsurgical assistant. CONCLUSIONS: Robotic assisted microsurgical varicocelectomy appears to be safe. There are advantages in decreasing operative duration and improving surgeon efficiency. The preliminary human results appear promising. Further evaluation and follow up is needed.
P-428 Wednesday, October 21, 2009 DIFFERENT SPERMATOZOAL PROTEIN PHOSPHORYLATION BETWEEN HEALTH MEN AND VARICOCELE OF SPERMATIC VEIN. C.-C. Chan, G. J. Wu, H.-A. Shui. ObGyn, Taipei City Hospital, Taipei, Taiwan; ObGyn, Tri-Service General Hospital, Taipei, Taiwan; Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan. OBJECTIVE: A clinical study targeting infertile males found that the majority of them suffer from varicocele of spermatic vein. Whenever spermatic vein dilates, it will adversely affect blood circulation. A current theory speculates that the expanded vein will cause elevation of testicle temperature and reduction of hormones inside the testicles, hence impeding the normal functions of the testicles. However, the exact mechanism linking male fertility and varicocele of spermatic vein is still not fully uncovered. DESIGN: This study involves 20 healthy males, and 20 varicocle of spermatic males. MATERIALS AND METHODS: Sperms from health men and varicocele of spermatic vein were isolated according to their motilities determined after liquefaction. Sperm protein samples were subjected to 2D electrophoresis. Western blot analyses were performed after 2D electrophoresis using to study protein phorphorylation. RESULTS: The research comparing normal males and those patients suffering from varicocele of spermatic vein, it was found that the spermatozoon of latter patients has more phosphorylation in 28 proteins included COPG2, Gamma-2 COP, exoribonulease 2, oxysterol-binding protein, Neutral alphaglucosidase AB precursor, Protocadherin alpha C1 precursor, Heat shock protein 1, hypothetical protein FJ, DNA-directed RNA-polymerases III, keratin cytokeratin-3, cytokeratin-9, FBXO21 protein, Heat shock protein 1L, HSV-1 stimulating-related protein, Myotonin-protein kinase, EH-domain protein 4, T-complex protein 1 alpha, Glyceraldehyde-3-phosphate dehydrogenase, protein disulfide-isomerase A3 precursor when comparing to the normal population. CONCLUSIONS: We have identified several proteins present at different amount in varicocele of spermatic vein sperm samples. These proteins could be candidates towards the development of diagnostic markers and open up the opportunity to gain further insight into the pathogenic mechanism involved in varicocele of spermatic vein. Supported by: The work were supported by grants from the TPECH and TSGH.
TESTIS P-429 Wednesday, October 21, 2009 THE ROLE OF VASCULAR ENDOTHELIAL GROWTH FACTOR IN ISCHEMIA-REPERFUSION INJURY OF TESTIS. H. Hashimoto, T. Ishikawa, K. Yamaguchi, K. Chiba, M. Shiotani, M. Fujisawa. Reproduction Center, Hanabusa Women’s Clinic, Kobe, Hyogo, Japan; Urology, Kobe Univercity Faculty of Medicine, Kobe, Hyogo, Japan. OBJECTIVE: Testicular torsion causes ischemia-reperfusion injury of testis and leads to male factor infertility. Its injury initiates a pathophysiologic cascade. In recent studies, protective effect of VEGF has been shown in various forms of ischemia- reperfusion injury. However, no study has investigated the role of VEGF in testicular ischemia-reperfusion injury. The aim of our study was to investigate the time course expression of VEGF, VEGF-receptor (R)1, VEGF-R2, inducible and endothelial nitric oxide synthase (iNOS and eNOS) and to evaluate the effect of VEGF in ischemiareperfusion injury of testis. DESIGN: Animal study. MATERIALS AND METHODS: Using 8-week-old SD rat, surgery for making unilateral experimental torsion was performed. After 1 hour of torsion, the testis was counter-rotated back to the natural position and reinserted into the scrotum. Bilateral castrations were performed in various time courses after reperfusion. Western bolt analysis and immunohistochemical
Vol. 92., No. 3, Supplement, September 2009