Age as a Limiting Factor for Successful Sperm Retrieval in Nonmosaic Klinefelter Syndrome

the sperm fraction used for the motility assay. This may also explain the inconsistent results seen in studies assessing the utility of SCSA to predict IVF and ICSI outcome. Another possible explanation is that factors other than oxidative damage may play a more critical role in determining the rate of motility loss. Supported by: None

P-154 Age as a Limiting Factor for Successful Sperm Retrieval in Nonmosaic Klinefelter Syndrome. H. Okada, O. Maruyama, S. Muto, Y. Yamamoto, M. Koshida, S. Horie. Teikyo University School of Medicine, Tokyo, Japan; Yamamoto Urological Clinic, Yonago, Japan; Koshida Clinic, Osaka, Japan. OBJECTIVE: Intracytoplasmic sperm injection (ICSI) now permits fertilization of eggs using very small numbers of spermatozoa. Improvements in testicular sperm extraction (TESE) techniques have expanded indications from oligozoospermia to nonobstructive azoospermia. Klinefelter syndrome, which accounts for 5% of the latter cases, is characterized by sterility caused by lack of spermatogenesis. Isolated foci of spermatogenesis have been found in involved testes, explaining the rare cases of sperm production with appearance in the ejaculate. Several institutions perform TESE using either ordinary testicular biopsy techniques or meticulous microdissection to retrieve spermatozoa for ICSI. The reported success rate for TESE in patients with Klinefelter syndrome is 56%, but no predictive factors have been identified given the paucity of cases in each report. We analyzed results of TESE to elucidate success determinants for TESE in patients with Klinefelter syndrome. DESIGN: Retrospective analysis of microdissection TESE in nonmosaic Klinefelter syndrome patients. A collaborative study with three institutions. MATERIALS AND METHODS: Eighty-four patients with nonobstructive azoospermia related to nonmosaic Klinefelter syndrome underwent TESE. To determine factors affecting successful sperm retrieval by TESE in patients with nonmosaic Klinefelter syndrome medical record were analyzed retrospectively. Correlations of patient characteristics, serum concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), and testosterone (T), as well as testicular volume with success in testicular sperm extraction were investigated. RESULTS: We succeeded in obtaining spermatozoa in 51 patients and failed in 33. LH, FSH, T, and testicular volume did not differ between patient groups defined by success and failure. Median ages for successful and failed TESE were 32.6 and 37.1, respectively (p⬍0.001). When we analyzed success rates of testicular sperm extraction for 5-year age brackets, the ratio of successful recovery of spermatozoa to failure decreased beyond 35 years. CONCLUSIONS: Testicular sperm extraction should be recommended before the critical age of 35 in nonmosaic Klinefelter syndrome patients. Supported by: None

previously produced sperm that responded normally in the HSAA was treated for an infection with 100 mg of doxycycline 2X daily. Sperm collected at day 5 of the treatment, and at days 10 and 20, after the treatment was finished were assayed in the HSAA evaluating the sperm activation events described above. RESULTS: No sperm in either the in vitro or in vivo studies synthesized DNA during the 15 min incubation in frog egg extract. Exposure of sperm to doxycycline results in irreversible hyperdecondensation in up to 30% of in vitro treated sperm, and 40% of in vivo treated sperm. Hyperdecondensed sperm are sperm with a 2 fold increase in size over that observed in the fully decondensed sperm. The hyperdecondensed sperm that did not recondense did not synthesize DNA. The in vivo doxycycline treated sperm had similar abnormal responses in the HSAA as were observed for the sperm treated in vitro with doxycycline (100 ␮g/ml). Only sperm that recondensed were able to synthesize DNA (2 hr time point). The abnormal responses in the HSAA observed in the sperm from the male treated with doxycycline for 5 days, were found to revert back to normal in sperm produced 10 days after finishing the treatment.

P-155 Doxycycline Exposure: Effects on Human Sperm Activation. D. B. Brown, E. E. Tirado. Repromedix Corp, Woburn, MA; The University of Texas Medical Branch, Galveston, TX. OBJECTIVE: Doxycycline is routinely used in fertility clinics to treat infections in males prior to their producing a semen sample for use in ART attempts at pregnancy. The objectives of this study are to determine if: 1) in vitro doxycycline treatment of sperm will induce abnormal response(s) when the sperm are analyzed in the human sperm activation assay (HSAA), 2) abnormal response(s) are observed in sperm from a male treated in vivo with doxycycline, when analyzed in the HSAA, 3) the abnormal response(s) are reversible, and the days required postantibiotic treatment for sperm to respond normally in the HSAA. DESIGN: In vitro dose response study, and an in vivo prospective case study. MATERIALS AND METHODS: In vitro study: Sperm ejaculates from fertile males were divided into 5 aliquots: control non-treated sperm, and sperm treated with doxycycline at a concentration of 2.5, 5, 10, and 100␮g/ml. After an exposure time of 1 hr, sperm were evaluated for sperm activation events in the HSAA (decondensation, DNA synthesis measured by determining 3H-TTP incorporation, and recondensation, at 15 min, and 2 hr). In vivo study: A fertile male who had

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Abstracts

CONCLUSIONS: These results are similar to those we previously reported for sperm exposed to 100␮g/ml reactive oxygen species (ROS), with the exception being no DNA synthesis occurred at the 15 min time point in the doxycycline treated sperm indicating that no DNA damage occurred.The abnormal hyperdecondensation is possibly a result of lipid soluble doxycycline changing the properties of the membrane. The clinical significance of these abnormal responses are not yet known. However, the results from this study suggest that males being treated with doxycycline should not provide samples for ART attempts at pregnancy until they have been off the drug at least 10 days. Supported by: CONACYT Mexico, Grant 90735 to E.E.T. P-156 Comparison of Sperm Motility Measurement Using SQA-V Automated Sperm Analyzer and Conventional Manual Methods. R. K. Sharma, T. M. Said, A. Agarwal. Cleveland Clinic Foundation, Cleveland, OH. OBJECTIVE: The accuracy of manual sperm motility assessment is affected by a variety of factors. Automated systems may be a potential

Vol. 84, Suppl 1, September 2005