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Is There a Place for Round and Elongated Spermatids Injection in Assisted Reproduction? 2 1 - 2 Spermatids are the earliest male germ cells with one set of haploid chromosomes. After experiments, mainly in rodents, the spermatid injection was introduced in human assisted reproduction to the treatment of men with non-obstructive azoospermia. Spermatid injection is a technique with particular difficulties that may negatively influence the outcome. The identification, isolation and the assessment of viability, especially for round spermatids, require intensive work and considerable experience. Up to date, it appears that the rates of fertilization and implantation with round spermatid injection are dramatically low and significantly less compared to the use of elongated spermatid injection. The extremely low fertilization potency of the round spermatids led to attempts for their in-vitro culture and maturation. The immaturity of round and elongated spermatids has raised concerns regarding the potential genetic risk for the offspring. Under these facts, a reconsideration of the use of spermatids in assisted human reproduction is necessary. 1 6 2017/10/1 1396/7/9 2003/01/15 1381/10/25 Byron Asimakopoulos Byron Asimakopoulos Nikos Nikolettos Nikos Nikolettos Safa Al-Hasani Safa Al-Hasani - Al-Hasani S, Ludwig M, Palermo I, Küpker W, Sandmann J, Johannisson R, Fornara P, Sturm R, Bals-Pratsch M, Bauer O, Diedrich K. (1999a) Intracytoplasmic injection of round and elongated spermatids from azoospermic patients: results and review. Hum Reprod 14: 97-107.##Al-Hasani S, Schِpper B, Küpker W, Sandmann J, Johannisson R, Fornara P, Sturm R, Bals-Pratsch M, Bauer O, Diedrich K. (1999b) Die intrazytoplasmatische Injektion von runden und elongierten Spermatiden bei Patienten mit Reifungsarrest der Spermatogenese. Geburtsh. Frauenheilk 59: 220-4.##Amer M, Soliman E, El-Sadek M, Mendoza C, Tesarik J. (1997) Is complete spermiogenesis failure a good##indication for spermatid conception? Lancet 350: 116.##Antinori S, Versaci C, Dani G, Antinori M, Selman HA. (1997a) The use of round or elongated spermatids for ICSI: which is more effective? In: Gomel V & Leung PCK "In vitro fertilization and assisted reproduction", Proc. 10th World Congr. In-Vitro Fertil.Ass. Reprod Vancouver (Canada), 547-51.##Antinori S, Versaci C, Dani G, Antinori M, Selman HA. (1997b) Successful fertilization and pregnancy using frozen-thawed round spermatids. In: Gomel V & Leung PCK "In vitro fertilization and assisted reproduction", Proc. 10th World Congr. In-Vitro Fertil Ass Reprod Vancouver (Canada), 699-703.##Araki Y, Motoyama M, Yoshida A, Kim S-Y, Sung H, Araki Sh. (1997) Intracytoplasmic injection with late spermatids: a successful procedure in achieving childbirth for couples in which the male partner suffers from azoospermia due to deficient spermatogenesis. Fertil Steril 67: 559-61.##Aslam L, Robins A, Dowell K, Fishel S. (1998) Isolation, purification and assessment of viability of spermatogenic cells from testicular biopsies of azoospermic men. Hum Reprod 13: 639-45.##Aslam I, Fishel S. (1999) Evaluation of the fertilization potential of freshly isolated, in-vitro cultured and cryopreserved human spermatids by injection into hamster oocytes. Hum Reprod 14: 1528-33.##Balaban B, Urman B, Isiklar A, Alatas C, Aksoy S, Mercan R, Nuhoglu A. (2000) Progression to the blastocyst stage of embryos derived from testicular round spermatids. Hum Reprod 15: 1377-82.##Barak Y, Kogosowski A, Goldman S, Soffer Y, Gonen Y, Tesarik J. (1998) Pregnancy and birth after transfer of embryos that developed from single-nucleated zygotes obtained by injection of round spermatids into oocytes. Fertil Steril 70: 67-70.##Barros A, Bernabeu R, Takahashi K, Oliveira C, Cremades N, Silva J, Sousa M, Tesarik J. (1998) Intracytoplasmic injection of ejaculate and testicle spermatids: report on 35 cycles. Hum Reprod 13 (s1): 154-5.##Bernabeu R, Cremades N, Takahashi K, Sousa M. (1998) Successful pregnancy after spermatid injection. Hum Reprod 13: 1898-1900.##Brannan CI, Bartolomei MS. (1999) Mechanisms of genomic imprinting. Curr. Opin Genet Dev 164-170.##Cox FG, Bürger J, Lip V, Mau UA, Sperling K, Wu B-L, Horsthemke B. (2002) Intracytoplasmic sperm injection may increase the risk of imprinting defects. Am J Hum Genet 71: 162-4.##Cremades N, Bernabeu R, Barros A, Sousa M. (1999) In-vitro maturation of round spermatids using co-culture on Vero cells. Hum Reprod 14: 1287-93.##De Kretser M, Kerr JB. (1969) The cytology of the testis. In Knobil E and Neill JD (eds), "The Physiology of Reproduction", Raven Press, New York, USA.##De Kretser DM, Loveland KL, Meinhardt A, Simorangkir D, Wreford N. (1998) Spermatogenesis. Hum Reprod 13: S1-8.##Edwards RG, Tarin JJ, Dean N, Hirsch A, Tan SL. (1994) Are spermatid injections into human oocytes now mandatory? Hum Reprod 9: 2217-9.##Fishel S, Green S, Bishop M. (1995) Pregnancy after intracytoplasmic injection of spermatid Lancet 345: 1641-2.##Gianaroli L, Selman HA, Magli MC, Colpi G, Fortini D, Ferraretti AP. (1999) Birth of a healthy infant after conception with round spermatids isolated from cryopreserved testicular tissue. Fertil Steril 72: 539-41.##Hiramoto Y. (1962) Microinjection of the live spermatozoa into sea urchin eggs. Exp Cell Res 27: 416-26.##Kahraman S, Polat G, Samli M, Sِzen E, ozgün OD, Dirican K, Ozbicer T. (1998) Multiple pregnancies obtained by testicular spermatid injection in combination with intracytoplasmic sperm injection. Hum Reprod 13: 104-10.##Khalili MA, Aflatoonian A, Zavos PM. (2002) Intracytolasmic Injection Using Spermatids and Subsequent Preganancies: Round Versus Elongated Spermatids. Journal of Assisted Reproduction and Genetics 19: 84-86.##Kimura Y, Yanagimachi Y. (1995) Mouse oocytes injected with testicular spermatozoa or round spermatids can develop into normal offspring. Development 121: 2397-2405.##Mansour RT, Aboulghar MA, Serour GI, Kamal A, Tawab NA, Fahmy I, Amin YM. (1996) Pregnancy and delivery after intracytoplasmic injection of spermatids into human oocytes. Middle East Fertil Soc J 1: 223-5.##Martin-du Pan R, Campana A. (1993) Physiopathology of spermatogenic arrest. Fertile Steril 60: 937-49.##Nikolettos N, Küpker W, Demirel LC, Schِpper B, Blasig C, Sturm R, Felberbaum R, Bauer O, Diedrich K, Al-Hasani S. (1999) Fertilization potentials of spermatozoa with abnormal morphology. Hum Reprod 14: 47-70.##Ogura A, Yanagimachi R. (1993) Round spermatid nuclei injected into hamster oocytes from pronuclei and participate in syngamy. Boil Reprod 48: 219-25.##Ogura A, Yanagimachi R, Usui N. (1993) Behaviour of hamster and mouse round spermatid nuclei incorporated into mature oocytes by electrofusion. Zygote 1, 1-18.##Ogura A, Matsuda J, Yanagimachi R. (1994) Birth of normal young after electrofusion of mouse oocytes with round spermatids. Proc Natl Acad Sci U.S.A 91, 7460-2.##Oppedisano L, Haines G, Hrabchak C, Fimia G, Elliott R, Sassone-Corsi P, Varmuza S. (2002) The rate of aneuploidy is altered in spermatids from infertile mice. Hum Reprod 17: 710-17.##Orstavik KH, Eiklid K, van der Hagen CB, Spetalen S, Kierulf K, Skjeldal O, Buiting K. (2003) Another case of imprinting defect in a girl with Angelman syndrome who was conceived by intracytoplasmic sperm injection. Am. J Hum Genet 72: 218-9,.##Palermo G, Joris H, Devroey P, Van Steirteghem AC. (1992) Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet 340, 17-8.##Preece MA, Moore GE. (2000) Genomic imprinting, uniparental disomy and foetal growth. TEM 11: 270-75.##Reik W, Walter J. (1998) Imprinting mechanisms in mammals. Curr Opin Genet Dev 8: 154-164.##Saremi A, Esfandiari N, Salehi N, Saremi MR. (2002) The first successful pregnancy following injection of testicular round spermatid in Iran. Arch Androl 48: 315-9.##Schoysman R, Vanderzwalmen P, Bertin G, Nijs M , van Damme B. (1999) Oocyte insemination with spermatozoa precursors. Curr. Opin. Urology 9: 541-5.##Schulze W, Thoms F, Knuth UA. (1999) Testicular sperm extraction: comprehensive analysis with simultaneously performed histology in 1418 biopsies from 766 subfertile men. Hum Reprod 14 (s1): 82-96.##Shamanski FL, Kimura Y, Lavoir M-C, Pedersen RA, Yanagimachi R. (1999) Status of genomic imprinting in mouse spermatids. Hum Reprod 14: 1050-6.##Silber SJ, Johnson L. (1998) Are spermatid injections of any clinical value? Hum Reprod 13: 509-23.##Silber SJ, Nagy Z, Devroey P, Tournaye H, Van Steirteghem AC. (1997) Distribution of spermatogenesis in the testicles of azoospermic men: the presence or absence of spermatids in the testes of men with germinal failure. Hum Reprod 12: 2422-28.##Silber SJ, Johnson L, Verheyen G, Van Steirteghem AV. (2000) Round spermatid injection. Fertil Steril 73: 897-900.##Sleutels F, Barlow DP, Lyle R. (2000) The uniqueness of the imprinting mechanism. Curr. Opin. Genet Dev 10: 229-33.##Sofikitis NV, Miyagawa I, Agapitos E, Pasyianos P, Toda T, Hellstrom WJ, Kawamura H. (1994) Reproductive capacity of the nucleus of the male gamete after completion of meiosis. J Assist Reprod Genet 11: 335-41.##Sofikitis NV, Yamamoto Y, Miyagawa I, Mekras G, Mio Y, Toda T, Antypas S, Kawamura H, Kanakas N, Antoniou N, Loutradis D, Mantzavinos T, Kalianidis K, Agapitos E. (1998) Ooplasmic. injection of elongating spermatids for the treatment of non-obstructive azoospermia. Hum Reprod 13: 709-14.##Sousa M, Barros A, Tesarik J. (1998) Current problems with spermatid conception. Hum Reprod 13: 255-8.##Sousa M, Cremades N, Silva J, Oliveira C, Ferraz L, Texeira da Silva J, Viana P, Barros A. (2002) Predictive value of testicular histology in secretory azoospermic subgroups and clinical outcome after microinjection of fresh and frozen-thawed sperm and spermatids. Hum Reprod 17: 1800-10.##Tesarik J, Mendoza C, Testart J. (1995) Viable embryos from injection of round spermatids into oocytes. N Engl J Med 333: 525.##Tesarik J, Mendoza C. (1996) Spermatid injection into human oocytes. I. Laboratory techniques and special features of zygote development. Hum Reprod 11: 772-9.##Tesarik J, Greco E, Rienzi L, Ubaldi F, Guido M, Cohen-Bacrie P, Mendoza C. (1998) Differentiation of spermatogenic cells during in-vitro culture of testicular biopsy samples from patients with obstructive azoospermia: effect of recombinant follicle stimulating hormone. Hum Reprod 13: 2772-81.##Tesarik J, Bahceci M, Ozcan C, Greco E, Mendoza C. (1999) Restoration of fertility by in-vitro spermatogenesis Lancet 353: 555-6.##Tesarik J, Balaban B, Isiklar A, Alatas C, Urman B, Aksoy S, Mendoza C, Greco E. (2000) In-vitro spermatogenesis resumption in men with maturation arrest: relationship with in-vivo blocking stage and serum FSH. Hum Reprod 15: 1350-4.##Urman B, Alatas C, Aksoy S, Mercan R, Nuhoglu A, Mumcu A, Isiklar A, Balaban B. (2002) Transfer at the blastocyst stage of embryos derived from testicular round spermatid injection. Hum Reprod 17: 741-3.##Vanderzwalmen P, Lejeune B, Nijs M, Segal-Bertin G, Vandamme B, Schoysman R. (1995) Fertilization of an oocyte microinseminated with a spermatid in an in-vitro fertilization programme. Hum Reprod 10: 502-3.##Vanderzwalmen P, Zech H, Birkenfeld A, Yemini M, Bertin G, Lejeune B, Nijs M, Segal L, Stecher A, Vandamme B, van Roosendaal E, Schoysman R. (1997) Intracytoplasmic injection of spermatids retrieved from testicular tissue: influence of testicular pathology, type of selected spermatids and oocyte activation. Hum Reprod 12: 1203-13.##Vanderzwalmen P, Nijs M, Schoysman R, Bertin G, Lejeune B, Vandamme B, Kahraman S, Zech H. (1998) The problems of spermatid microinjection in the human: the need for an accurate morphological approach and selective methods for viable and normal cells. Hum Reprod 13: 515-9.##Vanderzwalmen P, Nijs M, Stecher A, Zech H, Bertin G, Lejeune B, Vandamme B, Chatziparasidou A, Prapas Y, Schoysman R. (1998) Is there a future for spermatid injections? Hum Reprod 13: S71-84.##Verheyen G, Crabbe E, Joris H, Van Steirteghem A. (1998) Simple and reliable identification of the human round spermatid by inverted phase contrast microscopy. Hum Reprod 13: 1570-7.##Zech H, Vanderzwalmen P, Prapas Y, Lejeune B, Duba E, Schoysman R. (2000) Congenital malformations after intr acytoplasmic injection of spermatids. Hum Reprod 15: 969-71.## ##

Restoration of Spermatogenesis by Adenoviral Gene Transfer into Injured Spinal Cords of Rats 2 1 - 2 Materials and Methods: Young adult Sprague-Dawley rats (200-250g) were assigned into one of the three different groups of control, SCI, and adenovirus transfer (Ad) (n=3/ group). Control rats received no injury, nor any surgery. For SCI rats, SCI was produced by a 10g brass rod with a tip diameter of 2 mm which was dropped from a height of 12.5 mm onto exposed spinal cord at level of T10 with NYU impactor. Animals were perfused transcardially 43 days post SCI. Both spinal cord and testicular tissues were cryo-sectioned and ultra thin-sectioned, respectively. Cellular morphology and morphometry were done for spinal cord tissues. The testicular samples were processed for both light and transmission electron microscopy (TEM). The third group of rats underwent SCI first, followed by microinjection of LacZ adenoviral vectors (5x106 p.f.u./ �l) along the T6-T10 dorsal root entry zone bilaterally. The immune system of animals were suppressed before the Ad administration. Each Ad injection was done using a glass micropipet and a Nonoject injector. Rats were killed 43 days after Ad injections, and the tissues were studied as for other groups. 7 11 2017/10/12017/10/1 1396/7/9 2003/01/152003/01/15 1381/10/25 Mohammad A Khalili Mohammad A Khalili Alexander G Rabchevsky Alexander G Rabchevsky Spinal cord injury Gene therapy Spermatogenesis Rat Bors E, Comarr E. Neurological disturbances of sexual function with speical reference to 529 patients with spinal cord injury. Urol Surv 1960; 10: 191-222.##Hirsch IH, Huang B, Chancellor MB, Rivas DA, Salzman SK, Jost LK, Evenson DP. Spermatogenesis in early and chronic phases of experimental spinal cord injury in the rodent model. J Androl 1999; 20: 63-71.##Holstein AE, Saverwein D, Schirren U. Spermatogenesis in patients with traumatic transverse paralysis. Urologe 1985; 24: 208-11.##Linsenmeyer TA, Perkash I. Infertility in men with spinal cord injury. Arch Phys Med Rehabil 1991; 72: 747-54.##Linsenmeyer TA, Pogach L, Ottenweller JE, Huang HF. Spermatogenesis and the pituitary testicular hormone axis in rats during acute phase of spinal cord injury. J Urol 1994; 152: 1303-7.##Liu Y, Himes BT, Moul J, Huang W, Chow S, Tessler A, Fischer I. Application of recombinant adenovirus for in vivo gene delivery to spinal cord. Brain Research 1997; 768: 19-29.##Rabchevsky AG, Fugaccia I, Turner AF, Blades DA, Mattson MP, Scheff SW. Basic fibroblast growth factors enhances functional recovery following severe spinal cord injury to the rat. Exp Neurol 2000; 164: 280-91.##Rajasekaran M, Monga M. Cellular and molecular causes of male infertility in spinal cord injury. J Androl 1999; 20: 326-30.##Robbins PD, Ghivizzani SC. Viral vectors for gene therapy. Pharmacol Ther 1998; 80: 35-47.##Romero MI, Smith GM. Adenoviral gene transfer into the normal and injured spinal cord: enhanced transgene stability by combined administration of temperature-sensitive virus and transient immune blockade. Gene Therapy 1998; 5: 1612-21.##Stribley JM, Rehman KS, Niu H, Christman GM. Gene therapy and reproductive medicine. Fert Steril 2002; 77: 645-57.##Wickham Tj. Targeting adenovirus. Gene Therapy 2000; 7: 110-14.##Wood M, Charlton H, Wood K, Kajiwara K, Byrnes A. Immune responses to adenovirus vectors in the nervous system. Trends Neurosci 1996; 19; 497-501.## ##

The Effect of Murine Leukemia Inhibitory Factor on In Vitro Differentiation of Mouse Embryonic Stem Cells 2 1 - 2 Background: Embryonic stem cells (ESc) are pluripotent cells which have been used as a model to study the mechanism that control the embryogenesis and early mammalian development in vitro. The aim of this study was to isolate and produce embryonic stem cells from late blastocyst stage embryos in mice. Materials and Methods: Blastocyst stage embryos from pregnant NMRI mice were obtained and cultured for 24 h in DMEM medium. 4-6 days after hatching, the inner cell masses (ICM) formed colonies which were then collected mechanically and trypsinized. Several subcultures were prepared in the medium supplemented with 0.1 mM 2 Mercaptoethanol, 1000 U/ml Leukemia Inhibitory Factor (LIF) and 10% Fetal Bovine Serum (FBS). The (ESc) were recognized by alkaline phosphates histochemistry using azo-coupling method. Results: The results demonstrated that a highly pluripotent stem cell line was derived from the blastocyst stage embryos of NMRI mice; however, the rate of colonies was as low as 10%. Conclusion: The LIF is effective to culture and maintain the isolated ICM colonies in undifferentiated condition in the absence of feeder layer. 12 15 2017/10/12017/10/12017/10/1 1396/7/9 2003/01/152003/01/152003/01/15 1381/10/25 Ali A Movassagh-Pour Ali A Movassagh-Pour Mojdeh Salehnia Mojdeh Salehnia Ali A Pourfatollah Ali A Pourfatollah Sayed M Moazzeni Sayed M Moazzeni Embryonic stem cells Mouse embryo Leukemia Inhibitory Factor Anderson GB. (1992) Isolation and use of embryonic stem cells from livestock species. Anim Biotechnol 3: 165-175.##Bongso A, Fong CY, Ng SC, Ratnam S. (1994) Isolation and culture of inner cell mass cells from human blastocysts. Hum Reprod 9: 2119-2117.##Cole RJ, Edwards RC, Poul J. (1965) Cytodifferentiation in cell colonies and cell strains derived from cleaving ova and blastocysts of the rabbit. J Exp Cell Res 501-504.##Doetschman T, Williams P, Maeda N. (1988) Establishment of hamster blastocyst derived embryonic stem cells. Dev Biol 127: 224-227.##Donovan PJ, Stott D, Cairns LA, Heasman J, Wylie CC. (1986) Migratory and postmigratory mouse primordial germ cells behave differently in culture. Cell 44: 831-838.##Evans MJ, Kaufman M. (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 242: 154-156.##Martin GR. (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium contained with teratocarcinoma stem cells. Proc Nat Sci 78: 7634-7638.##Movassagh Pour A, Salehnia M, Pourfatollah M, Moazzeni SM. (2003) CFU-GM like colonies derived from embryonic stem cells cultured on the bone marrow stromal cells. Iranian Biomedical J (In press).##Odorico JS, Kaufman DS, Thomson JA. (2001) Multilineage differentiation from human embryonic stem cell lines. Stem Cells 19: 193-204.##Pera MF, Reubinoff BE, Truonson A. (2000) Human embryonic stem cells. Cell Science 113: 5-10.##Rathjeh PD, Lake J, Whyatt, Bettess MD, Rathjeh J. (1998) Properties and uses of embryonic stem cells: prospects for application to human biology and gene therapy. Reprod Fertil Dev 10: 31-47.##Reubinoff BE, Pera MF, Fong CY, Trounson A, Bongso A. (2000) Embryonic stem cell lines from human blastocyst: somatic differentiation in vitro. Nature Biotech. 18: 399-404.##Smith AG, Hooper L. (1987) Buffalo liver cell produce a diffusible activity which inhibits the differentiation of murine embryonal carcinoma and embryonic stem cells. Dev Biol 121: 1-9.##Smith AG, Heath JK, Donaldsmon DD, Wong GG, Moreau J, Stahl M, Rogers D. (1988) Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides. Nature 336: 688-690.##Smith AG. (1998) Cell therapy: In search of pluripotency. Current Biol 8; 802-804.##Smith AG. (1992) Mouse embryo stem cells: their identification, propagation and manipulation. Seminar in Cell Biol 3: 385-399.##Thomson JA, Marshal VS. (1998) Primate embryonic stem cells. Curr Topics Dev Biol 38: 133-165.##Thomson JA, Kalishman J, Golos TS, Durning M, Harris CP, Becker RA, Hearn JP. (1995) Isolation of a primate embryonic stem cell line. Proc Natl Acad Sci USA 92: 7844-48.## ##

Comparison between Transvaginal Sonography and Cytological results for detection of ovarian cysts 2 1 - 2 Background: Ovarian functional cyst is one of the most common pelvic mass in reproductive age which mostly resolves spontaneously. Sonography is a valuable tool for diagnosis of benign cyst with high accuracy. The objective of this cross sectional study was to evaluate the accuracy of transvaginal sonography in detecting type of ovarian cyst and compare the results wieh cytological results. Materials and Methods: 82 women in reproductive age who have had simple ovarian cysts with benign criteria which unresolved after taking contraceptive pills for 3 months were considered for this clinical study. Transvaginal ultrasound-guided aspiration of cysts were done and were then sent to the pathological evaluation. Also, all data regarding the size of the cysts and aspirated fliud were recorded in charts for further statistical analysis. Results: The accuracy of transvaginal ultrasound comparing with cytology on diagnosis for functional cysts was 94.9%, for epithelial ovarian cyst was 97.5% and for endometrioma was 97.5% (P= 0.0001).The size of cysts with diameter of <10cm was not related to the quality of cysts. Conclusion: The results showed that sonography is a valuable and reliable tool for diagnosis of benign ovarian cyst. It seems that if a mass appears benign by ultrasound morphologic criteria, probability of it being malignant is near to zero, which can be aspirated by transvaginal route without any fear from missing of malignancy or complication 16 19 2017/10/12017/10/12017/10/12017/10/1 1396/7/9 2003/01/152003/01/152003/01/152003/01/15 1381/10/25 Abass Aflatoonian Abass Aflatoonian Reza Aflatoonian Reza Aflatoonian Zahra Khashavi Zahra Khashavi transvaginal Sonography ovarian cyst cytology Brown DL, Doublet PM. (1998) Benign and malignant ovarian masses. Radiology 208: 103-110.##De crespingy L. CH. (1998) Ultrasound guided puncture for gynecology and pelvic lesion. Australian and newzealand J of OBGy; 25-227.##Diernaes E Rassmussen. (1987) Ovarian cyst management by puncture . Lancet 168-173.##Finkler et al. (1996) Management of ovarian cyst: BMJ 313-1103.##Geatije R, Poop L.W. (1994) Is differention of benign and mailgnant cystic adnexal masses . Acta OBGy scandinavica 73: 502-507.##Hansmann M. (1985) Ultrasound diagnosis in Gyncology. Radiology J 132: 1186-1190.##Kocak I, Auzel (1998) An evaluation of transvaginal ultrasound. guided aspiration of simple adnexal mass. Journal of OBGY 18: 103-108.##Jabra AA, Fishman Ek, Taylor G.A. (1993) Primary ovarian tumors evaluation clin Imaging 17: 199-203.##Kozlowski KJ. (1999) Ovarian Masses. Adoles Med 10: 335-37.##Kurjak A, Kupesic, (2002) Color dopller of malignant adnexal mass 21,22: 191-203.##Maylen M. silverberg M. (1990) Ovarian cyst aspiration. J of IVF-ET 7: 1536-1543.##Novak, S. (2002) Ovarian cancer, Jonatan. S.Br Gynecology Textbook. Lippincont Williams and Wilkins 32: 376.##Prasap kumar. (2001) Transvaginal ultra sonography in ovarian pathology, Rajan R, philadelphia W.B. textbook of ultrasound in OBGyn sandes company 30: 229.##Rajan R. (1993) Changing concept in gyncologycal diagnosis J. OBGYN 43: 1130-38.##Salat, Baroux. (1992) Manage of ovarian cysts. Biritish medical J l 313: 6031-37.##Zanetta, Gerards. (1996) Role of puncture and aspiration in expectant management- British Medical J 313: 7065-7069.## ##

Progesterone Shifts the Pinopodes Expression of Mouse Endometrium to Pre-Implantation Time After Ovarian Hyperstimulation 2 1 - 2 Background: The aim of this study was to determine the correlation between ultrastructural studies for pinopodes expression after ovarian hyperstimulation and progesterone injection in mice. Materials and Methods: Adult NMRI mice were superovulated using human menopasual gonadotropic (hMG) and human chorionic gonadotropic (hCG) hormones; after that, daily injection of progesterone (1 mg/mouse) was performed. Animals were sacrificed by cervical dislocation 3.5 and 4.5 days after hCG injection. Tissues of uterine horns were obtained and processed for scanning (SEM) and transmission (TEM) electron microscopy studies. The pseudopregnant control samples were studied same as experimental groups. Results: The SEM and TEM observations showed that in control groups on 3.5 days of pregnancy, there were some pinopodes. All apical cell surfaces expressed these projections on the forth day. In progestrone-injected group, well developed pinopods were expressed 3.5 days after hCG injection and they were transformed to small projections on the fourth day following hCG injection. Also, the life span of pinopods was limited to a short time. At the TEM levels, the pinopods were seen as swelling process on the apical surface, which were more pronounced on day 3.5 of hCG injection in hyperstimulated and progestrone injection. Conclusion: The progestrone may cause premature expression of pinopodes and the implantation failure after ovarian induction may be due to these timing changes. 20 23 2017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2003/01/152003/01/152003/01/152003/01/152003/01/15 1381/10/25 Mojdeh Salehnia Mojdeh Salehnia Implantation Window Pinopodes Progesterone Electron Microscopy Ertzeid G, Storeng R. (2001) The impact of ovarion stimulation on implantation and fetal development in mice. Hum Reprod 16: 221-225.##Kolb BA, Najmabadi S, Paulson RJ. (1997) Ultrastructural characteristics of luteal phase endometrium in patients undergoing controlled ovarian hyperstimulation. Feril Steril 67: 625-30.##Kramer B, De Wet G. (1994) Exogenous gonadotropin administration affects the glycocalyx of rat endometrial epithelial cells during the period of implantation. J Assist Reprod Gen 11: 504-509.##Kramer B, Magan A and De Wet G. (1993). Hyperstimulation affects vascular permeablility at implantation sites in the rat endometrium. J Assist Reprod Gen 10: 163-168.##Leesey B.A. (2001a) The role of the endometrium during embryo implantation. Hum Reprod 15: 36-50.##Lessey B.A. (2001b) Endometrial receptivity and the window of implantation. Bailieres Best. Pract. Res. Clin Obstet Gynaecol 14: 775-880.##Martel D., Monier M.N., Roche D., Psychoyos A. (1991) Hormonal dependence of pinopodes formation at the uterine luminal surface. Hum Reprod 6: 597-603.##Miller BG. (1979) Delayed interactions between progesterone a low doses of 17 β-estradiol in the mouse uterus. Endocrin 104: 26-33.##Navot D., Scott R.T., Droesch K., Veeck L.L., Liu H.C., Rosenwaks Z. (1991) The window of embryo transfer and the efficacy of human conception in vitro. Fertil Steril 55: 114-118.##Nikas G, Develioglu OH, Toner JP, Jones HWJR. (1999) Endometrial pinopodes indicate a shift in the window of receptivity in IVF cycles. Hum Reprod 14: 787-792.##Nikas G., Drakakis P., Loutradus D., Mara-Shoufari C., Koumantakis E., Michalas S., Psychoyos A. ( 1995) Uterine pinopodes as markers of the nidation window in cycling women receiving exogenous oestradiol and progesterone. Hum Reprod 10: 1208-1213.##Nikas, G, Drakakis P, Loutradus D, Mara-Shoufari C, Koumantakis E, Michala S. (1995) Uterine pinopodes as markers of the " nidation Window" in cycling women receiving exogenous oestradiol and progesterone. Hum Reprod 10: 1208-1213.##Nilsson O. (1958) Ultrastructure of mouse uterine surface epithelium under different estrogenic influences. 2. Early effect of estrogen administrered to spayed animals. J Ultrastruct Res 2: 73-95.##Paulson R.J., Saner M.V., Lobo R.A. (1990) Embryo implantation after human in vitro fertilization: importance of endometrial receptivity. Fertil Steril 53: 870-874.##Paulson RJ, Sauer MV and Lobo RA. (1997) Potential enhancement of endometrial receptivity in cycles using controlled ovarian hyperstimulation with antiprogestins: a hypthesis. Fertil Steril 67:321-325.##Psychoyos A, Nikas G. (1994) Uterine pinopodes as markers of uterine receptivity. Assist Reprod Rev 4: 26-32.##Simon C., Cano F., Valbuena D. (1995) Clinical evidence for a detrimental effect on uterine receptivity of high serum estradiol levels in high and normal responder patients. Hum Reprod 10: 2432-2437.##Singh M.M., Chaluhan S.C., Trivedi R. N., Maitra S.C, Kamboj V.P. (1996) Correlation of pinopod development on uterine luminal epithelial surface with hormonal events and endometrial sensitivity in rat. Eur Endocrinol 35: 107-117.##Stavreus-Evers A., Nikas G., Sahlin L., Eriksson H., Landgren B.M. (2001) Formation of pinopodes in human endometrium is associated with the concentrations of progesterone and progesterone receptors. Fertil Steril 76: 782-791.##Valbuena D, Jasper M, Remon J, Pellicer A, Sima C. (1999) Ovarian stimulation and endometrial receptivity. Hum Reprod 1: 2107-2111.## ##

Evaluation of the Predictive Value of Semen Parameters in Sperm Fertility Potential Using Intracellular Calcium Increase in Response to Progesterone 2 1 - 2 Background: While traditional semen parameters are of significant clinical value, total fertilization failure in IVF cycles is not uncommon. Sperm function testing such as; Hamster egg penetration test has severed limitations as a clinical test. The aim of this study was to evaluate the predictive value of semen parameters by using of intracellular calcium [Ca2+]I increase in response to progesterone. Materials and Methods: The [Ca2+]i response to progesterone was measured in spermatozoa of 86 patients referring to the Assisted Conception Unit for semen analysis. The patients were divided into 3 groups; according to their semen parameters and measured intracellular [Ca2+]i increasing in response to progesterone . Results: There was no significant correlation between each individual semen parameter and [Ca2+]i elevation in response to the progesterone, but most of the patients in each group had [Ca2+]i increasing as expected based on sperm parameters. However, there were cases in groups 1 and 2 (Normal and IVF) that demonstrated [Ca2+]i increases which were poor or lower than expected. Out of the 22 patients in the normal category, 8 cases had poor response to [Ca2+]I increase and out of the 47 patients in the IVF group, 9 patients were as well. In addition we measured [Ca2+]I increases in 6 fertile donor samples for comparison purposes. Conclusion: [Ca2+]i increase in response to progesterone is related to predicting value of sperm parameters in most cases. However, the response of sperm to progesterone could be different in some cases that are expected in normal or IVF category based on our semen analysis criteria. We suggest that the [Ca2+]i measurements may perfect the sperm fertility potential. 24 28 2017/10/12017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2003/01/152003/01/152003/01/152003/01/152003/01/152003/01/15 1381/10/25 Mir-Mehrdad Farsi Mir-Mehrdad Farsi Ca2+]i increases Progesterone Spermatozoa] Baldi. E., Krausz. C, and Forti. G. (1995) Nonegenomic actions of progesterone on human spermatozoa. Trends Endocrinol Metab 6: 198-205.##Barrat, C.L.R., Justin C.St John. (1998) Diagnostic tools in male fertility, Hum Reprod 13: 51-61.##Barrat.C.L.R. Naeeni, M., Clements.C, Cook ID. Clinical value of sperm morphology measurements for in vitro fertility: Comparisons between World Health Organisation (WHO) criteria of 1987 and 1992. Hum Repord 1995; 10: 587 - 593.##Barrat, C.L.R., McLeod, I.D., Dunphy, B.C. et al. prognostic significance of two putative sperm function tests: hypoosmotic swelling and bovine sperm mucus penetration test (penetraK). Hum Reprod 7: 1240-1244.##Bostofte, E., Bagger, P., Michael, A. et al. Fertility prognosis for infertile men: results of follow up study of semen analysis in infertile men from tow different populations evaluated by the Cox regression model. Fertil Steril 1990; 54: 1100-1106.##Bray C, Brown JCK, Publicover S, Barratt C.L.R. Progesterone Interaction with sperm plasma membrane, calcium influx and induction of the acrosome reaction. Reprod Med Review 1999; 7: 81-93. Cambridge University Press.##De Jonge CJ. The diagnostic significance of the induced acrosome reaction. Reprod Med Rev 1994; 3: 159-78.##Kirkman-Brown, J.C, Bray, C., Stewart, P.M. Barratt C.LR and Publicover SJ. Biphasic elevation of [Ca2+] in individual human spermatozoa exposed progesterone. Dev Biol 2000; 222: 326-335.##krausz C, Bonaccorsi L, Maggio P. Luconi M, Criscuoli L. Fuzzi B, Pellegrini S, Forti G, Baldi E.. Two functional assays of sperm responsiveness to progesterone and their predictive values in in vitro fertilization. Human Reprod 1996; 11: 1661-1667.##Krausz, C., Bonaccorsi, L., Luconi, M. Fuzzi B, Criscuoli L, Pellegrini S, Forti G, Baldi E.. intracellular calcium increase and acrosome reaction in response to Progesterone in human spermatozoa are correlated with in vitro fertizalation. Hum Reprod 1995; 10: 120-124.## ##

The Correlation Between Semen Parameters and Pregnancy Outcome after Intrauterine Insemination 2 1 - 2 Backgroud: Intrauterine insemination (IUI) is generally attempted before proceeding to more expensive and invasive assisted reproductive techniques such as invitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). This procedure is most commonly performed as a therapeutic method for couples with a wide variety of subfertility etiologies, such as low count or low motility of sperm, or an incompatibility between the sperm and the cervical mucus. The objective of this clinical trial study was to compare the correlation between the semen parameters and pregnancy rates in patients undergoing hyperstimulation and IUI. Materials and Methods: 336 infertile couples that underwent 336 cycles of IUI with washed husband�s semen were included in this study. All patients� charts were reviewed for age, etiology and duration of infertility, semen characteristics and pregnancy rates. The SPSS 9 software and Chi-square tests were applied for statistical analysis. P<0.05 was determined as statistical significance. Results: Total pregnancy rates were18.2% (61 out of 336 cycles). Postwash semen parameters including: sperm count ?10? 106 ,motility ?50% (grade III and IV >20%) had significant effect on pregnancy rates after IUI. The Outcome of this procedure was not significantly affected by female age, duration or etiology of infertility. Conclusion: Postwash semen quality was the most important factor for predication of successful pregnancy in this study. 29 32 2017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2003/01/152003/01/152003/01/152003/01/152003/01/152003/01/152003/01/15 1381/10/25 Marzieh Mehrafza Marzieh Mehrafza Nadia Nobakhti Nadia Nobakhti Zahra Atrkar Roushan Zahra Atrkar Roushan Havva Dashtdar Havva Dashtdar Mane Oudi Mane Oudi Ahmad Hosseini Ahmad Hosseini IUI pregnancy female age semenm parameters Aboulghar MA, Mansour RT, Serour GI, Amin Y, Abbas AM, Salah IM. (1993) Ovarian superstimulation and intrauterine insemination for the treatment of unexplained infertility. Fertil Steril 60: 303-6.##Berg U, Brucker C, Berg FD. (1997) Effect of motile sperm count after swim-up on outcome of intrauterine insemination. Fertil Steril 67: 747-50.##Brzechffa PR, Daneshmand S, Buyalos RP. (1998) Sequential clomiphene citrate and human menopausal gonadotrophin with intrauterine insemination: the effect of patient age on clinical outcome. Hum Reprod 13: 2110-4.##Burr RW, Siegberg R, Flaherty SP, Wang XJ, Matthews CD. (1996) The influence of sperm morphology and the number of motile sperm inseminated on the outcome of intrauterine insemination combined with mild ovarian stimulation. Fertil Steril 65: 127-32.##Campana A, Sakkas D, Stalberg A, Bianchi PG, Comte I, Pache T, Walker D. (1996) Intrauterine insemination: evaluation of the results according to the woman's age, sperm quality, total sperm count per insemination and life table analysis. Hum Reprod 11: 732-6.##Chaffkin LM, Nulsen JC, Luciano AA, Metzger DA. (1991) A comparative analysis of the cycle fecundity rates associated with combined human menopausal gonadotropin (hMG) and intrauterine insemination (IUI) versus either hMG or IUI alone. Fertil Steril 55: 252-7.##Dawood MY. (1996) In vitro fertilization, gamete intrafallopian transfer, and superovulation with intrauterine insemination: efficacy and potential health hazards on babies delivered. Am J Obstet Gynecol 174: 1208-17.##Dickey RP, Olar TT, Taylor SN, Curole DN, Rye PH, Matulich EM. (1991) Relationship of follicle number, serum estradiol, and other factors to birth rate and multiparity in human menopausal gonadotropin-induced intrauterine insemination cycles. Fertil Steril. 56: 89-92.##Dickey RP, Pyrzak R, Lu PY, Taylor SN, Rye PH. (1999) Comparison of the sperm quality necessary for successful intrauterine insemination with World Health Organization threshold values for normal sperm. Fertil Steril 71: 684-9.##Dodson WC, Haney AF. (1991) Controlled ovarian hyperstimulation and intrauterine insemination for treatment of infertility. Fertil Steril 55: 457-67.##Francavilla F, Romano R, Santucci R, Poccia G. (1990) Effect of sperm morphology and motile sperm count on outcome of intrauterine insemination in oligozoospermia and/or asthenozoospermia. Fertil Steril 53(5): 892-7.##Guzick DS, Sullivan MW, Adamson GD, et al. (1998) Efficacy of treatment for unexplained infertility. Fertil Steril 70: 207-213.##Harald Trummer, MD,Anthony Paul Caruso, MD,Larry I. Lipshultz, MD,Randall B. (2000) Meacham, MD .Recent Developments in the Evaluation and Treatment of Male Infertility. Infect Urol 13: 87-94.##Lee VM, Wong JS, Loh SK, Leong NK. (2002) Sperm motility in the semen analysis affects the outcome of superovulation intrauterine insemination in the treatment of infertile Asian couples with male factor infertility. BJOG 109: 115-20.##Miller DC, Hollenbeck BK, Smith GD, Randolph JF, Christman GM, Smith YR, Lebovic DI, Ohl DA. (2002) Processed total motile sperm count correlates with pregnancy outcome after intrauterine insemination. Urology 60: 497-501.##Pasqualotto EB, Daitch JA, Hendin BN, Falcone T, Thomas AJ Jr, Nelson DR, Agarwal A. (1999) Relationship of total motile sperm count and percentage motile sperm to successful pregnancy rates following intrauterine insemination.J Assist Reprod Genet 16: 476-82.##Sahakyan M, Harlow BL, Hornstein MD. (1999) Influence of age, diagnosis, and cycle number on pregnancy rates with gonadotropin-induced controlled ovarian hyperstimulation and intrauterine insemination. Fertil Steril 72: 500-4.##Shulman A, Hauser R, Lipitz S, Frenkel Y, Dor J, Bider D, Mashiach S, Yogev L, Yavetz H. (1998) Sperm motility is a major determinant of pregnancy outcome following intrauterine inseminationJ Assist Reprod Genet 15: 381-5.##Snick HK, Snick TS, Evers JL, Collins JA. (1997) The spontaneous pregnancy prognosis in untreated subfertile couples:the Walcheren primary care study. Hum Reprod 12: 1582-8.##van der Westerlaken LA, Naaktgeboren N, Helmerhorst FM. (1998) Evaluation of pregnancy rates after intrauterine insemination according to indication, age, and sperm parameters. J Assist Reprod Genet 15: 359-64.##Van Voorhis BJ, Barnett M, Sparks AE, Syrop CH, Rosenthal G, Dawson J. (2001) Effect of the total motile sperm count on the efficacy and cost-effectiveness of intrauterine insemination and in vitro fertilization. Fertil Steril 75: 661-8.##Yang JH, Wu MY, Chao KH, Chen SU, Ho HN, Yang YS. (1998) Controlled ovarian hyperstimulation and intrauterine insemination in subfertility. How many treatment cycles are sufficient? J Reprod Med 43: 903-8.##Zeyneloglu HB, Arici A, Olive DL, Duleba AJ. (1998) Comparison of intrauterine insemination with timed intercourse in superovulated cycles with gonadotropins: a meta-analysis. Fertil Steril 69: 486-91.## ##

The Chromosomal Abnormality of Failed Fertilized Human Oocytes in an In Vitro Fertilization Program 2 1 - 2 Background: The high fertilization failure after IVF treatment cycles could be related to chromosomal abnormalities. This study was carried out to assess the frequency of chromosomal abnormality on human oocytes lacking signs of fertilization 18-20 h after insemination . Materials and Methods: On day one, 18-20 h after insemination (IVF), fertilization was confirmed when two pronuclei (normal IVF) or more pronuclei (poly pronucleus FR) were present. Chromosomal analysis of unfertilized oocytes was carried out within 20-24 h of collection. All oocyte did not sign of pronuclei were collected from total fertilization failure, TFF (FR=0) or partial fertilization failure, PFF (FR=10-90%). Chromosomal preparation was carried out as described by Tarkowski�s techniques. The average of finding between two groups was compared by X2 test. Results: Chromosome spreading permitted adequate analyzing in 348 unfertilized oocytes. In 33.6% chromosomal aneulpoidy was observed with the following frequencies; hypo-hyploidy, 22/348 (6.4%), hyper-hyploidy, 42/348 (12.2%) and diploidy, 52/348 (15%). The frequency of aneuoplidy was significantly higher in TFF group 33/80 (41%) than PFF group 83/268 (31%), p<0.01, X2. The most frequent numerical aberration was observed in chromosome group, G of the human karyotyped. Conclusion: Since cytogentic analysis of failed fertilized oocytes and sperm function tests are very helpful for direct information on low success rate of fertilization, further studies analyzing on both gametes function in TFF cycles will be needed. 33 36 2017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/12017/10/1 1396/7/9 2003/01/152003/01/152003/01/152003/01/152003/01/152003/01/152003/01/152003/01/15 1381/10/25 Seyed M Kalnatar Seyed M Kalnatar Ahmad Ebrahimi Ahmad Ebrahimi Mehrdad Solimani Mehrdad Solimani Hossein Fazli Hossein Fazli chromosome abnormality cytogenetic unfertilized oocytes Total Fertilization Failure Almeida PA., and Bolton VN (1994) The relationship between chromosomal abnormalities in the human oocytes anf fertilization in vivo. Hum Reprod 9, 343-346.##Abruzzo, M.A. and Hassold, T.A. (1995) Etiology of nondisjunction in humans. Environ. Mol. Mutagen., 25 (Suppl. 26), 38-47.##Aflatounian., Karimzadeh., Dehghani., Kalantar SM., et al. 2000. The role of aspiration of half ovarian follicles prior to admistration of hCG or GnRH-a for prevention of severe OHSS in ART programs. Midd. East. Ferti. Soci.J. 5.1; 73-75##Angell, R.R., Ledger, W., Yong, E.L. et al. (1991) Cytogenetic analysis of unfertilized human oocytes. Hum Reprod 6, 568-573.##Barlow, P., Englert, Y., Puissant, F. et al.,. (1990). Fertilization failure in IVF: why and what next? Hum Reprod 5:451-456.##Bedford, J.M. and Kim, H.H. (1993) Sperm/egg binding patterns and oocyte cytology in retrospective analysis of fertilization failure in vitro. Hum Reprod 8, 453-463##Bongso, A., Ng, S.C., Ratnam, S. et al. (1988) Chromosome analysis in human oocytes failing to fertilize after insemination in vitro. Hum Reprod 3, 645-649.##Delhanty, D.A. and Penketh, R.J.A. (1990) Cytogenetic analysis of unfertilized oocytes retrieved after treatment with the LHRH analogue, buserelin. Hum Reprod 5, 699-702.##Dyban, A., De Sutter, P. and Verlinsky, Y. (1993) Preimplantation cytogenetic analysis. In Verlinsky, Y. and Kuliev, A.M. (eds), Preimplantation Diagnosis of Genetic Disease. Wiley-Liss, New York, pp. 98.##Edirisinghe, W.R., Murch, A.R. and Yovich, J.L. (1992) Cytogenetic analysis of human oocytes and embryos in an in-vitro fertilization programme. Hum Reprod 7, 230-236.##Kalnatar SM., Lenton EA., Barratt CLR (1995) Sperm function tests and fertilization failure following IVF. In Human sperm acrosome reaction, (Eds) Fenichel P & Parinaud J, Colloque INSERM/John Libbey Eurotext Ltd, 236, pp. 439-440.##Kalantar SM., Lenton EA (2000) Evaluating the ability of biological substancesfor induction of acrosome reaction in normospermic samples. Midd East Ferti. Soci J 5.2;71-78.##Kalantar SM., Lenton EA., Brewis I., et al (1998) Fertilization failure of IVF predictive power of SFT. Andrologia., 30:4-5, 180-181.##Kalantar SM., et al., Evaluation of a cut-off level for DNA normality of sperm to predict fertilization failure following IVF-ET. 5th Annual Meeting of MEFS. Amman Jordan, 1998.##Kamiguchi,Y., Rosenbusch, B., Sterzik, K. and Mikamo, K. (1993) Chromosome analysis of unfertilized human oocytes prepared by a gradual fixation-air drying method. Hum Genet 90, 533-541.##Lim, A.S.T., Ho, A.T.N. and Tsakok, M.F.H. (1995) Chromosomes of oocytes failing in-vitro fertilization. Hum Reprod 10, 2570-2575.##Ma, S., Kalousek DK., Zouves C., et al., (1989) Abnormal chromosome arrangements in human oocytes. Hum Reprod 5, 703-707.##Macas E., Floersheim Y., Hotz E. et al., (1990) Chromosome analyzing of human oocytes failing to fertilize in vitro. Fertil Steril 51, 992-997.##Mozdarani H & Aghdaei F (2001) Cytogenetics analysis of failed-fertilized oocytes from Iranian infertile women after in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) procedures. MEFSJ 6.3:216-225.##Nakaoka Y, Okamoto E, Miharu N, Ohama K. (1998), Chromosome analysis in human oocytes remaining unfertilized after in-vitro insemination: effect of maternal age and fertilization rate. Hum Reprod 13(2):419-24.##Pellestor F (1991) Frequency and distribution of aneuploidy in human female gametes. Hum Genet 86, 283-288.##Plachot, M. (1995) Oocyte - genetic aspects. In Grudzinskas, J.G. and Yovich, J.L. (eds), Gametes - The Oocyte. Cambridge University Press, Cambridge, pp. 95-107.##Sengoku K. ,.Tamate K,.Takuma N, Yoshida T.,.Goishi K and.Ishikawa M (1997) The chromosomal normality of unfertilized oocytes from patients with polycystic ovarian syndrome Human Reproduction 12, 3 pp.474-477,##Smith. R., Walker L., Cobo AC., et al., (1998) Contribution of chromosomal abnormalities to in vitro fertilization failures. Rev Med Chil 126(5):511-9.##Tarkowski, A.K. (1966) An air-drying method for chromosome preparation from mouse eggs. Cytogenetics, 5, 394-400.##Tarin, J.J., Gomez, E., Sampaio, M. et al. (1991) Cytogenetic analysis of human oocytes from fertile women. Hum Reprod 8, 1100-1103.##Wall, M. B., Marks, K., Smith, T. A. et al. (1996) Cytogenetic and fluorescent in-situ hybridization chromosomal studies on in-vitro fertilized and intracytoplasmic sperm injected 'failed-fertilized'##human oocytes. Hum Reprod 11, 2230-2238.##World Health Organization (1999) WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction. Cambridge University Press, Cambridge, UK.##Zenzes, M.T. and Casper, R.F. (1992) Cytogenetics of human oocytes, zygotes, and embryos after in vitro fertilization. Hum Genet 88, 367-37.## ##