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A bioassay procedure is described for quality control testing of various disposable items used in routine IVF procedures. This bioassay is performed over 4 days and uses the survival of human sperm in vitro at room temperature to assess which products are suitable for use. New products were tested for cytotoxicity using a general screening method and subsequent batches of every suitable item tested to detect interbatch variation. Products were considered suitable or unsuitable for use depending upon a calculated sperm survival index. Two main types of product were found to be cytotoxic, namely certain brands of syringe and surgical gloves, the common feature of both being the presence of rubber components. The bioassay was also used to investigate further the cytotoxic effect of the powdered and starch-free surgical gloves. The cytotoxic substances from both types of surgical glove were readily transferred to an embryo replacement catheter by touch, and washing of the gloves reduced this effect only moderately. The bioassay has proved inexpensive and convenient but more importantly it has been invaluable for detecting potential sources of cytotoxicity before they are introduced into a standard IVF protocol.

Between 1978 and 1990, 122 men underwent semen analysis before starting sterilising chemotherapy for Hodgkin's disease. Eighty-one (66%) had semen quality within the normal range, 25 were oligospermic (520610 6 sperm per ml) and five were azoospermic (no sperm in the ejaculate). Semen from 115 men was cryopreserved and after a median follow-up time of 10.1 years, 33 men have utilised stored semen (actuarial rate 27%) and nine partners have become pregnant resulting in 11 live births and one termination for foetal malformation. Actuarial 10 year rates of destruction of semen before death or utilisation and death before utilisation are 19% and 13% respectively. This retrospective cohort study demonstrates that approximately one-quarter of men utilising cryopreserved semen after treatment for Hodgkin's disease obtain a live birth. The high nonutilisation rate is intriguing and warrants further investigation. British Journal of Cancer (2002) Most patients with Hodgkin's disease can expect prolonged survival due to the success of chemotherapy regimens which have evolved since the first introduction of MOPP (mustine, vincristine, procarbazine and prednisolone) in 1964 (De Vita et al, 1972). Unfortunately, infertility is a common late effect of treatment and particularly important since most patients are young adults at diagnosis (Whitehead et al, 1982). Semen cryopreservation is currently the only available technique to preserve reproductive potential in men undergoing potentially sterilising treatments and in the UK is offered routinely to patients who are under 55 years of age. However, little information exists as to the benefits of this policy. In this study, semen quality before and after chemotherapy, rates of utilisation of cryopreserved semen and reproductive outcome have been analysed in a cohort of 122 men who presented to a regional cancer centre with newly diagnosed Hodgkin's disease between 1978 and 1990. PATIENTS AND METHODSNames of all male patients with newly diagnosed Hodgkin's disease registered at the Christie Hospital were cross-matched with the names of patients referred for semen cryopreservation at St Mary's Hospital, Manchester between January 1978 and December 1990. Permission to gather information from the files at St Mary's Hospital, Manchester was obtained from the Human Fertility and Embryology Authority. Details of patient age and stage of disease at diagnosis, treatment given, current disease status, dates and treatment of relapse, semen analysis before and after treatment, marital status and number of children fathered prior to diagnosis of Hodgkin's disease, use of cryopreserved semen, outcome of assisted reproduction and natural conceptions following treatment were obtained from case notes and the reproductive medicine data base. To compare semen quality between patients with multiple semen analyses a 'best' sample was selected by using a rank-sum procedure. Briefly, sperm concentration and motility were ranked independently and in ascending order, then the ranks of concentration and...

Advances in cancer treatment have led to significant improvements in the likelihood of reaching remission and long-term survival for men. Chemo- and radiotherapy-induced infertility are significant treatment side effects. Cryopreservation before the start of treatment enables sperm to be stored, thereby preserving the man's potential fertility. Here, we describe the successful use (with ICSI) of sperm cryopreserved prior to cancer treatment, for a total of 21 years. We believe this to be the longest period of sperm cryopreservation, resulting in a live birth, so far reported in the literature.

Ovarian hyperstimulation syndrome (OHSS) is a serious and potentially life-threatening complication following ovarian stimulation for in vitro fertilization (IVF). Coasting is the practice whereby the gonadotrophins are withheld and the administration of human chorionic gonadotrophin (hCG) is delayed until serum oestradiol (E2) has decreased to what is considered to be a safe level, to prevent the onset of OHSS. This study aimed to assess the length of coasting on the reproductive outcome in women at risk of developing OHSS. Coasting was undertaken when the serum E2 concentrations were > or = 17000 pmol/L but < 21000 pmol/L. Daily E2 measurements were performed and hCG was administered when hormone levels decreased to < 17000 pmol/L. Eighty-one women who had their stimulation cycles coasted were grouped according to the number of coasting days. Severe OHSS occurred in one case, which represented 1.2% of patients who underwent coasting because of an increased risk of developing the syndrome. No difference was found between cycles coasted for 1 - 3 days and cycles coasted for > or = 4 days in terms of oocyte maturity, fertilization and embryo cleavage rates. Women in whom coasting lasted for > or = 4 days had significantly fewer oocytes retrieved (P < 0.05) and decreased implantation rate (P < 0.05) compared to those coasted for 1 - 3 days. Pregnancy rate/embryo transfer and live birth rate did not differ between groups. In conclusion, coasting appears to decrease the risk of OHSS without compromising the IVF cycle pregnancy outcome. Prolonged coasting is, however, associated with reduced implantation rates, perhaps due to the deleterious effects on the endometrium rather than the oocytes.

PurposeGenetic variation may influence women’s response to ovarian stimulation therapy. The purpose of this study was to investigate any effects of genetic variants in the anti-Müllerian hormone (AMH) and AMH type II receptor genes on ovarian response/treatment outcomes and on current markers of ovarian reserve in individuals undergoing in vitro fertilisation (IVF) treatment.MethodsIn this prospective observational study, we genotyped the AMH c.146G>T, p.(Ile49Ser) and AMHR2 -482A>G variants in 603 unrelated women undergoing their first cycle of controlled ovarian stimulation for IVF and ICSI (intracytoplasmic sperm injection) using gonadotrophins at a tertiary referral centre for reproductive medicine. Pelvic ultrasound and blood hormone levels were taken on days 2–3 of the cycle. Genotypes were determined using TaqMan allelic discrimination assay. Regression analysis was performed to assess the relationship between the genotypes and the ovarian reserve markers (FSH, AMH, antral follicle count) and the early outcomes of response (number of oocytes retrieved and gonadotropin dose) as well as the treatment outcome (live birth).ResultsThere were no significant associations between the variants AMH c.146G>T and AMHR2 -482A>G with ovarian response in terms of number of oocytes retrieved (p = 0.08 and p = 0.64, respectively), live births (p = 0.28 and p = 0.52) and/or markers of ovarian reserve.ConclusionsGenotyping of the AMH c.146G>T and AMHR2 -482A>G polymorphisms does not provide additional useful information as a predictor of ovarian reserve or ovarian response and treatment outcomes.

A total of 364 consecutive patients requesting in-vitro fertilization (IVF) treatment were divided randomly into two groups. In the first group, two embryos in the original IVF cycle were allowed to divide prior to transfer, with any remaining embryos being cryopreserved at the pronucleate (PN) stage. In the second group, all the embryos were allowed to divide to the early cleavage (EC) stage, and the best two replaced; any suitable remaining embryos were frozen at the 2- to 4-cell stage. A total of 134 cycles (36.8%) fulfilled the study criteria for a fresh embryo replacement and supernumerary embryos cryopreserved. In the PN group, 72 out of 182 (39.6%) patients had a fresh embryo replacement accompanied by embryo cryopreservation, which was not significantly different from the EC group (62/182; 34.1%). The livebirth rate per fresh embryo transfer in the EC group (17/62; 27.4%) was significantly higher than that for the PN group (8/72; 11.1%; P < 0.05). Embryo survival following thawing was similar for the PN (96/129; 74.4%) and EC (79/102; 77.4%) stages. Although not significant, the livebirth rate following the transfer of thawed embryos was higher in the PN group (11/44; 25.0%) than in the EC group (4/38; 10.5%). Following one fresh and two freeze-thaw embryo replacements, the observed cumulative viable pregnancy rates were comparable for patients in both the PN (40.2%) and EC (41.1%) groups.