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The Swedish insemination act and the availability of donors.

Sweden was in 1985 the first country in the world to enact legislation concerned with the practice of donor insemination (DI). Included in this legislation was the requirement that information about the identity of the donors be kept and be made available to the resultant offspring upon 'maturity', should they seek this. This provision has attracted much interest, debate and criticism, and claims have been made that this has led to a decline in the numbers of men coming forward to donate semen in Sweden. This paper presents data covering the pre- and post-legislation period from one clinic, along with the only available data on all the Swedish clinics providing DI, which covers a more limited post-legislation period. This data shows that there has been an increase in number of donors being recruited, suggesting that the possibility of future contact by genetic offspring has not had the negative impact on the availability of donors predicted.

Confidentiality↗

The stages at which human fertilization arrests: microtubule and chromosome configurations in inseminated oocytes which failed to complete fertilization and development in humans.

The goal of fertilization is the union of one, and only one, sperm nucleus with the female pronucleus within the activated oocyte. For this to occur successfully, several events must transpire, including the incorporation of the entire spermatozoon into the oocyte, the completion of meiotic maturation with the extrusion of the second polar body, the metabolic activation of the previously quiescent oocyte, the decondensation of the sperm nucleus and the maternal chromosomes into the male and female pronuclei respectively, and the cytoplasmic migrations of the pronuclei, which bring them into apposition. Defects in any of these events are lethal to the zygote and might prove to be causes of infertility. In this study, the microtubules and DNA were imaged in inseminated human oocytes that had been discarded as unfertilized. The presence and number of incorporated sperm tails were also documented using a monoclonal antibody specific for the post-translationally modified acetylated-alpha-tubulin found in the tail, but not the oocyte, microtubules. An analysis of 211 oocytes from failed in-vitro fertilizations from 58 patient couples resulted in the determination of several previously undetectable phases at which fertilization arrests: (i) metaphase II arrest; (ii) arrest after the successful incorporation of the spermatozoon, (iii) arrest after the formation of the sperm aster; (iv) arrest during mitotic cell cycle progression; and (v) arrest during meiotic cell cycle progression. Data on polyspermy and arrested embryonic development are also presented. These results have implications for the diagnosis and treatment of female, as well as male, infertility.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Cycle↗

Gamete intra-Fallopian transfer or in-vitro fertilization after failed ovarian stimulation and intrauterine insemination in unexplained infertility?

A prospective randomized study was designed to compare gamete intra-Fallopian transfer (GIFT) and in-vitro fertilization (IVF) and embryo transfer in the treatment of couples who have failed to conceive after at least three cycles of ovarian stimulation and intrauterine insemination (IUI). A total of 69 couples with primary unexplained infertility of at least 2 years' duration plus at least three failed cycles of ovarian stimulation and IUI were randomly allocated to either GIFT or IVF/embryo transfer. The clinical pregnancy rate was 34% after GIFT treatment and 50% after IVF/embryo transfer. This difference was not statistically significant. The twin rate in the IVF/embryo transfer group was higher than in the GIFT group (53 versus 17%, P = 0.005). We conclude that patients with unexplained infertility and failed ovarian stimulation and IUI can still achieve encouraging pregnancy rates with IVF/embryo transfer or GIFT. Since IVF/embryo transfer is the least invasive of the two procedures and may yield diagnostic information, we would favour this therapy; however, the number of embryos transferred should be reduced to two to reduce the risk of twin pregnancy.

Adult↗

Effectiveness of various sperm processing methods in removing seminal plasma from insemination media.

The effectiveness of the wash and swim-up (1 and 2 wash cycle), two layer Percoll gradient, SpermPrep and underlay sperm preparation methods in removing seminal plasma from insemination suspensions was investigated. The number of wash cycles needed to rid sperm suspensions of seminal plasma (n = 15) was also determined. All sperm preparation methods were compared to the control wash and swim-up method, performed using Earle's buffered salt solution (EBSS), without serum supplementation so as not to mask seminal plasma concentrations. Control processing comprised 1 ml semen subjected to two wash cycles in EBSS followed by a swim-up period of 60 min under 5% CO2 in air. The Percoll method comprised 1 ml semen layered on a discontinuous 36 and 81% Percoll gradient (n = 14). In the SpermPrep method, 1 ml semen was run through a SpermPrep II column (n = 10), and underlay samples (n = 12) were processed by two wash cycles, after which sperm pellets were resuspended in the remaining medium, layered under 1.2 ml EBSS and allowed to swim up under 5% CO2 in air for 1 h. Seminal plasma and supernatant fractions obtained after each processing phase were stored at -20 degrees C and protein concentrations were determined by spectrophotometry. The wash and swim-up method was the most effective in removing seminal plasma from sperm suspensions, followed by the two-layer Percoll gradient, underlay and finally the SpermPrep II processing methods.

Chemical Fractionation↗

Induction of pre-ovulatory gonadotrophin surge with gonadotrophin-releasing hormone agonist compared to pre-ovulatory injection of human chorionic gonadotrophins for ovulation induction in intrauterine insemination treatment cycles.

The clinical outcome of intrauterine insemination (IUI) treatment cycles employing a gonadotrophin-releasing hormone agonist [GnRHa, triptorelin (Decapeptyl)] or human chorionic gonadotrophin (HCG) for ovulation induction was compared. A group of 48 patients presenting with amenorrhoea, oligomenorrhoea or unexplained infertility were all treated with human menopausal gonadotrophins (HMG) from day 5 of the cycle, on an individualized schedule. They were then randomly divided into two groups to receive either a single s.c. injection of 0.1 mg triptorelin or a single i.m. injection of 10,000 IU HCG after follicular maturation. IUI was performed approximately 24 and 48 h following the injection. A transitory increase in serum luteinizing hormone and follicle stimulating hormone concentrations was achieved following injection of GnRHa. A total of 24 patients received 72 treatment cycles with GnRHa, producing 11 conceptions (15.3%) and two abortions (18.2%), resulting in a term pregnancy rate of 13.6%. There were four cases of grade 3-4 ovarian hyperstimulation syndrome (OHSS), two of which were conception cycles. In all, 24 patients underwent 68 cycles treated with HCG, producing 18 conceptions (26.5%) and six abortions (33.3%), resulting in a term pregnancy rate of 19.0%. There were eight cycles of grade 3-4 OHSS, two of which were conception cycles. These results show that an s.c. injection of a relatively low dose of GnRHa can be as effective as HCG in producing pregnancy in IUI treatment cycles.

Abortion, Spontaneous↗

Post-operative artificial insemination--does it improve GIFT outcome?

Based on a retrospective study of our gamete intra-Fallopian transfer (GIFT) programme we have analysed 102 cycles where a standard GIFT procedure was implemented and 92 cycles where a post-operative intrauterine and intracervical insemination (post-operative IUI/ICI) was performed, in addition to GIFT. Our comparison suggests that the pregnancy outcome has been significantly improved (P less than 0.05) by this additional post-operative IUI/ICI. The standard GIFT group (102 cycles) yielded 38 clinical pregnancies and, ultimately, 29 delivered pregnancies (37.2/28.4%), whilst the post-operative IUI/ICI group (92 cycles) gave 48 clinical pregnancies initially and, currently, 38 pregnancies are ongoing or delivered (52.2/41.3%). The possible implications of the 'normal' presence of spermatozoa in the human female tract as an explanation for this improvement is discussed, and the adoption of IUI/ICI after GIFT is proposed, following a prospective controlled study.

Abortion, Spontaneous↗

Micro-centrifugation of human spermatozoa: its effect on fertilization of hamster oocytes after micro-insemination spermatozoal transfer.

Micro-centrifugation (MC) at 6500 r.p.m. (3352 g) has not been used previously for spermatozoal concentration and subsequent fertilization. We investigated MC for micro-insemination spermatozoal transfer (MIST) of human spermatozoa from normal donors into hamster oocytes. MC resulted in reduced penetration of hamster oocytes, both after MIST [77.4% (41/53) versus 87.8% (43/49) for control; NS] and after exposure to zona-free hamster oocytes [60.8% (79/130) versus 72.7% (88/121) for control; P less than 0.05]. However, MIST under the zona resulted in better incorporation of sperm nuclei when compared with zona-free hamster oocytes, for spermatozoa exposed to micro-centrifugation (77.4 versus 60.8%, P less than 0.05) and controls (87.8 versus 72.7%, P less than 0.05). Polyspermy was higher after MIST [22.0% (9/41) versus 13.9% (11/79); NS] for MC+, and [25.6% (11/43) versus 13.6% (12/88); NS] for MC-. We conclude that MC does have a negative, but minimal effect on the fertility of human spermatozoa with respect to hamster oocytes.

Animals↗

Intrauterine insemination and comparison of two methods of sperm preparation.

In a study of intrauterine inseminations (IUI) after clomiphene stimulation, a randomized comparison was made between a new method of sperm preparation, self migration in sodium hyaluronate (SH), and a traditional method, centrifugation and swim up (CS). After two IUI cycles with either SH or CS, the sperm preparation method was swapped and the patients received another two IUI cycles. Interjacent cycles of natural intercourse after clomiphene treatment served as the control. The SH method resulted in a significantly higher percentage recovery of progressive motile spermatozoa than the CS method, 17.7% versus 8.6% (P less than 0.01). The sperm samples were prepared by SH in 68 cycles and by CS in 57 cycles, resulting in six and five pregnancies, respectively. Pregnancies were obtained in 11 of 125 IUI cycles (8.8%) and in 3 of 124 control cycles (2.4%) (P less than 0.05). The pregnancy rate following IUI was highest in the patients with cervical factor (35%) and asthenozoospermia (23%), while none became pregnant in the group with oligozoospermia. In the unexplained infertility group, no difference between the pregnancy rates in IUI cycles and control cycles was seen. SH is a simple and rapid method of sperm preparation and it appears to give a high recovery of motile spermatozoa and a number of pregnancies which is comparable to that of CS. Treatment with IUI in cycles with a simple stimulation protocol seems to be valuable in cases involving either a cervical factor or asthenozoospermia.

Adult↗

Pregnancy after direct intraperitoneal insemination.

Direct intraperitoneal insemination (DIPI) and superovulation are simple procedures which may together represent a good alternative to gamete intra-Fallopian transfer (GIFT) in infertile women with patent Fallopian tubes. In the present study, pregnancy occurred in 25 of 96 couples (26%) and six (24%) of these aborted. The pregnancy rate for all cycles was 19.6% and multiple pregnancies were found in six of 25 (24%) patients. We observed no ectopic pregnancy. The combination of these techniques is concluded to be useful in achieving pregnancy in infertile women with patient Fallopian tubes.

Adult↗

Multiple follicular recruitment and intrauterine insemination outcomes compared by age and diagnosis.

We studied the outcome of our intrauterine insemination (IUI) programme, evaluating female age and diagnosis. One-hundred-and-twenty-six patients less than 36 years of age (mean 30.91 +/- 3.02 years) completed 306 cycles of multiple follicular recruitment (MFR) and timed IUI; 64 patients greater than or equal to 36 years of age (mean 38.36 +/- 2.08 years) completed 166 cycles (total 190 patients, 472 cycles). The male partners' semen was prepared for IUI with wash and swim-up techniques. Diagnostic groups were: male factor (n = 26), idiopathic (n = 33), endometriosis (n = 19), ovulatory disorder (n = 7), other (n = 19) and combined factors (n = 86). Pregnancy rates (% per couple, % per cycle) [overall (31.58, 12.7)] [less than 36 years (38.10, 15.69)] [greater than 36 years (18.75, 7.23)] were greater in the less than 36 years group (P less than 0.025). The probability of conception after three treatment cycles was 0.402 overall, 0.481 for age less than 36 years and 0.252 for age greater than or equal to 36 years. The probability of conception for male factor and idiopathic infertility patients was 0.469 and 0.411 respectively. An age effect was found on pregnancy rates in the idiopathic group only. In conclusion, MFR + IUI is a valuable treatment especially for male factor patients and patients less than 36 years old, with idiopathic infertility.

Adult↗

Antisperm antibody production following intrauterine insemination.

Fifty-one infertile women were studied prospectively to assess whether intrauterine insemination (IUI) led to the development of antisperm antibodies in cervical mucus and/or serum. All women were tested for the presence of antisperm antibodies in cervical mucus and serum before and after IUI treatment. Each woman underwent between 1 and 9 cycles of IUI (mean: 4.0 cycles). Five women (9.8%) developed serum antisperm antibodies after IUI treatment. Three of 49 women (6.1%) developed cervical mucus antisperm antibodies and three other women demonstrated disappearance of antibodies following IUI treatment. IUI did not increase mucus or serum antibody titres in women who presented with antisperm antibodies. The number of IUI cycles did not correlate with the development of antisperm antibodies. We conclude that IUI increases the risk of the female partner developing systemic antisperm antibodies but the clinical significance of this finding in unclear.

Adult↗

Induction of acrosome reaction in human spermatozoa used for subzonal insemination.

Human spermatozoa were injected into the perivitelline space of oocytes from 43 couples (44 cycles) in whom fertilization had failed in conventional in-vitro fertilization (IVF). The spermatozoa were treated to enhance the percentage of acrosome-free spermatozoa either by incubation for 24 h in T6 medium with 50% follicular fluid (v/v) or by incubation for 24 h in T6 medium followed by electroporation and incubation for a few hours in T6 medium with 3.5 mM pentoxifylline. After these two procedures, the mean percentage of acrosome-free spermatozoa increased to 35.5 and 53.9% respectively. Up to three spermatozoa were injected into the perivitelline space of metaphase II oocytes; few oocytes were damaged during the injection procedure. The overall fertilization rate was 30.9% of the 433 oocytes that were intact after subzonal insemination. Only 3% of the injected oocytes had more than two pronuclei. The cleavage rate of the fertilized oocytes was 80%. There was no difference in the fertilization and cleavage rates between the two sperm treatment procedures. One, two or three embryos were replaced in 34 cycles and seven patients became pregnant. In three of the four ongoing pregnancies, prenatal diagnosis by amniocentesis indicated a normal karyotype.

Acrosome↗

A prospective controlled study of in-vitro fertilization, gamete intra-fallopian transfer and intrauterine insemination combined with superovulation.

The relative effectiveness of in-vitro fertilization (IVF), gamete intra-Fallopian transfer (GIFT) and intrauterine insemination (IUI) combined with superovulation in the treatment of infertility were compared in 151 couples undergoing a single cycle of treatment. Treatment was selected as appropriate (IVF for tubal disease, GIFT or IUI/superovulation for nontubal infertility) but possible bias due to non-randomization was overcome by all couples having had favourable fertilization in a previous cycle of IVF. Furthermore, in a preliminary study of initial IVF treatment in 265 couples from whom the study patients were drawn, implantation and pregnancy rates in the diagnostic groups were similar. In the definitive study comparing IVF, GIFT and IUI/superovulation, the pregnancy rate observed with GIFT was highest (40%) but this was not significantly higher than with IVF (28%) or IUI/superovulation (20%). However, the implantation rate per egg transferred by GIFT (21%) was significantly higher than the implantation rate per embryo transferred by IVF (11%). Although the pregnancy rates with GIFT were not statistically greater than with IVF, a significant advantage is likely to be observed in larger groups in view of the better implantation rate. The lower pregnancy rates with IUI superovulation are to be expected because of limited ovarian stimulation, they are nevertheless of comparative interest.

Adult↗

Reliability of home urinary LH tests for timing of insemination: a consumer's study.

In this study, intrauterine insemination (IUI) was timed either after the detection of a urinary luteinizing hormone (LH) surge at home by the patient (group A), or following a positive LH test as interpreted by the gynaecologist (group B). Afterwards, samples tested by the patient were retested by the gynaecologist and vice versa. The gynaecologist also rechecked his own findings and the results were correlated with ultrasound data and charts of basal body temperature. Forty-seven cycles were evaluated. The patient's and the gynaecologist's readings agreed (+/- 12 h) in 42 of the cases (89%), and in five cycles (11%) a difference greater than 24 h was found. The intra-observer variation in the gynaecologist's results was +/- 12 h in four cycles (8.5%). These findings suggest that the LH test can be used as a reliable home device for the prediction of pending ovulation and timing of IUI.

Adult↗

Hormone and ultrasound parameters in ovarian stimulation cycles for direct intraperitoneal insemination.

To determine hormonal and ultrasound parameters associated with pregnancies, 115 women with unexplained infertility (n = 82), endometriosis (n = 22) or cervical factor (n = 11) were treated with direct intraperitoneal insemination (DIPI) after ovarian stimulation with clomiphene citrate and human menopausal gonadotrophins (HMG). Twenty women conceived and were compared with the remaining 95 non-pregnant women during one treatment cycle. Women with basal FSH levels less than or equal to 1.25 micrograms/l responded with higher oestradiol levels (P less than 0.0001), with the development of more follicles (P less than 0.05) and higher progesterone levels (P less than 0.05) than women with basal FSH levels greater than 1.25 micrograms/l, but the conception rates were similar. Women with miscarriages or biochemical pregnancies had a higher basal FSH value than both the women with term pregnancies and the non-pregnant women. Women with at least 3 preovulatory follicles greater than or equal to 15 mm had a higher pregnancy rate than those with fewer follicles, but a further increase was not observed above that number. The endometrium was thicker on the day of ovulation induction in cycles leading to a term pregnancy than in cycles without conception or with a biochemical pregnancy. No term pregnancy was observed when the endometrium was thinner than 8 mm. Women with a short luteal phase (less than 12 days) had a higher ratio of oestradiol/progesterone in the midluteal phase than women with a luteal phase of greater than or equal to 12 days and pregnant women.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Male infertility and donor insemination: couples' decisions, reactions and counselling needs.

This paper reports findings of a postal survey of 205 couples belonging to the National Association for the Childless, whose infertility had been attributed to the male partner. Spontaneous resolution was seldom reported, but 36 couples had become parents by donor insemination (DI) and 28 through adoption. Almost a third of respondents had managed to resolve their childlessness. Although reasons for rejecting DI or preferring other options were offered by 60 couples, 125 had taken active steps towards it. Most of the 80 couples who had been counselled had found this helpful. Regardless of counselling, of the 76 couples who had considered whether or not to tell their children of their mode of conception, 56 (74%) stated that they had no intention of doing so. Quantitative indices from two self-report scales confirm that male infertility is commonly a source of distress, especially for the female partner. However, the data also suggest that this is not necessarily associated with chronic marital discord. Couples who had already decided for or against DI reported less distress than others. The need for mutual support may well be enhanced until the couple's quest for a child has been either achieved or abandoned.

Counseling↗

Higher success rate by intracytoplasmic sperm injection than by subzonal insemination. Report of a second series of 300 consecutive treatment cycles.

Subzonal insemination (SUZI) and intracytoplasmic sperm injection (ICSI) were carried out in 300 treatment cycles in couples unable to be helped by conventional in-vitro fertilization treatment. More oocytes were damaged by ICSI (13.5%) than by SUZI (7.1%). The normal fertilization rate was substantially higher after ICSI (51.0%) than after SUZI (14.3%) and was related to the semen characteristics. The cleavage rate was similar for both procedures (77%). After 217 embryo transfers (72.3% of the treatment cycles) 66 pregnancies were established, i.e. pregnancy rates of 22.0% per started cycle and 30.4% per embryo transfer. So far, pregnancy loss has occurred in 27.3% of the pregnancies, nine healthy children have been born after eight deliveries and 41 clinical pregnancies are progressing uneventfully. Chorionic villus sampling or amniocentesis have been performed in 35 pregnancies and 39 normal fetal karyotypes have been obtained after cytogenetic analysis.

Female↗

Effect of sperm pre-treatments on the results of sub-zonal insemination (SUZI).

Follicular fluid and progesterone, which are present in the natural environment of oocytes, have been reported to induce the acrosome reaction and we compared their use in pretreatment of spermatozoa for human sub-zonal insemination (SUZI). Pre-treatment with follicular fluid (20% v/v) was associated with a higher fertilization rate than pre-incubation with progesterone (1 mmol/l) as assessed by both the embryos/injected oocytes rate (31.7 +/- 6.2% versus 13.5 +/- 5.9%, respectively; P < 0.01) and the male pronuclei/injected spermatozoon rate (10.5 +/- 3.3% versus 3.6 +/- 1.9%, respectively; P < 0.01). Since we have previously reported that pre-treatment with progesterone allowed a higher percentage of live-reacted spermatozoa to be obtained, these results suggest that either progesterone induces modifications of the plasma membrane, which prevent fusion with the oolema, or that follicular fluid not only induces the acrosome reaction but increases the fusion ability by compounds other than progesterone.

Female↗