Elevated basal estradiol levels have no negative prognosis in young women undergoing ART cycles.

The objective of this study was to determine the relationship between serum estradiol levels on day 2 of an in vitro fertilization and embryo transfer (IVF-ET) cycle and the chance of pregnancy and implantation rates according to age. Two hundred and forty-eight cycles of IVF-ET in patients treated with gonadotropin-releasing hormone analog (GnRHa, follicle-stimulating hormone (FSH) and human menopausal gonadotropin (hMG) were divided into several groups according to the basal serum concentrations of estradiol (E2):A < 25 pg/ml, B 26-50 pg/ml, C 51-75 pg/ml, D 76-100 pg/ml, E > 100 pg/ml. Furthermore, the patients were subdivided into two subgroups according to age: I younger; and II, older than 35 years. Pregnancy and implantation rates were analyzed for each subgroup according to basal E2 level and age. Statistical analysis was performed using analysis of variance, chi2 and Fisher's test. The number(s) of cycles for each subgroup were 164 and 84 for I and II, respectively. Even though there were no significant differences among groups for the pregnancy rates for individual groups, there was a tendency for a decreased rate with increasing levels of E2 only in women older than 35 years of age. When pregnancy rates of all women with E2 levels above 25 pg/ml were calculated, the differences between those observed in subgroup I (< or = 35 years) and II (> 35 years) was significant: p = 0.02 (38.7% vs. 18.7%, respectively). When implantation rates in the same groups and subgroups were analyzed, we found that again the differences were statistically significant: p = 0.001 (13.1%vs, 4.3% for I and II). The results of the present studies reveal that basal levels of E2 are a marker of poor prognosis for implantation and pregnancy in assisted reproduction (ART) cycles only in patients over the age of 35 years. These data cast doubt on the use of basal levels of E2 to screen women below the age of 35 prior to initiating an ART cycle. In addition, high levels of basal E2 per se may not be deleterious for reproductive efficiency in younger women, since pregnancy and implantation rates did not differ among patients younger than 35 years old (IA to E).

Spontaneous resolution of ectopic pregnancy in a surrogate after oocyte donation and frozen embryo transfer.

A case of tubal pregnancy in a young and healthy woman participating in a programme of in-vitro fertilization (IVF) gestational surrogacy is reported. The gestational surrogate was the 30 year old fertile sister of a 25 year old patient affected by stage 1 ovarian cancer. After mandatory oncological consultation, the donor was recommended to prospectively undergo controlled ovarian hyperstimulation cycles for embryo banking before being treated by total hysterectomy. Available embryos were cryopreserved and after adequate endometrial preparation using artificial cycles of hormone replacement therapy, three thawed frozen embryos were transferred to the surrogate. At 17 days following embryo transfer the surrogate was noted to have a negative beta-human chorionic gonadotrophin (HCG) serum concentration. All medication was suspended and a few days later normal menstrual bleeding occurred. After 2 weeks, the beta-HCG concentrations, performed as part of routine follow-up evaluation, were showing signs of trophoblast activity (236 mIU/ml). Taking into account the stable condition of the patient, a decision was made to undertake expectant management. At 43 days after embryo transfer, a complete tubal abortion was apparently seen in the posterior cul-de-sac by ultrasound associated with a subtle and short lasting pelvic pain. We stress that this ectopic gestation was able to maintain prolonged viability in conditions of absent corpus luteum and exogenous steroid supplementation.

Successful fertilization, pregnancy, and birth using epididymal sperm frozen 24 hours after conventional oocyte insemination.

OBJECTIVE: To assess if epididymal sperm cryopreserved 24 hours after exposure to oocytes in conventional IVF can be successfully used for intracytoplasmic sperm injection (ICSI) in a subsequent cycle. DESIGN: Case report. SETTING: University of California, Irvine, Center for Reproductive Health. PATIENTS: Two men with obstructive azoospermia requiring microsurgical epididymal sperm aspiration, IVF, and ICSI. INTERVENTIONS: Freezing of epididymal sperm 24 hours after egg exposure in conventional IVF and subsequent use for assisted fertilization in a new cycle. MAIN OUTCOME MEASURE: Frozen-thawed epididymal sperm survivability and maintenance of fertilization and pregnancy capacity. RESULTS: At the time of sperm aspiration procedure (cycle I) a total of 30 oocytes were available for insemination. Of these, 15 were used for conventional IVF resulting in 2 embryos (13%) and 15 were used for ICSI, resulting in 3 embryos (20%). Sperm was cryopreserved 24 hours after conventional IVF and thawed 6 months later in a new cycle. Upon thawing, sperm were still found to be motile and at this time (cycle II) only assisted fertilization was used. Of 27 oocytes injected, 12 (44%) produced normal, cleaving embryos. One singleton pregnancy with the birth of a healthy infant girl was achieved after the tubal transfer of 5 embryos. CONCLUSION: The birth of a normal, healthy infant girl with epididymal sperm frozen 24 hours after exposure to oocytes in conventional IVF emphasizes the value of freezing any aliquot of epididymal sperm, even if the motility is very low, to avoid additional surgery in the male. From a basic science standpoint, this observation may renew interest in the study of sperm cryopreservation after occurrence of acrosome reaction and hyperactivation.

The use of progestins for programming assisted reproductive cycles and gonadotropin-releasing hormone agonist flare-up protocols in older patients.

OBJECTIVES: To determine if the use of norethisterone acetate (NET) in the previous cycle affects the ovarian response to GnRH agonist (GnRH-a) in flare-up protocols and controlled ovarian hyperstimulation in older patients. DESIGN: Retrospective analysis of the outcome of the assisted reproductive technology (ART) cycle. PATIENTS: Eighty women > 37 years old undergoing controlled ovarian hyperstimulation (COH) for a ART cycle (GIFT, IVF, zygote intrafallopian transfer). Forty received NET during the luteal phase of the previous cycle for programming the procedure and 40 did not receive NET (control group). Gonadotropin-releasing hormone agonist in follicular phase (flare-up) protocols were administered to all the patients. MAIN OUTCOME MEASURES: Cycle outcome: amount of gonadotropins used, oocyte production and quality, fertilization, and pregnancy rates. Estradiol, FSH, and LH levels the first 3 days of COH in eight patients. RESULTS: There were no differences between both groups in the cycle outcome. Estradiol levels during the first 3 days of COH were higher in the patients that did not receive NET in the previous cycle. Follicle-stimulating hormone and LH levels were similar in both groups. CONCLUSIONS: The administration of NET in the previous cycle in patients > 37 years old does not affect the ovarian response to the combination of follicular phase GnRH-a and gonadotropins for COH.

The abnormally large follicle during controlled ovarian hyperstimulation: management and outcome of the cycle.

OBJECTIVE: To investigate if the presence of an abnormally large follicle during controlled ovarian hyperstimulation (COH) under pituitary suppression has any effect on the outcome of the cycle. DESIGN: Prospective, observational. SETTING: The reproductive endocrinology unit of a university hospital. PATIENTS: One hundred fifty patients undergoing COH for assisted reproductive techniques during a 6-month period (August 1990 to January 1991). INTERVENTIONS: Transvaginal follicular aspiration and IVF-uterine ET. RESULTS: A total of 19 cycles had abnormally large follicles identified on day 8 of the stimulation cycle after normal baseline ultrasound. Gonadotropins were continued and hCG injection was indicated when two or more follicles of the main cohort achieved a diameter of 20 to 22 mm. Twenty preovulatory oocytes were retrieved from 22 large follicles. Two were transferred for GIFT and 18 were inseminated in vitro, resulting in a 72.0% fertilization rate. The mean number of oocytes retrieved per patient was 10.9, 71.4% of which were mature with a fertilization rate of 67.7%. All these figures were comparable with the results obtained in the 131 patients of the control group undergoing IVF. No evidence of premature luteinization was observed in the study group, based on plasma P levels (x 0.83 ng/mL [conversion factor to SI unit, 3.180], range 0.31 to 1.40 ng/mL). The clinical pregnancy rate for the group with abnormally large follicles did not differ from the control group (27.8% versus 28.2%, respectively). CONCLUSIONS: The presence of an abnormally large follicle during COH under pituitary suppression does not affect the outcome of the cycle. Moreover, under these conditions, continuous gonadotropin stimulation of a follicle to diameters considerably larger than the standard ones does not have a detrimental effect on the oocyte contained in it, suggesting that oocyte aging is an independent process from follicular growth once LH surge is prevented.

Use of epididymal sperm for assisted reproduction in men with acquired, irreparable obstructive azoospermia.

Microsurgical epididymal sperm aspiration (MESA) and in vitro fertilization (IVF) is primarily offered to men with congenital absence of the vas deferens (CAVD). However, the IVF capacity of these epididymal sperm is low ( < 15%) and unpredictable. In this study, IVF and intracytoplasmic sperm injection (ICSI) results in patients with non-congenital, irreparable obstructive azoospermia were analysed. Thirty-three patients were evaluated for a total of 37 cycles of MESA and IVF. Most had obstruction secondary to failed vasectomy reversal and to epididymal blockage. The overall fertilization rate was 30% with regular IVF and 26% with ICSI, and six clinical pregnancies were obtained. Both rates are significantly higher than the IVF rate previously reported with sperm from men with CAVD (13%, P < 0.00001). In men with non-congenital obstructive azoospermia, a significant difference was found in the average sperm count (56.9 x 10(6) v. 12.3 x 10(6), P < 0.04) and total motile count (16.6 x 10(6) v. 1.6 x 10(6), P < 0.01) respectively for patients who achieved IVF and those who did not. It is concluded that (a) the real IVF capacity of human epididymal sperm is 30%, or 42%, if calculated only for patients who achieved fertilization, (b) this higher rate is an indirect support of the hypothesis that sperm from men with CAVD have intrinsic biochemical defects, related to cystic fibrosis mutations, responsible for their low and unpredictable IVF rate, and (c) MESA and IVF can be offered at the same time or as an alternative to patients requesting vasectomy reversal.

Comparison between nafarelin and leuprolide acetate for in vitro fertilization: preliminary clinical study.

OBJECTIVE: To evaluate the clinical effect of two different biochemical GnRH agonists (GnRH-a), nafarelin acetate and leuprolide acetate (LA), as adjunct to induction of ovulation in patients for IVF. DESIGN: Twenty-four women were assigned randomly to either nafarelin acetate or LA during IVF cycles. SETTING: University-affiliated clinics. PATIENTS: Infertile women undergoing IVF cycles in an academic research environment. INTERVENTIONS: Intranasal nafarelin at a dosage of 200 micrograms twice daily or LA at a dose of 1 mg/d SC was administered. Blood samples were collected on day 21 of previous cycle, days 2 and 8, and before hCG injection. MAIN OUTCOME MEASURE: Patient response as indicated by follicular phase serum levels of E2, FSH, and LH. RESULTS: Hormone profiles on cycle day 2 showed no statistical difference between both GnRH-a groups in FSH levels and a slight statistical difference for E2 levels. Patient response as demonstrated by follicular phase of E2, FSH, and LH measured on cycle day 8 and the day of hCG injection showed no statistically significant difference in both groups. Furthermore, the mean number of follicles, eggs retrieved, egg quality, fertilization rate, and number of embryos transferred and frozen were similar. The cycle cancellation rate and pregnancy rate per stimulation start were also not statistically different between the two groups. CONCLUSION: The study shows the comparable efficacy of these two drugs in controlled ovarian hyperstimulation (COH) protocols. The easy administration of nafarelin with prompt nasal absorption and the readily achieved blood level made nafarelin an option for use in COH in assisted reproductive technology.

Prospective randomized comparison of human chorionic gonadotropin versus intramuscular progesterone for luteal-phase support in assisted reproduction.

OBJECTIVE: The purpose of the study was to determine the impact of two forms of luteal-phase supplementation, human chorionic gonadotropin (hCG) and progesterone (P), during gonadotropin releasing hormone agonist (GnRh-a)/controlled ovarian hyperstimulation (COH) cycles. DESIGN AND PATIENTS: The study was a prospective, randomized evaluation of 77 patients. Group 1 patients (n = 38) received 2000 IU of hCG, injected subcutaneously, on days 3, 6, 9, and 12 after transvaginal aspiration of the oocytes (TVA = day 0). Group 2 patients (n = 39) received 50-mg daily injections of intramuscular (i.m.) P from days 2 to 14 after TVA. Blood tests were performed on days 0, 5, 8, and 12 after TVA. SETTING: The in vitro fertilization program of a tertiary care institution was the study setting. MAIN OUTCOME MEASURES: The main outcome measures were (1) pregnancy and implantation rates; (2) serum estradiol (E), P, and hCG levels; and (3) occurrence of side effects. RESULTS: Clinical pregnancy and implantation rates in group 1 versus group 2 were similar (36.7 vs 35.3 and 12 vs 14%, respectively). Regardless of pregnancy occurrence, on days 8 and 12 after TVA, serum E and P levels were higher in group 1 than group 2 but the resulting E/P ratios were similar. Five of 38 patients (group 1) developed moderate to severe ovarian hyperstimulation syndrome (OHSS) right after the first or second supplementary hCG injection. In these patients, the mean serum E level on the day of hCG trigger injection was about 3250 pg/ml and the number of follicles was between 9 and 17. In 6 of 39 patients (group 2) allergic reactions were observed at the P injection sites. CONCLUSIONS: Based on our data, hCG administration as a form of luteal supplementation did not translate, in comparison to P, into significant benefits for the patients. At the same time, it significantly increased the risk of ovarian hyperstimulation. We suggest that whenever, during COH cycles, serum E levels are over 2500 pg/ml and the number of follicles exceeds 10, luteal support with hCG should be excluded.

Pregnancy and implantation rates in normal replacement versus stimulated cycles.

The relative importance that endometrial development and embryo quality have on implantation rates achieved with assisted reproductive technology (ART) is the subject of controversy. Ovarian stimulation has been repeatedly mentioned as having a detrimental effect on endometrial receptivity (Paulson et al., 1990, Fertil. Steril., 53, 870-874). We compared pregnancy and implantation rates achieved with ART during stimulated cycles and hormonal replacement cycles, in patients matched for the following criteria: age < 35 years for the patient donating oocytes; transfer of at least two good quality embryos/oocytes and good quality transfer. All transfer cases performed during hormonal replacement cycles were done with donated oocytes. Comparison of results between techniques was not attempted due to potential differences in populations. The pregnancy and implantation rates achieved with each technique during stimulated and hormonal replacement cycles were not statistically different. In contradiction to previous results, our data suggest that differences in uterine receptivity between stimulated and hormonal replacement cycles in the age group studied are not of critical importance in embryo implantation. Good embryo quality appears to be the dominant factor in determining the success of ART.

Cystic fibrosis mutations impair the fertilization rate of epididymal sperm from men with congenital absence of the vas deferens.

One of the limiting factors for the successful treatment of male sterility due to congenital absence of the vas deferens (CAVD) is the low (< 20%) and extremely unpredictable rate of in-vitro fertilization of their epididymal spermatozoa. The recent demonstration that CAVD is a mild form of cystic fibrosis (CF) disease, almost exclusively involving the genital tract, has prompted us to investigate the hypothesis of whether there could be an association between particular CF genotype mutations and the in-vitro performance of epididymal sperm. In this study 63 patients with surgically confirmed diagnosis of CAVD undergoing microsurgical epididymal sperm aspiration (MESA) and in-vitro fertilization (IVF) were evaluated. The genetic screening was carried out on DNA extracted from peripheral lymphocytes and amplified by the polymerase chain reaction. A total of 12 mutations in the cystic fibrosis transmembrane regulator gene (CFTR), representing approximately 90% of the total known CF mutations, were tested. The presence or absence of mutations, as well as the type of mutation found, was correlated with the IVF and pregnancy rates. Of the 63 patients examined, 40 (64%) were positive to the CF screening and 23 were negative. In three cases no spermatozoa were found because of rete testis blockage. The difference in fertilization rates between patients testing positive and negative was highly significant (P = 0.000003), 13 versus 23%, respectively. In order to pinpoint which mutation could account for the most deleterious effect on the IVF results, we analysed these by keeping the patients separated by CF mutations.(ABSTRACT TRUNCATED AT 250 WORDS)

Can severe male factor infertility be treated without micromanipulation?

OBJECTIVE: To present data on IVF in severe male factor infertility using modified IVF laboratory methods and to compare them with the reported data of gamete micromanipulation. DESIGN: Retrospective, not randomized. SETTING: University of California, Irvine, Center for Reproductive Health. PATIENTS: Seventy-one patients with severe oligoasthenozoospermia defined as total motile count in the pretreatment sample of < 5 x 10(6). Two groups were identified, group I with total motile count between 1.5 and 5 x 10(6) and group II with total motile count < 1.5 x 10(6). INTERVENTIONS: Treatment of the semen samples with mini-Percoll and the modification of standard IVF techniques. These modifications include insemination with larger numbers of sperm, the use of culture tubes for insemination of oocytes, and pooling oocytes together in one or more culture tubes. MAIN OUTCOME MEASURES: Fertilization and pregnancy rates (PRs). Additionally, the number of patients achieving cryopreservation and patients with previous failure to fertilize are specified. RESULTS: Eighty percent of the patients achieved fertilization with an overall rate of 38% per oocyte. Fifty-three ETs were performed and 23 clinical pregnancies (43% per transfer and 35% per cycle) were achieved. Fifty percent of the patients had excess embryos for cryopreservation. In group I the fertilization rate was 54% with a PR of 56% per transfer and 48% per cycle. In group II the fertilization rate was 25% with a PR per transfer of 32% and 24% per cycle. Of 28 patients who had previous failure of fertilization, 25 fertilized and 8 pregnancies were established. CONCLUSIONS: These data demonstrate that by the modification of standard laboratory methods for IVF, monospermic fertilization, cleavage, and a high clinical PR can be achieved in cases of severe male factor infertility without having to resort to micromanipulation.

Effect of a gonadotropin-releasing hormone agonist on luteinizing hormone receptors and steroidogenesis in ovarian cells.

OBJECTIVE: To examine the effect of a gonadotropin-releasing hormone agonist (GnRH-a), leuprolide acetate (LA), on human chorionic gonadotropin/luteinizing hormone (LH) receptors content and progesterone (P) and estradiol (E2) production in cultured granulosa or luteal cells. DESIGN: Prospective. SETTING: Private Fertility Clinic and National Research Institute. PATIENTS: Twenty patients undergoing in vitro fertilization or gamete intrafallopian transfer programs. RESULTS: Human chorionic gonadotropin/LH receptors in human granulosa cells increased after 48 hours of culture, and LA inhibited such effect. Leuprolide acetate, 1 ng/mL, in the cultures produced an increase in P production. On the contrary, LA inhibited E2 production. Additionally, the in vivo effect of LA (2 micrograms/rat per 7 days) was studied in corpus luteum of superovulated rats. Luteal cells from LA-treated rats in culture produced lower P than the controls but showed an increase in aromatase activity. Luteal LH receptors declined after 48 hours of culture with LA. CONCLUSION: The high doses of gonadotropin necessary to induce ovarian hyperstimulation when GnRH-a is administered could be related with an inhibitory effect of these agonists on LH receptors and aromatase activity.

Aetiology of congenital absence of vas deferens: genetic study of three generations.

Bilateral congenital absence of the vas deferens (CAVD) is a form of male sterility (found in otherwise normal men) of unknown aetiology. Because males with cystic fibrosis (CF) almost invariably have CAVD as well, we investigated the hypothesis that men with isolated CAVD might share a common genetic background with males with CF. Genetic testing for CF was carried out in three generations of subjects: 44 patients with CAVD and their wives, 24 of their parents, and 13 of their offspring generated by microsurgical epididymal sperm aspiration (MESA) and in-vitro fertilization (IVF). DNA extracted from peripheral lymphocytes was amplified by the polymerase chain reaction (PCR) and then analysed for 12 mutations in the cystic fibrosis transmembrane conductance regulatory (CFTR) gene. Among 44 patients tested with CAVD, 26 (59%) were positive for at least one CF mutation, while the carrier frequency for CF mutations in the general population is only 4%. Four patients were found to be compound heterozygotes, three with genotypes Delta F-508/R117H, one with R553X/R117H. Among 24 parents tested, 15 (seven fathers, eight mothers) had sons with CAVD who were positive for CF mutations. Of these, nine (four fathers and five mothers) were found to be carriers for CF mutations. These four fathers, although carriers of CF mutations, were obviously fertile. Of the 13 offspring tested, six (three boys and three girls) had CF positive fathers. Of these, three (two girls and one boy) were found to be carriers for CF mutations. These MESA/IVF children are the first offspring to whom men with CAVD have been able to transmit CF mutations.(ABSTRACT TRUNCATED AT 250 WORDS)

Intrauterine release of oestriol in castrated rhesus monkeys induces local but not peripheral oestrogenic effects: a possible approach for the treatment and prevention of Asherman's syndrome.

Two types of oestrogen-medicated intrauterine devices (IUD) were studied in ovariectomized rhesus monkeys. An oestradiol (E2) fibre-wrapped IUD that released E2 at a rate of 3.57 micrograms/cm/day, or an oestriol (E3) fibre-wrapped IUD that releases E3 at a rate of 6.4 micrograms/cm/day, was inserted in eight animals and left in place for 4 weeks. Serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), E2 and E3 were determined by radioimmunoassay for 1 week before the IUD insertion, during the time the IUD was in place, and for 3 weeks after its removal. Uterine histology was performed at the time of IUD insertion and removal by light and transmission electron microscopy. Both E2 and E3 IUDs induced similar histological changes in the uterus, i.e. four- to five-fold increase in endometrial thickness, a shift of the gland/stroma ratio from 1:4 to 1:1, transformation to a marked pseudostratified epithelium with pronounced coiling of the glands, appearance of subnuclear and luminal secretion and, finally, change from spindle-dense stromal cells to plump eosinophilic cells. Oestradiol fibre-wrapped IUDs produced circulating E2 levels of 150-200 pg/ml during the entire 4 weeks. FSH and LH levels were decreased to an average of 55% and 65% from a castration baseline (P less than 0.001 and P less than 0.05, respectively). Oestriol fibre-wrapped IUDs produced circulating E3 levels of 100-250 pg/ml. However, FSH and LH levels were not altered in this group. The specific local oestrogenic effect of E3-IUDs without affecting the pituitary secretion of gonadotrophins, suggests their possible application in cases in which an exclusively oestrogenic effect at the uterine level, such as in Asherman's syndrome, is desired.

Severe ovarian hyperstimulation syndrome in assisted reproductive technology: definition of high risk groups.

In a retrospective analysis of 637 cycles of ovarian stimulation and transvaginal follicular aspiration for various assisted reproductive technologies, severe ovarian hyperstimulation syndrome (SOH) occurred in six (0.94%) cycles. The patients at a high risk of developing SOH in cycles of assisted reproduction were those who had excessive serum oestradiol levels on the day of human chorionic gonadotrophin (HCG) administration (oestradiol greater than 6000 pg/ml; 38% SOH) and a high number of oocytes obtained (greater than 30 oocytes; 23% SOH). In those patients with both oestradiol greater than 6000 pg/ml on the day of HCG administration and greater than 30 eggs retrieved, the chance of developing SOH was 80%. The higher the serum oestradiol levels and the more eggs retrieved, the higher the pregnancy rates observed. High oestradiol level did not appear to have a detrimental effect on pregnancy rates and outcome. Furthermore, our results are not consistent with suggestions that the addition of gonadotrophin-releasing hormone agonist to ovarian stimulation protocols, follicular aspiration and/or luteal support with progesterone may reduce the incidence of ovarian hyperstimulation syndrome.

Progesterone production in cultured human granulosa cells: correlation with follicular fluid hormone levels.

STUDY OBJECTIVE: To examine the progesterone (P) production by cultured granulosa cells and the hormonal content in the follicular fluid (FF) of ovarian-hyperstimulated women. DESIGN: Retrospective. SETTING: Private Fertility Clinic and National Research Institute. PATIENTS: Eighteen patients undergoing in vitro fertilization or gamete intrafallopian transfer programs. RESULTS: Progesterone levels Measured in the culture medium of granulosa cells decreased sixfold with culture time. Human luteinizing hormone (LH) increased P production only when basal P production was less than 1 microgram/mL. Granulosa cell P production in culture was negatively correlated with FF LH-human chorionic gonadotropin (hCG) levels. Follicular fluid follicle-stimulating hormone (FSH) levels were positively correlated with FF P and 17 beta-estradiol (E2) concentrations. Similar results were found between FF LH (hCG) and E2 levels, but there was no relationship between FF LH (hCG) and FF P values. CONCLUSION: The high dose of hCG administered during gonadotropin treatment could induce a decrease in the in vitro granulosa cell P production.

Mini-Percoll: a new method of semen preparation for IVF in severe male factor infertility.

Conventional methods of semen preparation for IVF, such as swim-up sedimentation, regular Percoll gradients, are not very effective in the treatment of severe oligoasthenozoospermia (total motile count less than 5 X 10(6)). For these cases, a new method of sperm preparation consisting of a reduced volume of a discontinuous Percoll gradient (mini-Percoll) has been utilized. The results obtained were analysed by comparing mini-Percoll versus resuspension in two groups of patients with severe male factor infertility. In the mini-Percoll group, a statistically significant improvement (P less than 0.001) was obtained in the post-treatment seminal parameters of motility, progression and proportion of normal forms. In the resuspension group (n = 12), only eight of 108 oocytes inseminated were fertilized (7%), while in the mini-Percoll group (n = 29), 124 out of 336 oocytes were fertilized (40%) (P less than 0.001). Furthermore, in the mini-Percoll group, a higher pregnancy rate (25 versus 7%) and low abortion rate (22 versus 33%) were observed. Although more studies are necessary to confirm these preliminary data, mini-Percoll seems to be a valid laboratory tool for semen samples of extremely poor quality.

Insulin-like growth factors and thecal-granulosa-cell function.

Insulin-like growth factors (IGFs) belong to a family of low mol. wt, single chain polypeptides inducing growth promotion and insulin-like metabolism effects and regulating both cell replication and differentiation. Recent studies in laboratory animals suggest that IGFs play an important role as intraovarian regulators in several mammalian species. The purpose of this article was to review current concepts of interactions between the IGFs and the thecal-granulosa-cell function. To provide a basic understanding of these interactions, we have first analysed data concerning biosynthesis, biochemical structure, pharmacokinetics, degradation, binding proteins and receptors for IGFs. Then we have discussed the specific interactions between IGFs and theca-granulosa-cell regulation, and analysed the significance of the relationship to the pathophysiology of some endocrine and reproductive disorders, including hyperandrogenism and female infertility.