Showing papers in "Human Reproduction in 1990"
TL;DR: It can be concluded that the method developed in the laboratory and which resulted in the use of stricter criteria for the evaluation of sperm morphology is a practical, reliable and repeatable method and has a good prognostic value for the prediction of expected IVF fertilization, the hamster test and hemizona assay.
Abstract: The evaluation of the morphology of human spermatozoa varies widely between and sometimes even within laboratories. The purpose of this study was to determine whether the method that has been developed in our laboratory and which resulted in the use of stricter criteria for the evaluation of sperm morphology is a practical, reliable and repeatable method and to establish the within and between observer variations. The criteria used for a 'normal' spermatozoon are based on the appearance of spermatozoa found in the mucus of the upper endocervical canal. The results of the morphological evaluations of 26 samples by four observers were statistically analysed by various methods. The method of Barnett showed a high degree of relative accuracy between observers with error variances of between 2.89 and 19.67 as well as high Spearman rank correlation coefficients of between 0.8675 and 0.6537 (P less than 0.0003). The Spearman correlation coefficient for 15 duplicate evaluations by one observer was 0.9650 (P less than 0.0001) while the coefficients of variation for repeated evaluations of single samples were also within acceptable limits. Based on these results, the method described in this article allows comparable and reliable results between and within observers to be obtained. From this and other studies it can be concluded that the method also has a good prognostic value for the prediction of expected IVF fertilization, the hamster test and hemizona assay.
614Â citations
362Â citations
TL;DR: It is demonstrated that removal of the first polar body has no effect on subsequent fertilization rates or embryonic growth to the blastocyst stage, and the accuracy of the polar body diagnosis was confirmed by polymerase chain reaction (PCR) analysis of an oocyte that failed to fertilize.
Abstract: In women who are heterozygous for a genetic disease, genetic analysis of the first polar body allows the identification of oocytes that contain the maternal unaffected gene. These oocytes can be fertilized and transferred to the mother without risk of establishing a pregnancy with a genetically abnormal embryo. We have demonstrated that removal of the first polar body has no effect on subsequent fertilization rates or embryonic growth to the blastocyst stage. We have developed a PCR technique to successfully analyze the PI type Z and PI type M genotypes of alpha-1-antitrypsin deficiency and applied this technique for a couple at risk for PI type ZZ alpha-1-antitrypsin deficiency. After standard IVF treatment to stimulate multiple follicle development, eight oocytes were aspirated transvaginally. Polar bodies were removed by micromanipulation from seven oocytes and fertilization occurred in six cases. PCR analysis was successful in five oocytes. One was PI type M, two were PI type Z and two were heterozygous MZ due to crossing over. Embryos from the two oocytes containing the unaffected gene (polar body PI type Z) were transferred in the same cycle 48 h after insemination. No pregnancy was established. The accuracy of the polar body diagnosis was confirmed by polymerase chain reaction (PCR) analysis of an oocyte that failed to fertilize.
338Â citations
TL;DR: It is concluded that removal of one or two cells at the 8-cell stage, while reducing the cellular mass, does not adversely affect the preimplantation/development of biopsied embryos in vitro and it is suggested that this approach could be used for preim implantation diagnosis of genetic defects.
Abstract: Normally fertilized human embryos biopsied 3 days after in-vitro fertilization (IVF) have been examined for effects on viability and development in vitro after removal of one or two cells at the 8-cell stage (1/8 and 2/8) from each embryo. A high proportion of 7/8 and 6/8 biopsied and unmanipulated embryos developed to the blastocyst stage between days 5 and 6 (79, 71 and 59%, respectively), and many biopsied embryos (56%) hatched from the zona pellucida in vitro. The viability of biopsied embryos which developed to the blastocyst stage was assessed by daily non-invasive measurement of the uptake of two energy substrates, glucose and pyruvate. Uptake of both substrates was generally lower in 7/8 and 6/8 biopsied embryos but only in proportion to the reduced cellular mass. The total cell number and the numbers of both trophectoderm (TE) and inner cell mass (ICM) cells in biopsied embryos at the blastocyst stage, counted by differential labelling of their nuclei, were also reduced in proportion but the ratio of ICM to TE cells was maintained in both 7/8 and 6/8 biopsied embryos. We conclude that removal of one or two cells at the 8-cell stage, while reducing the cellular mass, does not adversely affect the preimplantation/development of biopsied embryos in vitro and suggest that this approach could be used for preimplantation diagnosis of genetic defects.
328Â citations
TL;DR: A computerized database generated from the literature on cytogenetic studies in couples experiencing repeated pregnancy losses showed a relationship between the distribution of the chromosome abnormalities and the number of abortions, and it appeared that only translocations and inversions were associated with a higher risk of pregnancy wastage.
Abstract: A computerized database generated from the literature on cytogenetic studies in couples experiencing repeated pregnancy losses has been set up at the University of Quebec at Chicoutimi. At the present time, it contains data on 22,199 couples (44,398 individuals). The statistical analyses showed a relationship between the distribution of the chromosome abnormalities and the number of abortions. An uneven distribution of the chromosomal structural rearrangements according to the sex of the carrier was found (P less than 0.05). Overall, 4.7% of the couples ascertained for two or more spontaneous abortions included one carrier. It also appeared that only translocations (both reciprocal and Robertsonian) and inversions were associated with a higher risk of pregnancy wastage. Therefore, genetic counselling should be offered to these couples and investigations performed on their extended families.
245Â citations
TL;DR: There are complex temporal relationships between uterine blood flow, ovarian morphology, the concentrations of plasma oestradiol and progesterone and the thickness of the endometrium.
Abstract: Uterine arterial blood flow was studied at defined times during the ovarian or menstrual cycles. Transvaginal colour pulsed ultrasound was used to identify the vessels in 23 healthy women. Blood flow impedance as reflected by the pulsatility index (PI) was determined for both arteries on 132 occasions. There was no significant difference in the PI values between the right and left arteries. The average PI value was used for subsequent analysis (range 0.5-5.6; mean 2.8). Peak mean PI values occurred on day 1 of menses (4.6), the day of the plasma oestradiol peak (3.7) and the day of the LH peak plus 3 (2.9). The lowest mean PI values (indicating the least impedance to blood flow) occurred on the day of the LH peak minus 6 (2.6), and the day of the LH peak plus 9 (1.9). There are complex temporal relationships between uterine blood flow, ovarian morphology, the concentrations of plasma oestradiol and progesterone and the thickness of the endometrium.
199Â citations
TL;DR: An analysis of the influence of women's age on the results of in-vitro fertilization was performed, using 5590 attempts collected during the year 1986, and found increasing age was found to be related to decreased success rates.
Abstract: An analysis of the influence of women's age on the results of in-vitro fertilization was performed, using 5590 attempts collected during the year 1986. Increasing age was found to be related to decreased success rates from 19.8% per attempt below the age of 25 years, to 9% per attempt at 40 years or more. A cut-off point was found between the ages of 36 and 37 years, using a mathematical model. The decrease was related to a reduction in oocyte production (4.3 +/- 2.8 at 25 years or less and 3.3 +/- 2.1 at 40 years or more, P less than 0.001), and to a reduced implantation rate, whatever the number of transferred embryos. These findings were not due to spouse's age, rank of attempt, infertility diagnosis or oocyte stimulation regimen, since the effect of age remained significant when a logistical model including these confounders was applied. Finally, a woman's age must be considered as a prognostic factor when IVF is proposed to infertile couples.
189Â citations
TL;DR: It is proposed that the high production of lactate by human preimplantation embryos in vitro is an adaptation to the conditions of culture.
Abstract: The consumption of pyruvate and glucose and the production of lactate by 40 single human preimplantation embryos has been measured using a non-invasive technique. Twelve of the embryos showed abnormal fertilization. Of the 28 normally fertilized embryos, nine (32%) developed to the blastocyst stage in culture while the remainder degenerated or arrested during cleavage. In the normal embryos, pyruvate uptake exceeded that of glucose in the early developmental stages (days 2-5 post-insemination) before glucose became the predominant substrate in the blastocyst (day 6). Considerable quantities of lactate were formed throughout development, rising from a value of 43.6 pmol/embryo/h on day 2.5 to 95.4 pmol/embryo/h on day 5.5. The values of pyruvate and glucose uptake and lactate production of those embryos which arrested were below those which developed normally. On the basis that one mole of glucose can give rise to two moles of lactate, only 50% of the lactate produced could be accounted for in terms of glucose uptake from the medium. This figure rose to 90% in the blastocyst. The remaining lactate must be derived from endogenous sources, most probably glycogen. It is proposed that the high production of lactate by human preimplantation embryos in vitro is an adaptation to the conditions of culture.
184Â citations
TL;DR: It is demonstrated that CC results in a thinner endometrium than HMG alone, which may be of importance in improving pregnancy rates in IVF and possibly in other infertility therapy which involves the use of clomiphene citrate.
Abstract: We recently demonstrated, using transvaginal sonography, that conception cycles in in-vitro fertilization (IVF) are associated with a significantly thicker endometrium at midcycle than non-conception cycles, suggesting that endometrial growth may influence implantation. In the present study, to examine whether the type of stimulation protocol affects endometrial development, we compared the sonographic appearance of the endometrium in 22 patients randomized to receive clomiphene citrate and human menopausal gonadotrophin (CC/HMG) and in 19 who received HMG alone. A significantly thicker endometrium was observed in the HMG patients compared to the CC/HMG group (P less than 0.005) throughout the follicular phase of the cycle, although serum concentrations of oestradiol (E2) did not differ in the two groups. Twenty-three patients (13 in the HMG group and 10 in the CC/HMG group) had previous IVF cycles with CC/HMG stimulation in which endometrial thickness was measured. A thin endometrium recurred with subsequent CC/HMG cycles while increased growth occurred with HMG only compared to previous CC/HMG cycles. Therefore, ultrasound examination of the endometrium in this study demonstrated that CC results in a thinner endometrium than HMG alone. We believe these findings may be of importance in improving pregnancy rates in IVF and possibly in other infertility therapy which involves the use of clomiphene citrate.
182Â citations
TL;DR: It was concluded that the vaginal route for administering micronized progesterone can be advised as the treatment of choice in patients without ovarian function.
Abstract: In 43 patients without ovaries, endometrial biopsies at day 21 of 75 substituted cycles were studied by light and electron microscopy. The morphology of the endometrium was compared after oral, vaginal or intramuscular administration of progesterone, and correlated with the serum levels of 17-beta oestradiol and progesterone and the pregnancies obtained after oocyte donation. After vaginal application of micronized progesterone, endometrial morphology closely matched that of a natural cycle. This therapy was able to support two ongoing pregnancies. No adequate endometrial response was noted after oral ingestion of progesterone. The maturation of the endometrium after intramuscular injections of progesterone in oil was heterogeneous. It was concluded that the vaginal route for administering micronized progesterone can be advised as the treatment of choice in patients without ovarian function.
151Â citations
TL;DR: Preliminary data suggest that mini-Percoll seems to be a valid laboratory tool for semen samples of extremely poor quality in the treatment of severe oligoasthenozoospermia.
Abstract: 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.
TL;DR: The results demonstrate the feasibility of trophectoderm biopsy in human blastocysts and that sufficient extra-embryonic material can be obtained by this technique for preimplantation diagnosis of genetic disorders.
Abstract: Trophectoderm biopsy was carried out on 47 human blastocysts. A slit was made in the zona pellucida opposite the inner cell mass by micromanipulative techniques. The human blastocyst zona offered more resistance to slitting compared to that of the mouse. After 18-24 h, controlled herniation of the trophectoderm cells was observed. These cells were biopsied when the diameter of the herniation was approximately equal to that of the blastocyst. The size of the slit and the stage of embryonic development at which slitting was performed were important for successful herniation to occur. After slitting, 76% of day 5-6 blastocysts showed herniation whilst only 42% of day 7-8 blastocysts herniated. Further development of the manipulated embryos was not apparently impaired, as hatching occurred in 44% of the former and 20% of the latter, compared with 18.1% in non-manipulated controls. The biopsied cells (approximately 10-30) usually remained in a clump but 14% formed vesicles on the day after biopsy. There was, however, no evidence of adherence to the dish or formation of monolayers. These results demonstrate the feasibility of trophectoderm biopsy in human blastocysts and that sufficient extra-embryonic material can be obtained by this technique for preimplantation diagnosis of genetic disorders.
TL;DR: With the advent of modern techniques, e.g. monoclonal antibodies and sophisticated sperm function tests, many of the questions raised should be answered in the near future.
Abstract: The functional significance of white blood cells in the modulation of an anti-sperm antibody response, prevention of infection (including HIV) sperm transport/storage and sperm function is extensively discussed. A critical review of the existing literature is presented with future experimental lines of investigation outlined. A lack of controlled clinical studies in the human to validate data from animal species--for example--the involvement of white blood cells in the transport and storage of sperm in the female tract and the possible adverse effect of pathology (i.e. endometriosis) on these functions are presented. In conclusion, with the advent of modern techniques, e.g. monoclonal antibodies and sophisticated sperm function tests, many of the questions raised should be answered in the near future.
TL;DR: A new medium, BAT6, is described for the optimal in-vitro culture of mouse embryos after an adverse effect of iron was found and both the rate of embryonic development and the proportion of embryos reaching the blastocyst stage approached levels found in vivo.
Abstract: The effect of iron and iron chelators on the development of the mouse embryo in vitro from the 1-cell stage to the blastocyst has been investigated. An adverse effect of iron was found. The high affinity iron chelator, desferal, also blocked development, whilst transferrin (whether as apoprotein or saturated with iron), DETAPAC and EDTA promoted development. The addition of transferrin permitted development to the blastocyst stage of embryos from stains normally exhibiting the 2-cell block. Under such circumstances both the rate of embryonic development and the proportion of embryos reaching the blastocyst stage approached levels found in vivo. Based on these results, a new medium, BAT6, is described for the optimal in-vitro culture of mouse embryos.
TL;DR: The purpose of the present study was to investigate the effect of a single dose of RU 486 administered very early in the secretory phase on endometrial development and levels of progesterone receptors, on plasma levels of gonadotrophins and ovarian hormones and on the pattern of menstrual bleeding.
Abstract: This study investigated the effect of a single dose of RU 486 administered very early in the secretory phase on endometrial development and levels of progesterone receptors on plasma levels of gonadotropins and ovarian hormones and on the pattern of menstrual bleeding. 24 regularly menstruating subjects participated and were studied during a control a treatment and a follow-up cycle. In the treatment cycle a single dose of 200 mg of RU 486 was given in the evening of the 2nd day after urinary LH peak. Plasma was collected from cycle day 10 until menstruation in both control and treatment cycles. The lengths of the control treatment and follow-up cycles were equal. 3 of the subjects had slight vaginal bleeding in association with RU 486 intake which however did not disturb their normal menstrual rhythm. Plasma levels of estradiol progesterone and FSH were not affected in the treatment cycle whereas LH levels increased slightly. The elimination 1/2-life of RU 486 was 28.6 hours. An endometrial biopsy was taken 12 36 or 84 hours (LH + 3 LH + 4 and LH + 6) after drug intake (8 subjects in each group) and another biopsy was taken on the corresponding day in the control cycle. The specimens were assessed by morphometric analysis and for cytosolic progesterone receptor concentrations. Endometrial biopsies taken 12 hours (on LH + 3) after RU 486 intake contained significantly (p<0.001) lower levels of cytosolic progesterone receptors than in the control cycle but levels at 36 and 84 hours were similar. Morphometric analysis revealed a retardation of endometrial development as early as 12 hours after RU 486 intake. The degree of retardation was more marked after 36 and 84 hours. These results suggest that the effect on the endometrium of a single dose of RU 486 immediately after the LH peak may be sufficient to prevent implantation. Thus treatment with RU 486 in the early secretory phase of the menstrual cycle might be considered for use as a once-a-month pill for fertility control. (authors)
TL;DR: It is confirmed that the ovulatory HCG dose is the most important factor in inducing this severe complication and luteal supplementation with HCG and/or HCG production during implantation could exacerbate OHSS.
Abstract: In 1673 treatment cycles stimulated with buserelin and HMG, for IVF, GIFT or ZIFT, the severe ovarian hyperstimulation syndrome (OHSS) occurred in 10 cycles (0.6%). Eight patients were hyperandrogenic and showed an increased ovarian response to HMG. After replacement of a maximum of three embryos or zygotes, seven women became pregnant. Three women had a multiple gestation. All patients recovered uneventfully with conservative treatment. Support with progesterone or continuation of the agonist during the luteal phase did not prevent OHSS, confirming that the ovulatory HCG dose is the most important factor in inducing this severe complication. Luteal supplementation with HCG and/or HCG production during implantation could exacerbate OHSS.
TL;DR: IVF treatment may be a viable solution for polycystic ovarian syndrome patients resistant to an in-vivo protocol treatment regimes, and the fact that more oocytes are recovered from PCO patients, balances their lower fertilization and cleavage rates.
Abstract: Sixteen patients with polycystic ovarian syndrome (PCO) were treated by in-vitro fertilization (26 treatment cycles). The results were compared with 37 normo-ovulatory women with tubal disease (37 treatment cycles). The oestradiol and progesterone levels were higher during the follicular phase in the PCO patients, but were statistically significantly higher only on the day after human chorionic gonadotrophin administration. Although more oocytes were recovered per cycle from the PCO group (19.3 +/- 6.1) than from the control group (5.4 +/- 2.9) with P less than 0.004, the mean numbers of embryos per cycle were similar in both groups (3.7 +/- 2.4 versus 3.6 +/- 2.2, respectively). The pregnancy rate was also comparable in both groups (30.7 versus 29.7%, respectively). The fact that more oocytes are recovered from PCO patients, balances their lower fertilization and cleavage rates. We conclude, therefore, that IVF treatment may be a viable solution for PCO patients resistant to an in-vivo protocol treatment regimes.
TL;DR: Neither a genital tract origin, nor a hormonal contribution are strictly necessary for embryo co- culture, as already demonstrated by co-culture with trophoblastic tissue.
Abstract: The development of 1-cell mouse embryos in explanted oviducts, on mouse and bovine oviduct epithelial cells and on two established cell line supports is compared. The best rates of blastocyst formation were obtained using explanted oviducts; mouse and to a lesser extent, bovine oviduct epithelial cells allow good embryonic development, associated with high viability after transfer of the blastocysts obtained in co-culture. MDBK (from bovine kidney) and Vero (from Green monkey kidney) have been tested. MDBK allows high rates of blastocyst formation (67%) and the blastocysts obtained are viable. Vero does not allow the 2-cell block to be overcome. Maintenance of cell polarity for all the feeder layers did not improve embryo development. A preliminary study on the metabolic modifications induced by the feeder layers showed no modifications at all related to a decrease in glucose, an increase in lactate and early embryonic development. On the other hand, for the free amino acids, cellular supports with high embryotrophic activity seem to mimic tubal secretions, especially with a high level of glycine. Neither a genital tract origin, nor a hormonal contribution are strictly necessary for embryo co-culture, as already demonstrated by co-culture with trophoblastic tissue. Established cell lines, which are easy to handle and control, could be useful tools in embryo biotechnology.
TL;DR: The findings suggest that zygote cryopreservation should be initiated when pronuclear migration is completed, and this moment can be determined accurately by studying pronuclear association and nucleolar alignment.
Abstract: The effects of cryopreservation on human zygotes at various stages between the appearance of pronuclei and their close association were investigated. Pronuclear zygotes (n = 233) from 101 patients were frozen using propanediol 21-35 h following egg collection. The incidence of implantation of thawed pronuclear zygotes frozen 29-35 h following oocyte collection was significantly higher than that of younger pronuclear zygotes (28 versus 10%, respectively). Zygote age did not affect cell survival following cryostorage. The diameter and association of pronuclei and the number and distribution of nucleoli were determined from video tape recordings of 140 fresh zygotes. Pronuclear migration continued after pronuclear enlargement. The number of nucleoli remained constant during pronuclear migration, but their random distribution within the pronucleus diminished. Strongly adhered pronuclei had significantly more aligned nucleoli on adjacent sides than pronuclei which were still visually separated by ooplasm. This equatorial distribution of nucleoli was noted in the majority of zygotes older than 26 h. The findings suggest that zygote cryopreservation should be initiated when pronuclear migration is completed. This moment can be determined accurately by studying pronuclear association and nucleolar alignment.
TL;DR: Patients pretreated with the analogue had similar pregnancy and ovulation rates, needed larger doses and more days of gonadotrophin therapy and had more ovarian overstimulation than those receiving no pretreatment, and the role of superactive LHRH analogues for induction of a single ovulation for in-vivo fertilization is uncertain.
Abstract: This study was designed to compare the results of treatment with, firstly, exogenous gonadotrophins, with (57 cycles) and without (65 cycles) pretreatment with a superactive analogue of luteinizing hormone releasing hormone (LHRH) and, secondly, pure follicle stimulating hormone (FSH) (50 cycles) with those of human menopausal gonadotrophin (HMG) (72 cycles) in 46 women with clomiphene-citrate-resistant anovulation associated with polycystic ovaries. Patients randomly allocated to the analogue group received buserelin (Suprefact, Hoechst, UK, Ltd, Hounslow, Middlesex), 800 micrograms/day by nasal insufflation and when hypogonadism was achieved, patients were again randomly allocated for ovarian stimulation with either FSH or HMG. Controls received FSH or HMG alone. Patients pretreated with the analogue had similar pregnancy and ovulation rates, needed larger doses and more days of gonadotrophin therapy and had more ovarian overstimulation than those receiving no pretreatment. The role of superactive LHRH analogues for induction of a single ovulation for in-vivo fertilization is thus uncertain. Pure FSH had no advantages over HMG, the LH content of HMG having no deleterious effect on the ovary.
TL;DR: Treating infertile female patients with danazol hinders progression in a majority of patients, and conversely, there is a small but definite spontaneous regression rate.
Abstract: A group of 50 infertile female patients was assessed laparoscopically to study the natural history of endometriosis. They were treated either with danazol (n = 21), or diathermy ablation (n = 13) or had conservative surgery with postoperative danazol (n = 5) or were left untreated (n = 11). All these patients had second look laparoscopy performed after a mean interval of 12 months (range 9-18 months). Improvement was noted among 53% of those treated with danazol, 70% treated with diathermy, 80% treated with surgery and danazol and 27% of those left untreated. The status of endometriosis remained unchanged among 14% of those treated with danazol, 8% treated by diathermy and 9% of those left untreated. However, the disease became worse in the remaining women, 33% in the danazol treated group, 22% treated by diathermy, 20% treated with surgery and danazol and 64% of those left untreated. Thus treatment hinders progression in a majority of patients, and conversely, there is a small but definite spontaneous regression rate.
TL;DR: Eighty-two patients had 100 cycles of oocyte donation from 68 donors resulting in 27 clinical pregnancies, with the best results achieved using gametes from parous donors and transferring three to four oocytes fresh to the Fallopian tubes of a young recipient.
Abstract: Eighty-two patients had 100 cycles of oocyte donation from 68 donors resulting in 27 clinical pregnancies. The source of donated oocytes was 42 fertile volunteers and 26 patients from the assisted conception programme. The pregnancy rate was significantly higher when intra-Fallopian transfer was performed (36%; 21/59), compared with intrauterine transfer (15%; 6/41). The pregnancy rate following fresh gamete/embryo transfer (39%; 15/39) was slightly higher than for frozen embryo transfer (20%; 12/61). The age of the recipient significantly affected the pregnancy rate. The pregnancy rate was 50% in the 25-29 years age group and steadily dropped to 9.7% in the 45-49 age group. The pregnancy rate in patients with primary ovarian failure (50%; 8/16) was significantly higher than in patients with secondary ovarian failure (18%; 9/50). The pregnancy rate was significantly greater when parous donors (33%; 23/69) were used compared with non-parous donors (13%; 4/31). The number of gametes/embryos transferred significantly affected the pregnancy rate regardless of the treatment used. If one or two gametes/embryos were transferred, the pregnancy rate was 11% compared with 33% if three to four embryos were transferred. The age of the donors did not affect the pregnancy rate. The majority of the donors were under the age of 35 years. The best results (50% per cycle) were therefore achieved using gametes from parous donors and transferring three to four oocytes fresh to the Fallopian tubes of a young recipient.
TL;DR: A patient who developed severe OHSS and a potentially lethal complication, following ovulation induction for gamete intra-Fallopian transfer is reported.
Abstract: Despite advances in monitoring techniques, severe ovarian hyperstimulation syndrome still occurs in 1% of patients treated with ovulation induction for assisted conception techniques. The seriousness of this iatrogenic syndrome is reflected in the fact that deaths have been reported following it. We report a patient who developed severe OHSS and a potentially lethal complication, following ovulation induction for gamete intra-Fallopian transfer
TL;DR: Only the number of embryos replaced and the substitution protocol seemed to influence the implantation rate and the other parameters, and in particular the type of gonadal dysgenesis, seemed to have no effect on the results.
Abstract: Between February 1987 and February 1989, 13 women with primary ovarian failure due to gonadal dysgenesis were treated with embryo transfer following ovum donation in 22 cycles. Eight pregnancies were obtained (36.7% per transfer); four births of normal children, two spontaneous abortions and two other pregnancies currently ongoing (between 5 and 8 months). An association of percutaneous oestradiol, oestradiol valenate and intravaginal progesterone was used as hormone substitution and embryo transfer was only performed following assessment of the endometrium during a previous cycle. Apart from the day of embryo transfer, which was the same for all patients (the 2nd day after initiation of progesterone) various prognostic factors were analysed. These were the type of gonadal dysgenesis (45 XO, 46 XX or 46 XY), the number of embryos replaced, whether they had been frozen, whether the egg donor was anonymous and finally the influence of the hormone substitution protocol. Only the number of embryos replaced and the substitution protocol seemed to influence the implantation rate. The other parameters, and in particular the type of gonadal dysgenesis, seemed to have no effect on the results. The pregnancy rate per transfer was 30% for 45 XO (10 transfers), 25% for 46 XX (eight transfers) and 75% for 46 XY (four transfers).
TL;DR: The concentrations of glucose, lactate, progesterone and oestradiol have been measured in follicular fluids obtained from women attending a natural cycle IVF programme and a model is presented to account for these findings in terms of the movement of pyruvate, glucose and lactate from the vascular theca into follicular fluid, and of glycolysis occurring in the granulosa cells.
Abstract: The concentrations of glucose, lactate, progesterone and oestradiol have been measured in 122 follicular fluids obtained from women attending a natural cycle IVF programme. Some women received low-dose clomiphene. Fluids were recovered 28-35 h after the onset of the spontaneous LH surge. Mean follicular fluid volumes were greater for stimulated (4.46 ml) than spontaneous (3.11 ml) cycles. Mean glucose and lactate concentrations were similar in the two groups, with overall means of 3.29 mM (glucose) and 6.12 mM (lactate). Total glucose and lactate were greater in the fluids from stimulated (14.1 and 29.5 mol, respectively) than from the spontaneous cycles (11.5 and 20.2 mmol, respectively). There were negative correlations between follicular fluid volume and glucose concentration (r = -0.348) and between glucose concentration and lactate concentration (r = -0.367) but no relationship between follicular fluid volume and lactate concentration (r = 0.086). A model is presented to account for these findings in terms of the movement of pyruvate, glucose and lactate from the vascular theca into follicular fluid, and of glycolysis occurring in the granulosa cells.
TL;DR: Primordial follicles isolated from infant mouse ovaries by enzymatic disaggregation have been transferred to the ovaries of sterile adults and were capable of spontaneous ovulation and the majority of animals carrying them were sexually receptive to males.
Abstract: Primordial follicles isolated from infant mouse ovaries by enzymatic disaggregation have been transferred to the ovaries of sterile adults. They were transferred after being recombined and incubated in a plasma clot which was inserted either into ovaries which had been sterilized by X-irradiation or into vacant periovarian capsules immediately after ovariectomy. Irrespective of the type of operation, these grafts underwent morphogenesis to form replicas of the normal ovary, which were generally indistinguishable in respect of form, histological appearance and function. The numbers of follicles in these so-called transgerminative ovaries were highly variable, but all stages of follicular maturation up to Graafian stage were represented in most of them, while grafts that had been X-irradiated before transfer were sterile. Ovaries forming from fertile grafts were capable of spontaneous ovulation and the majority of animals carrying them were sexually receptive to males. Mating often resulted in pregnancies and those delivering at full-term produced normal offspring.
TL;DR: The cholesterol/phospholipid ratio was determined for spermatozoa from eight men with normal semen parameters and shows a close correlation between the C/PL ratio and the rate of sperm capacitation as assessed by the hamster egg penetration test, supporting the notion that the loss or reduction of membrane cholesterol constitutes an important step of capacitation in human spermatoza.
Abstract: The cholesterol/phospholipid (C/PL) ratio was determined for spermatozoa from eight men with normal semen parameters. There was a close correlation between the C/PL ratio and the rate of sperm capacitation as assessed by the hamster egg penetration test. A lower C/PL ratio correlated with a faster capacitation time. This supports the notion that the loss or reduction of membrane cholesterol constitutes an important step of capacitation in human spermatozoa.
TL;DR: An opinion is provided as to how the transfer of blastocysts produced via the co-culture system could enhance pregnancy rates and the various roles of human co-cultures in assisted reproduction.
Abstract: Assisted reproductive techniques have contributed significantly to alleviating subfertility in the childless couple. However, the take-home baby rates have been very low throughout the world. One of the contributory causes has been the reduced viability of replaced embryos brought about by suboptimal in vitro conditions. The culture of human embryos in the presence of passaged human tubal ampullary monolayers (co-cultures) is an attractive approach to improving the viability of embryos for assisted reproduction. Seventy per cent of blastocysts can be produced in human ampullary co-culture as compared to 33% in standard culture media. This paper discusses the various roles of human co-cultures in assisted reproduction and provides an opinion as to how the transfer of blastocysts produced via the co-culture system could enhance pregnancy rates. Particular emphasis is placed on human oviductal characteristics, the various co-culture systems, screening of co-cultures for microbes, freezing of ampullary cells, growth factors and embryonic blocks, specificity of co-cultures, sperm hyperactivation in co-culture and pregnancy rates. The first patient on a clinical trial who had four of her oocytes fertilized and grown in human ampullary co-culture and then replaced into her uterus became pregnant. The co-culture system may have tremendous potential in supporting human embryonic growth via embryotrophic factors.
TL;DR: The hyaluronate migration test effectively assesses the mucus-penetrating potential of a semen sample without the need for relatively large quantities of midcycle cervical mucus; it will therefore augment (as an internal control), although not necessarily replace, the homologous Kremer test and reduce the quantity of both patient and donor mucus needed for comprehensive crossed-hostility format testing of sperm-mucus interaction.
Abstract: Fifty-one comparisons were made of human sperm migration into capillary tubes containing either human cervical mucus ('Kremer test') or a synthetic mucus substitute consisting of a 5 mg/ml solution of sodium hyaluronate (average mol. wt 2 x 10(6)) in a phosphate-buffered medium. The results of these two tests were highly significantly correlated and dependent upon the same sperm characteristics reflecting sperm progressive ability (including the specific movement characteristic of lateral head displacement amplitude), morphological normality and cellular vitality as well as the concentration of these more functional cells in the semen. The result of the hyaluronate migration test, in conjunction with the mucus quality measures of Insler score and pH, allowed a 92.2% correct prediction of the Kremer test outcome (90.9% of normal tests and 93.1% of abnormal tests). In this data set, these values also corresponded to the sensitivity and specificity of the analysis, respectively. From these studies, we propose the hyaluronate migration test as a useful adjunct to routine semen analysis, sperm movement analysis and the more traditional in-vitro tests of sperm-cervical mucus interaction in the diagnostic investigation of infertile couples. It effectively assesses the mucus-penetrating potential of a semen sample without the need for relatively large quantities of midcycle cervical mucus; it will therefore augment (as an internal control), although not necessarily replace, the homologous Kremer test and reduce the quantity of both patient and donor mucus needed for comprehensive crossed-hostility format testing of sperm-mucus interaction.