Sequential use of clomiphene citrate, human menopausal gonadotropin, and human chorionic gonadotropin in human in vitro fertilization. II. Study of luteal phase adequacy following aspiration of the preovulatory follicles.

The 88 patients included in the in vitro fertilization program during 113 cycles were submitted to superovulation by sequential use of clomiphene citrate, human menopausal gonadotropin, and human chorionic gonadotropin. No correlation was found between estradiol and progesterone levels during the luteal phase and estradiol on the days preceding administration of human chorionic gonadotropin. Nineteen biopsies of the endometrium were carried out. The importance of the increase of estradiol between the day before and the day of administration of human chorionic gonadotropin is positively correlated with the quality of the endometrium.

Sequential use of clomiphene citrate, human menopausal gonadotropin, and human chorionic gonadotropin in human in vitro fertilization. I. Follicular growth and oocyte suitability.

Sequential treatment with clomiphene citrate, human menopausal gonadotropin, and human chorionic gonadotropin was successfully applied to 149 women in the framework of an in vitro fertilization and embryo transfer program and led to the birth of normal children. An average of 3.5 follicles was promoted at each cycle. Oocyte maturity was evaluated at recovery either histologically or with respect to cumulus-oocyte complexes (COC) and correlated to oocyte fertilizability. Three types of COC are described according to cell dissociation and mucification. The most expanded cumulus mass (type I) was highly fertilizable, compared with granular and poorly dissociated cumulus complexes (types II and III, respectively). No improvement in the type II and III oocyte fertilizability was obtained when type I follicular fluid was added to the incubation medium. Moreover, incubation of type III oocytes in medium containing type III follicular fluid seemed to decrease their fertilizability.

Chlamydia trachomatis associated with chronic inflammation in abdominal specimens from women selected for tuboplasty.

Chronic inflammation is a frequent cause of tuboplasty failure. Therefore, it would be useful for one to know the microbiologic agent of infection and to treat it before the tuboplasty. By laparoscopy, a search for Chlamydia trachomatis, Ureaplasma urealyticum, and other microbiologic agents was carried out in the peritoneum and tubes of 118 women divided into 3 groups. Sixty-nine had a checkup before tuboplasty, of which 30 were found to have a chronic inflammatory condition discovered during laparoscopy (group 1) and 39 to have no sign of inflammation (group 2). Forty-nine women with a completely normal pelvis, being followed for possible sterility, were used as a control group. Cultures and serodiagnosis show a significant difference for C. trachomatis between the pathologic groups and the control group. They show no noticeable difference for U. urealyticum. These findings, compared with those by other authors, indicate that C. trachomatis could be an important microbiologic agent in tubal sterility, strongly connected with a low-grade chronic inflammatory condition, and their presence at the time of tuboplasty is to be considered.

[Results of preparatory surgery for in vitro fertilization with embryo transfer]. / Résultats de la chirurgie préparatoire à la F.I.V.E.T.E.

68 cases out of a total of 407 patients who were destined to have IVF (16.7%) had most adhesions in the pelvis which made the ovaries inaccessible for laparoscopic recovery, in the two years between September 1981 and September 1983. Using very severe criteria for selection we rejected 46 cases as unsuitable for surgery at that time. Only 22 therefore remained to have preparatory surgery. This surgery was particularly aimed at improving the local conditions for laparoscopic recovery as well as improving the quality of ovulation. The results are encouraging because those women who were operated on had about the same level of success in the three stages of IVF as women who were not operated on. This three-stage protocol of exploratory laparoscopy, operation and recovery laparoscopy does seem to be a heavy one and probably will be able to be replaced by a transvaginal ultrasound technique when this has been developed far enough to show that it has advantages.

[A comparative study of salpingotomies with or without suture. An experimental microsurgical research carried out on rabbits' tubes (author's transl)]. / Etude comparée des salpingotomies avec ou sans suture. Recherche microchirurgicale expérimentale sur la trompe de lapine.

Two techniques of salpingotomy have been compared in two series of rabbit experiments with the aim of finding out whether, when carrying out conservative surgery in extra-uterine pregnancy with rupture of the tube, it is better to suture the salpingotomy or to leave it open without suture. The two series show that there is no difference as far as the frequency of large adhesions, as far as ovulation and the strength of the scar and the pregnancy rate g out conservative surgery in extra-uterine pregnancy with rupture of the tube, it is better to suture the salpingotomy or to leave it open without suture. The two series show that there is no difference as far as the frequency of large adhesions, as far as ovulation and the strength of the scar and the pregnancy rate g out conservative surgery in extra-uterine pregnancy with rupture of the tube, it is better to suture the salpingotomy or to leave it open without suture. The two series show that there is no difference as far as the frequency of large adhesions, as far as ovulation and the strength of the scar and the pregnancy rate following the two methods. On the other hand, implantation is less good when the tube is left open than when the salpingotomy is sewn up. In the first series there was a significant difference between the side that was operated on and the control side, whereas the second series there was very little difference. In spite of the fact that the scar seemed to be apparently of the same type, the eggs were able to descend more easily in the oviduct when it had been sutured.

[A microbiological study of swabs taken laparoscopically in cases of salpingitis and tubal sterility. Research for Chlamydia trachomatis and for mycoplasmas (author's transl)]. / Etude microbiologique des prélèvements coelioscopiques dans les annexites et les stérilités tubaires. Recherche de Chlamydia trachomatis et de mycoplasmes.

Research was made for chlamydia trachomatis and ureaplasma urealyticum in the peritoneum and the tubes of 99 women divided into 4 groups: 17 of them were being investigated because of acute salpingitis (Group A), 17 were being investigated for tubal sterility with chronic inflammation diagnosed laparoscopically (Group B), 29 were being investigated for tubal sterility without any laparoscopic evidence of inflammation (Group C) and 36 women had absolutely normal pelves and were being investigated for sterility. These were the control group (D). Swabs were also taken from the lower genital tracts as well as serological tests for chlamydia trachomatis and cytological samplings of the fluid from the Pouch of Douglas and the histology of the tubes. In the 17 women who had acute salpingitis the swabs 4 cases of C.T. and 4 of U.U. In the 46 women who had tubal sterility the laparoscopic swabs showed cases of C.T. and 7 of U.U. The swabs were most often positive in Group B. This group is characterised by a special appearence of the inflammation, with fluid present and viscous adhesions as well as peritoneal inflammatory cysts. These altogether help to make a presumptive diagnosis of C.T. infection on laparoscopy. In the control group of 36 cases there was no sign of C.T. in any case, although 2 swabs from the peritoneum showed U.U. So there is a statistically significant difference between the groups that were suspicious and the control group whether the results were obtained by cultures or by serological diagnosis. On the other hand there is no definitive difference as far as U.U. is concerned. These observations, which are similar to those published by other authors, lead us to think that micro-organisms and especially chlamydia trachomatis could be the bacteriological agent responsible for chronic inflammatory states found so frequently in women with tubal sterility.

Steroid profiles during cycles with HCG stimulation of luteal function during in-vitro fertilization cycles.

178 in-vitro fertilization cycles have been explored for luteal function. Clomiphene and human menopausal gonadotrophins were used for ovarian stimulation and some patients were supplemented with human chorionic gonadotrophin (HCG) (5000 IU) on the day of laparoscopy, and also 2 and 4 days later. An imbalance of luteal function was observed in non-supplemented stimulated cycles, with hyperoestrogenicity and a relatively low production of progesterone. The administration of HCG reduced this imbalance by improving the production of progesterone without affecting the production of oestradiol-17 beta.

Transvaginal sonographically controlled ovarian puncture for oocyte retrieval for in vitro fertilization.

Two hundred twenty-two patients took part in a trial of follicle puncture via the transvaginal route under sonographic control for the purpose of in vitro fertilization (IVF). Induction protocols were mainly human menopausal gonadotropin (hMG) + human chorionic gonadotropin (hCG) and clomiphene + hMC + hCG. In 79.7% oocyte aspiration could be achieved without difficulty via the transvaginal route. An average number of 4.7 oocytes per attempt was obtained; 10.7% evolutive pregnancies were obtained. No major incident was noted. This technique offers several crucial advantages: it reduces surgical risk, reduces the length of the patient's stay in hospital as well as the overall cost of the procedure, and it also makes possible puncture in some cases hitherto regarded as excluded.

Anonymous and non-anonymous oocyte donation preliminary results.

During the past year, we have developed an oocyte donation programme in 10 patients with complete absence of endogenous ovarian function (premature ovarian failure in seven cases, castration in two cases and Turner's syndrome in one case). In cases of anonymous donation, donors were volunteers devoid of any major genetical risk who were included in our IVF programme and who consented to donate one oocyte when at least seven oocytes were recovered, and two oocytes when at least 11 oocytes were recovered, to a recipient couple. As far as possible, morphological characteristics of both couples were paired. In cases of non-anonymous donation, donors were 'affective' donors, having at least one child. The resulting embryos after IVF of donated oocytes were either replaced directly in recipient women which required synchronization of the donor's and recipient's cycles, or cryopreserved and then thawed, usually at day 16 of recipient's artificial cycle, i.e. 2 days after introduction of the progestational compound. On the 10 patients entering this oocyte donation programme (20 cycles), 13 transfers were carried out resulting in four clinical pregnancies in three patients with premature ovarian failure and one with Turner's syndrome (20% pregnancy per cycle and 31% per transfer). Despite the small numbers, these good results prompted us to develop this protocol.

Cryopreservation of human embryos and oocytes.

The success rate of human embryo cryopreservation depends on technical and embryonic parameters. First of all, the cryoprotectant can affect embryo survival as we found by comparing two freeze-thaw procedures using propanediol (PROH) (1.5 mol) alone or with sucrose (0.1 mol). Embryo survival was significantly enhanced with sucrose (62 versus 32%). Embryo quality is another major parameter involved in the success of freezing; the rates of positive survival were found to be 67% for morphologically normal embryos versus 49% for embryos with fragments (P less than 0.001). The efficiency of embryo cryopreservation in an IVF programme could be estimated in 1986: a woman with extra embryos, stored after transfer of 3-4 fresh embryos (16% of all cycles), can expect a 22% pregnancy rate per transfer of fresh embryos and a 32% pregnancy rate per collection after transfer of the stored eggs. A comparative study of the cryopreservability of immature or mature oocytes was performed in humans. Human oocytes have a low survival rate (36%) whatever the cryopreservation protocol or the initial maturation stage. Immature human oocytes could survive freezing and thawing, mature and be fertilized in vitro, but with a very low efficiency.

Human embryo cryopreservation, extrinsic and intrinsic parameters of success.

Freezing and thawing (F - T) was applied to 490 early human embryos using propanediol as cryoprotectant. The survival rate of embryos frozen with propanediol alone did not exceed 31% (26/83). The combination of propanediol and sucrose, however, significantly increased the percentage of surviving (248/407 = 61%) and intact (188/407 = 46%) embryos and seemed to enhance embryo viability as suggested by the implantation rate (14.5 versus 8%) without, however, any statistical significance. Embryo survival, but not viability, was correlated with morphological features, whereas neither the age of embryos (1, 2 or 3 days post-insemination) nor the segmentation stage (regular or intermediate) were involved in F - T ability. Thirty-eight F - T embryos implanted when replaced in utero, representing 8% of all F - T embryos and 14% of the F - T replaced embryos. The pregnancy rate per transfer reached 19% (35/185) and was identical to the pregnancy rate per transfer of fresh embryos (253/1149 = 22%). In oocyte donation, too, embryo freezing did not impair the pregnancy rate (25%). In spontaneous cycles, synchronous transfer gave better results than asynchronous transfers (20 versus 10%), but spontaneous cycles had no significant advantage (16% pregnancy/transfer) as compared to stimulated (26%) and artificial (27%) cycles.

Results of planned in-vitro fertilization programming through the pre-administration of the oestrogen-progesterone combined pill.

The use of an oestrogen-progesterone combined pill permits the induction of ovulation in the absence of any developing follicle. Two treatments were compared. In the first, patients received no prior treatment before stimulation. In the second, combined oestrogen-progesterone treatment was given during approximately two menstrual cycles prior to stimulation. No differences between the two groups were found in relation to oocyte maturity, fertilization in vitro, cleavage, replacement and pregnancy. Fewer luteinizing hormone surges occurred in patients pre-treated with steroids. The utilization of the oestrogen-progesterone combined pill prior to induction of ovulation facilitates the forward planning of patients for in-vitro fertilization.

PIP: In order to facilitate the scheduling and management of cases for in vitro fertilization, a trial of pre-administration of a combined oral contraceptive was conducted. 103 women took 2 mg norethisterone acetate and 0.05 mg ethinyl estradiol for 45 to 70 days before induction of ovulation, allowing 5 days between treatments. This group then received either Clomid 100 mg for 6 days and HMG 225 IU on days 5, 7 and 9 (34 women), or HMG only (dose not specified, 69 women). The control groups were given identical ovulation induction regimens: Clomid and HMG (130 women) or HMG only (188 women). 70 of the patients selected for preliminary inhibition of ovulation had irregular menstrual cycles with normal prolactin; the others were picked for scheduling reasons only. No differences were documented between the groups in oocyte maturity, fertilization in vitro, cleavage, replacement and pregnancy. Pregnancy rates ranged from 10 to 16.6% in the 4 subgroups. Fewer LH surges occurred in patients pretreated with steroids. Thus the pretreatment of candidates for in vitro fertilization with ovulation blocking steroids has no quantitative effect on results, but does facilitate scheduling patients for the procedure.

Microbiology of specimens obtained by laparoscopy from controls and from patients with pelvic inflammatory disease or infertility with tubal obstruction: Chlamydia trachomatis and Ureaplasma urealyticum.

We cultured for Chlamydia trachomatis, Ureaplasma urealyticum, and Mycoplasma hominis and performed chlamydial serologic studies in 99 women undergoing laparoscopy. These women included patients with acute salpingitis, infertile women with and without mild pelvic inflammatory disease, and controls. C. trachomatis infection was significantly more common in patients than in controls. We also identified low-grade "silent" PID among women with infertility resulting from tubal obstruction and suggest this may be caused by chlamydiae.