Cryopreservation of human embryos at the pronucleate, early cleavage, or expanded blastocyst stages.

Supernumerary embryos following treatment by IVF or GIFT were cryopreserved at the pronucleate, early cleavage or expanded blastocyst stages. The success of embryo cryopreservation at these stages was evaluated in terms of (i) the proportion of embryos surviving the freeze/thaw procedure; (ii) the proportion of patients reaching embryo replacement; and (iii) the incidence of pregnancy per replacement. Significantly more embryos survived when frozen/thawed at the pronucleate (44/61; 72%) or early cleavage stages (48/80; 60%), than at the expanded blastocyst stage (13/34; 38%). A significantly higher proportion of patients had embryo replacements when embryos were frozen/thawed at the pronucleate (17/19; 89%) or early cleavage stages (21/24; 88%), than at the expanded blastocyst stage (9/17; 53%). Following replacement of frozen/thawed pronucleate and early cleavage stage embryos, clinical pregnancy rates of 8/17 (47%) and 3/21 (14%) clinical pregnancies were achieved, respectively. No pregnancies were achieved following replacement of frozen/thawed expanded blastocysts.

The cryopreservation of donor semen by a simplified method: use in an IVF and GIFT programme.

The cryopreservation of semen used in assisted reproduction procedures was carried out exclusively by a simplified method in which a mixture of semen and cryoprotectant was contained in 1-ml tuberculin syringes and plunged directly into liquid nitrogen. Donor semen samples halved and frozen in syringes and in straws in a controlled-rate freezer showed no significant difference in post-thaw motility (P = 0.217) or survival (P = 0.217) after 30 min. However, after 180 min the survival rate showed a significant reduction in syringes (P = 0.045). A significant difference (P less than 0.00008) in the rate of fertilization of oocytes was seen in IVF cycles using frozen-thawed donor sperm (58/142, 42%) when compared to fresh sperm from husbands (2315/3926, 59%). A significant reduction (P less than 0.00005) in fertilization rate was also observed in the case of supernumerary oocytes in GIFT cycles with the cryopreserved donor sperm (29/132, 22%) compared to the husbands' sperm (239/514, 46%). However, the pregnancy rate following IVF and embryo replacement was the same after fertilization with fresh sperm (75/351, 21%) as opposed to frozen sperm (3/14, 21%). Furthermore, a higher pregnancy rate was observed in GIFT with frozen donor sperm (9/19, 47%) than with fresh sperm from husbands (28/103, 27%), though this was not statistically significant (P = 0.079). These results show this simplified methods of semen cryopreservation to be effective when used in an IVF and GIFT programme, giving pregnancy rates comparable to fresh normospermic semen samples. The method is simple, quick and inexpensive.

Fertilization in vitro of supernumerary oocytes following gamete intra-fallopian transfer (GIFT).

Gamete intra-Fallopian transfer (GIFT) was performed in 130 treatment cycles over a 17-month period. In 91% (118/130) of the cycles one or more oocytes were available for insemination in vitro and only GIFT cycles with supernumerary oocytes were included in the present study. Pituitary and ovarian suppression was achieved with buserelin followed by stimulation of multifollicular development by human menopausal gonadotrophin (HMG). Failure of supernumerary oocytes to fertilize was associated with a significantly reduced pregnancy rate (3/23; 13%) compared to cycles where fertilization occurred in vitro (35/95; 37%). These findings demonstrate that the outcome of IVF of supernumerary oocytes may be of particular diagnostic value in couples where the female partner has not conceived following treatment by GIFT after pituitary down-regulation with buserelin and ovarian stimulation with HMG.

Endocrine changes in women conceiving during treatment with an LHRH agonist.

We report on eight patients who conceived during pituitary desensitization with buserelin in the luteal phase of the menstrual cycle. Pregnancy was diagnosed between day 12 and 21 of buserelin administration. Analysis of serum luteinizing hormone on day 12 showed that pituitary desensitization occurred in conjunction with increasing production of ovarian steroid hormones. Serum concentrations of human chorionic gonadotrophin (HCG) were less than 10 IU/l on day 1 of buserelin administration for seven of the eight patients. The serum concentration of HCG on day 12 showed a median value of 722 IU/l (range 14.6-798 IU/l). Five of the eight patients were given HCG support (10,000 IU) following the diagnosis of pregnancy--three of these patients have ongoing pregnancies and the remaining two had blighted ova on scan. Of the remaining three patients, one had a singleton pregnancy which miscarried at 9 weeks, one had a blighted ovum on scan and bled per vagina shortly after this, and one bled per vagina prior to a scan being carried out. Our results show that pregnancy can occur during pituitary desensitization with buserelin, despite patients being counselled not to have unprotected intercourse in the cycle during which administration commences. An HCG assay on day 1 of buserelin administration is not helpful. Pregnancy should be suspected when ovarian steroid production persists despite complete pituitary down-regulation.

Fertilization of human oocytes in vitro by spermatozoa from oligozoospermic and normospermic men.

Sperm were isolated from the semen of oligozoospermic (less than 20 x 10(6) sperm/ml) and normospermic (greater than or equal to 20 x 10(6) sperm/ml) men in an in-vitro fertilization (IVF) programme. Oocytes from the female partners were inseminated with either 75 or 100 x 10(3) motile sperm and checked for fertilization after 16-20 h. A significant reduction in the overall fertilization rate of oocytes was seen for the oligozoospermic group compared to the normospermic group, at both insemination concentrations. In the oligozoospermic group, a fertilization rate of 31% (19/61) was achieved when oocytes were inseminated with 75 x 10(3) sperm, and 38% (9/24) when inseminated with 100 x 10(3) sperm. This compared with rates of 57% (397/696) and 64% (650/1018), respectively, for normospermic cases at both insemination concentrations. No evidence of fertilization was seen in 36% (4/11) and 67% (4/6) of oligozoospermic cases when 75 or 100 x 10(3) sperm were used, compared with values of 13% (17/133) and 9% (20/212), respectively, in normospermic cases. After excluding zero cases, the fertilization rate of oocytes for the oligozoospermic group (75%; 9/12) was similar to the normospermic group (70% 650/935) when 100 x 10(3) sperm were used. However, when 75 x 10(3) sperm were used, the fertilization rate for the oligozoospermic group (41%; 19/46) was significantly lower than that of the normospermic group (62%; 397/645). Following the transfer of embryos into the female partner, clinical pregnancies were diagnosed in 2/7 (29%) oligozoospermic cases and 27/267 (10%) normospermic cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Prostacyclin-like, and kallikrein activity of amniotic fluid in pre-eclampsia.

Amniotic fluid from patients with pre-eclampsia was compared with samples obtained from normotensive controls with respect to the inhibiting effect on platelet aggregation (PGI2-like activity) and activating effect on the plasma kallikrein assay and Russell's viper venom test. After 39 weeks gestation, amniotic fluid from pre-eclamptic patients showed significantly less PGI2-like activity ( p less than 0.01) and significantly lower kallikrein levels (p less than 0.01) than that from normotensive controls. The study suggests that the biosynthesis and release of PGI2-like activity and kallikrein may be impaired in pre-eclampsia. In view of the association of pre-eclampsia with intravascular clotting, the highly significant reduction of PGI2-like activity seems important and appears to warrant a clinical trial of prostacyclin administration in this disorder.

A study of the procoagulant properties of amniotic fluid and their correlation with the lecithin/sphingomyelin ratio.

The procoagulant properties of amniotic fluid have been studied in samples taken throughout pregnancy and the results correlated with the lecithin/sphingomyelin ratio: acceleration effects on the prothrombin time. Russell's viper venom test, activated partial thromboplastin time and specific factor VII and X assays and contact acceleration. Although amniotic fluid is shown to function as a complete thromboplastin and to activate factor X directly, none of these tests of procoagulant activity of amniotic fluid is a reliable index of fetal lung maturity.

Benign and malignant struma ovarii.

Three cases of struma ovarii are reported, two of which were benign, and one which was malignant. Immunoperoxidase studies confirmed that the tumors secreted triiodothyronine and thyroxine, but no carcinoembryonic antigen nor alpha-fetoprotein. In the case of the malignant struma ovarii, the patient's thyroid was histologically normal, and thus the possibility of a metastasis to the ovary from a primary thyroid carcinoma was excluded.

Oral anticoagulants controlled by the British comparative thromboplastin versus low-dose heparin in prophylaxis of deep vein thrombosis.

The British comparative thromboplastin (BCT) was used to monitor the effectiveness of oral anticoagulants in preventing deep vein thrombosis (DVT) in patients undergoing major gynaecological surgery. All patients were screened for DVT with the use of the (125)I-fibrinogen scan.One hundred and forty-five patients aged 40 years or more were randomised into three groups. Group 1 received oral anticoagulant (nicoumalone) treatment, stabilised over five days before surgery and continuing into the second postoperative week. The other patients served as two contrast groups and were managed on a double-blind basis. Group 2 received a subcutaneous low-dose regimen of heparin calcium. Group 3 received subcutaneous saline. Eleven of 48 patients in the saline group, three of 49 patients in the heparin group, and three of 48 patients in the oral anticoagulant group developed DVT as judged by (125)I-fibrinogen scanning. The incidences in groups 1 and 2 were significantly lower than in the saline group. The falls in haemoglobin concentration and incidence of haemorrhage were similar in all three groups.The study showed that oral anticoagulant prophylaxis stabilised preoperatively and low-dose heparin were equally effective in preventing deep vein thrombosis in a moderate-risk group. Immediate preoperative prothrombin ratios of 2.0-2.5 and postoperative ratios of 2.0-4.0 with the BCT gave adequate protection without increased haemorrhagic risk.

Pregnancy following bilateral uterine artery ligation.

A successful pregnancy following bilateral uterine artery ligation performed in an attempt to relieve dysfunctional menorrhagia is reported. Serial biparietal diameters showed a normal growth curve and the newborn infant weighed 2640 g. The patient was normotensive throughtout the pregnancy. Prior to uterine artery ligation, she had had four pregnancies but there were no surviving children. It is suggested that uterine artery ligation may have a limited place in the management of patients with dysfunctional menorrhagia.

The termination of pregnancy by intravenous infusion of a synthetic prostaglandin F 2 analogue.

PIP: ICI 74 205, a synthetic analogue of prostaglandin F2 alpha (PGF2a) with an alkyl side chain extended by an ethyl group, was administered intravenously in 6 patients using normal saline as the vehicle. Starting dose was 5 mcg/minute in the 1st case and 10 mcg/minute in subsequent cases. Highest dose rate used in every case was 125 mcg/minute. All, except 1 patient, were in the second trimester of pregnancy. The other patient was in the first trimester of pregnancy. Diarrhea and vomiting, as well as a local cutaneous inflammatory reaction at the site of intravenous infusion, occurred in all patients during the infusion but disappeared after the infusion had been stopped. A fall in the plasma estradiol and progesterone levels was seen 2 hours after the start of the prostaglandin infusion. Five patients had a complete abortion. Abortion was incomplete in 1. Mean induction-abortion interval was 25 hours 15 minutes, with a range of from 7 hours 45 minutes to 49 hours 30 minutes. Although the results are encouraging, it is not possible to establish the effectiveness of ICI 74 205 because of the small series. Further research should be done to determine whether the initial uterine activity is due to a direct effect of prostaglandin on the uterine smooth muscle or whether the final outcome depends on changes reflected by the fall in plasma estrogen and progesterone level. Efficacy of ICI 74 205 in various dose regimens and other routes of administration should be investigated.^ieng