Formaldehyde in cryoprotectant propanediol and effect on mouse zygotes.

Cryopreservation of human zygotes and embryos has been routinely performed by in-vitro fertilization clinics for many years. Karran and Legge (1996) first reported that formaldehyde (FA) present in the cryoprotective solutions can have a deleterious effect on mouse oocytes. FA is a cytotoxic, carcinogenic and mutagenic chemical. The effect of FA on mouse zygotes was investigated. In addition, the concentrations of FA in propanediol (PROH) obtained from various sources were determined. Pooled 1-cell embryos were dispensed into droplets of modified Ham's F10 or human tubal fluid containing various concentrations of FA. Since bovine serum albumin (BSA) may minimize toxicity additional trials were done as above in the absence of BSA. FA concentration in the standard 1.5 M PROH, from different sources in water, was measured in the same assay using a standard curve of 0-100 microM FA. FA in a complex medium had a significant deleterious effect on embryo development and hatching but only at 1 mM concentration (P < 0.000001; see Tables I-III). There was no significant effect of FA at 100 microM. However, in a simple medium even 50 microM FA decreased embryo hatching. FA was present in 1.5 M PROH from different sources (range 1.0-35.3 microM concentration). It appears that FA concentrations do not increase with storage because FA concentrations were low even after opening and storage for 3 years on the shelf. This suggests that FA is a contaminant during the manufacturing process and may vary from manufacturer to manufacturer and batch to batch. Until further studies are done to confirm the lack of toxicity to embryos during cryopreservation (with or without FA scavengers) it may be prudent to screen all batches of cryoprotectants for FA as part of quality control.

Assisted hatching of embryos by micromanipulation for human in vitro fertilization: UAMS experience.

In vitro fertilization and embryo transfer (IVF) is utilized as a treatment for infertile couples who cannot conceive with standard therapy. Assisted hatching (AH) is a procedure whereby an opening is made in the zona pellucida of the embryos, thereby increasing the probability of implantation and pregnancy. AH is beneficial in patients with elevated FSH levels, older than age 38 or those who failed IVF repeatedly. Success rates after IVF with AH at the University of Arkansas for Medical Sciences (UAMS) compares favorably with rates achieved by other centers in the USA. Pregnancy rates after IVF with AH in patients older than 38 years is approximately 20% compared to a pregnancy rate of 10% in patients who did not have AH. This report summarizes the UAMS experience with IVF and AH.

Bilateral internal jugular venous thrombosis complicating severe ovarian hyperstimulation syndrome after prophylactic albumin administration.

OBJECTIVE: To report a case of bilateral thrombosis of the internal jugular veins in a patient after controlled ovarian hyperstimulation and IVF. DESIGN: Case report. SETTING: University-based IVF program. INTERVENTION(S): Ovulation induction with gonadotropins, IVF-ET, albumin administration, and therapeutic heparinization. MAIN OUTCOME MEASURE(S): Doppler ultrasound of neck veins. RESULT(S): Severe ovarian hyperstimulation syndrome and bilateral thrombosis of the internal jugular veins occurred despite prophylactic administration of albumin. This was treated successfully with therapeutic heparinization. CONCLUSION(S): Internal jugular venous thrombosis, a rare complication of ovulation induction with gonadotropins, should be considered in patients with neck pain and swelling.

Absence of glucose decreases human fertilization and sperm movement characteristics in vitro.

The effect of glucose in a modified Ham's F10 medium (MM) without hypoxanthine, phosphate and transition metals on human fertilization and sperm survival in vitro was determined. Mature human oocytes from in-vitro fertilization (IVF) patients or Percoll-washed human spermatozoa were randomly allocated to one of the treatment groups: normal Ham's F10, MM, MM with 5 mM glucose (HGMM) and MM with 0.5 mM glucose (LGMM). Oocytes were inseminated in one of the four media for 12-20 h and checked for fertilization. Sperm were incubated likewise for 4 and 24 h, and sperm motility and sperm movement characteristics including average path velocity (VAP), curvilinear velocity (VCL), straight line velocity (VSL), amplitude of lateral head displacement (ALH), beat cross frequency (BCF), straightness (STR), and linearity (LIN) were determined using computer-assisted semen analysis. Fertilization rates were significantly lower in oocytes cultured in MM (23.8%) compared to LGMM (75.5%), HGMM (73.6%) or Ham's F10 (71.1%). Sperm characteristics after 4 h incubation in all four media were similar, except VAP, VSL, VCL and ALH were significantly lower in MM (no glucose) in comparison with the other three media. After 24 h VAP, VSL, VCL, ALH, LIN and percentage rapid spermatozoa were significantly higher in spermatozoa incubated in HGMM or Ham's F10 compared with MM or LGMM. Also after 24 h, the percentage of spermatozoa which were highly motile was greater in HGMM than in Ham's F10. Absence of glucose significantly lowered fertilization rates and sperm movement characteristics in vitro.

Improved human zygote development in a modified Ham's F10 medium in vitro.

PURPOSE: Our purpose was to determine the effect of modified Ham's F10 media with and without glucose, hypoxanthine, phosphate, and transition metals (MM) on human embryo development. METHODS: Patients with at least one zygote were randomized by alternate assignment to one of the treatment groups. The treatment groups were as follows: normal Hams F10, MM, and MM with 0.5 mM glucose (LGMM). Patients were undergoing in vitro fertilization and embryo transfer or frozen-thawed embryo transfer. Zygotes (fresh and frozen-thawed) were incubated in one of three media for 24 hr before transfer. The groups were compared with respect to embryo quality, number of embryos transferred, age of the patients, clinical pregnancy rate, and implantation rate. RESULTS: The clinical pregnancy rate and the implantation rate were significantly higher in the group of patients whose zygotes were cultured in MM. LGMM did not appear to improve the pregnancy outcome compared to normal Ham's F10. However, the quality of the embryos cultured in MM or LGMM appears to be superior to the normal Ham's F10 with respect to the number of excellent and good-quality embryos and embryos without fragments. CONCLUSION: A modified Ham's F10 medium (MM) without added glucose appears to be superior to normal Ham's F10 for culture of human zygotes for embryo transfer.

Yeast infection of sperm, oocytes and embryos after intravaginal culture for embryo transfer.

OBJECTIVE: To report a case of an embryo culture infected with Candida albicans after intravaginal culture. DESIGN: Case report. SETTING: Private infertility practice and university medical center. PATIENT: A couple with tubal and male factor infertility. INTERVENTIONS: Superovulation, oocyte recovery, Percoll sperm preparation, and intravaginal culture of sperm and oocytes in a tissue culture tube. MAIN OUTCOME MEASURES: Yeast infection of sperm, oocyte, and embryo culture. RESULTS: Candida albicans infection occurred in the sperm, oocyte, and embryo culture when cultured in a sealed tube in the vagina. Candida albicans also was found in the prepared sperm suspension culture in a separate tube kept in a 37 degrees C incubator. CONCLUSIONS: Infection of the embryo culture with C. albicans probably occurred when contaminated sperm was added at the time of insemination. Sperm preparation by the Percoll gradient centrifugation failed to eliminate C. albicans in the semen.

The effect of monophasic combinations of ethinyl estradiol and norethindrone on gonadotropins, androgens and sex hormone binding globulin: a randomized trial.

The effects of different monophasic combinations of ethinyl estradiol and norethindrone on FSH, LH, sex hormone binding globulin, total testosterone, androstenedione, and dehydroepiandrosterone sulfate levels in non-obese, non-hirsute women were compared. Retrospective analysis of frozen serum from a prospective randomized trial in which women received one of three oral contraceptive pills containing ethinyl estradiol 50 micrograms/norethindrone 1 mg, ethinyl estradiol 35 micrograms/norethindrone 1 mg or ethinyl estradiol 35 g/norethindrone 0.5 mg for nine cycles was conducted. Blood samples were obtained prior to treatment and during the third, sixth and ninth pill cycles. Ethinyl estradiol 50 micrograms/norethindrone 1 mg and ethinyl estradiol 35 micrograms/norethindrone 1 mg suppressed FSH, LH, and total testosterone and increased sex hormone binding globulin to a similar degree. Ethinyl estradiol 35 micrograms/norethindrone 0.5 mg resulted in less suppression of FSH, LH, and total testosterone, but greater elevation of sex hormone binding globulin. Dehydroepiandrosterone sulfate was suppressed to a similar degree with ethinyl estradiol 35 micrograms/norethindrone 1 mg and ethinyl estradiol 35 micrograms/norethindrone 0.5 mg, but ethinyl estradiol 50 micrograms/norethindrone 1 mg resulted in the least suppression of dehydroepiandrosterone sulfate. Ethinyl estradiol 35 micrograms/norethindrone 1 mg caused greater suppression of androstenedione than did the other two oral contraceptives. Oral contraceptive-induced changes in gonadotropins, androgens, and sex hormone binding globulin can be predicted by considering the relative amounts of estrogen and progestin in the pill. When combined with 1 mg of norethindrone, 50 micrograms of ethinyl estradiol did not result in greater suppression of FSH, LH, or total testosterone or in greater elevation of sex hormone binding globulin than did 35 micrograms of ethinyl estradiol.

PIP: The effects of different monophasic combinations of ethinyl estradiol and norethindrone on follicle stimulating hormone (FSH), luteinizing hormone (LH), sex hormone binding globulin (SHBG), total testosterone, androstenedione, and dehydroepiandrosterone sulfate levels in nonobese, nonhirsute women were compared. Healthy young women were recruited from the Baltimore area who were 18-35 years old. 60 subjects were selected for the study. Retrospective analysis of frozen serum from a prospective randomized trial was conducted in which women received 1 of 3 oral contraceptive pills containing ethinyl estradiol 50 mcg/norethindrone 1 mg, ethinyl estradiol 35 mcg/norethindrone 1 mg, or ethinyl estradiol 35 g/norethindrone 0.5 mg for 9 cycles. Blood samples were obtained prior to treatment and during the 3rd, 6th, and 9th pill cycles. Ethinyl estradiol 50 mcg/norethindrone 1 mg and ethinyl estradiol 35 mcg/norethindrone 1 mg suppressed FSH, LH, and total testosterone and increased sex hormone binding globulin to a similar degree. Ethinyl estradiol 35 mcg/norethindrone 0.5 mg resulted in less suppression of FSH, LH, and total testosterone, but greater elevation of sex hormone binding globulin. Dehydroepiandrosterone sulfate was suppressed to a similar degree with ethinyl estradiol 35 mcg/norethindrone 1 mg and ethinyl estradiol 35 mcg/norethindrone 0.5 mg, but ethinyl estradiol 50 mcg/norethindrone 1 mg resulted in the least suppression of dehydroepiandrosterone sulfate. Ethinyl estradiol 35 mcg/norethindrone 1 mg caused greater suppression of androstenedione than did the other 2 oral contraceptives. Oral contraceptive-induced changes in gonadotropins, androgens, and sex hormone binding globulin can be predicted by considering the relative amounts of estrogen and progestin in the pill. When combined with 1 mg of norethindrone, 50 mcg of ethinyl estradiol did not result in greater suppression of FSH, LH, or total testosterone or in greater elevation of sex hormone binding globulin than did 35 mcg of ethinyl estradiol.

Effect of exogenous oestrogen on blood flow and quantitative histology of the corpora lutea of pseudopregnant rabbits.

The effect of exogenous oestrogen on blood flow and quantitative histology of the corpus luteum were studied in pseudopregnant rabbits. Pseudopregnancy was induced with human chorionic gonadotrophin and an oestradiol capsule was implanted on day 0 of pseudopregnancy. Regression of the corpus luteum was initiated in the mid-luteal phase by removal of the capsule. Thirty-two New Zealand white rabbits were randomly allocated to four groups (eight rabbits per group) for studies of blood flow--control: oestradiol capsule not removed: -24 h: oestradiol capsule removed 24 h before blood flow measurements; -48 h: oestradiol capsule removed for 48 h; and -48 h/+48 h: oestradiol capsule removed for 48 h and then reimplanted for an additional 48 h. All blood flow studies were performed on day 11 of pseudopregnancy with radioactive microspheres. Quantitative histology was performed on a separate group of 16 rabbits allocated to the same groups as above (four rabbits per group). Blood flow to the corpus luteum was unchanged in the -24 h group, but declined by 44% in the -48 h group. Blood flow returned to control values in the -48 h/+48 h group. Quantitative histology revealed no changes in the -24 h group, but a significant decrease in volume of the corpus luteum and volume of the luteal cell cytoplasm in the -48 h group. Corpus luteum volume was fully restored and luteal cell cytoplasm volume was partially restored in the -48 h/+48 h group. In contrast, the number of luteal cells, vessel space volume and capillary surface area did not change with oestrogen withdrawal.(ABSTRACT TRUNCATED AT 250 WORDS)

Lipoprotein alterations from a triphasic oral contraceptive containing ethinyl estradiol and gestodene. A 12-month trial.

To evaluate the lipid and lipoprotein changes induced by a triphasic oral contraceptive (OC) containing ethinyl estradiol and gestodene, 25 healthy women from the Baltimore metropolitan area were enrolled in an open-label, noncomparative study. Serum lipids were measured prior to starting the OCs and again during the 3rd, 6th and 12th treatment cycles. Mean lipid concentrations in each treatment cycle were compared to baseline levels using the t test for paired samples. Small but statistically significant (P < or = .05) increases in the mean concentrations of total cholesterol, total triglycerides, total high density lipoprotein (HDL) cholesterol, HDL3 cholesterol, apolipoprotein A1 and apolipoprotein B were noted. Although the increases were statistically significant, the mean lipid concentrations were still within the normal range. The mean HDL2 and low density lipoprotein cholesterol concentrations were unchanged, as was the mean total cholesterol/HDL ratio. Healthy women taking a triphasic OC containing ethinyl estradiol and gestodene have minimal changes in lipids and should not be at increased risk of atherosclerosis due to OC-induced lipid alterations.

A comparison of the reproductive outcome between women with a unicornuate uterus and women with a didelphic uterus.

OBJECTIVE: To compare the reproductive performance of women with a unicornuate uterus with the reproductive performance of women with a didelphic uterus. DESIGN: Retrospective. SETTING: Reproductive endocrinology service of a tertiary referral hospital. MAIN OUTCOME MEASURES: The proportion of pregnancies resulting in spontaneous abortion, preterm delivery, term delivery, and living children was determined for each group. RESULTS: Twenty-nine women with a unicornuate uterus and 25 women with a didelphic uterus were identified. Twenty women with a unicornuate uterus produced a total of 40 pregnancies, whereas 13 women with a didelphic uterus produced a total of 28 pregnancies. The 33% spontaneous abortion rate in the unicornuate group was not significantly different from the 23% rate in the didelphic group. The proportion of pregnancies resulting in preterm delivery, term delivery, and living children was similar in both groups. CONCLUSIONS: The reproductive performance of women with a unicornuate uterus is similar to the performance of women with a didelphic uterus.

Repeated pregnancy losses.

Our understanding of the causes of repeated pregnancy losses has been expanded during the past year as illustrated from the citations presented from the recent literature in this review. A novel contribution from genetics is the concept of parental gene imprinting, which could help us discuss differences between abnormal embryo and placental development. Advances in immunology, now enable us to use the general term antiphospholipid syndrome to describe pregnancy losses resulting from antibodies to cardiolipin and phosphatidylserine. In addition, other immune causes for pregnancy loss may be present as indicated by the observation that women with circulating antibodies to thyroglobulin or peroxidase are at increased risk for pregnancy loss. Finally, as our knowledge of the cytokines increases, their role in implantation and pregnancy maintenance will be delineated and their dysfunction identified. These disorders will thus become known as the immunodystrophic causes of repeated pregnancy losses.

Vaginal and bladder angiosarcoma after therapeutic irradiation.

Angiosarcoma involving the female genitourinary tract is a rare soft tissue malignancy of vascular origin. We have described probably the first reported case of postirradiation angiosarcoma involving the vagina and bladder, and have reviewed the existing literature on the subject of angiosarcoma resulting from previous therapeutic irradiation for gynecologic malignancy.

Contamination of enteral nutrition systems during prolonged intermittent use.

Two related studies were done to determine the incidence of bacterial contamination in enteral delivery systems that were used for 15 and 7.5 hr, rinsed after each use, and reused daily in vitro for 7 and 5 days, respectively. In the first study, systems infusing either a premixed formula (Ensure) or a hand-mixed formula (Vivonex) did not show bacterial growth until the 4th day, 1.0-2.0 X 10 colony-forming units per milliliter (CFU/ml) of Staphylococcus epidermidis. Thereafter there was sporadic growth of different organisms but never increasing growth during the 7 days of infusion. In the second study, systems with Ensure were initially contaminated with Staphylococcus aureus and Escherichia coli and reused for 5 days. S. aureus was eliminated by rinsing, but E. coli persisted in the delivery system at concentrations of 10(3)-10(6) CFU/ml. We conclude that clean enteral nutrition systems can be rinsed after short-infusion periods and reused up to 7 days in vitro without significant contamination; however, once a bag has become heavily contaminated some bacteria cannot be eradicated from the system by rinsing.