Identification of apolipoprotein A1 in the human embryonic secretome.

OBJECTIVE: To identify factors secreted by the human embryo and correlate levels with embryo morphology and pregnancy outcome. DESIGN: A laboratory-based study of human embryo protein synthesis and secretion and a retrospective analysis of spent embryo culture media as it relates to pregnancy outcome. SETTING: University-based academic IVF program. PATIENT(S): IVF patients who had donated cryopreserved human pronuclear-stage embryos. Patients undergoing fresh IVF cycles resulting in a blastocyst transfer who donated spent media drops. INTERVENTION(S): In vitro embryo culture and collection of spent media. MAIN OUTCOME MEASURE(S): Protein analysis and identification by two-dimensional gel electrophoresis and mass spectrometry, ApoA1 quantification by ELISA, and mRNA analysis by quantitative reverse transcriptase-polymerase chain reaction. RESULT(S): By protein gel electrophoresis, apolipoprotein A1 (ApoA1) was increased in the culture media from good-quality blastocysts (n = 6 embryos) compared to either cleavage-arrested embryos (n = 6 embryos) or poor-quality blastocysts (n = 6 embryos) using spent media from culture days 4 and 5, respectively. Apolipoprotein A1 concentrations were 23.1% greater in day 5 spent culture media from good-grade blastocysts (n = 30) when compared to poor-grade embryos (n = 30). However, in a group of patients (n = 20) with transfer of two good-quality blastocysts, ApoA1 levels from day 5 spent media did not correlate with embryo implantation and pregnancy. Quantitative reverse transcriptase-polymerase chain reaction confirmed the presence of ApoA1 mRNA transcripts in human blastocysts. CONCLUSION(S): Apolipoprotein A1 is produced by human preimplantation embryos, and increased levels are present in spent culture media containing blastocysts of higher morphologic grade. These results suggest a role for lipoproteins in early embryologic development.

Skewed X inactivation and IVF-conceived infants.

The objective of this study was to investigate whether skewed X chromosome inactivation (XCI) is associated with IVF. A retrospective cohort study was performed comprising 30 female infants conceived by IVF and 44 naturally conceived control infants matched for gestational age and sex. Cord blood DNA samples were obtained and XCI patterns were analysed using a methylation-sensitive assay. Eight IVF samples and 13 control samples were excluded from the study because they were either homozygous or alleles were too similar for the assay to determine skewing. Mildly skewed XCI (80-90% inactivation of one allele) was present in two of 22 (9.1%) IVF samples and two of 31 (6.5%) control samples. Extremely skewed XCI (>90% inactivation of one allele) was found in two of 22 (9.1%) IVF samples and none of 31 control samples. Neither difference was statistically significant. However, the mean degree of skewed XCI in the IVF group was 72.0% and in the control group was 62.4% (P=0.002). Larger studies are needed to clarify the relationship between IVF and skewed XCI.

17beta-hydroxysteroid dehydrogenase 3 deficiency in a male pseudohermaphrodite.

OBJECTIVE: To present the clinical, biochemical, and genetic features of a male pseudohermaphrodite whose condition was caused by 17beta-hydroxysteroid dehydrogenase 3 (17beta-HSD3) deficiency. DESIGN: Case report. SETTING: Gynecology practice in a university teaching hospital. PATIENT(S): A 15-year-old black American male pseudohermaphrodite with 17beta-HSD3 deficiency. INTERVENTION(S): Laboratory evaluation, genetic mutation analysis, bilateral gonadectomy, and hormone replacement. MAIN OUTCOME MEASURE(S): Endocrinologic evaluation and genetic analysis. RESULT(S): A diagnosis of 17beta-HSD3 deficiency made on the basis of hormone evaluation was confirmed through genetic mutation analysis of the HSD17B3 gene. Female phenotype was attained after gonadectomy, passive vaginal dilatation, and hormone therapy. CONCLUSION(S): Deficiency of 17beta-HSD3 was diagnosed in this patient on the basis of endocrinologic evaluation and was confirmed with genetic mutation analysis. The patient was able to retain her female sexual identity after surgical and medical treatment.

Perioperative morbidity and mortality from major gynecologic surgery in the elderly woman.

OBJECTIVE: To report perioperative morbidity and mortality rates in elderly women who underwent major gynecologic surgery. STUDY DESIGN: The charts of 110 women between 80 and 91 years of age who underwent major gynecologic surgery between July 1995 and May 2003 were retrospectively reviewed. RESULTS: The mean age was 83.1 years. Forty-nine procedures (44.1%) were performed for cancer, 32 (28.8%) for a mass and 23 (20.7%) for pelvic organ prolapse or urinary incontinence. Sixty-nine (62.7%) procedures were performed abdominally, 36 (32.4%) vaginally and 5 (4.5%) laparoscopically. Fifty (44.6%) patients experienced a postoperative complication, and 9 (8.1%) experienced a major one. Major complications included serious morbidity, in 5 (4.5%) patients, and mortality, in 4 (3.6%). Advanced age (> 85 years) was not associated with any of the outcomes of interest, while prior surgical history was significantly associated with a decreased hospital stay (p < 0.001). Increased hospital stay was associated with a moderate or severe medical history (p < 0.05) and laparotomy/laparoscopy vs. vaginal surgery (p < 0.01). CONCLUSION: Postoperative complications occurred frequently among women > 80 years of age who underwent gynecologic surgery. The increased perioperative morbidity in the elderly should be considered when performing surgery on women in that age group.

Serum total testosterone levels in a patient with late onset 21-hydroxylase deficiency and a twin gestation.

OBJECTIVE: To present serum androgen levels during pregnancy in a twin gestation complicated by maternal late onset 21-hydroxylase deficiency. DESIGN: Case report. SETTING: University teaching hospital reproductive endocrinology and infertility practice. PATIENT(S): A 27-year-old with nonclassic 21-hydroxylase deficiency and infertility, twin female fetuses, and elevated androgens. INTERVENTION(S): Steroid replacement. MAIN OUTCOME MEASURE(S): Serum T and 17-hydroxyprogesterone (17-OHP) levels. RESULT(S): Elevated androgen levels persisted throughout pregnancy in spite of aggressive steroid replacement. However, twin girls were born without any evidence of virilization. CONCLUSION(S): The changes associated with a twin gestation may result in excessive stimulation of androgens in mothers with nonclassic 21-hydroxylase deficiency. However, the increased placental aromatase provides protection.