In vitro differentiation of human endometrial stromal cells: effect of transforming growth factor beta 1 on prolactin secretion.

OBJECTIVE: To investigate the role of Transforming Growth Factor beta 1 on differentiation of human endometrial stromal cells. STUDY DESIGN: A prospective study. MATERIAL AND METHODS: Human endometrial cells obtained from 9 women were cultured in DMEM-HAMs F12 media to adequate cell confluence and then the flasks were cultured in the presence and absence of TGF b 1. The influence of TGF beta 1 were measured by prolactin production expressed as nanograms of prolactin/mg of total DNA in cells measured by diphenylamine reaction. RESULTS: The levels of prolactin in the culture medium with and without TGF beta 1 were 0.16 +/- 0.27 ng/microgram DNA/day and 0.24 +/- 0.41 ng/microgram DNA/day respectively. There was no significant difference between groups. CONCLUSION: There is no direct effect of TGF beta 1 on differentiation of human endometrial stromal cells. However it may involve in more complex relationship in this process.

Endometriosis and autologous lymphocyte activation by endometrial cells. Are lymphocytes or endometrial cell defects responsible?

OBJECTIVE: To determine whether the decreased mitogenicity of the endometrial cell for autologous lymphocytes noted in women with pelvic endometriosis is secondary to endometrial cell interaction with the lymphocytes or due to an inherent defect of the lymphocyte. STUDY DESIGN: The endometrial cells from 30 women with endometriosis and 30 matching controls were cultured. Autologous lymphocyte and endometrial cells were cocultured to observe the lymphocyte proliferative response to autologous endometrium in controls and to ectopic and eutopic endometrial cells from patients. The ability of lymphocytes to be stimulated by phytohemagglutinin (PHA) was simultaneously assayed. RESULTS: Ectopic and eutopic endometrial cells from women with endometriosis were less mitogenic for autologous lymphocytes than endometrial cells from controls. The lymphocytes from both patients and controls exhibited a similar stimulatory response to PHA. CONCLUSION: The diminished proliferative response of lymphocytes from women with endometriosis probably is not the consequence of an intrinsic lymphocyte abnormality. The fundamental defect in endometriosis may reside within the endometrial cell.

Human sperm bioassay has potential in evaluating the quality of cumulus-oocyte complexes.

Human sperm bioassay is routinely used as a quality control check for the culture media. This is one of the three bioassays chosen by the College of American Pathologists (CAP) for interlaboratory proficiency testing to assess the standards of in vitro fertilization (IVF) and andrology laboratories. This study utilized sperm bioassay to assess the quality of cumulus-oocyte complexes (COCs) retrieved in IVF procedures COCs, harvested from the female partner of IVF couples, undergoing identical ovarian stimulation protocols, were individually inseminated with the sperm of the corresponding male partner. Sperm motility in sperm-COC cocultures were compared. Cocultures were established by inseminating the 103 COCs, retrieved from 18 IVF couples with 1 x 10(5) to 2 x 10(5) sperm of the corresponding male partners of the couples. In all 18 cases, the sperm were prepared identically using the Percoll wash method. The cocultures were maintained for 48 h but the oocytes were removed immediately after the fertilization check (approximately 16 h). The motility of sperm in the cocultures and in the insemination stocks were noted and 17 of 18 sperm stocks used for insemination had similar high preinsemination motility (90.2 +/- 5.0%). At 48 h the sperm motility had significantly decreased in the cocultures compared to the insemination stocks; 52.7 +/- 19.9% versus 67.2 +/- 10.4%. There was no difference in the motility among the small, medium, and large COCs (56.4 +/- 24.6%, 52.5 +/- 17.9%, and 50.8 +/- 20.9%, respectively). In 45% of IVF cases, the motility in cocultures varied widely, falling below as well as above that of their corresponding insemination stocks. Furthermore, the sperm motility varied among the cocultures in both pregnant and nonpregnant patients but the extent of variation appears to be greater in the latter. The inter-COC coculture sperm motility variation most likely is due to the differences in the quality of cumulus-oocyte complexes.

Restoration of oocyte maturational competency during the nonbreeding season with follicle-stimulating hormone stimulation in squirrel monkeys (Saimiri boliviensis boliviensis).

The in vitro maturation potential of oocytes retrieved during the nonbreeding season with or without prior in vivo low-dose FSH stimulation was studied in adult squirrel monkeys (Saimiri boliviensis boliviensis). Additionally, the adequacy of various protein supplements in media used for oocyte maturation was investigated. Ovaries were removed from animals in the nonbreeding season (n = 9) with or without prior treatment with a low dose (1 mg) of FSH for four days before ovariectomy. Minimal estradiol elevation was observed in serum even with stimulation. For oocyte collection, ovaries were placed in warmed 21 mM HEPES-buffered Ham's F-10. Oocytes from unstimulated ovaries were retrieved and cultured (47 of 62 recovered) in bicarbonate-buffered Ham's F-10 medium containing 0.5% BSA as protein supplement. Negligible maturation was observed at 48 h (3 of 47; 6%), and no fertilization was seen after insemination. Immature oocytes from animals stimulated with a low dose of FSH were cultured (69 of 94 recovered). With prior FSH stimulation, oocytes placed in 0.5% BSA medium matured (13 of 24; 54%) and fertilized (7 of 21; 33%) in marked contrast to oocytes from the nonstimulated monkeys. Additionally, 20% monkey serum and 20% human follicular fluid were studied as alternative protein supplements for the FSH-pretreated oocytes; these produced similar maturation rates (10 of 22, 45%; 10 of 23, 43%, respectively) and fertilization rates (8 of 21, 38%; 6 of 21, 29%, respectively). In some cases, 2 pronuclei were observed at 16 h and 4 cells were observed at 40 h. Of those activated, 63% showed cleavage ranging from 2 to 8 cells by 96 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Premenstrual syndrome and related hormonal changes. Long-acting gonadotropin releasing hormone agonist treatment.

The results of long-acting, injectible gonadotropin releasing hormone agonist (GnRH-a) and placebo treatment of severe premenstrual syndrome (PMS) patients with regular menstrual cycles and without known psychiatric disorders are reported. Diagnosis was made according to the Menstrual Symptom Questionnaire and Menstrual Symptom Diary scores. In a placebo-controlled, crossover study, 12 subjects were given either normal saline or depot leuprolide acetate (7.5 mg) every 30 days, starting with the onset of menses. Each subject received the same agent twice before switching to the other and did not known which agent was given. A significant decrease in PMS symptoms was reported by all subjects in both treatment regimens. Biweekly venous blood sampling showed significant elevations of beta-endorphin levels and suppression of gonadotropin concentrations in subjects receiving depot leuprolide treatment. Short-term treatment of severe PMS with an injectible, long-acting GnRH-a may not treat the disease more than do saline injections in a group of women selected by certain criteria.

Operative laparoscopy and postoperative pain relief.

OBJECTIVE: To evaluate the immediate postoperative pain and discomfort in patients who underwent operative laparoscopy under general anesthesia with or without peritoneal lidocaine and incisional bupivacaine instillations. DESIGN: Fifty-four participants were prospectively randomized to three groups: group I, intraperitoneal (IP) lidocaine + intraincisional bupivacaine; group II, intraincisional bupivacaine; and group III, no additional drugs after general anesthesia (controls). SETTING: University Hospital, Reproductive Endocrinology and Infertility Clinics. PATIENTS: Private patients needing operative laparoscopy. INTERVENTIONS: One hundred milligrams of lidocaine were instilled with the irrigation device into the peritoneal cavity at the completion of surgery. Twenty-five milligrams of bupivacaine were injected into infraumbilical and suprapubic incisions. MAIN OUTCOME MEASURES: The analgesic use and modified McGill Present Pain Intensity scores were used for pain evaluation. RESULTS: McGill Present Pain Intensity scores for pain, and narcotic use in the recovery room were less in IP lidocaine-instilled group of patients (P less than 0.05). The mean maximum plasma lidocaine level achieved was 1.01 +/- 0.25 micrograms/mL. CONCLUSIONS: Peritoneal lidocaine and incisional bupivacaine use in operative laparoscopy as described after general anesthesia is safe and effective in reducing postoperative pain in the recovery room.

Mechanism of luteolysis: effect of estradiol and prostaglandin F2 alpha on corpus luteum luteinizing hormone/human chorionic gonadotropin receptors and cyclic nucleotides in the rhesus monkey.

Recent studies from our laboratory suggest that estrogen-induced luteolysis in the primate may be mediated through synthesis of prostaglandin F2 alpha (PGF2 alpha). To elucidate further the mechanism of luteolysis, normally cycling rhesus monkeys received intra-corpus luteum (CL) injections of estradiol (100 microgram), PGF2 alpha (500 microgram), or the appropriate vehicles on the seventh day after the preovulatory estradiol surge. Peripheral vein blood was drawn, at 30-minute intervals for 6 hours for progesterone and luteinizing hormone (LH) assays. Five hours after intra-CL injection, the CL was excised and the LH/human chorionic gonadotropin (hCG) receptor-binding activity and cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) were measured. PGF2 alpha significantly (p < 0.05) lowered progesterone within 1 hour, while the time required for estradiol to lower progesterone significantly was 3.5 hours; there was no significant change in LH, Estradiol and PGF2 alpha significantly (p < 0.05) decreased the LH receptor-binding capacity at 5 hours, without any change in the binding affinity. Also PGF2 alpha significantly (p < 0.05) increased cGMP in the CL, while cAMP remained unchanged; estradiol treatment resulted in a significant (p < 0.05) increase in cAMP with no change in cGMP. This study suggests that estradiol and PGF alpha cause a decrease in progesterone secretion by a loss of the LH/hCG receptor and the PGF2 alpha may act further through the cGMP system.

Ectopic pregnancy. An eight-year review.

Ruptured ectopic pregnancy constitutes a major gynecologic emergency that may result in death. From January 1968 through December 1975, 313 patients with ectopic pregnancy were treated at Chicago Lying-In Hospital. The historical and physical findings, diagnostic procedures, causative factors and patient management are reviewed and discussed. The most common symptoms were abdominal pain and amenorrhea. More than half the patients were misdiagnosed prior to admission. Only 30% had distinct adnexal masses. The treatment of choice was salpingectomy unless the opposite tube was absent or damaged. Three deaths occurred in this series. Only 31% of the patients gave histopathologic evidence of pelvic inflammatory disease.

PIP: 313 patients with ectopic pregnancy were treated at Chicago Lying-In Hospital in 1968-1975, for a frequency of 1 in 72 deliveries. Historical and physical findings, diagnostic procedures, causative factors, and patient management were reviewed for 284 of these patients. 97.5% of the pregnancies were tubal. 25% of the patients were nulliparas and 31.7% were primiparas. 34.9% had had 1 or more previous abortions. The mean age of the patients was 28. Most had been using no contraception, and only 3.9% had an IUD in situ. The most common symptoms were abdominal pain (96.7%) and amenorrhea (73.6%), while the most frequent abdominal findings were tenderness (83.4%), rebound pain (41.2%), and guarding (28.9%). Adnexal tenderness was found in 72.2% and 30% had distinct adnexal masses. An initial misdiagnosis of pelvic inflammatory disease was made in 132 (46.5%) cases. A culdocentesis and slide latex agglutination inhibition pregnancy test performed in 167 and 102 patients, respectively, gave 82.6 and 73.5% positive rates. Diagnostic laparoscopy was used routinely after 1970 on all nonacute patients in whom ectopic pregnancy was suspected, and this led to a significant drop in the rate of ruptured pregnancies (63% pre-1970 and 45% post-1970). 25% of patients were sterilized, but the treatment of choice was salpingectomy unless the opposite tube was absent or damaged. Gross evidence of pelvic inflammatory disease was noted in 36% of patients and 31% had salpingitis. The most common postoperative complication was fever (42.2%). 3 deaths occurred in the series (2 due to acute pulmonary edema resulting from fluid overload), for a maternal death rate from ectopic gestation of 13.83/100,000 live births. No fetuses survived.