Journal of the Society for Gynecologic Investigation最新文献

Objective: Nitric oxide (NO) plays a fundamental role in cervical ripening and it is synthesized in the human cervix. We studied the effect of the dinoprostone on cervical NO release in pregnant women, and we investigated the relationship between cervical NO metabolites, cervical length, and Bishop score.

Methods: Seventy-seven women underwent induction of labor at > or = 37 weeks of gestation, due to post-term pregnancy (23.8%), oligohydramnios (36.3) or preeclampsia (29.9%). Cervical fluid samples for NO metabolites (NOx), Bishop score, and cervical length were assessed immediately before (time 0 [T0]) and 6 hours after (T6) the local application of dinoprostone, a commercially available prostaglandin E2 (PGE2) analog.

Results: The mean patients' age was 34 +/- 3.2 years, mean gestational age at enrollment was 284 +/- 9.2 days, and nulliparous represented 31.2% of the study population. At time 0, Bishop score was less than 4 in 74% (57/77) of the subjects, mean cervical length was 28.6 +/- 5.8 mm, mean NOx concentration was 208.6 +/- 103.8 microM/mL; 6 hours later, at T6, the mean cervical length decreased to 19.5 +/- 8.8 mm, and the mean NOx concentration increased up to 316.7 +/- 240.9 microM/mL. Data were unaffected by parity or by regular uterine contraction patterns. A statistically significant positive correlation was found between changes in cervical NOx levels and Bishop score modification (P < .01; r = .494), as well as between the modification of NO metabolites concentration and cervical shortening (P < .01; r = .307).

Conclusions: Prostaglandin (PG)-induced cervical ripening is associated with local NO release. NO plays an active role in cervical remodeling since it positively correlates with both cervical shortening and Bishop score increase. NO oxide and PG are the two pathways that, cross activating each other, trigger the cascade of events responsible of cervical ripening.

Objectives: The cytokine milieu at the implantation site plays a role in human pregnancy. Th2 cytokines, such as interleukin (IL)-4 and IL-10, stimulate growth and development of placenta, whereas Th1 cytokines, such as tumor necrosis factor-alpha (TNF-alpha), are associated with pregnancy complications. Natural killer (NK) cells predominate at the implantation site. The aim of the present study is to investigate cytokine expression in NK cells when they are in close contact with JEG-3 trophoblast-like cells using an in vitro coculture model.

Methods: Female peripheral blood mononuclear cells (PBMCs) were cocultured with JEG-3 cells for 24 hours. PBMCs were harvested from the cocultures and stimulated with 25 ng/mL phorbol myristate acetate and 1 micromol/mL ionomycin in the presence of 2 micromol/mL monensin. NK cells were analyzed by flow cytometry for intracellular TNF-alpha, interferon-gamma (IFN-gamma), and IL-4 and IL-10 cytokines. Controls were PBMCs cultured without JEG-3 cells.

Results: The proportion of CD56+/TNF-alpha(+) NK cells was significantly decreased when they were in coculture with JEG-3 cells (26.1%) as compared to without JEG-3 cell coculture (40.8%) (P < .05). There was no difference in the proportion of CD56(+) NK cells expressing intracellular IFN-gamma, IL-4, and IL-10. Down-regulation of CD56+/TNF-alpha(+) NK cell levels was dependent on direct cell-to-cell contact between NK cells and JEG-3 cells. The expression of human leukocyte antigen (HLA)-G on trophoblast cell lines did not affect CD56+/TNF-alpha(+) NK cell levels under these experimental conditions.

Conclusion: We report that JEG-3 cells induce down-regulation of intracellular CD56+/TNF-alpha(+) NK cell levels. It is speculated that trophoblasts may secure themselves from NK cell cytotoxicity via this mechanism.

Objective: Cation channels comprised of transient receptor potential (TrpC) proteins may play a role in signal-regulated calcium entry and calcium homeostasis in myometrium. The objective of this study was to determine the relative abundance of specific TrpC mRNAs expressed in human myometrium and determine if TrpC mRNA and protein concentrations differ in fundal myometrium before and after the onset of labor.

Methods: A quantitative real-time polymerase chain reaction (Q-RT-PCR) procedure was developed for determining the concentration of TrpC mRNA expression in immortalized and primary human myometrial cells and myometrial fundus tissues from patients before and after the onset of labor. The corresponding TrpC proteins were detected by Western blot analysis and immunohistochemistry.

Results: hTrpC1, 3, 4, 5, 6, and 7 mRNAs were expressed in two lines of immortalized human myometrial cells and in primary human myocytes. In all of these cells, hTrpC1 and hTrpC4 mRNAs were the most abundant, followed by hTrpC6. A similar distribution was observed in fundal myometrium samples from patients before and after the onset of labor. hTrpC4 mRNA was significantly lower after the onset of labor; there were no significant changes in the concentrations of other TrpC mRNAs. Immunohistochemistry identified hTrpC1, 3, 4, and 6 proteins in myometrial smooth muscle cells. Western blot analysis of myometrial membranes demonstrated no statistically significant changes in hTrpC1, 3, 4, and 6 proteins between samples collected before and after the onset of labor.

Conclusions: We have demonstrated that hTrpC1 and hTrpC4 are the most abundant TrpC mRNAs in human myometrium, with TrpC6 being the next most abundant. There was no increase in TrpC mRNA or protein in fundal myometrium with the onset of labor. Nonetheless, these isoforms may play significant roles in signal regulated calcium entry in human myometrium.

Objective: To determine during mid-trimester amniocentesis if elevated concentrations of ADAM-8 (A Disintegrin And Metalloprotease 8) and/or cortisol can recognize women at risk for spontaneous preterm delivery.

Methods: The study involved 312 women who underwent mid-trimester amniocentesis. Thirteen patients, who progressed to preterm delivery, were matched with 21 controls for age, parity, gestational age at amniocentesis, and year of amniocentesis. ADAM-8 and cortisol levels were measured by enzyme-linked immunosorbent assay and radioimmunoassay, respectively.

Results: ADAM-8 mean amniotic fluid concentrations were significantly higher in women with preterm delivery than in women delivering at term (mean 1213.9 [SE 96.7] pg/mL [range, 780 to 1854 pg/mL] vs mean 937.2 [SE 50.3] pg/mL [range, 486 to 1508 pg/mL], P < .02). Amniotic fluid ADAM-8 concentrations higher than 1149 pg/mL had the highest specificity and odds ratio (OR) in the identification of the women with increased risk for preterm delivery (sensitivity 61.5%; specificity 81.7%; OR, 9.6 [95% confidence interval (CI), 1.8 to 50.3]). Women with preterm delivery had suggestively higher amniotic fluid concentrations of cortisol (mean 1.3 [SE 0.2] microg/dL [range, 0.4 to 2.2 microg/dL]) than women delivering at term (mean 1.0 [SE 0.09] microg/dL [range, 0.6 to 1.7 microg/dL], P < .07). Furthermore, cortisol levels were positively correlated with ADAM-8 levels (Spearman's r = .418, P < .014).

Conclusions: Elevated mid-trimester amniotic fluid ADAM-8 concentrations possibly are a risk factor for preterm delivery, particularly if ADAM-8 levels are greater than 1149 pg/mL. Potential intrauterine inflammation is also associated with suggestively increased amniotic fluid cortisol levels.

Objective: Matrix metalloproteinases (MMPs) play an important role in modeling and remodeling the extracellular matrix in leiomyomas. Hence, we investigated whether associations exist between leiomyomas and promoter polymorphisms in the MMP-1 and MMP-9 genes in a Japanese population.

Methods: We compared the distribution of polymorphisms in the promoter regions of MMP-1 (-1607 1G/2G) and MMP-9 (-1562 C/T) in 267 leiomyoma patients and 184 control patients using polymerase chain reaction-fragment-length polymorphism (PCR-RFLP) analysis.

Results: The allele frequencies of the MMP-1 -1607 2G and MMP-9 -1562 T polymorphisms were 74.6% and 18.6% in leiomyoma patients, and 71.3% and 18.6% in control patients, respectively. No significant differences in allele frequencies or genotype distributions were found between leiomyoma and control patients. Moreover, no associations were found between MMP-1 and MMP-9 genotypes and leiomyoma size or a family history of the condition.

Conclusion: These findings suggest that MMP-1 and MMP-9 promoter polymorphisms are unlikely to be associated with an increased risk of uterine leiomyomas in Japanese women.

Objective: Protein kinase B (Akt) and the mitogen-activated protein kinases (MAPKs) mediate hypertrophy in the adult heart, although their importance in the developing heart is poorly understood. The goal of the current study was to determine if volume loading the fetal heart resulting from chronic anemia affects regulation of Akt and the MAPKs and if this response is developmentally regulated.

Methods: Anemia was created by 7 days of isovolumic hemorrhage beginning at 101 days (early GA) or 129 days (late GA) gestational age (GA) in fetal sheep (term = 145 days), following which protein levels of total and active Akt, extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun NH2-terminal kinase (JNK), and p38 were determined in the right and left ventricle (RV and LV). RV protein-to-DNA ratios were also assessed.

Results: At both GAs, ventricular (RV + LV + septum) weight normalized to body weight was significantly increased in anemic fetuses. Anemia had no effect on expression of myocardial total or active Akt, JNK, or p38 at either GA. Levels of total ERK1/2 were also unchanged, although active ERK1/2 was significantly decreased in the late but not early GA anemic fetuses. Total JNK and total and active ERK1/2 and Akt were significantly greater in early versus late GA anemic fetuses. Protein-to-DNA ratios were unchanged in response to anemia at both GAs, but were greater in late GA compared to early GA anemic fetuses.

Conclusion: These results identify important developmental differences in the response of the MAPKs and Akt in the stressed fetal heart. Differences in protein-to-DNA ratios likely reflect the different populations of cardiomyocytes composing the fetal heart at these two GAs and their cell-dependent response to a hemodynamic load.

Objective: The cell membrane water channel protein aquaporins (AQPs) may be important in regulating the intramembranous (IM) pathway of amniotic fluid (AF) resorption. The objective of the present study was to determine whether aquaporin 3 (AQP3) is expressed in human fetal membranes and to further determine if AQP3 expression in primary human amnion cell culture is regulated by second-messenger cyclic adenosine monophosphate (cAMP).

Methods: AQP3 expression in human fetal membranes of normal term pregnancy was studied by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC). To determine the effect of cAMP on AQP3 expression, primary human amnion cell cultures were treated in either heat-inactivated medium alone (control), or heat-inactivated medium containing: (1) SP-cAMP, a membrane-permeable and phosphodiesterase resistant cAMP agonist, or (2) forskolin, an adenylate cyclase stimulator. Total RNA was isolated and multiplex real-time RT-PCR employed for relative quantitation of AQP3 expression.

Results: We detected AQP3 expression in placenta, chorion, and amnion using RT-PCR. Using IHC, we identified AQP3 protein expression in placenta syncytiotrophoblasts and cytotrophoblasts, chorion cytotrophoblasts, and amnion epithelia. In primary amnion epithelial cell culture, AQP3 mRNA significantly increased at 2 hours following forskolin or SP-cAMP, remained elevated at 10 hours following forskolin, and returned to baseline levels by 20 hours following treatment.

Conclusion: This study provides evidence of AQP3 expression in human fetal membranes and demonstrates that AQP3 expression in primary human amnion cell culture is up-regulated by second-messenger cAMP. As AQP3 is permeable to water, urea, and glycerol, modulation of its expression in fetal membranes may contribute to AF homeostasis.

Background: Recently we identified a weak zone in term, pre-labor (repeat Cesarean section) human fetal membranes (FM) overlying the cervix with biochemical characteristics suggestive of apoptosis and collagen remodeling. We suggested that this weak zone is the FM rupture initiation site. Vaginally delivered patients have a weak zone in their FM overlying the cervix; a comparable weak zone lies adjacent to the tear line in FM after spontaneous rupture (SROM).

Methods: FM from vaginally delivered patients with artificial rupture (AROM) and SROM were collected. FM of AROM patients were marked per vagina to identify the FM zone overlying the cervix. Postpartum FM were cut, strength tested, and piece strengths were remapped to their former location on a three-dimensional model. A 10-cm diameter zone centered on the marked area (AROM), or defined weak zone (SROM) was compared with the remaining FM.

Results: AROM FM exhibit a para-cervical weak zone. SROM FM exhibit a comparable zone on the tear line. The mean rupture strength within weak zones was 60% of the remaining membranes (P <.001). AROM and SROM FM weak zones both exhibit increased matrix metalloproteinase 9, increased poly (ADP-ribose) polymerase I cleavage, decreased tissue inhibitor of metalloproteinase 3 protein, and histology consistent with remodeling and apoptosis.

Conclusion: Vaginally delivered AROM FM contain a weak zone overlying the cervix. Vaginally delivered SROM FM contain a weak zone adjacent to the tear line that exhibits biochemical and mechanical characteristics suggestive of collagen remodeling and apoptosis comparable to those of the AR FM weak zone.

Objective: Human chorionic gonadotropin (hCG) has proliferative effects on uterine smooth muscle and leiomyoma tissue in vitro. We hypothesized that luteinizing hormone (LH) would have the same effect by activating the LH/hCG receptor, and it would follow that premenopausal women with higher basal LH levels would be more likely to have leiomyomata.

Methods: Randomly selected women, aged 35 to 49 years, from a prepaid health plan were screened for leiomyomata with pelvic ultrasound. Urine samples collected during the first or last 5 days of the menstrual cycle were analyzed for LH by immunofluorometric assay, and concentrations were corrected for creatinine (n = 523). Logistic regression and Bayes analyses were used to evaluate the association of LH with presence and size of leiomyomata, adjusting for age, and other risk factors.

Results: Women with higher LH were more likely to have leiomyomata (adjusted odds ratios for second and third tertiles were 1.7 and 2.0 compared with lower tertile; 95% confidence intervals, 1.0 to 2.7 and 1.2 to 3.4, respectively). The association was stronger for large leiomyomata. Bayes analyses designed to estimate LH effects on tumor onset separately from tumor growth showed significantly accelerated tumor onset but little evidence of effects on tumor growth. Age, an independent risk factor for leiomyomata, was not affected by inclusion of LH in the logistic models.

Conclusions: As hypothesized, women with higher LH were more likely to have leiomyomata, but this did not explain the age-related increase in leiomyomata during perimenopausal ages. Determining whether LH is causal or a marker for susceptibility will require further research.