Journal of the Society for Gynecologic Investigation最新文献

Objectives: To establish a multiplex amplification refractory mutation system (ARMS) in fluid and dried whole blood, and to perform a pilot study to examine the role for single-nucleotide polymorphisms (SNPs) of inflammation-associated genes (interleukin [IL]-1 and -10, tumor necrosis factor-alpha [TNFA], and toll-like receptor-4 [TLR4]) and their interaction with clinical chorioamnionitis (CAM) in prematurity.

Methods: We established a quadruplex ARMS to detect the four above SNPs. Fifty-four women delivered at gestational age less than 32 weeks and 83 healthy female volunteers were genotyped. We compared (1) mothers of preterm infants with volunteers, and (2) women delivered before 29 weeks' gestation (n = 29) with those delivered at 29 to 31 completed weeks (n = 25).

Results: Multiplex ARMS is feasible using both fluid and dried whole blood. We found no overall differences in genotype and allele frequencies between mothers of preterm infants and volunteers. Among women who had a preterm delivery, those with both CAM and IL10(-1082)*G allele, the risk for delivery before 29 weeks was markedly increased (odds ratio [OR] 22, 95% confidence interval [CI] 2.5 - 191).

Conclusion: The presence of both CAM and IL10(-1082)*G might play a role in extreme preterm delivery less than 29 weeks.

Objective: To test the hypothesis that plasma visfatin concentrations will be lower in women with gestational diabetes mellitus, we evaluated women with gestational diabetes mellitus and healthy pregnant women, and then correlated their plasma visfatin concentrations with body mass index (BMI) and various other parameters.

Methods: A total of 40 women were evaluated: 20 women with gestational diabetes mellitus and 20 healthy pregnant women to serve as control subjects. Plasma visfatin concentrations were analyzed using an enzyme-linked immunosorbent assay.

Results: Plasma visfatin concentrations were significantly lower in the gestational diabetes mellitus group (9.4 +/- 3.8 ng/mL) than in the healthy control group (12.6 +/- 4.5 ng/mL) (P = .023). A negative correlation was found between plasma visfatin concentrations and maternal age (r = -0.399, P = .011), first trimester body weight (r = -0.350, P = .027), and first trimester BMI (r = -0.336, P = .034). Multiple linear regression analysis revealed that maternal age (P = .017) and gestational diabetes mellitus/no gestational diabetes mellitus (P = .044) were independently related to plasma visfatin concentrations. However, no relationship was found with either gestational age at the time of sampling or first trimester BMI.

Conclusions: Our results show that there are decreased concentrations of plasma visfatin in gestational diabetes mellitus subjects and this may indicate that visfatin plays a role in the pathogenesis of gestational diabetes mellitus. However, further experiments are needed to clarify this role.

Objective: Nitric oxide (NO) is synthesized in the brain through the action of three isoforms of nitric oxide synthase (NOS). The local generation of NO in neurons, glia, and vasculature modulates neuronal activity, as well as regional cerebral blood flow. We propose that, in the fetal brain, cerebral hypoperfusion alters the expression of NOS isoforms, and that estrogen administration modulates the NOS response to hypoperfusion.

Methods: Sixteen chronically catheterized fetal sheep of known gestational age (124 to 128 days' gestation) were subjected to a 10-minute period of brachiocephalic occlusion (BCO) or to sham BCO; half of these fetuses were subjected to subcutaneous implant, which released 17beta-estradiol (E2; 0.25 mg/d) or placebo. Brain tissue was collected for mRNA and protein extraction 1 hour after the start of the BCO or sham BCO.

Results: All three isoforms of NOS were identified in fetal brain at both the mRNA and protein levels. BCO increased NOS1 (hippocampus, brainstem), NOS2 (hypothalamus), and NOS3 (hippocampus, cortex) at the protein level. Estradiol alone increased NOS1 (brainstem, cortex), NOS2 (hippocampus, hypothalamus), and NOS3 (brainstem, cerebellum) at the protein level, changes that were not mirrored at the mRNA level. The combination of BCO and estradiol produced smaller changes in NOS1 (brainstem, cortex), NOS2 (hippocampus, hypothalamus), and NOS3 (brainstem) protein levels than those produced by either stimulus alone.

Conclusions: We conclude that the fetal brain expresses all isoforms of NOS, and that NOS expression is altered by both BCO and estradiol, but that the most prevalent effect of estradiol is to reduce specific NOS responses to cerebral hypoperfusion. The present results suggest the possibility that the neuroendocrine responses to estradiol and BCO are modulated by central nervous system (CNS) NO biosynthesis.

Objective: To test the hypothesis that multiple courses of antenatal corticosteroids accentuate the decreases in blood-brain barrier permeability observed after a single course of corticosteroids in preterm ovine fetuses.

Methods: Chronically instrumented 106-day gestation ovine fetuses were studied after single and multiple courses of dexamethasone or placebo were given to ewes beginning at 104 to 106 or 76 to 78 days of gestation, respectively. In the single-course groups, the ewes received dexamethasone (6 mg, n = 6) or placebo (n = 6) as four intramuscular injections every 12 hours over 48 hours. In the multiple course groups, the ewes received the same treatment (dexamethasone, n = 9, or placebo, n = 8), once per week for 5 weeks starting at 76 to 78 days of gestation. Blood-brain barrier permeability was quantified with the blood-to-brain transfer constant (K(i)) for alpha-aminoisobutyric acid (AIB) in the brain regions of the fetuses 12 hours after the last injection of dexamethasone was given to the ewes at 106 to 107 days of gestation.

Results: Both single (analysis of variance [ANOVA]; main effects for dexamethasone treatment, F = 5.92, P <.04) and multiple (ANOVA; main effects for dexamethasone treatment, F = 4.74, P <.04) courses of antenatal corticosteroids were associated with decreases in blood-brain barrier permeability in the brain regions of the ovine fetus. However, the multiple courses did not accentuate (ANOVA; main effects for single versus multiple courses, F = 1.06, P = .32) the decreases in permeability observed after a single course.

Conclusion: Contrary to our hypothesis, antenatal treatment with a 5-week course of corticosteroids did not accentuate the reductions in blood-brain barrier permeability that we observed after a single course of corticosteroids in the fetus.

Objectives: Macrophage migration inhibitory factor (MIF), a multifunctional proinflammatory cytokine, has been recently involved in many aspects of reproduction including pregnancy. However, no evidence is available on the role of MIF in gestational tissues nor on factors regulating MIF production. This study, conducted on explants of human fetal membranes at term gestation, has been undertaken to investigate whether: (1) MIF is produced by fetal membranes; (2) nitric oxide (NO) can regulate local MIF production; and (3) MIF, in turn, can influence NO release in these tissues.

Methods: Tissues were obtained from 56 healthy women who underwent elective cesarean delivery. Fetal membranes have been incubated with either sodium nitroprusside (NP), a NO donor, or recombinant MIF (r-MIF), or a specific anti-MIF antibody (MIF-Ab). Reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot, enzyme-linked immunosorbent assay (ELISA), and colorimetric assay have been used to detect MIF mRNA and protein, inducible nitric oxide synthase (iNOS), and NO metabolites.

Results: Fetal membranes basally express MIF mRNA and protein and release MIF. Exposing tissues to NP results in an increase of MIF mRNA expression and protein release. Conversely, treatment of tissues with MIF is followed by a reduction in iNOS mRNA and protein expression as well as in NO release. These effects are reversed by adding MIF-Ab.

Conclusions: MIF is generated and released by human fetal membranes at term. MIF mRNA and protein expression and release are modulated by NO. MIF, in turn, can reduce iNOS expression and NO release by these tissues. NO could be a regulator of MIF production in pregnancy and labor.

Objective: Levosimendan is a calcium-sensitizing agent and inodilator working via potassium channels, which is under current investigation in the treatment of heart failure. We investigated the type of potassium channels that play a role on the dilatating effect of levosimendan on the contractile tones of the isolated human umbilical artery (HUA).

Methods: The response in the HUA was recorded isometrically by a force displacement transducer in isolated organ baths. Levosimendan was added to organ baths after precontraction with serotonin (5-HT, 1 microM). Levosimendan-induced relaxations were tested in the presence of the large conductance Ca2+-activated K+ channel inhibitor tetraethylammonium (TEA, 1 mM), the adenosine triphosphate (ATP)-sensitive K+ channel inhibitor glibenclamide (GLI, 10 microM), and the voltage-sensitive K+ channel inhibitor 4-aminopyridine (4-AP, 1 mM). All experiments were performed in solutions containing the cyclooxygenase inhibitor indomethacin (10 microM) and the nitric oxide synthase inhibitor L-NAME (100 microM).

Results: Levosimendan (10 nM to 3 microM) produced potent relaxation in the HUA. Vehicle had no significant relaxant effect. The relaxation to levosimendan was not affected by the K+ channel inhibitor, GLI. However, 4-AP (1 mM) and TEA (1 mM) inhibited levosimendan-induced relaxation significantly (P <.05).

Conclusion: These results show that levosimendan effectively and directly decreases the tone of the HUA. The mechanism of this levosimendan-induced relaxation in the HUA appears in part to be due to voltage-gated and large conductance Ca2+-activated K+ channel opening action.

Objective: Fetal brain injury is associated with chorioamnionitis, which is often present without signs of overt infection or fetal compromise. We aimed to determine if prolonged exposure to intrauterine inflammation caused by intra-amniotic infusion of lipopolysaccharide (LPS) would affect the fetal brain.

Methods: At 80 days of pregnancy ewes bearing singletons had osmotic pumps implanted intra-amniotically to infuse Escherichia coli LPS (055:B5; n = 8) or saline (n = 7) for 28 days. At delivery (110 days), umbilical arterial blood and chorioamnion were assessed for inflammation; cytokine concentrations (interleukin [IL]-6 and IL-8) in amniotic fluid and fetal and maternal plasma were measured. The fetal cerebral hemispheres were examined for gross anatomical changes and the number of activated microglia/macrophages, astrocytes, and oligodendrocytes estimated after immunohistochemical staining.

Results: Intra-amniotic administration of LPS caused chorioamnionitis, fetal leucocytosis, and a moderate to extensive infiltration of activated microglia/macrophages in the subcortical white matter in six of eight fetuses; the remaining two fetuses were less affected. Within these focal regions of damage there was an attenuation of astrocytic processes, axonal injury, and a reduction in the number of 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) immunoreactive oligodendrocytes in areas of extensive focal damage. In control fetuses there was mild (3/7) or no infiltration of activated microglia/macrophages in the subcortical white matter. Overall the infiltration of activated microglia/macrophages in the white matter was significantly greater in LPS-exposed fetuses compared to controls. In regions devoid of injury, the number of oligodendrocytes and astrocytes was not different between groups, nor was there a difference in the volume of cerebral white matter or density of blood vessels within the white matter. Amniotic fluid IL-6 and IL-8, and maternal plasma IL-8 concentrations were significantly increased by LPS infusion.

Conclusions: An increase in inflammatory cells and axonal disruption in the subcortical white matter of the fetal brain can accompany chorioamnionitis induced by intra-amniotic administration of LPS, but cystic lesions do not occur. Thus, the effect on the fetal brain is milder than that reported from animal models of acute fetal/intrauterine infection.

Objective: This study evaluated the secretions of zymogen and active forms of matrix metalloproteinase (MMP)-9 and MMP-2 and their specific inhibitors, TIMP-1 and TIMP-2 by fetal membranes stimulated with group B Streptoccocci (GBS).

Methods: We used an in vitro experimental model that allowed us to estimate the individual contribution of the amnion (AM) and the choriodecidua (CHD) to the microbial insult. Membranes were obtained after delivery by elective cesarean delivery from women at 37 to 40 weeks of gestation without evidence of either active labor or intrauterine infection. Membranes were mounted in Transwell devices (Costar, New York, NY), physically separating the upper and lower chambers; 1 x 10(6) CFU of GBS was added to either AM or CHD and the secretions and gelatinolytic activity of MMP-2 and MMP-9 were measured in both compartments by enzyme-linked immunosorbent assay (ELISA) and zymography. TIMPs secretion was measured by ELISA. Both MMPs were immunolocalized in tissue sections.

Results: The simultaneous stimulation at both sides was followed by increases of proMMP-9 (85.0 +/- 18.63 pg/mL) and proMMP-2 (4.10 +/- 1.90 ng/mL) in the CHD (P <.05). When only one side of the membrane was stimulated, the secretion level of proMMP-2 increased 2.3-fold and that of proMMP-9 2.5-fold in the CHD. The active forms of both enzymes did not change with any modality of stimulation. The secretion level of both TIMPs remained without significant changes. CHD and AM were positive for immunoreactive MMP-2 and MMP-9.

Conclusion: We propose that infection of fetal membranes with GBS is followed by active secretion of MMP and the CHD is the principal source of these mediators of extracellular matrix degradation.

Objective: The study's aim was to determine any association between choroid plexus cysts (CPCs) and trisomy 18 in a population of fetuses previously screened by nuchal translucency (NT) thickness measurement.

Methods: During the study period (May 1999 to December 2004), 7,795 fetuses had an NT scan and second-trimester fetal anatomical scan at our institution. The prevalence of trisomy 18 was determined among four types of pregnancies: 1) those with isolated CPCs, 2) those with CPCs and enlarged NT, 3) those with CPCs and other ultrasound markers, and 4) those with CPCs, enlarged NT, and other ultrasound markers. The fetal outcome according to NT and presence of CPCs was calculated. Incidence rates of enlarged NT and CPCs in fetuses with trisomy 18 and fetuses with normal chromosomes were also evaluated.

Results: For the entire population, ten trisomy 18 cases were diagnosed prenatally (prevalence, 0.13%). Among fetuses with enlarged NT, the likelihood ratio of trisomy 18 was significantly increased in fetuses with CPCs compared with fetuses without such cysts (333.6 versus 15.2, P = .002). However, among fetuses with normal NT, no significant difference was demonstrated for likelihood ratio of trisomy 18 between fetuses with and without CPCs.

Conclusion: In pregnancies complicated by isolated CPCs, fetal karyotyping is not indicated when no additional anomaly is detected on ultrasonographic examination and first-trimester NT results are normal.

Objective: Hemocidins are a novel class of antibacterial peptides generated proteolytically from hemoglobin. These peptides play a particularly important role in maintaining vaginal homeostasis during menstrual bleeding. To investigate the hemoglobin fragmentation process during the last stages of pregnancy, we examined uterine secretion (lochia) samples from a group of 22 healthy women who underwent cesarean delivery at term.

Methods: Patients were divided into three groups: (1) the elective cesarean deliveries without symptoms of spontaneous labor, (2) the nonelective cesarean deliveries with spontaneous beginning of labor, and (3) the nonelective cesarean deliveries during advanced labor. The samples were subjected to chromatographic estimation of free hemoglobin and peptides. In three representative patients the identification of all lochial peptides was performed.

Results: All samples contained a significant amount of free hemoglobin and its level increased with labor progression. The presence of peptide fractions was also detected in most lochia samples. They were confirmed to be human hemoglobin fragments, almost identical to the recently described bactericidal hemocidins from menstrual discharge. The level of peptides also increased during labor. The subgroup with advanced labor demonstrated the highest amount of hemocidins.

Conclusions: The presented results prove that proteolysis of free hemoglobin in the female upper reproductive tract begins together with the clinical symptoms of normal labor. We speculate that cesarean delivery affects molecular mechanisms involved in antibacterial hemocidins generation and, in effect, might be responsible for the increased risk of gynecologic infections in cesarean deliveries.