Matrix metalloproteinase-8 in cervical fluid in early and mid pregnancy: relation to spontaneous preterm delivery.

OBJECTIVE: To evaluate whether matrix metalloproteinase-8 (MMP-8) concentrations in cervical fluid in early and mid pregnancy are associated with subsequent preterm delivery (PTD) preceded by premature preterm rupture of membranes (PPROMs) or preterm labour (PTL) with intact membranes. METHODS: Cervical swab samples were collected from 5180 women in early and mid pregnancy. MMP-8 was determined by immunofluorometric assay (IFMA). The outcome measure was spontaneous PTD at < 37 weeks' gestation. RESULTS: The overall distribution and the median cervical fluid MMP-8 concentrations in early and mid pregnancy did not differ in women with term delivery and those with subsequent PTD. However, cervical fluid MMP-8 levels were lower in mid pregnancy in women with PTD preceded by PPROM at < 37 weeks as compared with women who delivered at term and women who had PTD initiated by spontaneous onset of labour (p = 0.016 and p = 0.023, respectively). CONCLUSION: Our data suggest that molecular mechanisms underlying PTL and PPROM differ and MMP-8 in cervical fluid may reflect different functions of this protease. Due to remarkable overlapping of cervical fluid MMP-8 values, this molecule may not have clinical applicability as a biomarker in cervical fluid at least among asymptomatic women in early and mid pregnancy.

Factors affecting matrix metalloproteinase-8 levels in the vaginal and cervical fluids in the first and second trimester of pregnancy.

BACKGROUND: Cervical ripening during pregnancy resembles an inflammatory process. Matrix metalloproteinases (MMPs), particularly MMP-8, have been linked to inflammatory processes. We studied the concentrations of, and factors associated with, MMP-8 in the lower genital tract fluids in the first and second trimesters. METHODS: In a prospective population-based cohort study, vaginal and cervical swab samples were obtained from 2130 unselected pregnant women undergoing their first and second trimester ultrasound screening. MMP-8 was determined by immunofluorometric assay. Use of antibiotics, history of vaginal bleeding, and history of sexual intercourse were recorded on both occasions. Vaginal smears were obtained for Gram-staining and leukocyte counting. Cervical length was measured by ultrasonography. The main outcome measures were MMP-8 concentrations in the vagina and cervix. RESULTS: The median (range) MMP-8 concentrations in vaginal and cervical samples were 107.4 microg/l (undetectable-2406.6 microg/l) and 318.3 microg/l (0.1-2074.6 microg/l), respectively, in the first trimester, and 112.5 microg/l (undetectable-2093.4 microg/l) and 344.8 microg/l (0.4-1783.5 microg/l), respectively, in the second trimester. Multiparity and vaginal leukocytosis were both associated with increased MMP-8 concentrations in vaginal and cervical samples in both trimesters. Bacterial vaginosis (BV) was associated with increased vaginal and cervical MMP-8 in the first trimester, but only with increased vaginal MMP-8 in the second trimester. A history of sexual intercourse (in the previous 48 h) was associated with lower MMP-8 concentrations in cervical samples in both trimesters. CONCLUSIONS: MMP-8 concentrations were lower in vaginal samples than in cervical samples, and no difference was found between the first and second trimesters. Multiparity, BV and an elevated leukocyte count in the vagina were associated with increased MMP-8 concentrations. Sexual intercourse had an opposite effect. The study suggests that MMP-8 is a physiologic constituent in lower genital tract fluids, where it may be involved in host response to inflammatory and infectious processes.

Cervical length measurement and cervical phosphorylated insulin-like growth factor binding protein-1 testing in prediction of preterm birth in patients reporting uterine contractions.

OBJECTIVES: To evaluate the performance of cervical phosphorylated insulin-like growth factor binding protein-1 (phIGFBP-1) testing and cervical length measurement separately and in combination with physician's clinical judgment in prediction of preterm birth among patients with self-reported uterine contractions and intact membranes. DESIGN: We enrolled a total of 246 women between 22 and 34 weeks of gestation. METHODS: The initial evaluation included cervical length measurement using transvaginal ultrasonography. Short cervix was defined as <25 mm. A swab sample was obtained from the cervix for phIGFBP-1. Admission was used as a clinical marker of an increased risk of preterm delivery

Intrauterine release of progesterone antagonist ZK230211 is feasible and results in novel endometrial effects: a pilot study.

BACKGROUND: Continuous administration of progesterone antagonists (PAs) results in endometrial suppression and amenorrhoea in several model systems. We compared the effects of intrauterine release of a highly specific PA, ZK230211, to those of a progestin using the levonorgestrel-releasing intrauterine system (LNG-IUS). METHODS: Forty-two women were randomly fitted with an IUS releasing either ZK230211 at a rate 1, 4 or 8 microg/24 h (ZK-IUS) or LNG (at 20 microg/24 h, LNG-IUS) at 4-8 weeks before hysterectomy. Bleeding patterns, endometrial morphology and content of ZK230211, and various immunohistochemistries (IHCs) were evaluated. RESULTS: Days of bleeding and spotting were unchanged by the use of ZK-IUSs but were increased by LNG-IUS (P < 0.01). ZK230211 was measurable in all endometrial specimens. Endometrium was partly suppressed in 9-30% of women following the use of ZK-IUSs, and in 67% after LNG-IUS. IHCs for Ki-67 and phosphorylated histone H3 were not suggestive of proliferative activity in any group. Compared to LNG, progesterone receptor (PR) was increased following ZK230211 in surface epithelium (all three doses P < 0.01-P < 0.05) and stroma at 4 microg/24 h (P < 0.05). Although low, androgen receptor staining was higher in endothelial epithelium following LNG than ZK230211 (P < 0.05). Insulin-like growth factor-binding protein-1 (IGFBP-1) was detectable only following LNG (P < 0.0001). CONCLUSIONS: Short-term intrauterine release of ZK230211 did not change bleeding patterns or result in endometrial suppression. Expression of proliferation markers was low following the use of both IUSs. Absence of IGFBP-1 and increase in PR reflect the PA effects of ZK230211.

The insulin-like growth factor system and Type 1 diabetic retinopathy during pregnancy.

AIMS/HYPOTHESIS: To find out whether the levels of insulin-like growth factor-I (IGF-I), IGF binding protein-1 (IGFBP-1), highly phosphorylated IGFBP-1 (hpIGFBP-1), and IGF binding protein-3 (IGFBP-3) are related to the progression of diabetic retinopathy (DR) during pregnancy and postpartum. METHODS: In a prospective study of 42 pregnant women with Type 1 diabetes and 9 nondiabetic controls, DR was graded from fundus photographs. Levels of serum total IGF-I and two different phosphoisoform patterns of IGFBP-1 and IGFBP-3 were measured during the first and third trimester of pregnancy and 3 months postpartum. RESULTS: Both the levels of serum total IGF-I (P<.0001) and IGFBP-3 (P=.003) were lower in the diabetic than in the nondiabetic women during pregnancy and postpartum (repeated-measures ANOVA between the groups). Additionally, the IGF-I and IGFBP-3 levels tended to be lower in the diabetic women with more severe DR at baseline than in those with less severe DR. There were no statistically significant differences in the levels of IGF-I and IGFBP-3 in the diabetic women with progression of DR compared with those without. No statistical differences appeared in the IGFBP-1 phosphoisoform patterns between the groups. CONCLUSIONS/INTERPRETATION: In diabetic women, mean serum levels of IGF-1 and IGFBP-3 are lower than in nondiabetic controls during pregnancy and/or postpartum. Because there was no clear connection between the IGF system and progression of DR during pregnancy, it is unlikely that these substances mediate the tendency of DR to progress during pregnancy.

Lack of effect of isoflavonoids on the vagina and endometrium in postmenopausal women.

OBJECTIVE: To determine the effects of soy-derived isoflavones on vaginal epithelium and the endometrium. DESIGN: Double-blind, randomized, placebo-controlled crossover trial. SETTING: Outpatient clinic of a university hospital. PATIENT(S): Sixty-four postmenopausal women with a history of breast cancer. INTERVENTION(S): The women took (in a randomized order) 114 mg of isolated isoflavonoids or placebo in tablets daily for 3 months; the treatment regimens were crossed over after a 2-month washout period. The subjects were studied before and on the last day of each treatment period. MAIN OUTCOME MEASURE(S): Vaginal dryness, maturation index (MI) of vaginal epithelium, endometrial thickness, histology, and expression of estrogen (E) and progesterone (P) receptors and the proliferation marker Ki-67 in the endometrium. RESULT(S): Isolated isoflavones did not relieve vaginal dryness. Maturation index values remained unchanged during the isoflavone regimen, but decreased during the placebo regimen. No changes were found in any of the variables measured in the endometrium. CONCLUSION(S): Daily administration of 114 mg of isolated isoflavones for 3 months had no effect on the subjective perception of vaginal dryness or on objective findings in the vagina or endometrium. This implies safety with regard to the endometrium.

Differential kinetics of serum and cervical insulin-like growth factor-binding protein-1 during mifepristone-misoprostol-induced medical termination of early pregnancy.

The kinetics of cervical and circulating phosphoisoforms of insulin-like growth factor-binding protein-1 (IGFBP-1) in normal and pathological early pregnancy are not well known. We investigated the profiles of IGFBP-1 in serum and in cervical secretion during medical termination of early pregnancy. Sixteen women requesting termination of pregnancy, with <63 days of amenorrhoea, received 200 mg of mifepristone on day 0, followed by either oral or vaginal administration of 0.8 mg of misoprostol on day 2. Serum and cervical swab samples, collected up to 6 weeks following the beginning of the treatment, were analysed for IGFBP-1 using two immunoenzymometric assays recognizing different patterns of IGFBP-1 phosphoisoforms. Serum mifepristone was also assayed. In the cervical samples, IGFBP-1 concentration, measured with both assays, increased substantially 2 days following administration of mifepristone. At 3 h after administration of misoprostol, IGFBP-1 had further increased several-fold in the cervix, but the increase was more pronounced as measured by the assay with preference for the amniotic fluid isoforms of IGFBP-1. A strong negative correlation was found between the time to abortion and the increase in cervical IGFBP-1 after administration of misoprostol, as measured by the assay preferring the phosphorylated isoforms of IGFBP-1. At 6 weeks, IGFBP-1 in the cervix had decreased to lower than pre-treatment levels, as measured by both assays. In serum, both assays showed a significant increase in IGFBP-1 concentrations after administration of mifepristone, and the highest values were measured on day 2, already before misoprostol administration. Thus, the kinetics of circulating and cervical IGFBP-1 differed from each other, indicating different sources and regulation of serum and cervical IGFBP-1.

Effects of ospemifene, a novel SERM, on hormones, genital tract, climacteric symptoms, and quality of life in postmenopausal women: a double-blind, randomized trial.

OBJECTIVE: Ospemifene, a novel selective estrogen receptor modulator, shows a potential for prevention and treatment of osteoporosis in postmenopausal women. We studied the effects of ospemifene on hormone levels, genital tract organs, climacteric symptoms, and quality of life. DESIGN: A double-blinded study in which 160 postmenopausal women were randomly allocated to receive either ospemifene at three different daily doses (30, 60, or 90 mg) or placebo for 3 months. RESULTS: No significant differences were observed among the study groups in clinical characteristics or parameters reflecting estrogen action at baseline. Ospemifene reduced follicle-stimulating hormone and insulin-like growth factor I levels, whereas estradiol failed to change at all, and luteinizing hormone was reduced only in the 90-mg group of ospemifene. In the vast majority of participants, the endometrium remained atrophic after 3 months of treatment with ospemifene. Although the rate of proliferative endometrium slightly increased in all groups, including placebo, no hyperplasia or bleeding occurred in any participant. Ospemifene had no effect on the appearance of proliferation marker Ki-67 in the endometrium as compared with placebo, and endometrial thickness increased by mean 0.4 to 0.6 mm (P < 0.01, P < 0.05 and P < 0.05 for 30, 60 and 90 mg ospemifene, respectively). Uterine volume slightly increased (8.4%-14.7%) in the ospemifene groups (P > 0.05), perhaps as a result of increased uterine blood flow. The most conspicuous finding was the significant estrogenic effect on vaginal epithelium, as evidenced by an increase in intermediate and superficial cells in repeat Pap smears. Ospemifene was not observed to aggravate climacteric symptoms or cause adverse events, nor did it suppress climacteric symptoms. CONCLUSIONS: Ospemifene at daily doses of 30 to 90 mg did not stimulate endometrium or aggravate hot flashes but clearly had a rather strong estrogenic effect on the vaginal epithelium during a 3-month treatment period. Such effects would be advantageous if ospemifene were found to be effective in the long-term prevention of osteoporosis.

Effects of ospemifene, a novel SERM, on vascular markers and function in healthy, postmenopausal women.

OBJECTIVE: Ospemifene, a novel selective estrogen receptor modulator (SERM), shows promise for bone preservation in postmenopausal women. This study examined the effects of ospemifene on different vascular surrogate markers. DESIGN: A double-blinded study was conducted in 160 healthy, postmenopausal women who used, in a randomized order, ospemifene (at daily doses of 30, 60, or 90 mg) or placebo for 3 months. RESULTS: Although ospemifene caused falls from basal levels in total cholesterol, low-density lipoprotein cholesterol, oxidized low-density lipoprotein cholesterol, and a rise in high-density lipoprotein cholesterol, the only statistically significant difference between ospemifene and placebo was an increase of triglyceride levels (11.3%) in the 90-mg group. Ospemifene caused no significant effect on endothelial markers or homocysteine. Of the markers reflecting coagulation and fibrinolysis, plasma fibrinogen was significantly reduced in the 60- and 90-mg groups of ospemifene (8.7% and 8.5%, respectively) when compared with the placebo group. No changes were seen in generation of thrombin or degradation of crosslinked fibrin D-dimer. The uterine or carotid arteries and 24-h ambulatory blood pressure were not affected by ospemifene. Ospemifene caused no changes in basal insulin or in a 2-h glucose tolerance test, suggesting unaltered insulin sensitivity. CONCLUSIONS: Neutral effects of short-term use of ospemifene on vascular surrogate markers imply no effect for ospemifene on the risk for cardiovascular disorders in healthy, postmenopausal women.

IGF-I, IGF binding protein (IGFBP)-3, phosphoisoforms of IGFBP-1, and postnatal growth in very low birth weight infants.

Impaired postnatal growth in very low birth weight (VLBW, <1500 g) infants is per se a major clinical challenge and may also serve as a model in studying the mechanisms of growth retardation in general. This study was undertaken to characterize the role of IGFs and their binding proteins (IGFBPs), key regulators of fetal and infant growth, during the postnatal period in VLBW infants. Forty-eight VLBW infants (gestational age 27.6 +/- 2.2 wk, birth weight 923 +/- 257 g) were studied. Blood samples were drawn at 1, 2, 4, and 8 wk of age for measurements of IGF-I, IGFBP-1 (lesser phosphorylated, lpIGFBP-1, and highly phosphorylated, hpIGFBP-1), IGFBP-3, and insulin, simultaneous growth velocities being assessed by a rigorous protocol of repeated, frequent lower leg length and body weight measurements. All regression analyses were adjusted for postnatal age and repeated measurements. Lower leg growth velocity showed a positive correlation with IGF-I (P = 0.01) and IGFBP-3 (P = 0.03), and weight growth velocity with IGFBP-3 (P = 0.057) and with lpIGFBP-1/hpIGFBP-1 ratio (P = 0.01). Moreover, concurrent glucocorticoid dose showed a negative correlation with both IGFBP-1 isoforms, observable, however, only in samples with high (>10 U/liter) insulin (lpIGFBP-1, P = 0.02; hpIGFBP-1, P = 0.007). In backward multiple regression analysis, the factor remaining significantly associated with lower leg growth velocity (R(2) = 0.63) was IGF-I, and factors associated with weight growth velocity (R(2) = 0.81) were IGFBP-3 and the lpIGFBP-1/hpIGFBP-1 ratio. In conclusion, circulating IGF-I and IGFBP-3, and the lpIGFBP-1/hpIGFBP-1 ratio, reflect short-term growth velocity in VLBW infants. lpIGFBP-1 isoforms, abundant in the circulation of these infants, may thus also have properties that are at least less inhibitory, if not promoting, on the growth-stimulating action of IGF-I. Finally, the regulation of IGFBP-1 by glucocorticoids may be divergent in situations with a high or low insulin concentration.

Immunohistochemical localization of the insulinlike growth factor binding protein-1 in female reproductive tissues by monoclonal antibodies.

Human insulinlike growth factor binding protein-1 (hlGFBP-1) is a secretory protein that modulates the receptor-binding and biological actions of the insulinlike growth factor I (IGF-I). Human endometrium expresses the mRNA for IGFBP-1, and this protein is secreted by the secretory phase and pregnancy endometrium as well as by ovarian granulosa cells in vitro. In this study, we examined the cellular localization of IGFBP-1 in female reproductive tissues by using a purified monoclonal antibody Mab 6303 with an immunoperoxidase technique. Proliferative- and early secretory-phase endometrium as well as all extrauterine tissues except decidualized cells at the implantation site on the ovaries of ovarian pregnancies were negative for IGFBP-1. In midsecretory-phase endometrium, focal staining was first observed in the cytoplasm of glandular epithelial cells, with weaker staining in the stromal cells. In late secretoryphase endometrium, strong immunostaining was observed in predecidualized stromal cells, with weak focal staining remaining in some of the glandular epithelial cells. In early pregnancy, intense staining was detected in the cytoplasm of decidualized stromal cells of zona compacta in each sample, whereas the nondecidualized stromal cells remained unstained. Strong to medium staining was detected simultaneously in the glandular epithelial cells in 70% of the early pregnancy specimens. In term pregnancy, IGFBP-1 was localized in decidual cells of placental bed and decidua parietalis. Immunolocalization of IGFBP-1 to both endometrial epithelial and stromal cells, although only stromal cells express the gene of IGFBP-1 [14], supports the hypothesis of paracrine actions between these cells. The localization of IGFBP-1 to decidualized cells at the extrauterine implantation sites implies its association with decidual differentiation.