CA 125 in seminal plasma: correlation with semen parameters

Ovarian cancer marker CA 125 was measured in human seminal plasma,and the concentrations ranged between 22 and 1149 U/ml, andbetween 39 and 4711 U/ejaculate. This very high patient-to-patientvariability was in contrast to a much lower within-patient variability,which was comparable to that of other semen parameters. No significantdifferences in CA 125 concentration were found in seminal plasmafrom normospermic patients, patients with male factors, vasectomizedmen, and in aliquots of samples which led to a pregnancy, viaartificial insemination or in-vitro fertilization. The seminalplasma CA 125 concentration was not correlated with sperm count,motility and morphology. In contrast, seminal plasma CA 125concentrations correlated with the age of the patient (P <0.001) and inversely with the volume of the ejaculate (P <0.001). These correlations were independent of each other. CA125 did not correlate with the prostatic marker zinc, but diddo so with the seminal vesicle marker fructose and the epididymalmarker carnitine.     

Effects of tumour necrosis factor-alpha, interleukin-1 alpha, macrophage colony stimulating factor and transforming growth factor beta on trophoblastic matrix metalloproteinases.

The aim of this study was to determine the effects of tumour necrosis factor alpha (TNF), interleukin-1 alpha (IL-1alpha), macrophage colony-stimulating factor (MCSF) and transforming growth factor beta (TGFbeta) on the secretion of matrix metalloproteinases (MMP), human chorionic gonadotrophin (HCG) and fetal fibronectin (fFN) by purified first trimester cytotrophoblastic cells (CTB) in vitro. CTB were obtained from legal abortions and cultured in vitro in the presence or absence of the different cytokines. Secreted gelatinases were analysed in the culture supernatants by zymography, by measurements of the total gelatinolytic activity and by enzyme immunoassays. HCG and fFN were measured by commercially available immunoassays. TNF increased the total gelatinolytic activity by increasing MMP-9 activity (P = 0.025-0.0177) but decreased MMP-2 activity (P < 0.03) and immunoreactivity (P < 0.05), fFN (P < 0.02) and HCG (P < 0.01). IL-1alpha significantly increased the secretion of fFN (P < 0.02), the activity (P < 0.02) and immunoreactivity (P < 0.05) of MMP-9 but had no effect on the other parameters. MCSF increased MMP-9 immunoreactivity (P < 0.05) and moderately decreased HCG. TGFbeta inhibited total gelatinolytic activity, MMP-9 activity and immunoreactivity, but was without effect on MMP-2 concentrations and activity. TGFbeta decreased HCG (P < 0.041) and increased fFN (P < 0.042). Our results indicate that TGFbeta, TNF and IL-1alpha are important regulators of trophoblastic MMP secretion.     

Status of p53 in first-trimester cytotrophoblastic cells

p53 has been called the cellular gatekeeper of the genome because it can induce cell-cycle arrest in G1, apoptosis or affect DNA replication in response to DNA damage. As p53 has been observed in first-trimester cytotrophoblastic cells (CTB), but its expression in normal cells is generally not detectable because of its short half-life, p53 could play an important role in cellular differentiation and/or in the control of the invasion of trophoblastic cells; therefore, p53 status was investigated in these cells. Using different antibodies recognizing different epitopes of p53 protein, abundant p53 expression was observed both in nuclear and in cytoplasmic compartments of first-trimester CTB. Whereas p53 was detected in the nuclei of few trophoblastic cells with an antibody recognizing the N-terminal epitope of the protein, high expression level of p53 in the cytoplasm of CTB was detected with an antibody recognizing the middle part of p53. The lack of immunoreactivity of p53 with antibodies recognizing the epitopes located at the N-terminus of p53 and the high level of p53 protein observed in the cytoplasm of CTB suggest that the N-terminus of p53 is involved in the formation of complexes. These cytoplasmic complexes were detected under non-reducing conditions in western blot analysis and had apparent molecular weights (MW) of 195, 167 or 125 kDa. These complexes could prolong the half-life of p53 in the cytoplasm of CTBs. By contrast, in the nuclei of CTBs, p53 seems to be present as a tetramer.     

Immunological heterogeneity of pregnancy-associated plasma protein-A (PAPP-A). Effects on the radioimmunoassay of PAPP-A

Pregnancy-associated plasma protein-A (PAPP-A) is a macromolecular glycoprotein produced by the trophoblast during pregnancy. Because the presence of PAPP-A in non-pregnant females is controversial, we re-evaluated our own radioimmunoassay technique for PAPP-A in the light of new observations about its immunological heterogeneity. Irrespective of the antibody (Geneva anti-PAPP-A or Dako anti-PAPP-A) the use of EDTA pregnancy plasma instead of pregnancy serum as a standard yielded different slopes of the standard curves and estimated significantly different amounts of PAPP-A in test samples. Moreover, highly purified tracers, isolated from maternal EDTA pregnancy plasma or pregnancy serum also produced significantly different results. So that the use of a tracer purified from EDTA plasma and EDTA pregnancy plasma as a standard will yield measurable levels of PAPP-A in non-pregnant female serum whereas the use of a tracer purified from maternal pregnancy serum and maternal pregnancy serum as a standard will not detect PAPP-A in the same samples. We conclude that, irrespective of the antiserum used, but depending on the biological origin of the tracer and the standard, different results will be obtained. PAPP-A is clearly immunologically heterogeneous, and the immunorecognition of PAPP-A will depend on whether or not blood coagulation has taken place.     

Placental protein 5 (PP5) inhibits thrombin-induced coagulation of fibrinogen

The effect of Placental Protein 5 (PP5) on thrombin-induced coagulation of diluted fibrinogen is described. In contrast to previous reports which failed to demonstrate antithrombin-like activity for PP5 on synthetic substrates, we show in this study that PP5 inhibits thrombin activity in a dose-dependent manner. PP5 acts thus in a very similar way to antithrombin III (ATIII). Although PP5 binds to heparin, it does not show any heparin-cofactor activity. Whereas catalytic amounts of heparin accelerate greatly the inhibitory effect of ATIII, no accelerating effect of heparin on PP5 could be observed under the same conditions. In spite of its anti-thrombin activity, PP5 cannot be considered as a pregnancy-analogue of ATIII, as it lacks the heparin-cofactor activity.     

Effects of interleukin-6 (IL-6) on cytotrophoblastic cells.

Tumour invasion and trophoblastic invasion share the same biochemical mediators: the matrix metalloproteinases (MMP) and their inhibitors. In contrast to tumour invasion of a host tissue, trophoblastic invasion during implantation and placentation is stringently controlled both in tissue localization and developmental stage. The factors responsible for these important regulatory processes are unknown, but in-vitro studies point to endometrial cytokines and growth factors as possible candidates. Here we examined the possibility that interleukin-6 (IL-6), a trophoblastic and endometrial cytokine, represents such a regulatory factor. Purified first trimester cytotrophoblastic cells (CTB) were cultured for 4 days in presence or absence of increasing concentrations of IL-6. MMP-2 and MMP-9 bioactivity (zymography) and immunoactivity were measured in the culture supernatants together with total human chorionic gonadotrophin (HCG), fetal fibronectin (FFN) and leptin. IL-6 did not change the cytotrophoblastic secretion of FFN or total HCG. In contrast, this cytokine induced a dose-dependent stimulation of the leptin secretion and increased the activity, but not the immunoreactivity, of MMP-9 and MMP-2. These results indicate that IL-6 could be considered as an endometrio-trophoblastic regulator of cytotrophoblastic gelatinases.     

Accuracy of single measurements of pregnancy-associated plasma protein-A, human chorionic gonadotropin and progesterone in the diagnosis of early pregnancy failure

Background: Circulating human chorionic gonadotropin (hCG) and progesterone are commonly used as markers of abnormal pregnancies. Previous studies have shown that pregnancy-associated plasma protein-A (PAPP-A) was also depressed in extrauterine pregnancies (EUP). Previously, PAPP-A was measured with polyclonal antibodies which were later shown to recognise also the pro-form of major basic protein (pro-MBP). Objective: To evaluate the clinical usefulness of PAPP-A measurements in early pregnancy. Study Design: Circulating PAPP-A, hCG and progesterone were measured in patients with EUP (n=68), abnormal intrauterine pregnancies (abIUP, n=31) and normal intrauterine pregnancies (nIUP, n=72). Gestational age was 30–70 days from the last menstruation. Results: For PAPP-A and hCG, a steep increase was observed from day 30 after last menstrual period onwards, this increase being much less important for abIUP and EUP. The values of PAPP-A and hCG were significantly decreased in abIUP and EUP, from 42 days after LMP onwards. There were no significant differences between abIUP and EUP. Progesterone concentration does not vary with amenorrhoea and was significantly lower in abIUP and EUP. Values in abIUP were significantly (P=0.02) lower compared with EUP for amenorrhoea above 42 days. ROC curves were constructed for amenorrhoea above 42 days. For a specificity of 99%, the sensitivity of PAPP-A, hCG and progesterone were 64.5, 93.3 and 76%, respectively. The threshold values were 14.3 mIU/l, 10,400 IU/l and 10.1 ng/ml for PAPP-A, hCG and progesterone. Conclusion: We confirm the decrease of PAPP-A concentrations in pregnancy failure, but hCG and progesterone remain the best clinical tools.     

Control of MMP-9 expression at the maternal-fetal interface

Cytotrophoblastic cells (CTB) from first trimester placenta form columns of invasive CTB. This invasive behaviour is due to the ability of CTB to secrete matrix metalloproteinases (MMPs) since tissue inhibitor of MMP (TIMP) inhibits their invasiveness. Although CTB behave like metastatic cells, in vivo they are only transiently invasive (first trimester) and their invasion is normally limited only to the endometrium and to the proximal third of the myometrium. This temporal and spatial regulation of trophoblast invasion is believed to be mediated in an autocrine way by trophoblastic factors and in a paracrine way by uterine factors. Several types of regulators have been investigated: hormones, extra-cellular matrix glycoproteins and cytokines or growth factors. This review is not intended to be an exhaustive catalogue of potential regulators of trophoblastic MMP-9 secretion but is aimed at summarising the most important signalling pathways involved in MMP-9 regulation.     

Can we improve the diagnosis of rupture of membranes? The value of insulin-like growth factor binding protein-1

Our objective was to assess the value of insulin-like growth factor binding protein-1 (IGFBP-1) and other tests for the diagnosis of rupture of the membranes (ROM). We included 49 women with suspected ROM. The gold standard for membranes status was defined based on clinical examination, ultrasonography, tests results (except IGFBP-1) and labour information. Sensitivity, specificity, positive predictive value and negative predictive value of each test were as follows, respectively: IGFBP-1 (86, 74, 73 and 87%); bromothymol (64, 100, 100 and 77%); fern test (62, 96, 93 and 75%) and ultrasonography (19, 100, 100 and 61%). The detection of IGFBP-1 in vaginal secretions has high sensitivity for the diagnosis of ROM.