.4bstraa 443

decidua both at term and in the first trimester. Immunohistology demonstrated a positive reaction with an&TNF antibodies in the decidua from first trimester and term samples, and strong binding to the cytotrophoblast layer of term amniochorion.

Analysis of cell dispersions of term decidua by flow cytometry using antibodies to TNF identified a small percentage of cells binding TNF. Similar experiments with dispersions of term amniochorion (n = 5) showed 8 per cent [range 4-14 per cent] of cells to bind TNF.

These results indicate that only a small number of cells at the maternofetal interface are able to bind TNF and thus be potential targets of its action.

ISOLATION AND CHARACTERIZATION OF BASIC FIBROBLAST GROWTH FACTOR FROM FIRST TRIMESTER TROPHOBLAST M. VuEkovid, V. Niketid” & 0. GenbaEev (Institute for the Application of Nuclear Energy, Zemun and “Faculty of Science, Department of Chemistry, University of Belgrade, Yugoslavia)

The presence of basic fibroblast growth factor (bFGF) was demonstrated in mid-gestation and term placenta by Gospodarowicz et al, 1985, Biochem. Biophys. Res. Commun., 128,554- 562. In this study, first trimester placenta was used because it is the purest available source of intact trophoblast tissue, and because the results might contribute to a better understanding of the role of bGFG in human placenta.

We directly applied first trimester placental cytosol fraction to heparin affinity chroma- tography. Molecular weight determination and identification were carried out using SDS gel electrophoresis and Western blottingwith commercially available antibodies to bGFG (R&D Systems Inc., USA). The presence of FGF was confirmed biochemically and biologically in a mitogenic assay on 3T3 cells and then compared with the commercially available FGF (Sigma Chemical Co., USA).

Polyclonal antibodies to bFGF in Western blots revealed immunoreactivity of higher molecular weight protein bands. These higher molecular weight forms did not show heparin binding affinity. Larger molecules of bFGF have also been isolated from other tissues.

Further studies will be directed towards purification and identification of biological activity of these higher molecular weight forms using first and third trimester placental tissue.

CORTICOTROPHIN-RELEASING FACTOR, GLUCOCORTICOIDS AND GLUCOCORTICOID RECEPTOR STATUS IN PATIENTS WITH PREGNANCY INDUCED HYPERTENSION J. Wacker, C. Maser-Gluth, G. Schreiber, P. Vecsei, W. Stolz & G. Bastert (Department of Obstetrics and Gynecology, University of Heidelberg, Germany)

In the present study we examined the role of corticotrophin-releasing factor (CRF), cortisol, and corticosterone, as well as the glucocorticoid-receptor (GR) status in patients with pregnancy induced hypertension (PIH). All parameters were determined in the last trimester of pregnancy. We studied 20 women with PIH with and without proteinuria.

Glucocorticoid receptors were determined by radioreceptor assay after isolation of mononuclear leucocytes (MNL). The corticotrophin-releasing factor-like immunoreactivity (CRF-LI) in 24 h urine was measured by radioimmunoassay after extraction and followed by reverse phase HPLC.

444 Placenta (1991), Vol. 12

The glucocorticoids and the corticotrophin-releasing factor-like immunoreactivity were markedly increased in patients with pregnancy induced hypertension (PIH). The number of glucocorticoid receptors (GR) of the mononuclear leucocytes was decreased in patients with PIH.

Many authors report that a large amount of CRF-LI in plasma is of placental origin. The high glucocorticoid concentration could be a consequence of high CRF-LI values which could lead to a decreased number of glucocorticoid receptors by down regulation.

INSULIN-LIKE GROWTH FACTOR-BINDING PROTEINS (IGFBPs) IN HUMAN PREGNANCY AND UMBILICAL CORD SERUM H. S. Wang & T. Chard (Department of Reproductive Physiology, St. Bartholomew’s Hospital Medical College, West Smithfield, London, EClA 7BE, UK)

In order to investigate the characteristics of insulin-like growth factor-binding proteins (IGFBPs), serum samples from normal males, non-pregnant females, pregnant women, and neonates were chromatographed on Sephadex G-200 at pH 8.2 after incubation with 1251-IGF-I for 30 min or overnight. Fractions corresponding to the 150 kDa and 40 kDa binding proteins were collected and pooled separately. The pooled fractions were fraction- ated on Sephadex G-50 (Fine) at pH 2.3 to dissociate insulin-like growth factors (IGFs) from IGFBPs. IGFBPs in crude serum or pooled fractions from gel filtration chromatography were cross-linked with ‘251-IGF-I using disuccinimidyl suberate (DSS), and separated on 15 % SDS-PAGE gel followed by autoradiography.

The chromatographic profile (U.V. absorbance at 280 nm and radioactivity) showed that the bulk of the IGF-I was associated with the 150 kDa binding proteins in both non-pregnant and pregnant serum. This was further supported by the appearance of band at 150 kDa on SDS-PAGE after cross-linking. Similar findings were observed in a mixture of non- pregnant serum and maternal serum at the time of delivery. These findings differ from previous studies and suggest that the 150 kDa binding proteins retain the ability to bind IGF-I in pregnant serum although some of their physicochemical characteristics are altered in such a way that they may split into small subunits on SDS-PAGE.

In umbilical cord serum (artery and vein), the majority of ‘2”I-IGF-I was bound to the 40 kDa binding proteins. This was observed both on chromatography and on SDA-PAGE after cross-linking. Thus the 40 kDa binding proteins appear to be the principal binding proteins for IGFs in the fetal circulation. It may be that the fetal liver, the main source of IGFBP, is unable to synthesize the 150 kDa binding proteins because of immaturity of GH receptors.

THE PLACENTA AND FETAL GROWTH IN TWIN PREGNANCY LATE IN GESTATION B. S. Ward (Department of Obstetrics and Gynaecology, University of Manchester, Saint Mary’s Hospital, Manchester Ml3 OJH, UK)

Conceptuses from 31 to 40 weeks’ gestation were studied from 40 twin pregnancies that produced separate and distinct placentas and whose precise gestational ages were known. Placental preparation was standardized and comparisons were restricted to conceptuses of the same gestational age.