detection of soluble tnf receptors in urine and amniotic: fluid from pregnant women

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4htrat7.7 369 PLACENTAL ROLE IN INTRAUTERINE GROWTH RATE. OBSERVATIONS ON PLACENTAL VASCULARISATION IN DIABETICS AND SMOKERS lnger Asmussen (Copenhagen, Denmark) The degree of vascularization of the placenta varies a lot. The richly vascularized diabetic placenta corresponding to the well nourished and big babies delivered by diabetic mothers is in contrast to the “dry” and leathery placenta from smokers giving birth to small babies and having an increased frequency of small-for-dates children. The study comprised placentae from the following categories: 30 healthy never-smokers (the control group), 22 healthy heavy smokers (smoking ten or more cigarettes per day), and nine diabetics (White group D, non-smokers). Placentae from controls and the smokers were full term, whereas the diabetics delivered 3 weeks prior to estimated term. The study was performed on fresh placental biopsies taken immediately at delivery and immersion tixed for transmission electronmicroscopy. The study was done double blind and the evaluation of the vasculature was restricted to terminal villi in order to ensure tissue identification. The study revealed that diabetics had extremely well vascularized placentae with many newly formed capillaries at the periphery of the villus. These capillaries had a very thin basal lamina in contrast to other capillaries in the villus which exhibited a thick multilaminal basal lamina suggestive of accelerated aging. The apparently newly formed capillaries were located in close apposition to cytotrophoblast cells. The smokers placentae contained markedly fewer capillaries in the terminal villi and these vessels often exhibited a thickened multilaminal basal membrane suggestive of accelerated aging process. Smokers placentae contained many fibroblasts and an increased amount of collagen. This was in contrast to the diabetics where the interstitial space only looked edematous. In conclusion, placental vascularizatiom’angiogenesis is accelerated in diabetics in contrast to smokers in whom vascular growth is markedly reduced. Smokers placentae exhibit increased fibroblast and collagen content identical to that which is found in scar formation. These differences in placental vascularization may account for the observed lowering in birthweight in smokers of[Tpring and the increase in birthweight in diabetic offspring. DETECTION OF SOLUBLE TNF RECEPTORS IN URINE AND AMNIOTIC FLClD FROM PREGNANT WOMEN K. Austgulen’, N.-B. Liabakk”, H. Scot?, M. Brockhaus” & T. Espevik” (” lnstitutt of Cancer Research, University of Trondheim, N-7006 Trondheim, Norway and ”Laborator? for lmmunohistochemistry and lmmunopathology, The Norwegian National Hospital, Oslo, Norway and ‘F. Hoffman-La Roche AG, 4002 Basel, Switzerland) lmmunoassays were established for detection of soluble 55 kD and 75 kD TNF receptor (R) fragments in biological fluids. These immunoassays were based on monoclonal antibodies against different epitopes of the TNFR and were specific for unoccupied TNFR. Urine collected from pregnant women contained high levels of 55 kD TNFR fragments, whereas such TNFR fragments were not detected in normal control urines. The 75 kD fragments, howe\ er, were observed in urine in pregnancy as well as in urine from non-pregnant women. The secretion of 55 kD TNFR fragments in urine seemed to increase with gestational age until spontaneous delivery. Amniotic fluid was collected from amniocentesis at 18/l 9 weeks

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Page 1: Detection of soluble tnf receptors in urine and amniotic: Fluid from pregnant women

4htrat7.7 369

PLACENTAL ROLE IN INTRAUTERINE GROWTH RATE. OBSERVATIONS ON PLACENTAL VASCULARISATION IN DIABETICS AND SMOKERS lnger Asmussen (Copenhagen, Denmark)

The degree of vascularization of the placenta varies a lot. The richly vascularized diabetic placenta corresponding to the well nourished and big babies delivered by diabetic mothers is in contrast to the “dry” and leathery placenta from smokers giving birth to small babies and having an increased frequency of small-for-dates children.

The study comprised placentae from the following categories: 30 healthy never-smokers (the control group), 22 healthy heavy smokers (smoking ten or more cigarettes per day), and nine diabetics (White group D, non-smokers). Placentae from controls and the smokers were full term, whereas the diabetics delivered 3 weeks prior to estimated term. The study was performed on fresh placental biopsies taken immediately at delivery and immersion tixed for transmission electronmicroscopy. The study was done double blind and the evaluation of the vasculature was restricted to terminal villi in order to ensure tissue identification. The study revealed that diabetics had extremely well vascularized placentae with many newly formed capillaries at the periphery of the villus. These capillaries had a very thin basal lamina in contrast to other capillaries in the villus which exhibited a thick multilaminal basal lamina suggestive of accelerated aging. The apparently newly formed capillaries were located in close apposition to cytotrophoblast cells.

The smokers placentae contained markedly fewer capillaries in the terminal villi and these vessels often exhibited a thickened multilaminal basal membrane suggestive of accelerated aging process.

Smokers placentae contained many fibroblasts and an increased amount of collagen. This was in contrast to the diabetics where the interstitial space only looked edematous.

In conclusion, placental vascularizatiom’angiogenesis is accelerated in diabetics in contrast to smokers in whom vascular growth is markedly reduced. Smokers placentae exhibit increased fibroblast and collagen content identical to that which is found in scar formation. These differences in placental vascularization may account for the observed lowering in birthweight in smokers of[Tpring and the increase in birthweight in diabetic offspring.

DETECTION OF SOLUBLE TNF RECEPTORS IN URINE AND AMNIOTIC FLClD FROM PREGNANT WOMEN K. Austgulen’, N.-B. Liabakk”, H. Scot?, M. Brockhaus” & T. Espevik” (” lnstitutt of Cancer Research, University of Trondheim, N-7006 Trondheim, Norway and ” Laborator? for lmmunohistochemistry and lmmunopathology, The Norwegian National Hospital, Oslo, Norway and ‘F. Hoffman-La Roche AG, 4002 Basel, Switzerland)

lmmunoassays were established for detection of soluble 55 kD and 75 kD TNF receptor (R) fragments in biological fluids. These immunoassays were based on monoclonal antibodies against different epitopes of the TNFR and were specific for unoccupied TNFR. Urine collected from pregnant women contained high levels of 55 kD TNFR fragments, whereas such TNFR fragments were not detected in normal control urines. The 75 kD fragments, howe\ er, were observed in urine in pregnancy as well as in urine from non-pregnant women.

The secretion of 55 kD TNFR fragments in urine seemed to increase with gestational age until spontaneous delivery. Amniotic fluid was collected from amniocentesis at 18/l 9 weeks

Page 2: Detection of soluble tnf receptors in urine and amniotic: Fluid from pregnant women

370 Placenta (1994, Vol. 12

of gestational age as well as at term. The 55 kD TNFR fragments were also detected in amniotic fluid. The monoclonal antibodies against the TNFR were also used for immuno- histochemical staining of frozen sections from placental tissue at term. Scattered 55 kD as well as 7.5 kD TNFR positive stromal cells were detected. The 55 kD positive cells were most abundant in the placental tissue, and the villous syncytiotrophoblasts reacted positive towards the 55 kD TNFR antibody exclusively. These data suggest that soluble TNFRs, and especially the 55 kD fragment, might contribute to the immunological balance between mother and fetus in pregnancy.

CARCINOGEN-INDUCED CHANGES IN THE SECRETION OF HCG IN EARLY PLACENTAL EXPLANTS E. R. Barnea SC R. Shurtz-Swirski (Feto-Placental Unit, Rappaport Institute, Rambam Medical Centre, Technion, Haifa, Israel)

Polycyclic aromatic hydrocarbons (PAH) are considered as a major class of carcinogens/ mutagens. Their presence in the environment is widespread and have been demonstrated to cause increased morbidity and mortality in humans. Early pregnancy is particularly liable to environmental influences among them cigarette smoking.

Benzo(a)pyrene (BP), a PAH, is a major constituent of cigarette smoke. Using explant model cultures of young placenta we have examined BP effects on hCG secretion in both static and dynamic cultures (superfusion). In this model we have recently reported that hCG is spontaneously pulsatile (Barnea & Kaplan,JCEM, 1989). Results show that BP 5-50 pM exerts a time and dose-dependent stimulatory effect upon hCG secretion in static cultures. In addition, pre-exposure to the compound overnight causes a significant increase in pulse characteristics: increase in the area under the curve and amplitude of significant pulses (PULSAR program). The effect of another major carcinogen, methylcholantherene, was similar. Our data suggests that there is a strong interaction between adverse environment and placental well being in vitro. However, since there was a maintained response to stimulatory effect of GnRH(a) 10-l” M in superfusion (used as determinant of placental functional integrity), the increase in hCG was not due to a toxic effect of the PAH but perhaps to events related to genome expression.

EFFECTS OF DIFFERENT UMBILICAL FLOW RATES ON PLACENTAL TRANSFER OF COMPOUNDS DURING IN SITU PERFUSION IN THE RABBIT Maria Monica Barzago, Daniela Omarini, Jose Aramayona, Angela Bortolotti, Giorgio Lucchini & Maurizio Bonati (Perinatal Clinical Pharmacology Unit Istituto di Ricerche Farmacologiche “Mario Negri” Via Eritrea 62, 20157 Milano, Italy)

To optimize an in situ placental perfusion model in the rabbit the effects of different umbilical flow rates on the placental transfer of compounds were assessed. The study set out