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Introduction: Currently freeze-dried, gamma-sterilized, or glycerol-preserved amniotic membranes are widely used. However, it is not clear whether this devitalized state is the optimal application form. Therefore within this study the ideal condition for midterm storage of human amniotic membranes was assessed to ensure the availability of vital amniotic membranes, in particular for burn wounds. Methods and Materials: For this purpose mothers were serologically tested and term placentae were collected and washed. After the amniotic membrane was peeled off and further washed, biopsies were taken for microbiological testing and various storage experiments (different media and temperatures). • cell culture medium, 37 °C • glycerol, 4 °C • 10% DMSO, −80 °C Viability of fresh and stored amniotic membranes was determined with the MTT-based EZ4U- Assay (Biomedica, Vienna, Austria). Results and Discussion: Best results were obtained while storing the membranes in cell culture medium at 37 °C, whereas storage in glycerol at 4 °C resulted in cell death within the first 4 days. To our knowledge this is the first study investigating the viability of amniotic membrane under different storage conditions. The influence on wound healing is currently under investigation.

Objective The purpose of the study was to assess the feasibility of analysis of fetal nucleated red blood cells (NRBC) present in the maternal circulation by laser-scanning cytometry. Methods CD71-positive cells were obtained by magnetic cell sorting of peripheral blood of pregnant women after density centrifugation. Immunofluorescence for the Hbγ-chain was combined with fluorescent staining of DNA (TO-PRO-3) and fluorescence in situ hybridization (FISH) with a Y-chromosome specific probe. The cells were scanned on a slide using a laser-scanning cytometer (LSC). Events double positive for Hbγ and TO-PRO-3 were relocated and their morphology and FISH reactivity were visually assessed. Determination of male fetal sex with LSC was compared with findings from amniocentesis. Results In 8/15 pregnancies with male fetuses and in 0/9 with females (apart from one case with a male/female twin pregnancy), we detected Y-chromosome-positive NRBC. In pregnancies with female fetuses, Y-chromosome-positive cells other than NRBC were found in all women who had previously given birth to male babies, whereas women with no abortion and no male babies in their history did not present with Y-chromosome-positive non-NRBC. Conclusion On the basis of automatic relocation of once-defined cells of fetal origin from the current pregnancy, laser-scanning cytometry is likely to facilitate repeated (poly-)FISH analysis and single-cell PCR for noninvasive prenatal diagnosis. Copyright © 2003 John Wiley & Sons, Ltd.

We investigated the contracting actions of the isoprostanes (isoPs), 8-iso-prostaglandin (PG) F(2alpha) and 8-iso-PGE(2), in comparison to the effects of the thromboxane (TX) A(2)-mimetic U 46619 and the traditional prostaglandin PGE(2) in the isolated rat aorta, isolated rat gastric fundus and the isolated guinea-pig ileum. U 46619 and 8-iso-PGF(2alpha) caused contractions in the rat aorta and rat gastric fundus in a concentration-dependent manner, whereas these agonists showed no effects in the guinea-pig ileum. However, 8-iso-PGE(2) and PGE(2) caused contractions in all isolated organs used. The prostanoid TP-receptor antagonist SQ 29,548 (10 nM) significantly antagonized vasoconstrictions induced by the agonists used in the rat aorta. SQ 29,548 at a final concentration of 3 microM, but not at lower concentrations, significantly inhibited contractions induced by U 46619, 8-iso-PGF(2alpha) and 8-iso-PGE(2) in the rat fundus. Responses to PGE(2) were unchanged. The prostanoid EP(1)-receptor antagonist SC 51089 (3 microM) significantly inhibited contractions induced by 8-iso-PGE(2) and PGE(2) in the rat fundus and in the guinea-pig ileum. SC 51089 had no effect on responses to any of the agonists tested. Our results show that 8-iso-PGE(2), in contrast to 8-iso-PGF(2alpha), can also cause contractions by activation of the EP(1)-receptors in the rat gastric fundus and the guinea-pig ileum. The findings of the present study do not support the existence of a unique isoP-receptor in the tissues used.