Placental apoptosis is increased in vivo in preeclampsia (PE) and intrauterine growth restriction (IUGR). The cause and pathological implications of this phenomenon are unknown. This study considers the apoptotic susceptibility of villous trophoblasts from normal, PE, and IUGR pregnancies. Cultured cytotrophoblasts (CTs) and an in vitro model of syncytialization were used. CTs were isolated from term placentas of 12 normal, 12 PE, and 12 IUGR pregnancies. Apoptosis was determined by terminal dUTP nick-end labeling (TUNEL), Annexin V binding, and ADP:ATP ratios. Cells were stimulated with tumor necrosis factor-alpha/interferon-gamma or reduced oxygen (<5 KPa). For CTs, ADP:ATP <1 correlates with Annexin V binding. For normal pregnancy, tumor necrosis factor-alpha and depleted oxygen significantly increased TUNEL, Annexin V binding and ADP:ATP in CTs and syncytiotrophoblasts (STs). Spontaneous apoptosis was similar between groups for both cell types. After stimulation, TUNEL and Annexin V binding of CTs were significantly raised in PE and IUGR as compared with normal pregnancy. After oxygen reduction, ADP:ATP in CTs and STs were significantly elevated in IUGR. TUNEL was also increased in STs in PE after oxygen depletion and was significantly raised in STs from IUGR pregnancies after stimulation with both agonists. This is the first description of enhanced apoptosis in isolated villous trophoblasts in PE and IUGR. These intrinsic differences may represent an important factor in the pathophysiology of these conditions.
This report describes the development of a portable spirometer capable of collecting primarily alveolar breath into 1.8-L canisters for subsequent gas chromatographic/mass spectrometric (GC/MS) analysis. Based on CO2 measurements, greater than 97% of the breath collected is alveolar in origin. Sample collection takes place in approximately two minutes. Clean air for inhalation is provided by two organic vapor respirator cartridges. Studies of the breakthrough volume of test compounds at both the 5- and 50-micrograms/m3 levels indicate that each cartridge filter can be used to sample over 300 L of air and that this volume is not altered by intermittent use and storage of the filter for up to five days. In experiments designed to mimic human breathing, recoveries of test compounds through the device at the 5-mu/m3 level ranged from 87 to 112%. Essentially no volatile organic compound (VOC) memory (i.e., adsorption carryover by the device) was measured at the 50-micrograms/m3 level. The data suggest that the device can be used successfully for organic compounds with volatilities greater than that of p-dichlorobenzene.
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