A method is described for the rapid determination of phospholipid phosphorus in samples containing less than 0.5 μg phosphorus. Phospholipid phosphorus is first converted to inorganic phosphate by heating a dried lipid extract briefly over a Bunsen flame in the presence of magnesium nitrate, then dissolving the resulting residue in dilute hydrochloric acid at 95 C. The determination of the inorganic phosphate content of the sample then requires only the addition of Triton X‐100 and an acidic ammonium molybdate‐malachite green reagent. Absorbance is measured at 650 nm after 5 min at room temperature.
The RNA-P and DNA-P content of the nucleus and the RNA-P content of the whole cell of the livers of 8-to 20-day chick embryos and of adult fowls have been determined. The DNA-P content of the liver nuclei was slightly higher in the 8-and 10-day embryo than in all the other stages examined. A significant decrease in the RNA content of the cell occurred during embryonic development. The RNA content of the adult cell was the same as that of the 14-to 16-day embryo. The proportion of the cellular RNA contributed by the nucleus also decreased during development. In respect to both nuclear RNA content and distribution of RNA between nucleus and cytoplasm, the adult resembled the 8-to 12-day embryo. Examination of the fine structure of the cell showed that, as development progressed, free ribosomes decreased in number and the rough membranes increased. Slices of 8-, 14-, and 20-day embryonic livers and of adult livers were incubated with 14C-leucine, and the amount of labeled amino acid incorporated into whole tissue protein and into the proteins of the subcellular fractions was measured. Embryonic liver incorporated 14 C-leucine 15 to 30 times more rapidly than adult liver. The microsomal protein was always more highly labelled than the protein in any other subcellular fraction; however, in the 8-day embryonic and the adult liver the proportion of total counts found in the nuclear fraction was considerably higher than in the 14-or 20-day embryonic liver. The significance of an apparent correlation between the proportion of the cell's RNA contributed by the nucleus and the proportion of total counts in the nuclear fraction is discussed.
Respiratory distress syndrome occurs in infants born with immature lungs. The immature fetal lung lacks an adequate supply of surfactant, a phospholipid-rich substance which is produced in the type II cells of the alveolar epithelium. In the fetus, surfactant is secreted into the potential air spaces of the lung and passes into the amniotic fluid as gestation proceeds. It is now clear that most methods currently in use for assessing fetal lung maturity depend on the detection of a sudden release of surfactant into the amniotic fluid as the lung reaches a critical stage of maturity. These methods, which include the lecithin/sphingomyelin ratio, the lung profile, total phospholipid or lecithin concentration, fluorescence depolarization, lamellar body phospholipid concentration, and the "shake" test, are reviewed in the light of recent understanding of the nature of surfactant. In assessing each method, we have examined possible sources of error in performing the test in the laboratory, factors which could theoretically limit its ability to reflect the state of fetal lung maturity and current information regarding its reliability, in terms of clinical performance. Guidelines for future research in this area are also suggested.
SUMMARYLamellar bodies, produced by secretory cells in the alveolar epithelium, are the major source of surfactant phospholipid. As the fetal lung matures, the membranous content of the lamellar bodies is secreted into the alveolar spaces and passes into the amniotic fluid, from which it can be isolated in a morphologically recognisable form. A method is described for the rapid isolation of a lamellar body fraction from amniotic fluid using a small air-driven clinical ultracentrifuge. The lamellar body phospholipid content of amniotic fluid increases towards the end of gestation, but the time of onset and the rate of this increase show wide individual variation. Preliminary results suggest that the lamellar body phospholipid content of amniotic fluid may be a useful index of fetal lung maturity.
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