Procoagulant activity in normal human urine associated with subcellular particles

Procoagulant activity (PCA) in normal human urine was found to be sedimented by centrifugation at X 100,000g. Therefore, studies were done to identify the structures associated with the procoagulant activity. Transmission electron microscopy of the X 100,000g pellet revealed numerous membrane-bound vesicles as well as fibrous material. Filtration of normal urine through a 0.2-micron filter removed more than 90% of the procoagulant activity. Scanning electron microscopy of the filter surface revealed 0.1 to 1.1 micron particles and fibrous material. By centrifugation at pH 3 and 5 the fibrous material and particles were separated. The procoagulant activity remained with the particles in each case. The fibrous material was shown to be Tamm-Horsfall protein by SDS-PAGE and Western blotting using anti-Tamm-Horsfall protein serum. Purified Tamm-Horsfall protein itself was not procoagulant. Therefore, PCA in normal human urine is associated with particles 0.1 to 1.1 micron in diameter which appear to be lipid membranes in various arrangements.     

Oligohydramnios in each sac of a triplet gestation caused by Motrin--fulfilling Kock's postulates

A triplet gestation was treated with Motrin for oral tocolysis. A significant reduction of amniotic fluid was observed, but this was corrected by stopping the drug. A cause-and-effect relationship was established by restarting the Motrin and observing the same development of oligohydramnios, which again returned to normal with discontinuation of the drug.     

Differentiation Dependent Activation of the Myelin Genes In Purified Oligodendrocytes is Highly Resistant to Hypoglycemia

We have previously demonstrated that the developmental upregulation of myelin-specific genes in mixed glial cultures is strongly attenuated by hypoglycemia. The present study was designed to evaluate the effect of hypoglycemia on differentiation-dependent upregulation of myelin genes in purified oligodendrocyte cultures. The expression of major myelin protein genes, i.e., proteolipid protein (PLP), basic protein (BP) and myelin associated glycoprotein (MAG) were monitored by Northern blot analysis. In control cultures maintained at 6 mg/ml of glucose, the expression of all the genes upregulated rapidly, and plateaued at approximately day 4. A similar pattern of differentiation-dependent upregulation was observed for the gene encoding a lipogenic enzyme, i.e., malic enzyme (ME). In contrast to mixed glial cultures, however, this developmental gene upregulation was not significantly affected by severe hypoglycemia (approximately 0.02 mg/ml). The results indicate that the effect of glucose deprivation on oligodendrocyte genes observed in mixed glial cultures is mediated by other cells. The upregulation of the genes in differentiating oligodendrocytes was accompanied by the production of myelin-related membrane that was isolated by density gradient fractionation. In contrast to the effect on gene expression, this anabolic activity was highly dependent on glucose, as seen from a profound suppression by severe hypoglycemia.     

Video endoscopy by nurse practitioners: a model for colorectal cancer screening

The use of paramedical personnel to perform sigmoidoscopy as a screening test for colorectal cancer has been advocated as a means of increasing the availability of this test to the population at risk. A model system has been developed utilizing flexible videosigmoidoscopy performed by nurse practitioners with videotape review by physician endoscopists. Of the 100 patients studied, 36 were found to have polyps. Near excellent concordance (k = 0.72) was observed between the nurse practitioner's findings and those of the physician. Using the physician's review as the standard, overall sensitivity and specificity of the nurse practitioner's examinations were 75% and 94%, respectively. In conclusion, videosigmoidoscopy performed by nurse practitioners and reviewed by physician endoscopists is a feasible approach to colorectal cancer screening since it is safe, provides videotape documentation to ensure quality control, and expands available resources for the performance of this examination.     

Neutrophil elastase produces 52-kD and 30-kD glucocorticoid receptor fragments in the cytosol of human leukemia cells

Characterization of glucocorticoid receptors in leukemia cells is important to understand mechanisms of glucocorticoid resistance but has been impeded by receptor fragmentation in cytosol extracts. We recently found that formation of 52- and 30-kilodalton (kD) glucocorticoid receptor fragments in cytosol of leukemia cells is due to proteolysis and is blocked by diisopropylfluorophosphate (DFP). In the present study, we identify a 28-kD serine protease in cytosol of leukemia cells that binds [3H]DFP and correlates with the formation of 52- and 30-kD receptor fragments. This protease is immunoprecipitated by antiserum to neutrophil elastase. Limited digestion of [3H]dexamethasone-21-mesylate- labeled receptors by purified neutrophil elastase produces 52- and 30- kD receptor fragments. Receptor fragmentation in the cytosol of leukemia cells in inhibited by methoxysuccinyl-alanyl-alanyl-prolyl- valyl-chloromethylketone, a highly specific inhibitor of neutrophil elastase. The addition of as few as 5% neutrophils to a lymphoid cell suspension provides sufficient elastase to produce receptor fragmentation. Our findings indicate that neutrophil elastase is responsible for receptor fragmentation in the cytosol of leukemia cells. The neutrophil elastase may be endogenous to the leukemia cells or may come from neutrophils that contaminate leukemia cell suspensions.