Characterization of glucose transport activity reconstituted from heart and other tissues

We examined several aspects of glucose transport reconstituted in liposomes, with emphasis on transporters of rat heart (mostly GLUT4) compared to those of human erythrocytes (GLUT1), and on effects of agents that modulate transport in intact cells. Several types of samples gave higher reconstituted activity using liposomes of egg lipids rather than soybean lipids. Diacylglycerol, proposed to activate transporters directly as part of the mechanism of insulin action, increased the intrinsic activity of heart transporters by only 25%, but increased the size of the reconstituted liposomes by 90%. The dipeptide Cbz-Gly-Phe-NH2 inhibited GLUT4 with a Ki of 0.2 mM, compared to 2.5 mM for GLUT1, which explains its preferential inhibition of insulin-stimulated glucose transport in adipocytes. Verapamil, which inhibits insulin- and hypoxia-stimulated glucose transport in muscle, had no effect on reconstituted transporters. Heart transporters had a higher Km for glucose uptake (13.4) than did GLUT1 (1.6 mM), in agreement with a recent study of GLUT1 and GLUT4 expressed in yeast and reconstituted in liposomes. Transporters reconstituted from heart and adipocytes were 40-70% inactivated by external trypsin, suggesting the presence of trypsin-sensitive sites on the cytoplasmic domain of GLUT4. NaCl and KCl both reduced reconstituted transport activity, but KCl had a much smaller effect on the size of the liposomes.     

In vivo modulation of iododeoxyuridine metabolism and incorporation into cellular DNA by 5'-amino-5'-deoxythymidine in normal mouse tissues and two human colon cancer xenografts

This in vivo study examines the ability of 5'-amino-5'-deoxythymidine (5'-AdThd) to modulate 5-iododeoxyuridine (IdUrd) cellular metabolism in two human colon cancer xenografts (HT 29 and HCT-116), two actively proliferating normal mouse tissues (bone marrow and intestine), and a quiescent normal mouse tissue (liver). 5'-AdThd is a thymidine analogue that at low concentrations (<30 micrometer) can increase thymidine kinase activity, which is the rate-limiting enzyme for activation of IdUrd. We reported recently that the in vitro incubation of HT 29 and HCT-116 cells in 5'-AdThd + IdUrd resulted in an enhancement of 5-iodo-2'-dUTP pools, IdUrd DNA incorporation, and subsequent radiosensitization compared with incubation with IdUrd alone (Clin. Cancer Res., 1: 407-416, 1995). These in vitro effects were more significant in the radioresistant cell line HT 29. Using a 6-day continuous infusion of IdUrd (50 or 100 mg/kg/day) and/or 5'-AdThd (200 mg/kg/day), no increase in systemic toxicity (percentage of body weight loss) was observed in athymic nude mice with 5'-AdThd alone or when combined with IdUrd. There was significant dose-dependent, systemic toxicity with IdUrd, which was reversible within 3 days of completing the lower-dose IdUrd infusion. However, a comparison of plasma levels during the 6-day continuous infusion of IdUrd +/- 5'-AdThd showed a significant interaction of IdUrd and 5'-AdThd, resulting in higher plasma levels by day 6 of both compounds and the principal metabolites, iodouracil and deoxyuridine, which is consistent with nonlinear saturating effects on dihydrouracil dehydrogenase. Coadministration of IdUrd and 5'-AdThd resulted in an increase in the percentage of IdUrd DNA incorporation in the two proliferating normal tissues, which was significant only with the lower IdUrd dose. No effect on IdUrd DNA incorporation was found in normal liver at either IdUrd dose +/- 5'-AdThd. Similar to our in vitro data, the continuous infusion of IdUrd and 5'-AdThd showed a significant effect by increasing the percentage of IdUrd DNA incorporation in HT-29 xenografts at both IdUrd doses, whereas coadministration of 5'-AdThd had no such effect in HCT-116 xenografts.     

Intrapreneurial nursing: the Comprehensive Lower Extremity Assessment Form

The Comprehensive Lower Extremity Assessment Form was developed in response to the need for a screening tool in a nurse-managed foot care clinic. It differs from other such tools because it includes clinical measures that identify the potential for foot pathology. The Comprehensive Lower Extremity Assessment Form also serves as an assessment teaching guide in a foot care course and is included as part of a home-study program. The authors demonstrate how the Comprehensive Lower Extremity Assessment Form has generated revenue as part of an intrapreneurial outgrowth of their foot clinic and provides a comprehensive approach to lower extremity assessment. The form can be tailored to meet the needs of the advanced practice nurse, the clinical setting, or patient population.     

Indicine-N-oxide: the antitumor principle of Heliotropium indicum

Extracts of Heliotropium indicum Linn. (Boraginaceae) showed significant activity in several experimental tumor systems. The active principle is isolated and shown to be the N-oxide of the alkaloid, indicine, previously isolated from this plant. Supporting structural data and anti-tumor data are provided.     

Removal of HBSAg from blood in vitro. I. Effects of washing alone, glycerol addition and removal, and glycerolization, freezing, and washing

Red blood cells from HBSAg-positive blood were washed in the Fenwal Elutramatic, Haemonetics Processor 15, or the IBM Blood Processor with sodium chloride solutions, or in the Huggins Cytoglomerator with sugar solutions. The Fenwal Elutramatic and IBM Blood Processor were the most efficient washing systems, the Haemonetics Processor 15 was less efficient, and the Huggins Cytoglomerator was the least efficient in removing the HBSAg. Washing to remove the HBSAg from red blood cells containing 40 per cent W/V glycerol in an ionic medium was more efficient than washing HBSAg from liquid-stored red blood cells or red blood cells containing 20 per cent W/V glycerol. The original and modified dilution/agglomeration wash cycles used in the Huggins Cytoglomerator were not able to remove the HBSAg from units of blood that were radioimmune assay (RIA) positive and counterelectrophoresis (CEP) negative. Freezing had no effect on the removal of the HBSAg in vitro, whereas the concentration of 40 per cent W/V glycerol in the red blood cells that were washed did. HBSAg was not found in the amorphous debris remaining in the polycarbonate disposable bowl used in the Haemonetics Processor 15 or in the microaggregates remaining in washed red blood cells.     

Synovial-type (group II) phospholipase A2 human seminal plasma

Phospholipase A2 (PLA2) was analysed in seminal plasma of fertile, subfertile, and vasectomized men as well as in prostatic secretion and tissue. Immunological cross-reactivity was observed between synovial-type PLA2 antiserum and the enzyme present in seminal plasma. There was a highly significant correlation between the concentration of the synovial-type PLA2, as measured by a time-resolved fluoroimmunoassay and the catalytic activity of the PLA2. The results show that the PLA2 content in human seminal plasma is very high (approximately 1000 times of that present in blood plasma) and that the enzyme belongs to the synovial-type group II phospholipase A2. The results also indicate that the enzyme is secreted by the prostate.