Transport of putrescine across duodenal, jejunal and ileal brush-border membrane of chicks (Gallus domesticus).

Luminal polyamines and their absorption are essential for proliferation of the enterocytes and, therefore, nutrition, health and development of the animal. The transport systems that facilitate the uptake of putrescine were characterized in chick duodenal, jejunal and ileal brush-border membrane vesicles prepared by MgCl2 precipitation from three-week-old chicks. An inwardly-directed Na+ gradient did not stimulate putrescine uptake and, therefore, putrescine transport in chick intestine. In the duodenum, jejunum and ileum, kinetics of putrescine transport fitted a model with a single affinity component plus a non-saturable component. The affinity (Kt) for [3H]putrescine transport across the brush-border membrane increased along the length of the small intestine. A model of intermediate affinity converged to the data obtained for [3H]putrescine transport with Kt approximating 1.07 and 1.05 mM or duodenum and jejunum, respectively; and high affinity with a Kt of 0.35 mM for the ileum. The polyamines cadaverine, putrescine, spermidine and spermine strongly inhibited the uptake of [3H]putrescine into chick brush-border membrane vesicles, more so for the jejunum and ileum than the duodenum. The kinetics of cadaverine, spermidine and spermine inhibition are suggestive of competitive inhibition of putrescine transport. These uptake data indicate that a single-affinity system facilitates the intestinal transport of putrescine in the chick; and the affinity of transporter for putrescine is higher in the ileum than in the proximal sections of the small intestine. In addition, this study shows that the ileum of chicks plays an important role in regulating cellular putrescine concentration.

Dextran sulfate protects porcine but not bovine cultured endothelial cells from free radical injury.

Previous studies demonstrated that the polyanion dextran sulfate (DS) protects rat coronary and porcine aortic endothelium (PAE) from oxygen-derived free radical (OFR) injury due to hydrogen peroxide (H2O2) or xanthine/xanthine oxidase (X/XO). To determine if DS has a similar protective effect in bovine aortic endothelium (BAE) and bovine brain microvascular endothelium (BBME), H2O2 or X/XO was added to confluent cultures. Cell injury was assessed 1 d later by measuring the percentage of viable cells (by trypan blue exclusion) and the release of lactate dehydrogenase (LDH) into the medium. After H2O2 doses of 6.0 mM for BAE and BBME and 0.8 mM for PAE, and after X doses of 10 microM and XO doses of 0.3 U/mL for all cell types, approximately 50% of cells were viable. Cultures were pretreated with DS (0.001 to 500 microg/mL) 24 to 26 h prior to H2O2 or X/XO exposure. Pretreatment at concentrations of 0.5, 5, and 50 microg/mL significantly increased the percentage of viable cells and reduced LDH release in cultures of PAE, but not BAE or BBME, treated with H2O2. Similarly, pretreatment with DS concentrations of 5 and 50 microg/mL significantly increased the percentage of viable cells and reduced LDH release in cultures of PAE, but not BAE or BBME, treated with X/XO. Thus, DS protected porcine but not bovine endothelium. Catalase (10 U/mL) increased the percentage of viable cells and reduced LDH release in H2O2-treated BAE and BBME, suggesting that DS likely acts by a different mechanism and does not neutralize H2O2. These results suggest that the protective effect of DS on OFR-injured endothelium is species-dependent.