No abstract

No abstract

The physicochemical effects of a series of alkanols, alkanediols and glycerol on erythrocyte shape and hemolysis at 4 and 20 degrees C were examined. We calculated the dielectric constant of the incubation medium, Ds, and the dielectric constant of the erythrocyte membrane Dm in the presence of organic solutes. The ratio Ds/Dm = -38.48 at 20 degrees C defines the normal biconcave shape in a medium without hemolytic agents. A decrease in Ds/Dm favors externalization or internalization with consequent hemolysis. Alkanols and alkanediols convert biconcave erythrocytes into echinocytes, which is accompanied by an increase in the projected surface area. Glycerol converts biconcave erythrocytes into stomatocytes, which was accompanied by a marginal decrease in the projected surface area. Progressive externalization in alkanols and alkanediols or internalization in glycerol resulted in a decrease in the projected surface area and the formation of smooth spheres. The degree of shape change induced was related to the degree of hemolysis and the ratio Ds/Dm. A decrease in temperature reduced both the degree of shape change and hemolysis. Our results suggest that physicochemical toxicity may be a result of a temperature dependent hydrophobic interaction between the organic solutes and the membrane and is best interpreted by the ability of the solutes to change Ds and Dm. These results are discussed with respect to the physicochemical constants of the organic solutes.

Two primary explosives involved in groundwater contamination, 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), were detected on-site at low ppb levels with a semiautomated fiber optic biosensor. Validation of the Analyte 2000 (manufactured by Research International Inc., Woodinville, WA) for TNT and RDX detection was performed at two Superfund sites, Umatilla Army Depot and Naval Surface Weapons Center Crane. Samples from monitoring wells were split for analysis using the fiber optic biosensor on-site and using U.S. EPA SW-846 Method 8330 (reverse-phase high performance liquid chromatography) in an offsite laboratory. The Analyte 2000, a multifiber probe fluorimeter, was coupled to a fluidics unit for semiautomated operation. The fiber optic biosensor assay is based on a competitive fluorescent immunoassay performed on the silica core of a fiber probe. From these studies, the limit of detection was determined to be 5 μg/L for both TNT and RDX. In addition to the field samples, extensive laboratory analyses were performed to determine cross-reactivity, matrix effects, and false positive/negative rates.

We have measured freeze-thaw damage to isolated spinach (Spinacia oleracea 1.) chloroplast thylakoid membranes in the presence of different galactose-specific seed lectins to determine whether the binding of proteins to the membrane surface can lead to cryoprotection. O f the seven lectins investigated, five were protective to different degrees and two showed no measurable effect. Protection was afforded by a reduction of the solute permeability of the membranes. This reduced the solute influx during freezing and thereby osmotic rupture of the thylakoid vesicles during thawing. Using model membranes and fluorescently labeled lectins, we could show that the proteins bound exclusively to the digalactosyl lipids in the membranes. Binding was a prerequisite for the protective effect, because the presence of up to 5 mM galactose in the samples completely inhibited both binding of the lectins to thylakoid and model membranes and cryoprotection. The degree of binding was, in contrast, not related to the cryoprotective efficiency of different lectins; cryoprotection was a function of the hydrophobicity of the proteins.