Cellular response to oxidative stress at sulfhydryl group receptor sites on the erythrocyte membrane

Ellman's reagent was used to induce an oxidative stimulus on the exofacial membrane sulfhydryl groups of the human erythrocyte. Thiol-disulfide exchange occurring extracellularly was monitored using resonance Raman spectroscopy, and intracellular changes were observed by 1H spin echo nuclear magnetic resonance spectroscopy of the intact cell. The stimulus caused oxidation and depletion of the glutathione pool, which was followed at higher concentrations of Ellman's reagent by a depletion of intracellular ergothioneine levels. Larger changes are induced intracellularly than would be expected from the stoichiometry of the reaction at the exofacial surface. A mechanism is proposed which links exofacial sulfhydryl receptor sites via the transport proteins to spectrin and glutathione. The consequences for the cellular redox balance of an extracellular stimulus of this type are discussed.     

Studies on the effective size of phospholipid headgroups in bilayer vesicles using lectin-glycolipid interaction as a steric probe

An experimental approach is described which provides information about the relative, effective size of phospholipid headgroups in bilayer vesicles. It is based on determination of the binding of lectins (Ricinus communis agglutinin or concanavalin A) to synthetic glycolipids inserted in such vesicles, using a vesicle agglutination assay. It is shown that the ability of a glycolipid containing a shorter (4-member) spacer arm to bind the appropriate lectin is highly sensitive to the headgroup structure of the surrounding phospholipid in mixed glycolipid-phospholipid vesicles. Furthermore, when the phospholipid was phosphatidate a change in protonation or in monovalent counter-ion species (Li+, NH+4, N(CH3)+4 or Na+) significantly influenced lectin binding. The interference with lectin binding described above was reduced when the glycolipid spacer arm was extended from a 4- to a 6-member length. Furthermore, the sensitivity to phospholipid headgroup structure or to changes in the ionic environment was completely eliminated when the glycolipid contained a longer (10- or 12-member) spacer arm between the hydrophobic part and the lectin-binding group. It is concluded that the modulation of lectin binding in the former case is due to steric inhibition determined by the effective (hydrated) size of the various phospholipid headgroups.     

Sedimentation in fluvial and lacustrine environments

Sedimentation in rivers is dominated by a complex set of physical processes, associated with the unidirectional flow of water. Variations in these processes give rise to different fluvial channel types, whose character can commonly be recognised in the ancient record. Chemical and biological processes are comparatively unimportant in fluvial sedimentation. In contrast, physical, chemical or biological processes can each dominate sedimentation in lakes. Physical (clastic) deposition dominates in high-latitude and mountain lakes (in which chemical and biological activity are low), and in lakes with high relief of the drainage basin and lake floor. Its variety reflects a range of processes influenced by river inflow, wave and current action, thermal and density effects. Economic benefits from the study of lake and river sedimentation include both resource and environmental aspects. An example is given of a mercury pollution study in a fluvial ecosystem. It shows that return to background levels can take place within a relatively short interval after cessation of pollutant input.     

Arsenic Bioaccumulation in Freshwater Fishes

The arsenic ambient water quality criterion (AWQC) for protection of human health via ingestion of aquatic organisms is currently 0.14 μ g/L. This AWQC is derived using a bioconcentration factor (BCF) of 44, which is a consumption-weighted average based on two data points for oysters and fish that was proposed by the U.S. Environmental Protection Agency in 1980 for broad application to freshwater and marine environments. This BCF is based on the assumption that bioaccumulation is a simple linear function of the exposure concentration. In the nearly quarter of a century since this BCF was promulgated, there have been additions to the arsenic bioaccumulation database and a broader scientific understanding of bioaccumulation mechanisms and how they can be applied to estimating tissue concentrations in aquatic organisms. From this database, we identified 12 studies of arsenic bioaccumulation in freshwater fishes in order to explore differences in laboratory-generated BCFs and field-generated bioaccumulation factors (BAFs) and to assess their relationship to arsenic concentrations in water. Our analysis indicates that arsenic concentrations in tissue and arsenic BAFs may be power functions of arsenic concentration in water. A power function indicates that the highest BCF values may occur at low background levels and may decrease as environmental concentrations increase above the ambient range.     

Evaluation of laser-Doppler perfusion imaging for measurement of blood flow in cortical bone.

Most techniques currently available to measure blood flow in bone are time consuming and require destruction of the tissue, but laser-Doppler technology offers a less invasive method. This study assessed the utility of laser-Doppler perfusion imaging (LDI) to measure perfusion in cortical bone. Twelve mature New Zealand White rabbits were assigned to one of three groups: normal control, constriction (norepinephrine), or dilatation (nitroprusside). The left and right medial tibiae were consecutively scanned at red (634-nm) and near-infrared (810-nm) wavelengths to examine the repeatability of LDI output. The pharmacological intervention groups were injected with the respective drug, and LDI measurements at 810 nm were obtained concurrently with colored microsphere-determined flow in all of the groups. LDI effectively quantified blood flow in cortical bone and detected physiologically induced changes in perfusion. A significant positive correlation was found between microsphere-determined flow and LDI output (r = 0.6, P < 0.05). Repeatability of consecutive LDI measurements was within 5%. The effectiveness of LDI to measure perfusion in bone suggests this method has potential for investigating the role of blood flow in bone metabolism and remodeling.     

Determination of 8-oxoguanine in human plasma and urine by high-performance liquid chromatography with electrochemical detection

A highly sensitive and selective method for determining 8-oxoguanine in plasma and urine was developed by high-performance liquid chromatography with electrochemical detection. The compound was separated by gradient elution on a C₁₈ reversed-phase column with a mobile phase of acetonitrile and 0.1 M sodium acetate, pH 5.2. 8-Hydroxy-2′-deoxyguanosine was used as internal standard. 8-Oxoguanine was detected electrochemically by setting the potential to +300 mV vs. Pd reference. The sensitivity of the assay was 22 ng/ml with a signal-to-noise ratio of 7:1. The within-day relative standard deviations for 8-oxoguanine quality control samples with concentrations of 3340, 1340 and 84 ng/ml were 3.6, 4.3 and 5.7% for plasma, and 4.1, 4.6 and 6.2% for urine, respectively. The day-to-day relative standard deviations for the same samples were 3.8, 6.8 and 7.1% for plasma, and 3.9, 7.0 and 7.9% for urine, respectively. The method is designed to study the pharmacokinetics and metabolic fate of O⁶-benzylguanine in a phase I clinical trial. Previously, O⁶-benzyl-8-oxoguanine was identified as the primary metabolite of O⁶-benzylguanine in humans. We now demonstrate that 8-oxoguanine is a further metabolite of O⁶-benzylguanine.