Alterations of human erythrocyte membrane fluidity by oxygen-derived free radicals and calcium

Two possible reasons for the structural alterations of cell membranes caused by free radicals are lipid peroxidation and an increase in the intracellular calcium ion concentration. To characterize the alterations in membrane molecular dynamics caused by oxygen-derived free radicals and calcium, human erythrocytes were spin-labeled with 5-doxyl stearic acid, and alterations in membrane fluidity were quantified by electron spin resonance oxidase (0.07 U/mL) decreased membrane fluidity, and the addition of superoxide dismutase and catalase inhibited the effect on membrane fluidity of the hypoxanthine-xanthine oxidase system. Hydrogen peroxide (0.1 and 1 nM) also decreased membrane fluidity and caused alterations to erythrocyte morphology. In addition, a decrease in membrane fluidity was observed in erythrocytes incubated with 2.8 mM CaCl₂. On the other hand, incubation of erythrocytes with calcium-free solution decreased the changes in membrane fluidity caused by hydrogen peroxide.

These results suggest that changes in membrane fluidity are directly due to lipid peroxidation and are indirectly the result of increased intracellular calcium concentration. We support the hypothesis that alterations of the biophysical properties of membranes caused by free radicals play an important role in cell injury, and that the accumulation of calcium amplifies the damge to membranes weakened by free radicals.     

Topography and functions of sulfhydryl groups of the human erythrocyte glucose transport mechanism

Summary Membrane-impermeant and -permeant maleimides were applied to characterize the location and function of the sulfhydryl (SH) groups essential for the facilitated diffusion mediated by the human erythrocyte glucose transport protein. Three such classes have been identified. Type I SH is accessible to membrane-impermeant reagents at the outer (exofacial) surface of the intact erythrocyte. Alkylation of this class inhibits glucose transport; D-glucose and cytochalasin B protect against the alkylation. Type II SH is located at the inner (endofacial) surface of the membrane and is accessible to the membrane-impermeant reagent glutathione maleimide only after lysis of the erythrocyte. D-glucose enhances, while cytochalasin B reduces, the alkylation of Type II SH by maleimides. Reaction of Types I and II SH with an impermeant maleimide increases the half-saturation concentration for binding of D-glucose to erythrocyte membranes. By contrast, inactivation of Type III SH markedly decreases the half-saturation concentration for the binding of D-glucose and other transported sugars. Type III SH is inactivated by the relatively lipid-soluble reagents N-ethylmaleimide (NEM) and dipyridyl disulfide, but not by the impermeant glutathione maleimide. Type III SH is thus located in a hydrophobic membrane domain. A kinetic model constructed to explain these observations indicates that Type III SH is required for the translocation event in a hydrophobic membrane domain which leads to the dissociation of glucose bound to transport sites at the membrane surfaces.     

Kinetics of channelized membrane ions in magnetic fields

The cyclotron resonance model for channel ion transport in weak magnetic fields is extended to include damping losses. The conductivity tensor is obtained for different electric field configurations, including the circuital field E phi normal to the channel axis. The conductivity behavior close to the cyclotron resonance frequency omega c is compared to existing Ca2+-efflux data in the literature. A collision time of .023 s results from this comparison under the assumption that K+ ions are transiting in a 0.35 G field. We estimate a mean kinetic energy of 3.5 eV for this ion at resonance. This model leads to discrete modes of vibration (eigenfrequencies) in the ion-lattice interaction, such that omega n = n omega c. The presence of such harmonics is compatible with recent results by Blackman et al. [1985b] and McLeod et al. [1986] with the interesting exception that even modes do not appear in their observations, whereas the present model has no restriction on n. This harmonic formalism is also consistent with another reported phenomenon, that of quantized multiple conductances in single patch-clamped channels.     

Measurement of Free Intracellular Calcium in the Brain by 19F-Nuclear Magnetic Resonance Spectroscopy

We report the first measurement of the free intracellular calcium level in an actively metabolising intact cerebral tissue preparation. To this end, we applied the recently developed 19F-nuclear magnetic resonance calcium chelator, 5,5'-F2-1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (5FBAPTA), in superfused cerebral cortical slices to give values for the intracellular Ca2+ concentration of 350 and 480 nM, at external calcium concentrations of 1.2 and 2.4 mM, respectively. Under both conditions, the intracellular Ca2+ concentration was increased by depolarisation using a high external K+ concentration. Interleaved 31P spectra showed that the presence of the 5FBAPTA had a deleterious effect on the metabolic state of the tissue with an external Ca2+ concentration of 1.2 mM, but normal viability was maintained using 2.4 mM.     

部分氧化的铁钼蛋白的圆二色散谱和电子顺磁共振谱的研究

棕色固氮菌固氮酶FeMo蛋白与过量（5－6个当量）的酸性靛蓝保温30－60分钟后,蛋白中的P－金属原子族全部氧化,然而蛋白中的FeMoCo全都处于还原状态。Na2S2O4使这种部分氧化的FeMo蛋白中的P-ciuster重新不,甲基紫精可加快这种还原,而亚甲蓝等氧化剂则使这种蛋白中的FeMoCo受到氧化,对这种部分氧化的FeMo蛋白分别进行CD还原滴定和测定氧化过程中的EPR／ABS的变化已经得到p-Cluster和FeMoCo的氧化还原当量数目。     

Phosphorus-31 magnetic resonance spectroscopy of forearm flexor muscles in student rowers using an exercise protocol adjusted for differences in cross-sectional muscle area

To assess exercise energy metabolism of forearm flexor muscles in rowers, six male student rowers and six control subjects matched for age and sex were studied using phosphorus-31 magnetic resonance spectroscopy (31P-MRS). Firstly, to adjust for the effect of differences in cross-sectional muscle area, the maximal cross-sectional area (CSAmax) of the forearm flexor muscles was estimated in each individual using magnetic resonance imaging. Multistage exercise was then carried out with an initial energy production of 1 J.cm-2 CSAmax for 1 min and an increment of 1 J.cm-2 CSAmax every minute to the point of muscle exhaustion. A series of measurements of 31P-MRS were performed every minute. The CSAmax was significantly greater in the student rowers than in the control subjects [19.8 (SD 2.2) vs 17.1 (SD 1.2) cm2, P less than 0.05]. The absolute maximal exercise intensity (J.min-1) was greater in the rowers than in the control subjects. However, the maximal exercise intensity per unit of muscle cross sectional area (J.min-1.cm-2) was not significantly different between the two groups. During mild to moderate exercise intensities, a decrease in phosphocreatine and an increase in inorganic phosphate before the onset of acidosis were significantly less in the rowers, indicating a requirement of less adenosine 5'-diphosphate to drive adenosine 5'-triphosphate production. The onset of acidosis was also significantly delayed in the rowers. No difference was observed in forearm blood flow between the two groups at the same exercise intensity (J.min-1.cm-2).(ABSTRACT TRUNCATED AT 250 WORDS)     

Volatile production by Aspergillus versicolor as a possible cause of odor in houses affected by fungi

Aspergillus versicolor, which has been isolated from several mould affected houses was shown by laboratory studies under axenic conditions to produce several specific volatile compounds on water agar. These compounds were not produced by the fungus when grown on a rich malt extract medium or on several synthetic media. The volatile compounds were analysed by GC-MS. The majority of the peaks represented aromatic compounds. A non-aromatic substance which previously has been revealed by us among prominent volatile compounds sampled from building materials from so-called mould houses was produced only on water agar. According to a comparison with the mass spectrum and retention time of pure reference compound this compound is ethylhexanol, a compound not previously reported as a mould metabolite. The presence of this compound was correlated with pungent odor in the cultures.     

磁场对大豆共生固氮的效应

恒定磁场处理慢生大豆根瘤菌“005”和接种后的大豆植株,发现磁场可以提高根瘤的固氮活性。在一定的磁场强度(70—100mT)下,固氮活性平均可以提高4—5倍,植株的结瘤数和根瘤重量平均提高2—3倍。从这样的根瘤中所分离出的根瘤菌,由慢生型转变成快生型,在100植株中有17株的根瘤分离出快生菌。生长世代时间和培养溶液中的pH值与慢生型不同,而与快生型相同。     

What limits nitrate uptake from soil?

Abstract. An accepted view, that unless nitrate concentrations in the soil solution are very low (e.g. below 0.1–0.2 mol m^?3) the growth of high-yielding crops is not limited by the availability of nitrogen, is challenged. Conventional analyses of nutrient supply and demand, based on calculations of apparent inflow rates (uptake rates per unit total root length) are invalid. Apparent inflow rates are inversely proportional to root length. The convention of using total root length grossly overestimates the fraction of the root system active in nutrient uptake. Consequently, inflow rates based on total root lengths underestimate the true values, indicating unrealistically low nutrient concentration differentials between bulk soil and root surfaces required to drive uptake. An alternative method of analysis is suggested. This is based on total nutrient uptake rather than on inflow rate. Measurements of the former do not depend on estimates of active root length and can be made directly and reliably. The method was applied to data obtained from a pot experiment using spring wheat (Triticum aestivum L., cv. Wembley) grown in soil without nitrogen fertilizer (N₀) or with nitrogen fertilizer equivalent to 200kg N ha^?1 (N⁺). Soil nitrate concentrations calculated using the conventional method based on total root length, suggested that any increases in concentration above those measured in the N⁰ treatment should not have resulted in increased uptake and growth. However, the N⁺ plants were always bigger than those in the No treatment, refuting this suggestion. Theoretical uptakes of nitrogen (calculated initially on the basis of a fully active root system) were adjusted, by reducing the effective root length incrementally, until the theoretical uptake matched the measured net uptake of nitrogen. The mean fractions of the root systems likely to have been involved in nitrate uptake were 11% and 3.5% of the total lengths of root in the N⁰ and N⁺ treatments, respectively.     

A method for estimating oxygen availability of stationary plant cell cultures in liquid media

A method for estimating the oxygen availability in plant cell cultures grown in stationary liquid media (e.g. many protoplast cultures) was developed. The method is based on short-term measurements of respiration rate versus oxygen concentration on a sample of cells, suspended in liquid media. From such data it is possible to estimate the oxygen concentration at the bottom of a stagnant liquid culture, by calculating the amount of oxygen reaching the cells by diffusion. As an example, rape (Brassica napus L. cv. Omega) hypocotyl protoplasts were grown with different oxygen concentrations at the site of the cells, obtained by varying the cell density, the height of the liquid layer and the oxygen content of the gas phase. The number of surviving calli was positively correlated with the estimated oxygen availability in the range between 60 and 350 M O₂, below 60 M all cells died. This indicates that oxygen availability can be a limiting factor in the range usually encountered in protoplast cultures, and that the method can be useful when designing optimal growth conditions for stationary cultures of plant cells.Abbreviations C₁ bulk oxygen concentration in agitated medium - C_o oxygen concentration in medium at the gas-liquid interface, in equilibrium with the gas - C_x oxygen concentration at cell level - D diffusion constant of oxygen in water - K_La oxygen transfer rate - l height of liquid above cells - n number of cells per ml - R_x respiration rate per cell