Effect of hydrogen peroxide on the binding of Merocyanine 540 to human erythrocytes

The fluorescent dye Merocyanine 540 (MC540) is often used as a probe to monitor the molecular packing of phospholipids in the outer leaflet of biomembranes. In a previous study we showed that the increased staining of erythrocytes with a perturbed membrane structure was mainly due to an increase in the fluorescence yield of cell-bound MC540, rather than to an increase of the number of bound molecules. Erythrocytes and ghosts exposed to continuous fluxes of H₂O₂ exhibited pronounced lipid peroxidation. Further, red blood cells subjected to this form of oxidative stress also showed increased staining with MC540. It appeared that this was caused by a strong increase in binding of MC540, together with a slight red shift of the fluorescence emission maximum and a small increase in the fluorescence yield of bound MC540. The changed MC540 binding characteristics were not observed when lipid peroxidation was suppressed by the presence of the antioxidant BHT in the incubation medium. However, open ghosts exposed to H₂O₂ showed no increase of MC540 binding, excluding a direct involvement of lipid peroxidation. Measurement of fluorescence emission spectra and gel filtration studies showed that MC540 can bind to H₂O₂-exposed hemoglobin. Experiments with erythrocytes lysed in hypotonic medium after exposure to H₂O₂ revealed that peroxidation of lipids with H₂O₂ induced a non-specific permeabilization of the plasma membrane to MC540, thereby allowing MC540 to bind to the oxidatively denatured, more hydrophobic hemoglobin. These results indicate that conclusions about packing of phospholipids in the outer leaflet of the membrane based on increased MC540-staining should be drawn with care. Received 27 September 1996; received after revision 5 November 1996; accepted 27 November 1996     

平台用钢微观组织对其分层面上解理断裂应力的影响

利用专门设计的厚度方向缺口四点弯曲试样，结合相应的弹塑性有限元计算结果，在液氮温度下测取了一种海洋平台用钢的６种微观组织分层面上的解理断裂应力σ＊ｆ．结果显示，均匀细小的铁素体＋贝氏体组织具有比铁素体＋带状珠光体为高的σ＊ｆ值．对于同一终轧温度，轧后冷速愈高，其得到的微观组织之σ＊ｆ值愈高．而对于同一轧后冷却制度，则终轧于Ａｒ３与再结晶停止温度之间者其得到的微观组织具有最高的σ＊ｆ值．讨论了各微观组织组成相的作用     

Spectrum characteristics and pump wavelengths of Nd3+-doped double-clad fiber

Conclusion The optimum wavelength of Nd³⁺-doped double-clad fiber laser or amplifier made in China is near 830 nm. The efficiency of Nd³⁺-doped double-clad fiber devices pumped by 820-nm laser is low due to ESA. In order to increase the photochemical stability of the outer cladding, the material of the outer cladding must be advanced.     

Two classes of metabolites fromTheonella swinhoei are localized in distinct populations of bacterial symbionts

The marine spongeTheonella swinhoei (lithistid Family Theonellidae, Order Astrophorida) has yielded many important, bioactive natural products, most of which share structural features with bacterial natural products. The presence of microbial symbionts inT. swinhoei has been reported, and it was originally suggested that the cytotoxic macrolide swinholide A and many of the bioactive cyclic peptides fromT. swinhoei were all produced by symbiotic cyanobacteria. By transmission electron microscopy, we found four distinct cell populations to be consistently present inT. swinhoei: eukaryotic sponge cells, unicellular heterotrophic bacteria, unicellular cyanobacteria and filamentous heterotrophic bacteria. Purification and chemical analyses of each cell type showed the macrolide swinholide A to be limited to the mixed population of unicellular heterotrophic bacteria, and an anti-fungal cyclic peptide occurred only in the filamentous heterotrophic bacteria. Contrary to prior speculation, no major metabolites were located in the cyanobacteria or sponge cells.     

Effect of various stressors on the levels of lipid peroxide,antioxidants and Na+, K+-ATPase actrivity in rat brain

The level of malondialdehyde (MDA), an index of lipid peroxidation, and the antioxidants superoxide dismutase (SOD) and glutathione (GSH), as well as the activity of Na⁺, K⁺-ATPase, were assessed in whole rat brain after immobilization, anemic hypoxia (NaNO₂) and 72 h starvation. The effect of these stressors on plasma glucose and corticosterone levels was also observed. Hypoxia and starvation stimulated the lipidj peroxide formation in braini as indicated by an increase in the level of MDA, being higher after starvation than hypoxia. Brain SOD activity was also increased in response to hypoxia and starvation while GSH content was only diminished ini hypoxia. However, neither MDA nor antioxidants were affected by immobilization. On the other hand, the activity of brain Na⁺, K⁺-ATPase was significantly increased by immobilization and hypoxia but decreased in starvation. A similar pattern of change was also observed in plasma glucose and corticosterone levels in response to these stressors. These results elucidate differences in the biochemical response of animals towards various types of stress, with increased lipid peroxide formation in hypoxia and starvation.     

Antioxidant survey to assess antagonism to redox stress using a prokaryotic and an eukaryotic system

Using a prokaryote (Escherichia coli) and a metazoa-resembling eukaryote (Ochromonas danica), we surveyed antioxidants which might overcome redox stress imposed by menadione sodium bisulphite (MD) and buthionine sulphoximine (BSO). BSO oxidant stress was evident only inO. danica; MD oxidant stress was evident in both organisms. Glutathione, its precursors, e.g. cysteine, homocysteine, and 2-oxo-4-thiazolidine carboxylic acid, and red blood cells, emerged as prime antioxidants for relieving BSO and MD oxidant stress. BSO and MD oxidant activity and antioxidant-annulling effect inO. danica were judged comparable to those found in animal cells whereas the resultsE. coli were not entirely equivalent. TheO. danica system emerged as a practical, rapid, and useful system for pinpointing oxidant stressors and antioxidants, and shows promise for studies with mammalian systems.     

(−)-Epigallocatechin gallate,a polyphenolic tea antioxidant,inhibits peroxynitritemediated formation of 8-oxodeoxyguanosine and 3-nitrotyrosine

Reaction with peroxynitrite at pH 7.4 and 37°C was found to increase the 8-oxodeoxyguanosine levels in calf thymus DNA 35-38-fold. This oxidation of deoxyguanosine, as well as the peroxynitrite-mediated nitration of tyrosine to 3-nitrotyrosine, was significantly inhibited by ascorbic acid, glutathione and (–)-epigallocatechin gallate, a polyphenolic antioxidant present in tea. For 50% inhibition of the oxidation of deoxyguanosine to 8-oxodeoxyguanosine, 1.1, 7.6 of 0.25 mM ascorbate, glutathione or (–)-epigallocatechin gallate, respectively, was required. For 50% inhibition of tyrosine nitration, the respective concentrations were 1.4, 4.6 or 0.11 mM. Thus, (–)-epigallocatechin gallate is a significantly better inhibitor of both reactions than either ascorbate or glutathione. Reaction of (–)-epigallocatechin gallate with peroxynitrite alone resulted in the formation of a number of products. Ultraviolet spectra of two of these suggest that the tea polyphenol and/or its oxidation products are nitrated by peroxynitrite.     

从高放废液中去除超铀元素的TRPO流程热实验

阐述了从高放废液中去除超铀元素的ＴＲＰＯ流程热实验。热实验是用真实动力堆的 高放液进行的，取得了很好的效果。超铀元素镅、镎、钚和铀的去污系数分别大于 ３ ０００，４ ０００， ９５０和 ７ ０００。 ＴＲＰＯ流程在去除超铀元素的同时，还能去除对放射性废物长期危害起决定作用 的９９Ｔｃ。实验结果表明ＴＲＰＯ流程具有去污效率高，反萃效果好，锕系组分之间交叉污染小和试 剂成本低等诸多优点。ＴＲＰＯ流程是一个有效的从高放废液中去除超铀元素的流程。