软X射线对血液有形成分表面电荷的影响——2、淋巴细胞和血小板膜电荷的变化

软X射线不仅能引起红细胞表面电荷的变化,同时也能导致淋巴细胞和血小板表面电荷下降,表现为照射后它们的电泳率下降。低剂量范围内,这种电荷的变化是暂时性的,照后4小时降到最低点,24小时后恢复到对照的水平。细胞电泳率的下降与辐射剂量相关。淋巴细胞是一个复杂的细胞群,正常状态下,按细胞在电场中泳动速度的快慢,可分为两个组分:快峰为T细胞,慢峰为B细胞。软X射线照射以后,T和B细胞的电泳率皆减慢,频数分布峰值下降,离散度加大。血小板成分单一,电泳率较一致。 从照射浓集的血小板再加回自身血浆中电泳率的下降较照射血浆再加到血小板中的电泳率下降大得多;受照射的血小板在磷酸缓冲液中电泳率下降较在血浆悬液中严重得多;2000 rad照后,悬浮于血浆中的血小板电泳率能恢复,而悬浮于磷酸缓冲液中则不能恢复,三个方面来看,血浆中可能存在抗辐射因子。超氧化物岐化酶能有效地预防血小板电泳率的下降,从而可阻止血小板的凝聚。     

Superoxide Dismutase Activities in Senescing Apple Fruit (Malus domestica Borkh.)

Total superoxide dismutase (SOD:superoxide: superoxide oxidoreductase; EC 1.15.1.1) activity and individual activities of its three different metalloenzymes, CuZn-SOD, Fe-SOD, and Mn-SOD, were investigated during senescence of apples (Malus domestica Borkh.). Total SOD activity and relative activities of its three forms varied greatly among cultivars. Activities underwent considerable change during senescence of fruit, but changes were different among cultivars. Whether fruit senesced at 0°C or 20°C had little effect on SOD activities, and application of the antioxidant diphenylamine (DPA) did not alter activities. SOD activities increased with occurrence of the physiological disorder “senescent breakdown,” but did not increase with occurrence of the disorder “superficial scald.” In a given apple cultivar, changes in total SOD activity generally paralleled activities of the different SOD forms. Such activity may reflect changes affecting food and nutritive quality of the fruit.     

仙人掌超氧化物歧化酶的纯化及其部分性质研究

10 0 0g仙人掌茎经匀浆、硫酸铵分级沉淀、SephadexG - 75凝胶层析和DEAE - 5 2离子交换层析 3个步骤 ,将仙人掌SOD纯化到SDS -PAGE均一纯 ,得白色粉状SOD 0 8985g ,比活 910U/mg,活力回收 4 3 1% ,纯化倍数 76。该酶相对分子质量 37 1kD ,H2 O2 和KCN的抑制实验结果表明为Cu、Zn -SOD。该酶的最大紫外吸收在 2 75nm处。 5 0℃保温 15 0min ,SOD的活性不变 ,在pH =5 0～ 9 0时SOD有较好的稳定性     

Shape design of cerium oxide nanoparticles for enhancement of enzyme mimetic activity in therapeutic applications

Cerium oxide nanoparticles (CONPs), widely used in catalytic applications owing to their robust redox reaction, are now being considered in therapeutic applications based on their enzyme mimetic properties such as catalase and super oxide dismutase (SOD) mimetic activities. In therapeutic applications, the emerging demand for CONPs with low cytotoxicity, high cost efficiency, and high enzyme mimetic capability necessitates the exploration of alternative synthesis and effective material design. This study presents a room temperature aqueous synthesis for low-cost production of shape-selective CONPs without potentially harmful organic substances, and additionally, investigates cell viability and catalase and SOD mimetic activities. This synthesis, at room temperature, produced CONPs with particular planes: {111}/{100} nanopolyhedra, {100} nano/submicron cubes, and {111}/{100} nanorods that grew in [110] longitudinal direction. Enzymatic activity assays indicated that nanopolyhedra with a high concentration of Ce⁴⁺ ions promoted catalase mimetic activity, while nanocubes and nanorods with high Ce³⁺ ion concentrations enhanced SOD mimetic activity. This is the first study indicating that shape and facet configuration design of CONPs, coupled with the retention of dominant, specific Ce valence states, potentiates enzyme mimetic activities. These findings may be utilized for CONP design aimed at enhancing enzyme mimetic activities in therapeutic applications.

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Proteome alterations induced in human white blood cells by consumption of Brussels sprouts: Results of a pilot intervention study

Epidemiological studies indicate a correlation of cruciferous vegetables consumption with reduced incidence of cancer. This study was designed to investigate molecular mechanisms, which may help to understand the beneficial effects of Brussels sprout consumption. In order to avoid the limitations of in vitro model systems, we performed a dietary intervention study with five participants. We investigated, whether sprout consumption affects the proteome profile of primary white blood cells. In order to achieve maximal sensitivity in detecting specific adaptive proteome alterations, we metabolically labelled freshly isolated cells in the presence of ³⁵S‐methionine/cysteine and performed autoradiographic quantification of protein synthesis. Proteins were separated by 2‐DE and spots of interest were cut out, digested and identified by MS. After the intervention, we found a significant up‐regulation of the synthesis of manganese superoxide dismutase (1.56‐fold) and significant down‐regulation of the synthesis of heat shock 70 kDa protein (hsp70; 2.27‐fold). Both proteins play a role in malignant transformation of cells. Hsp‐70 is involved in the regulation of apoptosis, which leads to elimination of cancer cells, while SOD plays a key role in protection against reactive oxygen species mediated effects. Our findings indicate that the alteration of the synthesis of these proteins may be involved in the anticarcinogenic effects of cruciferous vegetables, which was observed in earlier laboratory studies with animals.     

锰酶及其模拟物研究进展

对生物体内和模拟的含锰金属酶，即含锰超氧化物歧化酶(Mn—SOD)、含锰的过氧化氢酶(Mn—catalase)及含锰的光合成氧释放配合物(Mn—OEC)的结构、活性部位、催化作用机理及其模型化合物的研究进展进行了综述。     

Superoxide Dismutase as a Novel Macromolecular Nitric Oxide Carrier: Preparation and Characterization

Nitric oxide (NO) is an important molecule that exerts multiple functions in biological systems. Because of the short-lived nature of NO, S-nitrosothiols (RSNOs) are believed to act as stable NO carriers. Recently, sulfhydryl (SH) containing macromolecules have been shown to be promising NO carriers. In the present study, we aimed to synthesize and characterize a potential NO carrier based on bovine Cu,Zn-superoxide dismutase (bSOD). To prepare S-nitrosated bSOD, the protein was incubated with S-nitrosoglutathione (GSNO) under varied experimental conditions. The results show that significant S-nitrosation of bSOD occurred only at high temperature (50 °C) for prolonged incubation time (>2 h). S-nitrosation efficiency increased with reaction time and reached a plateau at ~4 h. The maximum amount of NO loaded was determined to be about 0.6 mol SNO/mol protein (~30% loading efficiency). The enzymatic activity of bSOD, however, decreased with reaction time. Our data further indicate that NO functionality can only be measured in the presence of extremely high concentrations of Hg²⁺ or when the protein was denatured by guanidine. Moreover, mildly acidic pH was shown to favor S-nitrosation of bSOD. A model based on unfolding and refolding of bSOD during preparation was proposed to possibly explain our observation.     

Copper(I)-Dioxygen Adducts and Copper Enzyme Mechanisms

Primary copper(I)-dioxygen (O₂) adducts, cupric-superoxide complexes, have been proposed intermediates in copper-containing dioxygen-activating monooxygenase and oxidase enzymes. Here, mechanisms of C−H activation by reactive copper-(di)oxygen intermediates are discussed, with an emphasis on cupric-superoxide species. Over the past 25 years, many synthetically derived cupric-superoxide model complexes have been reported. Due to the thermal instability of these intermediates, early studies focused on increasing their stability and obtaining physical characterization. More recently, in an effort to gain insight into the possible substrate oxidation step in some copper monooxygenases, several cupric-superoxide complexes have been used as surrogates to probe substrate scope and reaction mechanisms. These cupric superoxides are capable of oxidizing substrates containing weak O−H and C−H bonds. Mechanistic studies for some enzymes and model systems have supported an initial hydrogen-atom abstraction via the cupric-superoxide complex as the first step of substrate oxidation.