UV-B辐射对蚕豆叶膜脂过氧化的影响及其机制

温室种植的吞豆在0（CK）,8．82kJ/m2(T1)和12．6kJ/m2(T2)3种剂量的紫外线B（UV－B）辐射引起膜脂变化及其机制的研究结果表明,UV0B处理后,蚕豆叶片中丙二醛（MDA）和H2O2含量升高,膜脂肪酸不饱和度指数（IUFA）降低,脂氧合酶（LOX）活性升高,超氧歧化酶（SOD）活性稍有波动,而3种多胺－腐胺（Put)精胺（Spd)和尸胺（Spn)在照射7天后均有积累,但在处理后期（21d)有所回落,推测由LOX主导的酶促膜过氧化作用和氧自由基引起的非酶促过作用在膜结构的破坏中起重要作用,SOD活性和多胺含量的变化蚕豆对UV－B胁迫的一种适应性生理反应。     

射频电磁场生物学效应的某些研究进展

当今无线电波及其通讯被广泛应用,人类无时无刻不处在由电台、电视台、移动电话、紧急通讯设备及雷达等产生的电磁辐射中。射频辐射（Ｒａｄｉｏ－ｆｒｅｑｕｅｎｃｙ Ｒａｄｉａｔｉｏｎ ＲＦＲ）的致热效应已肯定,然而其非热效应仍不清楚。就后者的可能作用机理及目前较为热点的问题,即移动电话等产生的电磁辐射的一些研究现状做一综述。     

太赫兹辐射及其生物效应研究进展

随着太赫兹源和探测技术的不断进步，太赫兹技术迅速发展并在众多领域有着广泛的应用前景. 特别是在生物医学领域，太赫兹技术有望成为一种新型治疗手段. 本文首先介绍了太赫兹的电磁波特点及3种太赫兹波产生方式. 其次介绍了太赫兹辐射在生物上的两大效应：热效应和非热效应. 最后从细胞和生物体两大层面上，详细介绍了太赫兹辐射对不同细胞的生物效应和一些相关分子通路改变，以及太赫兹辐射在不同生物体上的作用效果，为太赫兹生物相关研究人员提供参考.     

简化的真空冷冻干燥器

一种简化的真空冷冻干燥器,只需用一个冷阱,用丙酮干冰作为混合冷却剂,真空冷冻干燥的效果与以往一般用两只冷阱的真空冷冻干燥器完全一样,而且干冰的用量仅为以往的1/3。真空冷冻干燥装置见图1。其操作流程是保温瓶内先装2/3     

特定电磁辐射增强大豆种子超弱光子辐射

生物超弱光子辐射(简称PE)是生物代谢过程的一种普遍现象,它控制细胞内和细胞间的新陈代谢、功能调节以及信息传递。PE光强度与细胞活力、环境因素以及化学物质的作用有关。 红外辐射(包括特定电磁辐射)能产生广泛和显著的生物效应,国内外已有报道,化学     

蛋白质二级结构的真空紫外圆二色性研究

利用同步辐射真空紫外圆二色谱仪和特制的样品池,测定溶液中蛋白质的真空紫外圆二色谱,测定波长低至175nm,并应用一种新的计算法分析计算了蛋白质5种二级结构的含量,所得结果与用X射线衍射法测定的结果一致.讨论了获得好的真空紫外圆二色谱的几个重要因素.结果表明,真空紫外圆二色法是目前测定溶液中蛋白质二级结构的较好方法之一.     

中子低剂量刺激生长技术的研究

中子低剂量刺激生长技术是一种具有巨大潜力,并具有较高经济效益和社会效益的新兴实用技术,经多家单位对淡水鱼,对虾,扇贝,青虾和鸡等多方面实验都具有提高孵化率,成活率,抗病能力和加快生长速度提高产量的作用。     

介绍两种简便的生化样品浓缩方法

在生理生化实验中,经常要将某些样品进行浓缩。现在介绍我们实验室中曾经用过的两种简便方法,以供大家参考。一真空减压透析浓缩法真空减压透析浓缩法的基本原理是利用浓缩装置内外的压力差,将所欲浓缩样品中     

一种在豌豆植物中瞬时表达外源蛋白的新方法

以豌豆植物为实验材料建立了一种瞬时表达外源蛋白的新方法-发芽种子真空侵染法。以绿色荧光蛋白作为报告基因对该体系进行优化的结果表明其最佳工作条件为:菌体工作液浓度OD600=1.0~1.5,真空压力0.08 MPa,真空侵染时间1 min。该方法操作简单,可以同时侵染大量的豌豆植物材料,并将实验周期缩短为15 d,植物侵染后12~14 d可收获外源蛋白,外源蛋白表达量与叶片注射法相当,是一种在豌豆植物中批量生产外源蛋白的新方法。     

细胞衰老在非酒精性脂肪性肝病发生发展中的作用

细胞衰老是细胞在受到内外环境的损伤作用后被激活的一种细胞应答状态,可作为一种固有的细胞保护机制用于抵抗这些损伤刺激。然而,长时间持续的细胞衰老状态则与炎症、纤维化以及恶性肿瘤的发生有着密切的关系,其中在非酒精性脂肪性肝病的发生发展中细胞衰老扮演着重要的角色。因此,探讨细胞衰老在非酒精性脂肪性肝病发生发展中的作用,并发现其可能用于临床治疗的分子靶点及其作用机制具有重要意义。     

Two-component structure of the current pulse of a ranaway electron beam generated during electric breakdown of elevated-pressure nitrogen

Conditions are investigated at which two current pulses of ranaway electron beams are generated in elevated-pressure nitrogen during one voltage pulse. It is shown that the regime with two runaway electron beam current pulses takes place at decreased values of the electric field strength E in the gap (or decreased values of the parameter E/p, where p is the gas pressure). The regime with two runaway electron beam current pulses is observed both at high (1500?C3000 Torr) and low (below 100 Torr) pressures. It is shown that, for the second runaway electron beam current pulse to form, the voltage across the gap should be partially reduced during the first pulse. At low nitrogen pressures (~10 Torr), the regime in which two runaway electron beams are generated can be implemented by increasing the breakdown strength of the gap and/or increasing the value of E/p. In experiments carried out in atmospheric-pressure air with a picosecond time resolution, a rather complicated structure of the beam current pulse is observed at a voltage rise time of ~300 ps.     

Determination of the rate of reduction of oxyferrous cytochrome P450 2B4 by 5-deazariboflavin adenine dinucleotide T491V cytochrome P450 reductase

The use of 5-deazaFAD T491V cytochrome P450 reductase has made it possible to directly measure the rate of electron transfer to microsomal oxyferrous cytochrome (cyt) P450 2B4. In this reductase the FMN moiety can be reduced to the hydroquinone, FMNH(2), while the 5-deazaFAD moiety remains oxidized [Zhang, H., et al. (2003) Biochemistry 42, 6804-6813]. The rate of electron transfer from 5-deazaFAD cyt P450 reductase to oxyferrous cyt P450 was determined by rapidly mixing the ferrous cyt P450-2-electron-reduced 5-deazaFAD T491V reductase complex with oxygen in the presence of substrate. The 5-deazaFAD T491V reductase which can only donate a single electron reduces the oxyferrous cyt P450 and oxidizes to the air-stable semiquinone, with rate constants of 8.4 and 0.37 s(-1) at 15 degrees C. Surprisingly, oxyferrous cyt P450 turns over more slowly with a rate constant of 0.09 s(-1), which is the rate of catalysis under steady-state conditions at 15 degrees C (k(cat) = 0.08 s(-1)). In contrast, the rate constant for electron transfer from ferrous cyt b(5) to oxyferrous cyt P450 is 10 s(-1) with oxyferrous cyt P450 and cyt b(5) simultaneously undergoing spectral changes. Quantitative analyses by LC-MS/MS revealed that the product, norbenzphetamine, was formed with a coupling efficiency of 52% with cyt b(5) and 32% with 5-deazaFAD T491V reductase. Collectively, these results suggest that during catalysis a relatively stable reduced oxyferrous intermediate of cyt P450 is formed in the presence of cyt P450 reductase but not cyt b(5) and that the rate-limiting step in catalysis follows introduction of the second electron.     

Membrane-associated cytochrome cy of Rhodobacter capsulatus is an electron carrier from the cytochrome bc1 complex to the cytochrome c oxidase during respiration.

We have recently established that the facultative phototrophic bacterium Rhodobacter capsulatus has two different pathways for reduction of the photooxidized reaction center during photosynthesis (F.E. Jenney and F. Daldal, EMBO J. 12:1283-1292, 1993; F.E. Jenney, R.C. Prince, and F. Daldal, Biochemistry 33:2496-2502, 1994). One pathway is via the well-characterized, water-soluble cytochrome c2 (cyt c2), and the other is via a novel membrane-associated c-type cytochrome named cyt cy. In this work, we probed the role of cyt cy in respiratory electron transport by isolating a set of R. capsulatus mutants lacking either cyt c2 or cyt cy, in the presence or in the absence of a functional quinol oxidase-dependent alternate respiratory pathway. The growth and inhibitor sensitivity patterns of these mutants, their respiratory rates in the presence of specific inhibitors, and the oxidation-reduction kinetics of c-type cytochromes monitored under appropriate conditions demonstrated that cyt cy, like cyt c2, connects the bc1 complex and the cyt c oxidase during respiratory electron transport. Whether cyt c2 and cyt cy are the only electron carriers between these two energy-transducing membrane complexes of R. capsulatus is unknown.     

Mobile cytochrome c2 and membrane-anchored cytochrome cy are both efficient electron donors to the cbb3- and aa3-type cytochrome c oxidases during respiratory growth of Rhodobacter sphaeroides

We have recently established that the facultative phototrophic bacterium Rhodobacter sphaeroides, like the closely related Rhodobacter capsulatus species, contains both the previously characterized mobile electron carrier cytochrome c2 (cyt c2) and the more recently discovered membrane-anchored cyt cy. However, R. sphaeroides cyt cy, unlike that of R. capsulatus, is unable to function as an efficient electron carrier between the photochemical reaction center and the cyt bc1 complex during photosynthetic growth. Nonetheless, R. sphaeroides cyt cy can act at least in R. capsulatus as an electron carrier between the cyt bc1 complex and the cbb3-type cyt c oxidase (cbb3-Cox) to support respiratory growth. Since R. sphaeroides harbors both a cbb3-Cox and an aa3-type cyt c oxidase (aa3-Cox), we examined whether R. sphaeroides cyt cy can act as an electron carrier to either or both of these respiratory terminal oxidases. R. sphaeroides mutants which lacked either cyt c2 or cyt cy and either the aa3-Cox or the cbb3-Cox were obtained. These double mutants contained linear respiratory electron transport pathways between the cyt bc1 complex and the cyt c oxidases. They were characterized with respect to growth phenotypes, contents of a-, b-, and c-type cytochromes, cyt c oxidase activities, and kinetics of electron transfer mediated by cyt c2 or cyt cy. The findings demonstrated that both cyt c2 and cyt cy are able to carry electrons efficiently from the cyt bc1 complex to either the cbb3-Cox or the aa3-Cox. Thus, no dedicated electron carrier for either of the cyt c oxidases is present in R. sphaeroides. However, under semiaerobic growth conditions, a larger portion of the electron flow out of the cyt bc1 complex appears to be mediated via the cyt c2-to-cbb3-Cox and cyt cy-to-cbb3-Cox subbranches. The presence of multiple electron carriers and cyt c oxidases with different properties that can operate concurrently reveals that the respiratory electron transport pathways of R. sphaeroides are more complex than those of R. capsulatus.     

Output power variations in a plasma relativistic microwave amplifier during a 500-ns relativistic electron beam current pulse

A relativistic plasma microwave amplifier with a gain of about 30 dB and an output power of about 60–100 MW in the frequency range from 2.4 to 3.2 GHz is studied experimentally. The total duration of the output microwave pulse is equal to the duration of the current pulse of the driving relativistic electron beam (500 ns); however, the maximum output power is observed only within 200 ns. It is shown that variations in the output microwave power during the current pulse of the annular relativistic electron beam are caused by variations in the beam radius and thickness. Analysis of the experimental data and results of numerical simulations show that the thickness of the electron beam is determined by the density of the cathode emission current.     

Double mutant studies identify electrostatic interactions that are important for docking cytochrome c2 onto the bacterial reaction center

Cytochrome c2 (cyt) is the mobile electron donor to the reaction center (RC) in photosynthetic bacteria. The electrostatic interactions involved in the dynamics of docking of cyt onto the RC were examined by double mutant studies of the rates of electron transfer between six modified Rhodobacter sphaeroides RCs in which negatively charged acid residues were replaced with Lys and five modified Rhodobacter capsulatus Cyt c2 molecules in which positively charged Lys residues were replaced with Glu. We measured the second-order rate constant, k2, for electron transfer from the reduced cyt to the oxidized primary donor on the RC, which reflects the energy of the transition state for the formation of the active electron transfer complex. Strong interactions were found between Lys C99 and Asp M184/Glu M95, and between Lys C54 and Asp L261/Asp L257. The interacting residues were found to be located close to each other in the recently determined crystal structure of the cyt-RC complex [Axelrod, H., et al. (2002) J. Mol. Biol. (in press)]. The interaction energies were approximately inversely proportional to the distances between charges. These results support earlier suggestions [Tetreault, M., et al. (2001) Biochemistry 40, 8452-8462] that the structure of the transition state in solution resembles the structure of the cyt-RC complex in the cocrystal and indicate that specific electrostatic interactions facilitate docking of the cyt onto the RC in a configuration optimized for both binding and electron transfer. The specific interaction between Asp M184 and Lys C99 may help to nucleate short-range hydrophobic contacts.     

Formation of engineered intersubunit disulfide bond in cytochrome bc1 complex disrupts electron transfer activity in the complex

Protein domain movement of the Rieske iron-sulfur protein has been speculated to play an essential role in the bifurcated oxidation of ubiquinol catalyzed by the cytochrome bc1 complex. To better understand the electron transfer mechanism of the bifurcated ubiquinol oxidation at Qp site, we fixed the head domain of ISP at the cyt c1 position by creating an intersubunit disulfide bond between two genetically engineered cysteine residues: one at position 141 of ISP and the other at position 180 of the cyt c1 [S141C(ISP)/G180C(cyt c1)]. The formation of a disulfide bond between ISP and cyt c1 in this mutant complex is confirmed by SDS-PAGE and Western blot. In this mutant complex, the disulfide bond formation is concurrent with the loss of the electron transfer activity of the complex. When the disulfide bond is released by treatment with beta-mercaptoethanol, the activity is restored. These results further support the hypothesis that the mobility of the head domain of ISP is functionally important in the cytochrome bc1 complex. Formation of the disulfide bond between ISP and cyt c1 shortens the distance between the [2Fe-2S] cluster and heme c1, hence the rate of intersubunit electron transfer between these two redox prosthetic groups induced by pH change is increased. The intersubunit disulfide bond formation also decreases the rate of stigmatellin induced reduction of ISP in the fully oxidized complex, suggesting that an endogenous electron donor comes from the vicinity of the b position in the cytochrome b.     

Soluble variants of Rhodobacter capsulatus membrane-anchored cytochrome cy are efficient photosynthetic electron carriers

Photosynthetic (Ps) electron transport pathways often contain multiple electron carriers with overlapping functions. Here we focus on two c-type cytochromes (cyt) in facultative phototrophic bacteria of the Rhodobacter genus: the diffusible cyt c2 and the membrane-anchored cyt c(y). In species like R. capsulatus, cyt c(y) functions in both Ps and respiratory electron transport chains, whereas in other species like R. sphaeroides, it does so only in respiration. The molecular bases of this difference was investigated by producing a soluble variant of cyt c(y) (S-c(y)), by fusing genetically the cyt c2 signal sequence to the cyt c domain of cyt c(y). This novel electron carrier was unable to support the Ps growth of R. capsulatus. However, strains harboring cyt S-c(y) regained Ps growth ability by acquiring mutations in its cyt c domain. They produced cyt S-c(y) variants at amounts comparable with that of cyt c2, and conferred Ps growth. Chemical titration indicated that the redox midpoint potential of cyt S-c(y) was about 340 mV, similar to that of cyts c2 or c(y). Remarkably, electron transfer kinetics from the cyt bc1 complex to the photochemical reaction center (RC) mediated by cyt S-c(y) was distinct from those seen with the cyt c2 or cyt c(y). The kinetics exhibited a pronounced slow phase, suggesting that cyt S-c(y) interacted with the RC less tightly than cyt c2. Comparison of structural models of cyts c2 and S-c(y) revealed that several of the amino acid residues implicated in long-range electrostatic interactions promoting binding of cyt c2 to the RC are not conserved in cyt c(y), whereas those supporting short-range hydrophobic interactions are conserved. These findings indicated that attaching electron carrier cytochromes to the membrane allowed them to weaken their interactions with their partners so that they could accommodate more rapid multiple turnovers.     

Ca2+参与茉莉酸诱导蚕豆气孔关闭的信号转导

以Fluo-3AM为Ca~(2 )荧光探针,结合激光共聚焦扫描显微技术,观察到在处理后数十秒内,气孔关闭之前,茉莉酸(JA)可引起[Ca~(2 )]cyt的迅速上升;叶照和JA的前体物亚麻酸(LA)几乎不能引起[Ca~(2 )]cyt的明显变化;钙的螯合剂EGTA预处理可完全阻断JA诱导气孔关闭的效应,并且JA不再引起保卫细胞[Ca~(2 )]cyt增加;质膜Ca~(2 )通道的抑制剂硝苯吡啶(nifedipine,NIF)可减弱JA诱导气孔关闭的效应,也使JA诱导保卫细胞[Ca~(2 )]cyt增加的幅度有所下降;胞内Ca~(2 )释放的抑制剂钌红不能明显改变JA诱导气孔关闭的趋势,但使JA引起的保卫细胞[Ca~(2 )]cyt增加有所降低。实验结果表明:Ca~(2 )参与JA诱导气孔关闭的信号转导;推测JA引起的[Ca~(2 )]cyt升高可能主要来源于胞外,但不能完全排除胞内Ca~(2 )的释放。     

REDUCTION OF HEATING ARTIFACTS IN THIN SECTIONS EXAMINED IN THE ELECTRON MICROSCOPE

Certain phenomena affecting contrast obtained from tissue sections with the electron microscope have been investigated and a technique is described for reducing destruction by the electron beam of fine details in sections. It has been concluded that loss of embedding material is slightly higher at exposed surfaces of sections than it is at surfaces covered by substrate film. Covering of both surfaces of sections with thin films of formvar, collodion, or carbon materially improves the general appearance, reduces distortion, and sometimes reduces loss of tissue mass from the section as result of exposure to the electron beam. This improvement is considered to result from the relatively high melting-point of the covering films which serve to eliminate or reduce surface-tension or other forces operating in methacrylate softened by the electron beam.