Crystal structure and functional characterization of selenocysteine-containing glutathione peroxidase 4 suggests an alternative mechanism of peroxide reduction

Glutathione peroxidases (GPX) are anti-oxidative enzymes that reduce organic and inorganic hydroperoxides to the corresponding alcohols at the expense of reduced glutathione. The human genome involves eight GPX genes and five of them encode for selenocysteine-containing enzymes. Among the human GPX-isoforms, GPX4 is unique since it is capable of reducing complex hydroperoxy ester lipids such as hydroperoxy phospholipids and hydroperoxy cholesterolesters. Using a number of genetically modified mouse strains the biological role of GPX4 has comprehensively characterized but the molecular enzymology is less well explored. This lack of knowledge is partly related to the fact that mammalian selenoproteins are not high-level expressed in conventional overexpression systems. To explore the structural and functional properties of human GPX4 we expressed this selenoprotein in a cysteine-auxotrophic E. coli strain using a semi-chemical expression strategy. The recombinant enzyme was purified in mg amounts from the bacterial lysate to electrophoretic homogeneity and characterized with respect to its protein-chemical and enzymatic properties. Its crystal structure was solved at 1.3?Å resolution and the X-ray data indicated a monomeric protein, which contains the catalytic selenium at the redox level of the seleninic acid. These data suggest an alternative reaction mechanism involving three different redox states (selenol, selenenic acid, seleninic acid) of the catalytically active selenocysteine.     

Genes encoding acidic and basic class III β-1,3-glucanases are expressed in tomato plants upon viroid infection

-1,3-glucanases are hydrolytic enzymes considered to constitute part of the general array of defense genes induced by pathogen infection in higher plants. We have isolated and characterized two complementary DNA clones, corresponding to new -1,3-glucanases from tomato plants (Lycopersicon esculentum) which are expressed upon challenge with citrus exocortis viroid. Amino acid sequence comparison revealed that they are most similar to -1,3-glucanases from tobacco, particularly to PR-Q, the unique component of the class III -1,3-glucanase. The deduced amino acid sequences of the two tomato -1,3-glucanases indicate that, although being highly similar in amino acid sequence, they have different isoelectric points: pI 10.5 for the basic isoform (Tom PR-Q b) and pI 5.2 for the acidic one (Tom PR-Q a). The expression of these two -1,3-glucanase messenger RNAs (mRNAs) in response to viroid infection and ethephon treatments was examined. mRNAs for these two isoforms are coordinately expressed and induced similarly to mRNAs for other PR proteins, indicating that they are part of a general and coordinate mechanism of response of tomato plants susceptible to viroid infection.     

Structurally unrelated algal oligosaccharides differentially stimulate growth and defense against tobacco mosaic virus in tobacco plants

Tobacco plants were treated with structurally unrelated oligosaccharides obtained from Chilean marine macroalgae. These oligosaccharides were prepared by chemical depolymerization of native polysaccharides extracted from brown and red algae and correspond to pure polymers of around 20 units of guluronic acid (Poly-Gu), mannuronic acid (Poly-Ma) and sulphated galactan (Poly-Ga). These oligosaccharides were solubilized in water, at a final concentration of 500 μg mL⁻¹, and sprayed on tobacco leaves, once a week for a month. Their effects on the stimulation of growth and defense against tobacco mosaic virus (TMV) were determined 7 and 15 days after the final spraying, respectively. The activities of several defense and antioxidant enzymes and the levels of water-soluble antioxidant compounds were determined. Plants treated with Poly-Ga and Poly-Ma showed an increase in height of 23% and 49%, respectively, whereas Poly-Gu did not stimulate growth. Plants treated with Poly-Gu, Poly-Ma and Poly-Ga showed an increase in defense against TMV corresponding to decreases in the number of necrotic lesions of 9%, 22% and 74%, respectively. The stimulation of plant growth correlates with activation of the antioxidant enzyme ascorbate peroxidase (AP) and with a decrease in ascorbate level. On the other hand, the stimulation of defense against TMV is correlated with the activation of the defense enzyme phenylalanine ammonia-lyase (PAL). These results indicate that algal oligosaccharides differentially stimulate growth and defense against TMV in tobacco plants and that these processes involve the activation of the enzymes AP and PAL, respectively.     

Influence of waterlogging on some anti-oxidative enzymatic activities of two barley genotypes differing in anoxia tolerance

The experiment was conducted to investigate the formation of oxidative stress and the development of anti-oxidative enzymes in two barley genotypes differing in anoxia tolerance. Waterlogging led to significant reduction in root and shoot weight, green leaf area and tillers per plant, but tolerant Xiumai 3 was much less reduced than sensitive Gerdner. Malondialdehyde (MDA) content, an indicator of membrane lipid peroxidation, significantly increased in Gerdner when the plants were subjected to waterlogging, but remained little changed in Xiumai 3. Superoxide dismutase (SOD) activity was increased with waterlogging treatment and the sensitive cultivar had higher activity than the tolerant one during the experimental duration. At early stage of waterlogging treatment, both peroxidase (POD) and catalase (CAT) activities significantly increased in Xuimai 3, while obviously decreased in Gerdner. Moreover, both cultivars showed substantial increase in both POD and CAT with the progress of waterlogging exposure. Glutathione reductase (GR) activity was increased in both tolerant- and sensitive cultivars under waterlogging. It may be assumed from the current results that SOD activity appears to be not a constraining factor limiting the scavenging of ROS, and it is the change of POD and CAT activity under waterlogging that determine the status of oxidative stress. The difference between genotypes in waterlogging tolerance could be distinguished from the changed patterns of these enzymatic activities.     

沙漠化过程中四个共有种的生长和抗氧化系统酶类变化

探讨了自然条件下沙质草原沙漠化过程中4个共有植物种群的形态、生长和抗氧化系统酶类变化规律．结果表明,沙漠化过程中共有种种群株高、密度、密度百分比总体上呈降低趋势,其中扁蓿豆中度沙漠化阶段前生长渐趋旺盛,中度沙漠化阶段后生长受到限制,未达到显著水平(P>0．05)．羊草受损最重,中度沙漠化阶段株高下降至原生植被阶段的57．19％,密度和密度百分比仅为原生植被阶段的2．50％和6．22％．糙隐子草和冷蒿在潜在沙漠化、轻度沙漠化或中度沙漠化阶段株高的增加与种群所处的阶段性优势地位及其抗逆性增强有关．共有种群SOD、POD活性普遍在潜在沙漠化、中度沙漠化阶段增加,轻度沙漠化、重度沙漠化阶段降低．羊草过氧化氢酶(CAT)活性较高,对沙漠化的响应不显著(P>0．05);扁蓿豆CAT活性在潜在沙漠化、重度沙漠化阶段明显升高(P≤0．01)．在重度沙漠化阶段,共有种3种酶活性普遍下降,只有扁蓿豆CAT活性上升．共有种群丙二醛含量从原生植被到中度沙漠化阶段均呈“先升后降再升”的变化,不同阶段之间差异显著(P≤0．05)．综合分析表明,4个共有种中,羊草对沙漠化较敏感,扁蓿豆则生命力最强。     

病毒感染引起的炎症反应：宿主对抗病毒的“双刃剑”

先天性免疫系统作为宿主抵抗外来病原入侵的第一道防线,也是最迅速的防御系统。宿主先天性免疫系统中的模式识别受体识别入侵信号并激活炎症信号通路,诱导产生大量促炎性细胞因子,引起炎症反应。病毒感染是激活炎症反应的条件之一,诱导机体产生强烈的免疫应答,强大的炎症反应调控网络在宿主抗病毒过程中发挥关键作用,以维持机体的平衡。本文综述了病毒感染引起的炎症反应,重点介绍了宿主对炎症反应的调控网络,以及DNA和RNA病毒对炎症反应的调节机制,为病毒感染引起的免疫性疾病的治疗提供参考。     

近40年沈阳城市森林春季物候与全球气候变暖的关系

1960-2005年期间,沈阳气候受全球气候变暖的影响,年平均气温总体呈上升趋势,气温升幅为0.96℃,但以1982年为界,1960-1982年为偏冷阶段,气温降幅最大值为1.95 ℃:1983-2005年为偏暖阶段,气温升幅最大值为1.60 ℃.一年四季气温处于上升阶段,以冬、春季气温升幅最大,分别为2.3 ℃和1.35℃.城市森林主要树种的春季物候期,在气候偏冷阶段,春季物候期出现较晚,而在偏暖阶段,春季物候期提前发生.同时,物候春季开始日期与结束日期有密切的同步相关性,并与物候季节节奏的长短呈负相关.树木萌动期早晚与冬季和早春气温高低呈显著的负相关,冬春季气温越高,芽萌动越提前.展叶早晚主要受展叶前的春季气温高低的影响,与冬季气温相关性不显著.始花前2-8旬,特别是2-14旬气温对始花期影响最显著.寒冷指数(CI)与树木芽萌动期、展叶始期呈显著正相关,而与开花始期相关性不显著.预测了CO2倍增和气温升高条件下,沈阳城市森林主要树种的萌动期、展叶始期和开花始期分别提前40.41、43.08和24.13 d.     

硒对水稻的生态毒理效应及临界指标研究

通过土壤添加硒盆栽试验,研究了黄棕壤不同浓度Se对水稻生物量、叶绿素含量、抗氧化酶系统的影响.结果表明,低浓度Se(<8mg·kg^-1)对水稻根和地上生物量、叶绿素a含量、叶绿素a/b有促进作用,并整体提高了作物抗氧化酶系统,使MDA下降,SOD、CAT、POD、GSHPx活性相应提高.而高浓度Se(>16mg·kg^-1)对水稻根和地上生物量、叶绿素a含量、叶绿素a/b有明显的抑制作用,对抗氧化酶系统产生胁迫效应,使GSHPx明显上升,MDA上升,SOD、CAT、POD酶活性明显下降.各项生理指标与土壤Se处理浓度之间相关分析表明,土壤Se浓度与地上部干重相对百分率、根干重百分率、叶绿素a/b、MDA、SOD、CAT、POD之间有显著相关性.比较胁迫效应10%值的临界指标EC10值表明,作物地上部干重抑制率作为土壤临界值制定的依据更为可靠,EC10为21mg·kg^-1.     

Iron withholding: A defense against viral infections

A variety of laboratory and clinical investigations during the past 15 years have observed that one of the dangers of excessive iron is its ability to favor animal viral infections. The metal is essential for host cell synthesis of virions and can also impair defense cell function and increase oxidative stress. In both animal models and humans, viral infections cause upregulation of the iron withholding defense system. Factors that suppress the system enhance viral progression; factors that strengthen the system augment host defense. Procedures designed to reinforce the system are being developed and tested; some of these may become useful adjuncts in prevention and management of viral diseases.     

Antioxidative protection and proteolytic activity in tolerant and sensitive wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) varieties subjected to long-term field drought

Field drought studies were performed in order to assess oxidative stress, proteolytic activity and yield loss under natural stress conditions. Flag leaves of two drought-tolerant (Yantar and Zlatitsa) and two drought-sensitive (Miziya and Dobrudjanka) winter wheat varieties were analyzed. Stress intensity was assessed by relative electrolyte leakage and proline accumulation. Senescence progression was followed by loss of chlorophyll and protein. Lipid peroxidation, H₂O₂ content, activities of superoxide dismutase (SOD), catalase (CAT), and non-specific peroxidase (GPX) isoforms, as well as proteolytic activities were analyzed from heading throughout grain filling. Weakening of membrane integrity and oxidative damage to lipids were more pronounced in the sensitive varieties under field drought. The activities of Fe- and Cu/Zn SOD isoforms decreased in the controls, but remained high in drought-treated plants. The activities of MnSOD isoforms and CAT were enhanced towards grain filling, especially in the sensitive varieties under drought. GPX activities were rised under drought but progressively diminished. Accelerated senescence under field drought was linked to higher proteolytic activity with variety specific differences in the protease response, but without a clear correlation to drought resistance or sensitivity. Field drought led to higher oxidative stress more pronounced for drought sensitive varieties, especially during the grain filling period.     

牲畜粪便对草地生态系统影响的研究进展

粪便归还是放牧影响草地生态系统的主要途径之一。本文对粪便分解过程和粪便对草地生态系统的土壤理化性质、微生物活性、植物群落结构和生物量、牲畜取食格局影响等方面的研究进展进行了综述,最后提出了草地生态系统牲畜粪便研究的未来趋势。     

AM真菌和镰刀菌对西瓜根系几种酶活性的影响

在温室盆栽条件下研究了丛枝菌根(Arbuscular Mycorrhiza, AM)真菌Glomus versiforme和西瓜枯萎镰刀菌Fusarium oxysporum f.sp. niveum对西瓜根系中过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、β-1,3-葡聚糖酶和几丁质酶活性的影响。结果表明,接种AM真菌的西瓜根系中4种酶的活性均高于对照,先接种G. versiforme,后接种F. oxysporum f.sp. niveum处理的4种酶的活性均高于只接种F. oxysporum f.sp. niveum 的处理,且酶的活性峰值出现较早。表明接种G. versiforme 能预先诱导这4种酶的产生,提高其活性,从而提高西瓜对F. oxysporum f.sp. niveum侵染的抗性。接种G. versiforme的感枯萎病西瓜品种“郑杂5号”酶的增加幅度大于抗病品种“京欣1号”的接种处理,说明G. versiforme对提高感病西瓜品种酶活性的作用更大。     

水稻对稻负泥虫取食的生理响应

以水稻负泥虫-水稻互作为研究对象,分析了水稻负泥虫Oulema oryzae Kuwayama取食过程中抗、感2个品种水稻植株内保护性酶POD、SOD、CAT和MDA活性的变化动态.结果表明:在水稻负泥虫取食为害的过程中,SOD活性持续增强,POD、CAT和MDA活性呈现出先升高后降低的趋势,而且抗、感水稻植株中4种酶...     

AM真菌和镰刀菌对西瓜根系几种酶活性的影响*

在温室盆栽条件下研究了丛枝菌根(Arbuscular Mycorrhiza, AM)真菌Glomus versiforme和西瓜枯萎镰刀菌Fusarium oxysporum f.sp. niveum对西瓜根系中过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、β-1,3-葡聚糖酶和几丁质酶活性的影响。结果表明,接种AM真菌的西瓜根系中4种酶的活性均高于对照,先接种G. versiforme,后接种F. oxysporum f.sp. niveum处理的4种酶的活性均高于只接种F. oxysporum f.sp. niveum 的处理,且酶的活性峰值出现较早。表明接种G. versiforme 能预先诱导这4种酶的产生,提高其活性,从而提高西瓜对F. oxysporum f.sp. niveum侵染的抗性。接种G. versiforme的感枯萎病西瓜品种“郑杂5号”酶的增加幅度大于抗病品种“京欣1号”的接种处理,说明G. versiforme对提高感病西瓜品种酶活性的作用更大。     

环境条件对芽孢杆菌B-9987菌株防御酶诱导效应的影响

研究室内外条件下,海洋芽孢杆菌不同处理物对番茄防御酶系的诱导作用。分别使用不同浓度芽孢杆菌B-9987发酵液和无细胞滤液喷施番茄植株,于室内外两种环境条件下养殖,测定了处理11d内番茄5种防御酶的变化趋势。两种处理诱导抗性结果显示,在两种环境条件下处理植株苯丙氨酸解氨酶（PAL）、多酚氧化酶（ppo）、过氧化物酶（POD）、过氧化氢酶（CAT）较空白对照植株酶活性均有不同程度增加。除超氧化物歧化酶（SOD）外,室内环境较室外更利于B-9987菌株处理液对番茄防御酶系的诱导作用发挥;相同环境条件下,发酵液的处理诱导活性较强。B-9987菌株对番茄防御酶系有较好的诱导作用,但诱导作用的发挥受环境条件影响明显。     

Cross-talking between autophagy and viral infection in mammalian cells

Autophagy is a cellular process in degradation of long-lived proteins and organelles in the cytosol for maintaining cellular homeostasis, which has been linked to a wide range of human health and disease states, including viral infection. The viral infected cells exhibit a complicated cross-talking between autophagy and virus. It has been shown that autophagy interacts with both adaptive and innate immunity. For adaptive immunity, viral antigens can be processed in autophagosomes by acidic proteases before major histocompatibility complex (MHC) class II presentation. For innate immunity, autophagy may assist in the delivery of viral nucleic acids to endosomal TLRs and also functions as a part of the TLR-or-PKR-downstream responses. Autophagy was also reported to suppress the magnitude of host innate antiviral immunity in certain cases. On the other hand, viruses has evolved many strategies to combat or utilize the host autophagy for their own benefit. In this review we discussed recent advances toward clarifying the cross-talking between autophagy and viral infection in mammalian cells.     

Technical enzymes produced in transgenic plants

Technical enzymes are used in many industrial applications. Nowadays technical enzymes are often produced in transgenic host organisms. The use of transgenic plants with respect to high level of expression at low costs as a prerequisite for successful commercial production of technical enzymes is discussed. This review summarises recently published examples for production of technical enzymes in plants. In addition, plastid transformation and viral vectors are discussed as methods which might be useful for obtaining high expression level of recombinant proteins in plants.     

降钙素原和超敏C-反应蛋白在病毒和细菌感染早期鉴别中的临床应用

为探讨降钙素原(PCT)和超敏C-反应蛋白(hs-CRP)在病毒感染和细菌感染患者鉴别诊断中的价值,对我院276例患者,包括病毒感染组(64例)、细菌感染组(164例)和非感染组(48例)进行降钙素原和超敏C-反应蛋白水平的检测,并对三组间的差异进行分析。结果显示,细菌感染组和病毒感染组PCT、hs-CRP水平显著性高于非感染组(p<0.05);细菌感染组PCT、hs-CRP水平显著性高于病毒感染组(p<0.05)。工作特征曲线及Logistic回归分析结果显示,PCT在鉴别诊断细菌感染和病毒感染的效果高于hs-CRP。研究结果表明,降钙素原、超敏C-反应蛋白可对病毒感染和细菌感染进行早期的鉴别诊断,而降钙素原的效果较好。