Electrochemical evaluation of cellular physiological status under stress in Escherichia coli with the rpoS-lacZ reporter gene

We developed an electrochemical detection method for evaluating cellular physiological status based on the stringent response as a means to monitor cell viability. A reporter plasmid was constructed by inserting the beta-galactosidase gene (lacZ) under the control of the rpoS promoter, and then used to transform E. coli cells. Electrochemical responses from the products catalyzed by beta-galactosidase expressed by these E. coli cells were detected using the chronoamperometric technique in a nondestructive manner. Comparisons of response currents between the relA-positive strain and relA-negative strain revealed that increases in these currents were caused by the stringent response due to the stressful alcoholic environment, and thus as a model of stressful cultivating conditions. The current was proportional to the beta-galactosidase activity assayed by a conventional method that required the destruction of cells. The cellular physiological status, which depends on the stringent response as a viability marker, therefore, could then be evaluated online with a current using the rpoS-lacZ reporter gene in the relA-positive strain without pretreatment.     

信号分子ppGpp与微生物环境适应性

微生物能感知环境胁迫信号,通过触发严谨反应对生长速率进行调节,并通过一系列代谢调控,使细胞能在不利环境中生存。高度磷酸化的鸟苷四/五磷酸ppGpp/pppGpp（文中以ppGpp统称）作为信号分子对微生物生理具有广泛的调节作用,至今仍是微生物学研究热点之一。ppGpp对于微生物适应高温、高压等环境起到了积极的作用。综述了信号分子ppGpp合成降解机制及其调控微生物适应性方面的研究进展。     

The role of the omega subunit of RNA polymerase in expression of the relA gene in Escherichia coli

The rpoZ gene for the omega subunit of Escherichia coli RNA polymerase constitutes single operon with the spoT gene, which is responsible for the maintenance of stringent response under nutrient starvation conditions. To identify the physiological role of the omega subunit, we compared the gene expression profile of wild-type Escherichia coli with that of an rpoZ deleted strain by microarray analysis using an E. coli DNA chip. Here we report on a set of genes which show changes in expression profile following the removal of rpoZ. We have seen that relA, which is responsible for the synthesis of the stringent factor ppGpp and many ribosomal proteins, exhibited noticeable changes in mRNA levels and were therefore further analyzed for their expression using a GFP/RFP two-fluorescent protein promoter assay vector. In the absence of rpoZ, the promoter for the relA gene was severely impaired, but the promoters from the ribosomal protein genes were not affected as much. Taking these results together we propose that the omega subunit is involved in regulation of the relA gene, but induction of the stringently controlled genes in the absence of rpoZ is, at least in part, attributable to a decrease in ppGpp level.     

Studies on the Degradation Mechanisms of Proteins and their Derivatives in the Foods

The volatile sulfur components produced by boiling soybean meal hydrolyzates (AMINOSAN-EKI) have been identified as dimethyl sulfide and hydrogen sulfide. No mercaptan or disulfides were detected.

The main precursor of dimethyl sulfide is supposed to be methionine methylsulfonium compound derived from methionine and pectin substances (–COOCH₃) during the hydrolysis of soybean meal by hydrochloric acid.     

重复遭遇战对雌性黑线仓鼠生理状态的影响

以独居型啮齿动物——黑线仓鼠(Cricetulus barabensis)为对象,主要测定了与优势-从属关系有关的体重、生理状态和粪便激素变化规律相关的参数。野生成年仓鼠捕获于鲁西南山区农田并于实验室内单独饲养。按照陌生和体重匹配的原则配对,实验动物在中立竞技场中每天进行15 m in的遭遇战并收集粪便,连续28 d。以攻击行为、防御行为、社会行为、非社会行为确定个体的优势-从属地位。用于实验的动物共33只。行为实验结束后,将动物解剖,确定其繁殖及生理状态。结果表明:4周重复遭遇战期间,(1)优势雌体体重显著增加;(2)肾上腺指数显著降低;(3)雌体肾上腺指数与卵巢指数无显著的相关关系;(4)雌体粪便皮质醇及雌二醇含量随时间表现出明显的波动,但优势与从属个体间差异不显著;(5)优势雌体的粪便雌二醇含量与皮质醇含量呈显著的正相关关系。研究结果提示,重复遭遇战未影响雌体的存活及潜在繁殖能力,这可能与黑线仓鼠的独居生活方式有关。     

The A-factor regulatory cascade and cAMP in the regulation of physiological and morphological development in Streptomyces griseus

In the A-factor regulatory cascade leading to the onset of streptomycin biosynthesis and aerial mycelium formation in Streptomyces griseus, the A-factor receptor protein (ArpA) serves as a DNA-binding repressor and A-factor releases the repression by binding to ArpA and dissociating it from the DNA. Mutants defective in arpA therefore produce streptomycin and aerial hyphae in the absence of A-factor. A gene that inhibits streptomycin production and aerial hyphae formation in an arpA mutant was cloned on a high-copy-number plasmid and found to encode a eukaryotic-type adenylate cyclase (CyaA). Consistent with this, an exogenous supply of cAMP at high concentration almost abolished streptomycin production and aerial hyphae formation. On the other hand, cAMP at lower concentrations stimulated or accelerated these developmental processes. The effects of cAMP were detectable only in arpA mutants, and not in the wild-type strain; an exogenous supply of cAMP or cyaA disruption in the wild-type strain caused almost no effect on these phenotypes. Thus the effects of cAMP became apparent only in the arpA-defective background. cAMP at high concentrations inhibited stringent response factor ppGpp production, which is important for the onset of antibiotic biosynthesis. cAMP also influenced the timing of tyrosine phosphorylation of more than nine proteins. These findings show that a cAMP regulatory relay for physiological and morphological development functions in a concerted and interdependent way with other signal transduction pathways. Journal of Industrial Microbiology & Biotechnology (2001) 27, 177–182. Received 21 September 1999/ Accepted in revised form 14 September 2000     

Effect of physiological status on olfactory and visual responses of female Hypothenemus hampei during host plant colonization

The response of Hypothenemus hampei (Ferr.) (Coleoptera, Scolytidae) (coffee berry borer, CBB) females to olfactory stimuli was demonstrated to be related with the physiological status; especially with age, whether they were mated or not, and whether they have already oviposited. A special behaviour, the abandonment of the native berry, was also found to play a major role. In contrast, response to visual stimuli (false coffee berries) was found to be independent of the physiological status of CBB females. Variations in behavioural responses to visual and olfactory stimuli are discussed in relation to colonization strategy.     

基于光谱信息的作物氮素营养无损监测技术

氮素是作物生长发育和产量品质形成所必需的营养元素。快速、无损、准确地监测作物氮素状况,对于诊断作物生长特征、提高氮肥运筹水平和利用效率、降低过量施氮带来的农田环境污染,深入开展精确农业和数字农业的研究与应用具有重要意义。本文围绕作物氮素特征光谱产生的机制、反射光谱对氮素营养的响应及光谱指数的生理意义等解析了作物氮素营养光谱无损监测的技术机理,阐明了作物氮素监测的光谱学和生理生态学基础,进而概述了国内外有关作物反射光谱获取及叶片、冠层和空间水平上氮素营养光谱监测的研究进展。针对作物氮素营养监测亟待解决的问题和作物生产需求,提出今后进一步研究的领域应重点围绕氮素生化组分的监测方法、氮素监测模型的普适性、氮素监测仪的开发、地空遥感信息的融合、遥感与其他技术的集成等方面。     

Structure of the PPX/GPPA phosphatase from Aquifex aeolicus in complex with the alarmone ppGpp

The crystal structure of the prototype exopolyphosphatase/guanosine pentaphosphate phosphohydrolase protein family member from Aquifex aeolicus in complex with the intracellular second messenger guanosine tetraphosphate was determined at 2.7-Å resolution. The hydrolytic base is identified as E119. The dual specificity established for the Escherichia coli homolog is shown to be compatible with a common active site for guanosine pentaphosphate and polyphosphate hydrolysis. Distinct and different degrees of closure between the two domains of the enzyme are associated with substrate binding. The arginines R22 and R267, residing in different domains, are crucial for guanosine pentaphosphate specificity as they interact with the unique 3′-ribose phosphorylation.     

锌对镉胁迫下马蔺生长、镉积累及生理抗性的影响

通过溶液培养研究了不同浓度锌(Zn)对镉(Cd)胁迫下马蔺(Iris lacteavar. chinen-sis)生长、Cd积累及抗氧化酶活性等的影响.结果表明:加入1~100mg.L-1Zn后,Cd胁迫下马蔺地上部Cd含量变化不大,而地下部Cd含量显著增加并呈先增后降的趋势;1mg.L-1Zn处理下马蔺地下部Cd含量最高,比对照显著增加51.4%;Zn浓度高于1mg.L-1后Cd含量均出现不同程度的下降,但仍高于对照.与单独Cd处理(10mg.L-1)相比,添加低浓度Zn(1~10mg.L-1)后,马蔺地上部生物量和叶绿素含量呈增加趋势,丙二醛(MDA)含量降低;在10mg.L-1Zn处理下,叶绿素a、b含量达到峰值,分别增加5.21%和22.27%,MDA含量降低25.46%,表明低浓度Zn缓解了Cd对马蔺的毒害.随Zn浓度的增加,Zn对Cd毒害的缓解作用逐渐降低,当溶液中Zn达到一定浓度(100mg.L-1)时,马蔺毒害加重,其生物量、叶绿素含量均下降,MDA含量显著增加.在试验胁迫浓度范围内超氧化物歧化酶(SOD)、过氧化物酶(POD)活性始终呈增加趋势,表明马蔺在受到Cd胁迫及Cd-Zn共存的条件下SOD和POD起着重要的抗氧化保护作用.     

矿井废水灌溉对小麦生理特性及重金属积累的影响

以井水灌溉为对照 (CK),采用盆栽试验研究了矿井废水灌溉对小麦生理特性和重金属积累的影响.设置了3个矿区废水灌溉处理:洗煤废水(T₁)、经沉淀处理的洗煤废水 (T₂) 和煤矸石淋溶水 (T₃),于返青期开始进行矿区废水灌溉处理.结果表明： 矿井废水灌溉处理对小麦的生长发育和产量均有不同程度的负面影响.到开花期时,T₁、T₂和T₃处理小麦的单茎质量和叶面积、根系活力和光合速率均显著低于对照(P＜0.05),T₃处理小麦株高和叶绿素含量(SPAD值)显著降低(P＜0.05);T₁、T₂和T₃处理的籽粒产量分别比对照下降15.4%、9.8%和17.8%.此外,矿井废水灌溉处理小麦籽粒中Cr、Pb、Cu和Zn的含量均显著高于对照,表明矿井废水灌溉导致重金属在小麦籽粒中积累.
     

叶片结构的水力学特性对植物生理功能影响的研究进展

叶片是植物进行光合、呼吸、蒸腾作用的主要器官, 早期的研究主要集中于水分在叶片中的运输路径, 而对叶脉结构及其生态学意义研究甚少。近年来关于叶片叶脉结构、气孔结构的功能及叶片水力学特性的意义研究已经成为植物生理生态的研究热点。该文综述了叶脉的结构性状的指标(叶脉密度、直径、间距等), 叶片水力学结构特性对植物生长、水分运输、气体交换、光合作用等生理功能的影响, 及其与植物对干旱适应性之间的关系。叶脉结构是决定叶片生理功能的基础, 因此在未来的工作中应分析比较不同种类植物叶脉结构形态与导水、光合、呼吸、同化作用之间的关系, 建立植物茎干-枝-叶系统水力传导的机理性模型, 用以探索不同植物功能结构和高效用水生理生态学机制, 据此评估不同种类植物在未来气候情景下的地位。     

Integrating cellular senescence with the concept of damage accumulation in aging: Relevance for clearance of senescent cells

Understanding the aging process and ways to manipulate it is of major importance for biology and medicine. Among the many aging theories advanced over the years, the concept most consistent with experimental evidence posits the buildup of numerous forms of molecular damage as a foundation of the aging process. Here, we discuss that this concept integrates well with recent findings on cellular senescence, offering a novel view on the role of senescence in aging and age‐related disease. Cellular senescence has a well‐established role in cellular aging, but its impact on the rate of organismal aging is less defined. One of the most prominent features of cellular senescence is its association with macromolecular damage. The relationship between cell senescence and damage concerns both damage as a molecular signal of senescence induction and accelerated accumulation of damage in senescent cells. We describe the origin, regulatory mechanisms, and relevance of various damage forms in senescent cells. This view on senescent cells as carriers and inducers of damage puts new light on senescence, considering it as a significant contributor to the rise in organismal damage. Applying these ideas, we critically examine current evidence for a role of cellular senescence in aging and age‐related diseases. We also discuss the differential impact of longevity interventions on senescence burden and other types of age‐related damage. Finally, we propose a model on the role of aging‐related damage accumulation and the rate of aging observed upon senescent cell clearance.     

On the use of cellular telephony for audio interaction with animals

Playback is an important method of surveying animals, assessing habitats and studying animal communication. However, conventional playback methods require on-site observers and therefore become labour-intensive when covering large areas. Such limitations could be circumvented by the use of cellular telephony, a ubiquitous technology with increasing biological applications. In addressing concerns about the low audio quality of cellular telephones, this paper presents experimental data to show that owls of two species (Strix varia and Megascops asio) respond similarly to calls played through cellular telephones as to calls played through conventional playback technology. In addition, the telephone audio recordings are of sufficient quality to detect most of the two owl species' responses. These findings are a first important step towards large-scale applications where networks of cellular phones conduct real-time monitoring tasks.     

神经肽Y(NPY)的生理功能研究进展

神经肽Y(NPY)是机体内的一种重要且保守的神经递质,一般以前体形式存在,释放的有活性的NPY主要通过与其受体结合发挥作用。NPY受体包含了亚型Y1、Y2、Y3、Y4、Y5、Y6、Y7、Y8。Y1和Y2是NPY发挥收缩血管作用的关键受体;Y1、Y2和Y5是NPY调节动物摄食行为的关键受体;Y1、Y2和Y4是NPY调控动物焦虑、沮丧行为的必要受体。着重对NPY与其各种受体结合后如何行使动物的相关生理功能的情况进行了阐述。     

Dynamics in the bacterial community-level physiological profiles and hydrological characteristics of constructed wetland mesocosms during start-up

The objective of this work was to study the effect of plant presence (Phragmites australis) and inoculant origin on wetland mesocosm start-up dynamics. Eight mesocosms were studied based on a duplicated 2² factorial design tracking bacterial community and hydrological changes during an 8 month start-up period. The mesocosms were characterized in terms of their hydrological character based on evapotranspiration (ET), porosity, and a dispersion coefficient. The microbiological regime was characterized using a microbial activity measure and community-level physiological profiling (CLPP) employing BIOLOG™ ECO plates. CLPP-related indices such as substrate richness, substrate diversity, over-all community profile, and community divergence are also presented. It was found that mesocosm porosities decreased over time as a result of media-related biofilm development. This biofilm development also contributed to a substantial increase in the dispersion coefficient in the mesocosms over the start-up period. Dispersion coefficients in planted systems reached values of ∼50-55 cm²/min whereas in the unplanted systems values of ∼30-35 cm²/min were observed. Bacterial community divergence in the mesocosms was quantified using a Euclidean-based divergence metric. All mesocosms showed a sharp increase in community divergence until day 75, at which point a steady state was reached. The interstitial communities were also characterized in terms of similarity based on the experimental design treatments. Four stages of mesocosm development were identified that can be described by an initial community state based on the origins of the initial inoculum [days 0-6]; a dynamic period where adjustments and shifts in the bacterial community occurred in all mesocosms [days 7-26]; a period where all interstitial CLPPs were quite similar [days 27-73]; and finally a shift towards unplanted and planted mesocosm CLPP groupings [days 74-232].     

盐胁迫对玫瑰生长和生理特性的影响

以野生玫瑰和3个玫瑰栽培品种一年生扦插苗为试材,对不同浓度NaCl胁迫下玫瑰的生物量、光合作用、渗透调节物质含量、根活力和离子含量进行了研究.结果表明:盐胁迫抑制了玫瑰的生长,与地上部相比,根系对盐胁迫更加敏感;盐胁迫下,野生玫瑰的游离脯氨酸和可溶性糖含量显著高于栽培玫瑰;品种‘紫雁’高于‘紫枝’和‘中科二号’;盐胁迫对野生玫瑰光合性能和根活力的影响明显小于栽培玫瑰.野生玫瑰的耐盐性优于栽培玫瑰;而栽培玫瑰中‘紫雁’优于‘紫枝’和‘中科二号’.上述生理指标可作为玫瑰耐盐性评价的参考指标.     

Restoration of the cellular thiol status of peritoneal macrophages from CAPD patients by the flavonoids silibinin and silymarin

During continuous ambulatory peritoneal dialysis (CAPD) the peritoneal immune cells, mainly macrophages, are highly compromised by multiple factors including oxidative stress, resulting in a loss of functional activity. One reason for the increase of inflammatory reactions could be an imbalance in the thiol-disulfide status. Here, the possible protective effects of the antioxidant flavonoid complex silymarin and its major component silibinin on the cellular thiol status were investigated. Peritoneal macrophages from dialysis fluid of 30 CAPD patients were treated with silymarin or silibinin up to 35 days.

A time-dependent increase of intracellular thiols was observed with a nearly linear increment up to 2.5-fold after 96 hours, reaching a maximum of 3.5-fold after 20 days of culture. Surface-located thiols were also elevated. The stabilization of the cellular thiol status was followed by an improvement of phagocytosis and the degree of maturation as well as significant changes in the synthesis of IL-6 and IL-1ra. Furthermore, the treatment of peritoneal macrophages with flavonoids in combination with cysteine donors resulted in a shortened and more efficient time course of thiol normalization as well as in a further increased phagocytosis. In addition, GSH-depletion in thiol-deficient media simulating CAPD procedures led to intracellular thiol deficiency similar to the in vivo situation.

It is concluded that treatment with milk thistle extracts silymarin and silibinin alone or, more effectively in combination with cysteine donors, provide a benefit for peritoneal macrophages of CAPD-patients due to a normalization and activation of the cellular thiol status followed by a restoration of specific functional capabilities.