Control of glutathione and phytochelatin synthesis under cadmium stress. Pathway modeling for plants

Glutathione (GSH) plays several roles in cell metabolism such as redox state regulation, oxidative stress control, and protection against xenobiotics and heavy metals. GSH is synthesized in two steps catalysed by gamma-glutamylcysteine synthetase (gamma-ECS) and glutathione synthetase. gamma-ECS is feedback inhibited by GSH, which has led to the proposal that this enzyme acts as the rate-limiting step in the pathway. Thus far, the study of GSH metabolism has been confined to GSH synthesis (GSH supply), without considering the GSH-consuming enzymes (GSH demand). Several works have shown that the demand block of enzymes may have a significant control on a pathway; therefore, we hypothesize that GSH-consuming enzymes may exert some control on GSH synthesis. A kinetic model of GSH and phytochelatin synthesis in plants was constructed using the software GEPASI and the kinetic data available in the literature. The main conclusions drawn by the model concerning metabolic control analysis are (1) gamma-ECS is indeed a rate-limiting step in GSH synthesis, but only if GSH-consuming enzymes are not taken into account. (2) At low demand, GSH-consuming enzymes exert significant flux-control on GSH synthesis whereas at high demand, supply and demand blocks share the control of flux. (3) In unstressed conditions, flux to GSH is controlled mainly by demand, so that gamma-ECS determines the degree of homeostasis of the GSH concentration. Under cadmium exposure, the GSH demand increases and flux-control is re-distributed almost equally between the supply and demand blocks. (4) To enhance phytochelatins synthesis without depleting the GSH pool, at least two enzymes (gamma-ECS and PCS) should be increased and/or, alternatively, a branching flux (GSH-S-transferases) could be partially diminished.     

基于生态水文调节服务的石家庄雨涝灾害供需匹配分析与街区规划干预

在雨涝灾害威胁日益凸显的背景下,基于生态系统水文调节服务供需匹配分析的规划防涝优先干预区识别,为高效降低城市雨涝风险、利用规划措施防涝提供了新的研究思路与科学支撑。运用数据叠置、水文模拟及公式计算法,将城市原生雨涝供给能力与雨涝需求水平统一至同一评估体系中。结合降雨量、下垫面类型,运用径流曲线模型对街区地表径流调节率进行计算,生成供给能力评估结果;以GIS水文模拟结果计算得出的危险性、暴露性和脆弱性指数生成需求水平评估结果。根据石家庄中心四区3a一遇、50a一遇情景供需匹配结果,划定城市街区尺度的规划干预等级,并分类提出相应规划策略。研究结果表明:石家庄市中心四区低供给高需求街区在3a一遇降雨情景下以点状聚集分布;50a一遇降雨情景下,在京广铁路线沿线呈现纵向聚集形态。同时,在两种降雨情景下,规划干预高等级街区均集中出现于桥西区苑东街道、彭后街道、东华街道,长安区长丰街道、建北街道,裕华区裕翔街道、建华南街道。根据规划干预等级、供需相对关系及供需失衡原因,分别提出3a一遇、50a一遇降雨情景下的九类规划干预策略,为城市雨涝灾害的规划应对提供优化思路。     

Experimental supply-demand analysis of anaerobic yeast energy metabolism

Experimental supply-demand analysis of yeast fermentative energy metabolism shows that control of the glycolytic flux is shared between supply and demand. In glucose limited chemostat cultures the supply block was modulated in a dilution rate change and demand block via a benzoic acid titration. Under these conditions the supply block had a flux control of 0.90 and the demand block a flux control of 0.10.     

城市内涝调节服务供需关键区识别与优先级划分

在全球气候变化和城市化背景下,城市内涝灾害频发,严重危及城市居民的人身安全、财产安全及公共安全。如何精准施策,针对关键区域进行城市内涝调节治理已成为社会各界关注并亟需解决的重要问题。基于生态系统服务供需视角,以径流调节率表征城市内涝调节的供给水平,以胁迫性、暴露性和脆弱性指标表征城市内涝调节的需求水平,构建了完整的技术路线和指标体系,从而识别城市内涝调节供需失衡的关键区,并确定规划干预的优先级。基于此,以岛屿型城市厦门岛为例,使用遥感数据和人口、社会、经济数据,运用ArcMap10.8、ENVI5.3和GeoDA平台,模拟评估了城市街区尺度下城市内涝调节服务的供需水平及空间分布特征。通过供需匹配将研究单元划分为:高供-高需,低供-高需,低供-低需和高供-低需4种类型,并识别出114个供需严重失衡的关键区,该类关键区面临城市内涝调节服务供给短缺的严峻挑战,是未来城市规划过程中需要重点优化的对象。在此基础上,使用优先级指数将关键区划分为5个规划干预优先级,明确了城市内涝治理的优先次序。研究结果为开展具有针对性的城市内涝调节治理实践提供了新的科学依据,同时在解决城市内涝灾害问题,推进以人为核...     

Mapping ecosystem services: The supply and demand of flood regulation services in Europe

Ecosystem services (ES) feature highly distinctive spatial and temporal patterns of distribution, quantity, and flows. The flow of ecosystem goods and services to beneficiaries plays a decisive role in the valuation of ES and the successful implementation of the ES concept in environmental planning. This is particularly relevant to regulating services where demands emerge often spatially separated from supply. However, spatial patterns of both supply and demand are rarely incorporated in ES assessments on continental scales. In this paper, we present an ES modeling approach with low data demand, fit to be employed in scenario analysis and on multiple scales. We analyze flood regulation services at a European scale by explicitly addressing the spatial distribution of ES demand. A flood regulation supply indicator is developed based on scenario runs with a hydrological model in representative river catchments, incorporating detailed information on land, cover, land use and management. Land use sensitive flood damage estimates in the European Union (EU) are employed to develop a spatial indicator for flood regulation demand. Findings are transferred to the EU territory to create a map of the current supply of flood regulation and the potential supply under conditions of natural vegetation. Regions with a high capacity to provide flood regulation are mainly characterized by large patches of natural vegetation or extensive agriculture. The main factor limiting supply on a continental scale is a low water holding capacity of the soil. Flood regulation demand is highest in central Europe, at the foothills of the Alps and upstream of agglomerations. We were able to identify areas with a high potential capacity to provide flood regulation in conjunction with land use modifications. When combined with spatial patterns of current supply and demand, we could identify priority areas for investments in ES flood regulation supply through conservation and land use planning. We found that only in a fraction of the EU river catchments exhibiting a high demand, significant increases in flood regulation supply are achievable by means of land use modifications.     

应对暴雨洪涝灾害的平原城市生态水文调节服务供需评估——以天津市为例

生态水文调节服务是生态系统服务的重要组成,能够有效调节地表径流,缓解暴雨洪涝灾害。但由于城市建设扩张、生态环境退化、极端降雨频发等因素,导致生态水文调节服务出现供需失衡。平原城市中地形水动力较弱及城市发展需求高的特征更是加剧了这种现象。基于生态系统供需视角,以生态水文调节率表征城市生态水文调节服务的生态供给,以暴雨洪涝风险表征城市生态水文调节服务的社会需求,构建应对暴雨洪涝灾害的平原城市生态水文调节服务供需研究框架。以典型平原城市天津市为例,应用SWAT模型、随机森林模型和ArcGIS定量测度城市生态水文调节服务供需水平,划分四种供需空间匹配类型,识别供需失衡关键区域,并进行五级规划干预等级分区。结果表明：(1)供给能力呈现“东南沿海高,西北近山低”的空间分布;需求水平表现出“多中心聚集,圈层向外递减”的分布规律。(2)供需空间分布呈现正相关关系,低供-低需在四种供需匹配类型中占据主导,同时空间聚集性最明显。(3)规划干预分区存在明显的空间差异,优先干预区占比5.41%,整体与建成区分布一致,大部分集中在市内六区,其次分布在滨海新区核心区,是未来规划治理的重点。研究结果为城市规划管理从...     

Extracellular glycerol regulates the cardiac energy balance in a working rat heart model

We reported previously that glycerol is a substrate for energy production in cardiomyocytes. Increasing glycerol availability results in increased glycerol uptake and its involvement in complex lipid biosynthesis and energy production. This study evaluated the relationship between glycerol supply, energy demand, and intermediary metabolism leading to energy production. The work was performed on isolated rat heart perfused in the working mode. Glycerol concentrations modeled the fasting (0.33 mM) and fed (3.33 mM) states. Cardiac energy demand was modeled by increasing heart rate from 350 to 450 beats/min (bpm). Increasing glycerol supply increased glycerol uptake from 1.4 (350 bpm) to 3.8 (450 bpm) and from 9.7 (350 bpm) to 34.2 (450 bpm) micro mol glycerol/heart in 30 min at 0.33 and 3.33 mM glycerol, respectively. At low glycerol supply, increasing heart rate did not influence the complex lipid synthesis. Conversely, high glycerol concentration increased the complex lipid synthesis by 5- and 30-fold at 350 and 450 bpm, respectively. Increasing glycerol supply and heart rate significantly increased glycerol oxidation rate. Moreover, increasing glycerol supply did not affect glucose oxidation but increased palmitate uptake and significantly decreased its beta-oxidation. Physiological concentrations of glycerol contribute to the cardiac intermediary metabolism, both for energy production and glycerolipid synthesis. Increasing energy demand enhances the requirement and use of glycerol. Glycerol contributes to the regulation of cardiac metabolism and energy balance, mainly by decreasing the contribution of fatty acid oxidation, and may thus represent a new factor in cardiac protection through the reduction of oxygen demand.     

绿色基础设施的洪水调节服务供需测度研究进展

绿色基础设施(green infrastructure, GI)可提供高效且可获取的洪水调节服务,其供需测度是提升服务水平的基础与前提,可为城市土地利用与绿色基础设施规划提供决策依据。以GI洪水调节服务供需为研究对象,梳理GI洪水调节服务供需的概念内涵、主体构成和尺度关系。回顾了GI洪水调节服务供需测度方法,包括土地利用测度法、生态系统测度法和洪水调蓄模型法等供应水平测度方法,以及风险评估法、洪水淹没模型法和经济损失法等需求水平测度方法。基于生态系统服务的供需关系,构建了GI洪水调节服务供需分析框架,通过供需数量均衡和空间匹配将供需关系分为四种情景,对应服务动态平衡、调整服务供需水平和服务流动机制3种调控手段。从服务测度重点、动态变化分析、供需阈值效能、服务流动机制等四个方面提出了未来研究方向,以期对绿色基础设施规划研究和生态系统服务管理与决策研究有所启示。     

Mismatches between ecosystem services supply and demand in urban areas: A quantitative assessment in five European cities

Assessing mismatches between ecosystem service (ES) supply and demand can provide relevant insights for enhancing human well-being in urban areas. This paper provides a novel methodological approach to assess regulating ES mismatches on the basis of environmental quality standards and policy goals. Environmental quality standards (EQS) indicate the relationship between environmental quality and human well-being. Thus, they can be used as a common minimum threshold value to determine whether the difference between ES supply and demand is problematic for human well-being. The methodological approach includes three main steps: (1) selection of EQS, (2) definition and quantification of ES supply and demand indicators, and (3) identification and assessment of ES mismatches on the basis of EQS considering certain additional criteria. While ES supply indicators estimate the flow of an ES actually used or delivered, ES demand indicators express the amount of regulation needed in relation to the standard. The approach is applied to a case study consisting of five European cities: Barcelona, Berlin, Stockholm, Rotterdam and Salzburg, considering three regulating ES which are relevant in urban areas: air purification, global climate regulation and urban temperature regulation. The results show that levels of ES supply and demand are highly heterogeneous across the five studied cities and across the EQS considered. The assessment shows that ES supply contributes very moderately in relation to the compliance with the EQS in most part of the identified mismatches. Therefore, this research suggests that regulating ES supplied by urban green infrastructure are expected to play only a minor or complementary role to other urban policies intended to abate air pollution and greenhouse gas emissions at the city scale. The approach has revealed to be appropriate for the regulating ES air purification and global climate regulation, for which well-established standards or targets are available at the city level. Yet, its applicability to the ES urban temperature regulation has proved more problematic due to scale and user dependent constraints.     

Regulation of ATP supply in mammalian skeletal muscle during resting state-->intensive work transition

In the present debating paper, the problem how the rate of ATP supply by oxidative phosphorylation in mitochondria is adjusted to meet a greatly increased demand for ATP during intensive exercise of skeletal muscle is discussed. Different experimental results are collected from different positions of the literature and confronted with five conceptual models of the regulation of the oxidative phosphorylation system. The previously performed computer simulations using a dynamic model of oxidative phosphorylation are also discussed in this context. The possible regulatory mechanisms considered in the present article are: (A) output activation: an external effector activates directly only the output of the system (ATP turnover); (B) input/output activation: an external effector activates directly the output (ATP usage) and input (substrate dehydrogenation) of the system; (C) removal of substrate shortage: only ATP consumption and substrate supply by blood are directly activated; (D) removal of oxygen shortage: only ATP consumption and oxygen supply by blood are directly activated; (E) each step activation: an external effector activates both the ATP-consuming subsystem and all the steps in the ATP-producing subsystem (particular enzymes/carriers/blocks of oxidative phosphorylation, substrate supply, oxygen supply). The performed confrontation of the considered mechanisms with the presented results leads to the conclusion that only the each step activation model is quantitatively consistent with the whole set of experimental data discussed. It is therefore postulated that a universal effector/regulatory mechanism of a still unknown nature which activates all steps of oxidative phosphorylation should exist and be discovered. A possible nature of such an effector is shortly discussed.     

Acidotic alterations in oxidative metabolism influencing rat renal slice ammoniagenesis

We believe that two findings are interconnected and help to comprehend a major mechanism behind the regulation of renal ammonia production during acidosis. First, slices from acidotic compared to control and alkalotic rats produce more ammonia from glutamine. Second, inhibition of renal oxidative metabolism at various points by metabolic inhibitors augments slice ammoniagenesis. Based on this, our purpose was to determine whether enhanced renal ammoniagenesis during acidosis could occur through the same mechanism as the metabolic inhibitors. However, metabolic inhibitors (malonate; arsenite; 2,4-dinitrophenol) usually decrease while acidosis increases slice gluconeogenesis. There is one known exception. Fluorocitrate, which blocks citrate metabolism, simulates the acidotic condition by enhancing both ammonia and glucose production. Accordingly, a block of oxidative metabolism if located prior to citrate oxidation in the tricarboxylic acid cycle could theoretically augment ammoniagenesis during acidosis. Lactate, is a major renal fuel whose oxidative metabolism would be blocked by fluorocitrate. There, we concentrated on the effects of acidosis on lactate as well as glutamine metabolism. Lactate decarboxylation decreases in the face of increased glucose production during acidosis, and lactate inhibition of glutamine decarboxylation decreases in slices from acidotic rats. Also, we found lesser oxygen consumption in the presence of lactate by kidney slices from acidotic rats compared to control and alkalotic rats. We postulate that relatively less incorporation of lactate into the TCA cycle, causing decreased citrate formation and citrate oxidation during acidosis, contributes, at least in part, to acidotic adaptation of ammoniagenesis.     

The regulatory design of an allosteric feedback loop: the effect of saturation by pathway substrate

Aspects of metabolic regulation can be fruitfully studied with a combination of generic modelling, control analysis and graphical analysis using rate characteristics. This paper analyses a prototypical supply-demand system consisting of a biosynthetic subsystem subject to allosteric inhibition by its product and a demand process that consumes this product. The effect of changes in affinity of the committing supply enzyme for the pathway substrate on the regulatory properties of the supply subsystem is compared for the Monod-Wyman-Changeux and the reversible Hill allosteric enzyme models. We found that the Hill model has a distinct advantage in that the steady-state concentration at which it maintains the product is set by the half-saturating product concentration and is independent of changes in the degree of saturation for substrate. In contrast, with the Monod-Wyman-Changeux model this set point varies with affinity for substrate. Explicitly incorporating reversibility in all rate equations made it possible to distinguish between kinetic and thermodynamic aspects of regulation. Combining the supply and demand rate characteristics allows us to explore both the control distribution at steady state and the regulatory performance of the system over a wide range of demand activities.     

Regulation of muscle carbohydrate metabolism during exercise.

This review examines the mechanisms that regulate muscle carbohydrate metabolism during exercise. Muscle carbohydrate utilization is regulated primarily by two factors, namely, delivery of substrate to the glycolytic pathway either from glycogenolysis or from transport of extracellular glucose into the fibers, and formation of triosephosphate by phosphofructokinase. The regulation involves the integration of the glycolytic controls with other metabolic controls and the needs of the whole muscle in meeting the physiological demand. The controls operating in the glycolytic sequence in vivo appear to couple glycolytic recruitment to signals from the rate of energy demand, the TCA cycle state, and the mitochondrial redox state so as to satisfy the major regulatory goal of maintaining the supply of ATP for tension development.     

Dual regulation of the AMP-activated protein kinase provides a novel mechanism for the control of creatine kinase in skeletal muscle.

The AMP-activated protein kinase (AMPK) is activated by a fall in the ATP:AMP ratio within the cell in response to metabolic stresses. Once activated, it phosphorylates and inhibits key enzymes in energy-consuming biosynthetic pathways, thereby conserving cellular ATP. The creatine kinase-phosphocreatine system plays a key role in the control of ATP levels in tissues that have a high and rapidly fluctuating energy requirement. In this study, we provide direct evidence that these two energy-regulating systems are linked in skeletal muscle. We show that the AMPK inhibits creatine kinase by phosphorylation in vitro and in differentiated muscle cells. AMPK is itself regulated by a novel mechanism involving phosphocreatine, creatine and pH. Our findings provide an explanation for the high expression, yet apparently low activity, of AMPK in skeletal muscle, and reveal a potential mechanism for the co-ordinated regulation of energy metabolism in this tissue. Previous evidence suggests that AMPK activates fatty acid oxidation, which provides a source of ATP, following continued muscle contraction. The novel regulation of AMPK described here provides a mechanism by which energy supply can meet energy demand following the utilization of the immediate energy reserve provided by the creatine kinase-phosphocreatine system.     

Matching the supply of bacterial nutrients to the nutritional demand of the animal host

Various animals derive nutrients from symbiotic microorganisms with much-reduced genomes, but it is unknown whether, and how, the supply of these nutrients is regulated. Here, we demonstrate that the production of essential amino acids (EAAs) by the bacterium Buchnera aphidicola in the pea aphid Acyrthosiphon pisum is elevated when aphids are reared on diets from which that EAA are omitted, demonstrating that Buchnera scale EAA production to host demand. Quantitative proteomics of bacteriocytes (host cells bearing Buchnera) revealed that these metabolic changes are not accompanied by significant change in Buchnera or host proteins, suggesting that EAA production is regulated post-translationally. Bacteriocytes in aphids reared on diet lacking the EAA methionine had elevated concentrations of both methionine and the precursor cystathionine, indicating that methionine production is promoted by precursor supply and is not subject to feedback inhibition by methionine. Furthermore, methionine production by isolated Buchnera increased with increasing cystathionine concentration. We propose that Buchnera metabolism is poised for EAA production at certain maximal rates, and the realized release rate is determined by precursor supply from the host. The incidence of host regulation of symbiont nutritional function via supply of key nutritional inputs in other symbioses remains to be investigated.     

Insig-dependent ubiquitination and degradation of 3-hydroxy-3-methylglutaryl coenzyme a reductase stimulated by delta- and gamma-tocotrienols

Sterol-regulated ubiquitination marks 3-hydroxy-3-methylglutaryl coenzyme A reductase, a rate-determining enzyme in cholesterol synthesis, for endoplasmic reticulum (ER)-associated degradation by 26 S proteasomes. This degradation, which results from sterol-induced binding of reductase to ER membrane proteins called Insigs, contributes to the complex, multivalent feedback regulation of the enzyme. Degradation of HMG-CoA reductase is also stimulated by various forms of vitamin E, a generic term for alpha-, beta-, delta-, and gamma-tocopherols and tocotrienols, which are primarily recognized for their potent antioxidant activity. Here, we show that delta-tocotrienol stimulates ubiquitination and degradation of reductase and blocks processing of sterol regulatory element-binding proteins (SREBPs), another sterol-mediated action of Insigs. The gamma-tocotrienol analog is more selective in enhancing reductase ubiquitination and degradation than blocking SREBP processing. Other forms of vitamin E neither accelerate reductase degradation nor block SREBP processing. In vitro assays indicate that gamma- and delta-tocotrienol trigger reductase ubiquitination directly and do not require further metabolism for activity. Taken together, these results provide a biochemical mechanism for the hypocholesterolemic effects of vitamin E that have been observed in animals and humans.     

Hierarchical protein folding pathways: a computational study of protein fragments

We have previously presented a building block folding model. The model postulates that protein folding is a hierarchical top-down process. The basic unit from which a fold is constructed, referred to as a hydrophobic folding unit, is the outcome of combinatorial assembly of a set of "building blocks." Results obtained by the computational cutting procedure yield fragments that are in agreement with those obtained experimentally by limited proteolysis. Here we show that as expected, proteins from the same family give very similar building blocks. However, different proteins can also give building blocks that are similar in structure. In such cases the building blocks differ in sequence, stability, contacts with other building blocks, and in their 3D locations in the protein structure. This result, which we have repeatedly observed in many cases, leads us to conclude that while a building block is influenced by its environment, nevertheless, it can be viewed as a stand-alone unit. For small-sized building blocks existing in multiple conformations, interactions with sister building blocks in the protein will increase the population time of the native conformer. With this conclusion in hand, it is possible to develop an algorithm that predicts the building block assignment of a protein sequence whose structure is unknown. Toward this goal, we have created sequentially nonredundant databases of building block sequences. A protein sequence can be aligned against these, in order to be matched to a set of potential building blocks.