Evaluation of the Correlation between the Content of Alkaloids and Chemical Elements in Washington Lupine (Lupinus polyphyllus Lindl.) Based on Statistical Analysis and Mathematical Simulation

A considerable number of linear and nonlinear correlations between concentrations of quinolizidine alkaloids and concentrations of K, Ca, Mg, Ba, Zn, Cu, Ni, and V were found in lupine seeds and leaves by using methods of correlation and regression analysis. Mathematical models of regulation of alkaloid metabolism by these chemical elements were constructed. The models of regulation were considered separately for individual plant organs.     

Investigation of some theoretical models relating the concentrations of glucose and insulin in plasma

In this survey 24 possible theoretical models for the regulation of plasma glucose concentration by plasma insulin concentration have been proposed and tested. A set of data, consisting of plasma glucose concentrations at various times after an intravenous injection of glucose, was selected from the literature. An attempt was made to fit each model to the chosen data using a digital computer program and ten of these were successful. The behaviour of these ten models under different experimental conditions was compared, using a simulation technique, with further published data. At this stage six models were consistent with a steady state at the start or the end of an experiment, but only three were reasonably good at predicting all aspects of the chosen experiments. One was particularly good. Two other sets of data were taken from the literature and the six best models were fitted in order to calculate replicate values of the physiological parameters.     

The human hepatocyte cell lines IHH and HepaRG: models to study glucose, lipid and lipoprotein metabolism

Metabolic diseases reach epidemic proportions. A better knowledge of the associated alterations in the metabolic pathways in the liver is necessary. These studies need in vitro human cell models. Several human hepatoma models are used, but the response of many metabolic pathways to physiological stimuli is often lost. Here, we characterize two human hepatocyte cell lines, IHH and HepaRG, by analysing the expression and regulation of genes involved in glucose and lipid metabolism. Our results show that the glycolysis pathway is activated by glucose and insulin in both lines. Gluconeogenesis gene expression is induced by forskolin in IHH cells and inhibited by insulin in both cell lines. The lipogenic pathway is regulated by insulin in IHH cells. Finally, both cell lines secrete apolipoprotein B-containing lipoproteins, an effect promoted by increasing glucose concentrations. These two human cell lines are thus interesting models to study the regulation of glucose and lipid metabolism.     

Genetic Aspects of the Relationship between Isoquinoline Alkaloids and Mineral Elements in Greater Celandine (Chelidonium majus L.)

Interrelations between the total content of isoquinoline alkaloids, the concentrations of quaternary protoberberines and benzophenanthridines, and the amount of K, Cu, Co, Al, Ba, and Zn in aerial parts of individual celandine plants were revealed, within a single cenopopulation, using correlation analysis and regression analysis. Mathematical models describing the regulation of isoquinoline metabolism by some of the mineral elements were obtained in analytical form. The results suggest that this process is genetically determined.     

Genetic aspects of the relationship between isoquinoline alkaloids and mineral elements in greater celandine (Chelidonium majus L.)

Interrelations between the total content of isoquinoline alkaloids, concentrations of quaternary protoberberines and benzophenanthridines, and the amount of K, Cu, Co, Al, Ba, and Zn in aerial parts of individual celandine plants were revealed, within a single cenopopulation, using correlation analysis and regression analysis. Mathematical models describing the regulation of isoquinoline metabolism by some of the mineral elements were obtained in the analytical form. The results suggest that this process is genetically determined.     

Mathematical modeling and analysis of glucose and glutamine utilization and regulation in cultures of continuous mammalian cells

A number of factors have been shown to affect the metabolism of glucose and glutamine in mammalian cells and their mechanisms have been partially elucidated. Despite these efforts, a quantitative knowledge of the significance of these factors, the regulation of glucose and glutamine utilization, and particularly the interactions of these two nutrients is still lacking. Controversies exist in the literature. To clarify some of these controversies, mathematical models are proposed in this work which enable to separate and identify the effects of individual factors. Experimental data from five cell lines obtained in batch, fed-batch, and continuous cultures, both under steady-state and transient conditions, were used to verify the model formulations. The resulting kinetic models successfully describe all these cultures. According to the models, the specific consumption rate of glucose (Q(Glc)) of continuous animal cells under normal culture conditions can be expressed as a sum of three parts: a part owing to cell growth; a part owing to glucose excess; and a part owing to glutamine regulation. The specific consumption rate of glutamine (q(Glc)7) can be expressed as a sum of only two parts: a part owing to cell growth; and a part owing to glutamine excess. Using the kinetic models the interaction and regulation of glucose and glutamine utilizations are quantitatively analyzed. The results indicate that, whereas q(Glc) is affected by glutamine, q(Gln) appears to be not or less significantly affected by glucose. It is also shown that the relative utilizations of glucose and glutamine by anabolism and catabolism are mainly affected by the residual concentrations of the respective compounds and are less sensitive to growth rate and the nature of growth limitation.(c) 1995 John Wiley & Sons, Inc.     

Streptozotocin-induced increase in cholesterol ester transfer protein (CETP) and its reversal by insulin in transgenic mice expressing human CETP

High plasma triacylglycerol and low high-density lipoprotein levels are risk factors for cardiovascular disease in diabetes. Plasma high-density lipoprotein levels are regulated by cholesterol ester transfer protein (CETP). The regulation of CETP under diabetic conditions is not clear, and this is due to a lack of appropriate models. We used transgenic mice expressing human CETP to study the regulation of this protein under type-1 diabetic conditions and further investigated whether insulin reverses the effect of diabetes. Mice expressing human CETP under the control of its natural flanking region and age-matched littermates not expressing this protein were made diabetic by injecting streptozotocin, and the reversal of diabetes was assessed by injecting insulin. The plasma total cholesterol, low-density lipoprotein-cholesterol, and triacylglycerol concentrations were elevated, whereas high-density lipoprotein-cholesterol concentrations were reduced after the onset of diabetes. Insulin injection partially recovered this effect. The plasma cholesterol ester transfer activity, CETP mass, and hepatic CETP mRNA abundance were significantly higher in diabetic mice that were partially restored by insulin administration. There was a strong correlation between high-density lipoprotein-cholesterol concentrations and cholesterol ester transfer activity. These results suggest that an increase in CETP under diabetic conditions might be a major factor responsible for increased incidence of diabetes-induced atherosclerosis.     

Increased thirst and vasopressin secretion after myocardial infarction in rats

Impaired regulation of salt and water balance in left ventricular dysfunction and heart failure can lead to pulmonary and peripheral edema and hyponatremia. Previous studies of disordered water regulation in heart failure have used models of low cardiac output with normal cardiac function (e.g., inferior vena cava ligation). We investigated thirst and vasopressin (AVP) secretion in a rat myocardial infarction model of chronic left ventricular dysfunction/heart failure in response to a 24-h water deprivation period. Thirst (implied from water drunk), hematocrit, plasma renin activity, and plasma AVP concentrations increased with water deprivation vs. ad libitum water access. Thirst and plasma AVP concentrations were significantly positively correlated with infarct size after 24-h water deprivation but not under ad libitum water access conditions. The mechanism by which this occurs is unclear but could involve increased osmoreceptor sensitivity, altered stimulation of baroreceptors, the renin-angiotensin system, or altered central neural control.     

Modeling of Galpha(s) and Galpha(i) regulation of human type V and VI adenylyl cyclase

We examined the kinetics of Galpha(s) and Galpha(i) regulation of human type V and type VI adenylyl cyclase (AC V and AC VI) in order to better model interactions between AC and its regulators. Activation of AC VI by Galpha(s) displayed classical Michaelis-Menten kinetics, whereas AC V activation by Galpha(s) was cooperative with a Hill coefficient of 1.4. The basal activity of human AC V, but not that of AC VI, was inhibited by Galpha(i). Both enzymes showed greater inhibition by Galpha(i) at low Galpha(s) concentrations; however, human AC V was activated by Galpha(i) at high Galpha(s) concentrations. Neither regulator had an effect on the K(m) for Mg-ATP. Mutations made within the Galpha(s) binding pocket of AC V (N1090D) and VI (F1078S) displayed 6- and 14-fold greater EC(50) values for Galpha(s) activation but had no effect on Galpha(i) inhibition of basal activity or K(m) for Mg-ATP. Galpha(s)-stimulated AC VI-F1078S was not significantly inhibited by Galpha(i), despite normal inhibition by Galpha(i) upon forskolin stimulation. Mechanistic models for Galpha(s) and Galpha(i) regulation of AC V and VI were derived to describe these results. Our models are consistent with previous studies, predicting a decrease in affinity of Galpha(i) in the presence of Galpha(s). For AC VI, Galpha(s) is required for inhibition but not binding by Galpha(i). For AC V, binding of two molecules of Galpha(s) and Galpha(i) to an AC dimer are required to fully describe the data. These models highlight the differences between AC V and VI and the complex interactions with two important regulators.     

皮质酮诱导的PC12细胞梯度应激损伤模型的构建及评价

目的:建立皮质酮诱导的PC12细胞梯度应激损伤模型,为细胞应激水平的评估和细胞应激损伤调控研究提供实验基础和对象。方法:通过检测不同浓度皮质酮(0～1 000μmol/L)在经过不同干预时间(8～48 h)后PC12细胞活力,观察皮质酮对细胞活力的影响,筛选最佳干预条件的细胞模型。分光光度法和微量法检测细胞模型的关键应激指标(MDA、SOD、NADH、LDH),对模型进行评价。结果:当皮质酮浓度在200μmol/L以下且干预时间为12 h时,细胞活力在半数失活率以下,可减少各组由于细胞活力下降而产生的混杂因素。与空白对照组比较,皮质酮浓度依赖性地升高模型组的MDA、NADH和LDH水平,降低SOD水平(P<0.01),符合梯度应激模型的构建要求。结论:成功建立了PC12细胞梯度应激损伤模型,在干预时间为12 h的情况下,干预浓度为0μmol/L、25μmol/L、50μmol/L、100μmol/L、150μmol/L、200μmol/L,使得细胞模型应激损伤程度梯度增加,可作为开展细胞应激损伤评估及调控实验的基础和对象。     

Xenopus laevis larvae (Amphibia,Anura) as model suspension feeders

Larvae of the South African clawed frog, Xenopus laevis (Daudin), are efficient, obligate suspension feeders. We examine the relationship between the ambient particle concentration offered these larvae as food and their filtering, ingestion, and buccal pumping rates. We demonstrate that: (i) the larvae can sense and respond to a broad range of particle concentrations, down to 0.2 mg 1^–1 (dry weight); (ii) their metabolic needs theoretically can be met by particle concentrations as low as 5 mg 1^–1; and (iii) their patterns of regulation of filtering and ingestion fit predictions from certain models used to describe zooplankton feeding dynamics. Two such models are discussed: the modified Monod (Michaelis-Menten) model, with a lower threshold below which the tadpoles do not feed, and an energy optimization model. Both the models and the observed behavior of the tadpoles allow for stability of populations of food organisms. Tadpole feeding dynamics apparently are compatible with both the predictions and assumptions of these models, suggesting similar regulation of feeding by tadpoles and zooplankton. However, the size, morphology, and behavior of X. laevis larvae make their feeding regulation uniquely accessible to direct observation.Contribution No. 223, Center for Great Lakes Studies, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin.Contribution No. 223, Center for Great Lakes Studies, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin.     

Genetic aspects of the interrelation between alkaloids and chemical elements in Atropa belladonna L. and Glaucium flavum Grantz. plants

The variability of the contents of tropane and isoquinoline alkaloids, ashes, Na, K, Ca, Mg, Fe, Mn, Cu, Zn, Co, Mo, Cr, Al, Ba, V, Ni, Sr, Cd, Pb, J, and Ag was studied in individual plants of the industrial population of belladonna (Atropa belladonna L.) and yellow horned poppy (Glaucium flavum Crantz.). Numerous linear and nonlinear correlations of isoquinoline and tropane alkaloids with ashes and mineral elements were revealed by means of correlation and regression analyses. Alkaline earth elements (especially Sr and Ba) were shown to have a major role in the regulation of tropane alkaloid accumulation in belladonna leaves. K and Ni were of particular importance in the aerial part of yellow horned poppy. These elements at the suboptimal concentrations were most favorable for isoquinoline alkaloid accumulation in yellow horned poppy. Analytical mathematical models were derived for the regulation of alkaloid metabolism in test plants by some mineral elements (Ba, Mg, Al, Sr, Ni, Mn, and K). Our results indicate that the interrelation between alkaloids and elements in these plants is genetically determined.     

Differential regulation of cardiac ANP and BNP mRNA in different stages of experimental heart failure

Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) are cardiac hormones that are involved in water and electrolyte homeostasis in heart failure. Although both hormones exert almost identical biological actions, the differential regulation of cardiac ANP and BNP mRNA in compensated and overt heart failure is not known. To study the hypothesis that cardiac BNP is more specifically induced in overt heart failure, a large aortocaval shunt of 30 days duration was produced in rats and compared with compensated heart failure. Compensated heart failure was induced either by a small shunt of 30 days duration or by a large shunt of 3 days duration. Both heart failure models were characterized by increased cardiac weight, which was significantly higher in the large-shunt model, and central venous pressure. Left ventricular end-diastolic pressure was elevated only in the overt heart failure group (control: 5.7 +/- 0. 7; small shunt: 8.6 +/- 0.9; large shunt 3 days: 8.5 +/- 1.7; large shunt 30 days: 15.9 +/- 2.6 mmHg; P < 0.01). ANP and BNP plasma concentrations were elevated in both heart failure models. In compensated heart failure, ANP mRNA expression was induced in both ventricles. In contrast, ventricular BNP mRNA expression was not upregulated in any of the compensated heart failure models, whereas it increased in overt heart failure (left ventricle: 359 +/- 104% of control, P < 0.001; right ventricle: 237 +/- 33%, P < 0.01). A similar pattern of mRNA regulation was observed in the atria. These data indicate that, in contrast to ANP, cardiac BNP mRNA expression might be induced specifically in overt heart failure, pointing toward the possible role of BNP as a marker of the transition from compensated to overt heart failure.     

In Vivo Regulation of Threonine and Isoleucine Biosynthesis in Lemna paucicostata Hegelm. 6746

Little, if any, regulation of threonine synthesis was observed in Lemna paucicostata Hegelm. 6746 supplemented with concentrations of threonine and/or isoleucine that allow for uptake of these amino acids in amounts sufficient for total plant requirements, and that increase tissue concentrations of soluble threonine manyfold. High tissue concentrations of soluble threonine generated endogenously in isoleucine-supplemented plants were no more effective in regulation than a similar concentration of threonine accumulated from the medium. These studies exclude also major regulation of threonine biosynthesis by bivalent repression by threonine plus isoleucine. Isoleucine biosynthesis was severely inhibited by supplementation with isoleucine, but not with threonine or methionine. The fivefold increase in soluble threonine in isoleucine-supplemented plants suggests that threonine dehydratase is a major locus for feedback regulation of isoleucine synthesis. It is concluded that regulation of threonine biosynthesis differs from that of the other amino acids of the aspartate family (isoleucine, methionine, and lysine), each of which strongly feedback regulates its own synthesis. Methionine supplementation had a negligible effect on the tissue concentration of soluble threonine, indicating that threonine is not important in balancing changes of flux into methionine by equivalent changes of flux through the step catalyzed by aspartokinase.     

Role of phosphate in the central metabolism of two lactic acid bacteria--a comparative systems biology approach

Lactic acid-producing bacteria survive in distinct environments, but show common metabolic characteristics. Here we studied the dynamic interactions of the central metabolism in Lactococcus lactis, extensively used as a starter culture in the dairy industry, and Streptococcus pyogenes, a human pathogen. Glucose-pulse experiments and enzymatic measurements were performed to parameterize kinetic models of glycolysis. Significant improvements were made to existing kinetic models for L. lactis, which subsequently accelerated the development of the first kinetic model of S. pyogenes glycolysis. The models revealed an important role for extracellular phosphate in the regulation of central metabolism and the efficient use of glucose. Thus, phosphate, which is rarely taken into account as an independent species in models of central metabolism, should be considered more thoroughly in the analysis of metabolic systems in the future. Insufficient phosphate supply can lead to a strong inhibition of glycolysis at high glucose concentrations in both species, but this was more severe in S. pyogenes. S. pyogenes is more efficient at converting glucose to ATP, showing a higher tendency towards heterofermentative energy metabolism than L. lactis. Our comparative systems biology approach revealed that the glycolysis of L. lactis and S. pyogenes have similar characteristics, but are adapted to their individual natural habitats with respect to phosphate regulation.     

Simple kinetic models explaining critical phenomena in enzymatic reactions with isomerization of the enzyme and substrate

Kinetic models for enzyme reactions are considered which take into account enzyme and substrate isomerization. Application of graph-theoretic methods allows to reveal fragments in schemes which may induce multiple stead-states or concentrational selfoscillations. The role of substrate isomers in the inhibition of enzyme isomers to produce critical phenomena is considered. The boundaries of parameter domains for critical phenomena are estimated. It is shown that the controlled change in concentrations of substrate and enzyme isomers may be important in regulation of enzyme systems, if different enzyme isomers are inhibited mainly by different substrate isomers. The models are used for interpretation of possible critical phenomena in the open reaction catalyzed by lactate dehydrogenase. It is shown that lactate dehydrogenase may act as a trigger in carbohydrate metabolism by changing "critically" its activity in relation to changes in pH and pyruvate fluxes. Slow enzyme inhibition by enolpyruvate is suggested as a possible reason for glycolytic oscillations.     

大肠杆菌苏氨酸合成途径动力学模型的构建与分析

动力学模型分析有利于理解生物系统的调控机制,从而为高效细胞工厂的理性设计提供指导。基于以往发表的相关途径动力学模型和测量的酶动力学数据,开发了大肠杆菌苏氨酸合成途径的动力学模型。模型包含从天冬氨酸至苏氨酸的合成途径及葡萄糖开始的为合成途径提供前体以及能量的代谢途径。与以往模型不同的是新模型中考虑了能量和还原力的平衡,从而使模型模拟的系统自身成为一个不需要从外界提供能量和还原力的自洽系统。模型稳态分析的结果表明PTS、G6PDH和HDH等反应对苏氨酸合成反应的通量控制系数较大,通过过表达这些反应的酶可以有效增加苏氨酸合成反应的通量。