Membrane-traversing mechanism of thyroid hormone transport by monocarboxylate transporter 8

Monocarboxylate transporter 8 (MCT8) mediates thyroid hormone (TH) transport across the plasma membrane in many cell types. In order to better understand its mechanism, we have generated three new MCT8 homology models based on sugar transporters XylE in the intracellular opened (PDB ID: 4aj4) and the extracellular partly occluded (PDB ID: 4gby) conformations as well as FucP (PDB ID: 3o7q) and GLUT3 (PDB ID: 4zwc) in the fully extracellular opened conformation. T₃-docking studies from both sides revealed interactions with His192, His415, Arg445 and Asp498 as previously identified. Selected mutations revealed further transport-sensitive positions mainly at the discontinuous transmembrane helices TMH7 and 10. Lys418 is potentially involved in neutralising the charge of the TH substrate because it can be replaced by charged, but not by uncharged, amino acids. The side chain of Thr503 was hypothesised to stabilise a helix break at TMH10 that undergoes a prominent local shift during the transport cycle. A T503V mutation accordingly affected transport. The aromatic Tyr419, the polar Ser313 and Ser314 as well as the charged Glu422 and Glu423 lining the transport channel have been studied. Based on related sugar transporters, we suggest an alternating access mechanism for MCT8 involving a series of amino acid positions previously and newly identified as critical for transport.     

Homologous recombination in plants

In plants three different approaches have been used to study homologous DNA recombination; extrachromosomal recombination (ECR) between transfected DNA molecules, intrachromosomal recombination (ICR) between repeated genes integrated into and resident at the genome and recombination between introduced DNA and homologous sequences in the genome (gene targeting). ECR is efficient (10^–1 to 10^–3) and occurs mainly during a limited time period early after transfection. It proceeds predominantly via nonconservative single-strand annealing. ICR, which in most cases is described best by the double-strand break repair model of recombination, occurs at frequencies of one event in 10⁵ to 10⁷ cells. ICR takes place throughout the whole life-cycle of a plant, in all organs and at different developmental stages. As there exists no predetermined germline in plants, somatic recombination events can be transferred to the next generation. Recombination frequencies are enhanced by DNA damage. Gene targeting, like ICR, occurs at low rates in plant cells. Almost nothing is known about the enzymes involved in homologous recombination in plants.     

Subcellular trafficking of the substrate transporters GLUT4 and CD36 in cardiomyocytes

Cardiomyocytes use glucose as well as fatty acids for ATP production. These substrates are transported into the cell by glucose transporter 4 (GLUT4) and the fatty acid transporter CD36. Besides being located at the sarcolemma, GLUT4 and CD36 are stored in intracellular compartments. Raised plasma insulin concentrations and increased cardiac work will stimulate GLUT4 as well as CD36 to translocate to the sarcolemma. As so far studied, signaling pathways that regulate GLUT4 translocation similarly affect CD36 translocation. During the development of insulin resistance and type 2 diabetes, CD36 becomes permanently localized at the sarcolemma, whereas GLUT4 internalizes. This juxtaposed positioning of GLUT4 and CD36 is important for aberrant substrate uptake in the diabetic heart: chronically increased fatty acid uptake at the expense of glucose. To explain the differences in subcellular localization of GLUT4 and CD36 in type 2 diabetes, recent research has focused on the role of proteins involved in trafficking of cargo between subcellular compartments. Several of these proteins appear to be similarly involved in both GLUT4 and CD36 translocation. Others, however, have different roles in either GLUT4 or CD36 translocation. These trafficking components, which are differently involved in GLUT4 or CD36 translocation, may be considered novel targets for the development of therapies to restore the imbalanced substrate utilization that occurs in obesity, insulin resistance and diabetic cardiomyopathy.     

Effects of photoperiod, temperature and testosterone-treatment on plasma T3 and T4 levels in the Djungarian hamster, Phodopus sungorus

The effects of photoperiod, temperature and testosterone treatment on plasma T3 and T4 levels were investigated in the Djungarian hamster. Plasma T3 level was affected by temperature (25 degrees C less than 7 degrees C) but not by photoperiod. Plasma T4 level was affected by photoperiod (short day less than long day) at 25 degrees C. Administration of testosterone increased plasma T4 level under short photoperiod at 25 degrees C. Thus, higher plasma T4 level under long photoperiod at 25 degrees C might be induced by testosterone.     

Effects of photoperiod,temperature and testosterone-treatment on plasma T3 and T4 levels in the Djungarian hamster,Phodopus sungorus

Summary The effects of photoperiod, temperature and testosterone treatment on plasma T₃ and T₄ levels were investigated in the Djungarian hamster. Plasma T₃ level was affected by temperature (25°C<7°C) but not by photoperiod. Plasma T₄ level was affected by photoperiod (short day < long day) at 25°C. Administration of testosterone increased plasma T₄ level under short photoperiod at 25°C. Thus, higher plasma T₄ level under long photoperiod at 25°C might be induced by testosterone.     

Mise en évidence de l'induction de la coagulation plasmatique par les hémocytes chezLocusta migratoria

Summary When performed at low temperature (4°C), centrifugation of hemolymph fromLocusta migratoria separates a plasma which does not exhibit any coagulation when it is preserved from the hemocyte pellet. Blood cells added to plasma, induce a rapid and normal coagulation (precipitation of coagulogen).

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Mise en évidence de l'induction de la coagulation plasmatique par les hémocytes chezLocusta migratoria

     

不同方法制备的Pt对电极染料敏化太阳能电池的光电性能研究

采用磁控溅射（sputtering）、高温热分解（high-temp）H2Pt Cl6·6H2O溶液和低温化学还原（low-temp）H2Pt Cl6·6H2O溶液等方法在FTO导电基片上制备了三类不同的Pt对电极,通过改变磁控溅射时间或旋涂退火循环次数,对FTO上不同对电极的载Pt量进行了调节.分析了三种不同方法制备的对电极的晶相结构、表面形貌以及透光率等特性.基于纳米管阵列和不同条件制备的对电极组装了系列染料敏化太阳能电池.测量了正照射和背照射条件下的电池光电性能,并分析了制备方法对对电极微结构和形貌的影响,以及对组装电池光电性能的影响,并对其各自的适用性和优缺点进行了比较分析.     

Temperature acclimation of Mg2+Ca2+-myofibrillar ATPase from a cold-selective teleost, Salvelinus fontinalis: a compromise solution

Brook trout (Salvelinus fontinalis, Mitchill) were acclimated over 15 weeks to either +4 degrees C or +24 degrees C. The effects of temperature on myofibrillar Mg2+Ca2+-ATPase activities were investigated. In contrast to goldfish, temperature acclimation does not alter the kinetic properties of the brook trout myofibrillar ATPase. Activation energy (delta G not equal to) is lower and substrate turnover number is higher than values previously reported for cold-adapted stenotherms. Properties of brook trout ATPase appear to be a compromise enabling function across a broad temperature range. The different strategies of adapting to seasonal temperature variations are briefly discussed.     

Quantitative determination of the activity of acid peptidases of industrial origin]

Reagent ninhydrine-Cd++, reacts with free alpha and epsilon amino groups of proteins. Horse-heart apomyoglobin was subjected to exhaustive succinylation, rendering the product non reactive to ninhydrine. The succinylglobin was submitted to enzyme digestion at pH 2.0, 4.0, 4.7 and 6.0. The commercially available enzymes contain mainly pepsin-like and chymosin-like enzymes. The enzymatic digests of succinyl-globin contain new free alpha-amino groups reacting with ninhydrin. Enzymatic digestion was performed under various condition (ratio E/S, pH). The results were compared to those obtained with synthetic substrate: PRO-HIS-LEU-SER-PHE(NO2)-NLEU-ALA-LEU-OME. The price of the synthetic substrate used, was more than 100 times the cost of succinyl-globin, thus the use of this substrate is a valuable tool for the quantitative estimation of peptidase activity in commercially available (pepsin, chymosin-like) enzymes.     

Process optimization of reaction of acid leaching residue of asbestos tailing and sodium hydroxide aqueous solution

Silica is the major component of the acid leaching residue of asbestos tailing.The waterglass solution can be prepared by the reaction of the residue with sodium hydroxide aqueous solution.Compared to the high temperature reaction method,this process is environmental friendly and low cost.In this paper,the reaction process of the residue and the sodium hydroxide aqueous solution is optimized.The optimum reaction process parameters are as follows:the usage of sodium hydroxide is 26.4 g/100 g acid leaching residue,the reaction temperature is 90℃,the reaction time is 1 h,and the ratio of the liquid/solid is 2.0.The significance sequence of the process parameters to the alkali leaching reaction effect is the usage of sodium hydroxidethe ratio of the liquid/solidthe reaction timethe reaction temperature.The significance sequence to the leaching ratio of SiO2 is the ratio of the liquid/solidthe usage of sodium hydroxidethe reaction timethe reaction temperature.The significance sequence to the modulus of the sodium silicate is the ratio of the liquid/solidthe usage of sodium hydroxidethe reaction timethe reaction temperature.Under the optimum conditions,the leaching ratio of the SiO2 is 77.5%,and the modulus of the sodium silicate is 3.15.The XRD analysis result indicates that the major components of the alkali leaching residue are serpentine,talc,quartz and some albite.     

Two-state model ofParamecium bursaria thigmotaxis

A theoretical framework has been developed for analysis of the interaction betweenParamecium bursaria and a glass surface. Adhesion to and detachment from a solid substrate were considered in the model as transitions between alternative states in cell behavior: (a) swimming, and (b) motionless (positive thigmotactic) state. According to the model and experimental data, a change in the fraction of swimming cells is described by a negative exponential course. The proposed model allows positive thigmotaxis, generally referred to in the literature simply as thigmotaxis, to be considered as the rate-constant of transition into the motionless state. This approach permits quantitative determination of thigmotaxis, and reveals its dependence on the phase of culture growth and the type of medium surrounding the cells. In the mineral maintenance solution, paramecia from the stationary phase of growth swim more slowly than those in the logarithmic phase of growth, and show enhanced thigmotaxis. However, a general relationship between thigmotaxis and swimming speed was not established.     

Two-state model of Paramecium bursaria thigmotaxis.

A theoretical framework has been developed for analysis of the interaction between Paramecium bursaria and a glass surface. Adhesion to and detachment from a solid substrate were considered in the model as transitions between alternative states in cell behavior: (a) swimming, and (b) motionless (positive thigmotactic) state. According to the model and experimental data, a change in the fraction of swimming cells is described by a negative exponential course. The proposed model allows positive thigmotaxis, generally referred to in the literature simply as thigmotaxis, to be considered as the rate-constant of transition into the motionless state. This approach permits quantitative determination of thigmotaxis, and reveals its dependence on the phase of culture growth and the type of medium surrounding the cells. In the mineral maintenance solution, paramecia from the stationary phase of growth swim more slowly than those in the logarithmic phase of growth, and show enhanced thigmotaxis. However, a general relationship between thigmotaxis and swimming speed was not established.     

Melatonin in the plasma of growing sheep subjected to short and skeleton long photoperiods

Summary The levels of melatonin in plasma were measured at hourly intervals for 24 h in 8 sheep, 4 under 8L:16D (short day) and 4 under 7L:10D:1L:6D (skeleton long day) after 38 days of exposure. Mean concentrations did not differ significantly between treatments (52 pg/ml under short days; 91 pg/ml for skeleton long days), but the levels were more stable during 24 h in the SD treatment. Under skeleton long days there were 3 peaks during the 10D scotophase, with low levels during the 6D scotophase and the 7L photophase.     

Phytanic acid impairs mitochondrial respiration through protonophoric action

Refsum disease is a rare, inherited neurodegenerative disorder characterized by accumulation of the dietary branched-chain fatty acid phytanic acid in plasma and tissues caused by a defect in the alphaoxidation pathway. The accumulation of phytanic acid is believed to be the main pathophysiological cause of the disease. However, the exact mechanism(s) by which phytanic acid exerts its toxicity have not been resolved. In this study, the effect of phytanic acid on mitochondrial respiration was investigated. The results show that in digitonin-permeabilized fibroblasts, phytanic acid decreases ATP synthesis, whereas substrate oxidation per se is not affected. Importantly, studies in intact fibroblasts revealed that phytanic acid decreases both the mitochondrial membrane potential and NAD(P)H autofluorescence. Taken together, the results described here show that unesterified phytanic acid exerts its toxic effect mainly through its protonophoric action, at least in human skin fibroblasts. Received 4 August 2007; received after revision 26 September 2007; accepted 10 October 2007 J. C. Komen, F. Distelmaier: These authors contributed equally to this work.     

One lipid, multiple functions: how various pools of PI(4,5)P2 are created in the plasma membrane

Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P₂] is a minor lipid of the inner leaflet of the plasma membrane that controls the activity of numerous proteins and serves as a source of second messengers. This multifunctionality of PI(4,5)P₂ relies on mechanisms ensuring transient appearance of PI(4,5)P₂ clusters in the plasma membrane. One such mechanism involves phosphorylation of PI(4)P to PI(4,5)P₂ by the type I phosphatidylinositol-4-phosphate 5-kinases (PIP5KI) at discrete membrane locations coupled with PI(4)P delivery/synthesis at the plasma membrane. Simultaneously, both PI(4)P and PI(4,5)P₂ participate in anchoring PIP5KI at the plasma membrane via electrostatic bonds. PIP5KI isoforms are also selectively recruited and activated at the plasma membrane by Rac1, talin, or AP-2 to generate PI(4,5)P₂ in ruffles and lamellipodia, focal contacts, and clathrin-coated pits. In addition, PI(4,5)P₂ can accumulate at sphingolipid/cholesterol-based rafts following activation of distinct membrane receptors or be sequestered in a reversible manner due to electrostatic constrains posed by proteins like MARCKS.     

Effect of ambient cold on urine concentration by the intact and denervated dog kidney

Summary In dogs transferred from room temperature to a low temperature of 1±(SD) 3.5°C, the plasma norepinephrine (NE) level increased significantly. Urine concentration decreased in the cold, in the intact and similarly in the chronically denervated kidney. After return to a warm environment urine concentration improved in both kidneys while plasma NE remained elevated. The data speak against an essential role of the renal nerves in the adaptation of renal function to ambient cold.Acknowledgments. We are greatly indebted to Dr J. Kurkus and Dr R. Gellert who performed the operations of surgical bladder division, and to Mrs W. Radziszewska for determination of plasma norepinephrine. The study was supported within the national research problem No. 10.4.     

电子封装热应力数值分析

针对目前电子封装的封装密度越来越高、封装厚度越来越薄、封装体在基板上所占面积越来越大,发热引起的失效越来越严重等问题,以晶体管瞬态热应力分析为例,建立热力耦合力学模型.利用ansys研究电子封装热失效问题,得到温度场、应力场和变彤场的分布规律.温度和应力的主要规律包括两点,一是温度和应力都在角点处变化明显,应力比较集中.温度从上到下逐层变化,逐渐减小,并且层与层之间温度变化不大,模型中间部分温度层厚度几乎相同,下半部分同一温度层有规律的变化.二是当温度较高时,在受约束面上和角点处应力值较大,并且在模型的角点和中部出现应力集中现象.     

Binding of centchroman--a nonsteroidal antifertility agent to human plasma proteins

Centchroman, a non-steroidal antifertility agent showed a low affinity (Kd = 13.19 X 10(-6) M) and nonsaturable binding to human plasma. Centchroman did not compete either with sex hormone binding globulin or corticosteroid binding globulin. Polyacrylamide gel electrophoresis and temperature dependent binding characteristics revealed that the protein responsible for centchroman binding to human plasma resembles albumin.     

Temperature-dependent activity of mosquito larvicidal factor(s) present inBacillus sphaericus 1593–4 and 1691

Summary The larvicidal factor(s) ofBacillus sphaericus 1593–4 and 1691 was more active at high temperature (33–35 °C) as compared to low temperature (23–25 °C) for the larvae ofCulex fatigans andAnopheles subpictus.     

An analysis of the effects of urethane on cardiovascular responsiveness to catecholamines in terms of its interference with Ca++ mobilization from both intra and extracellular pools

Urethane (1 X 10(-2) - 1 X 10(-1) M) reduced, in a concentration-dependent manner, both intra and extracellular Ca++ dependent noradrenaline-induced contractions of perfused rabbit ear artery as well as the tonic contractions produced by perfusion with high K+ solution. However, a quantitative analysis of the data indicated that for urethane concentrations similar to those found in plasma during anesthesia urethane antagonism is confined to noradrenaline-induced contractions which depend upon the mobilization of Ca++ from intracellular storage sites. In KCl-contracted arteries, urethane enhanced the relaxant effects of isoprenaline. - Urethane reduced the amplitude of contractions of spontaneously beating guinea-pig right atrium at concentrations which have only a limited effect on frequency. In addition, it decreased in a concentration-dependent manner the amplitude of isoprenaline-activated electrically driven, and K+ depolarized guinea-pig right ventricular strips. Urethane had no effect on the chrono and inotropic actions of isoprenaline on cardiac preparations. In in vivo experiments the chronotropic response to low doses of isoprenaline was significantly higher in urethane-treated as compared to unanesthetized rats. The higher dose of isoprenaline tested produced a significant fall in systolic blood pressure in urethane-anesthetized rats. A significant correlation exists between the chronotropic response to isoprenaline and resting heart rate values in urethane-anesthetized rats. These results indicate that urethane, at concentrations similar to those found in plasma during anesthesia selectively interferes with mobilization of Ca++ from intracellular storage sites. In addition, the interference of urethane anesthesia with the isoprenaline chronotropic effect 'in vivo' cannot be explained by a direct interference of urethane with beta-adrenoceptors at cardiac level.