Inhibition of rat liver transglutaminase by nucleotides.

Tissue-type transglutaminase (TGase) was purified from rat liver, and the effects of nucleotides on its activity were examined. The enzyme activity is inhibited by ATP in a concentration-dependent way, with complete inhibition by 3 mM ATP. Partially-purified TGase from human brain was inhibited by ATP in a manner similar to that observed with the rat liver enzyme. This suggests that the inhibition is a common phenomenon for tissue-type TGase in all species and tissues. The inhibition is reversible since full activity is restored by lowering the ATP concentration. CTP has a TGase-inhibitory potency equivalent to that of ATP, whereas GTP and UTP possess about 50% of the inhibitory activity of ATP. ADP inhibits TGase activity to the same extent as ATP, but AMP causes much less inhibition, and there is no inhibition by adenosine or adenine. The inhibition by ATP is insensitive to ionic strength and is non-competitive with the substrate putrescine. Since ATP levels in cells are of mM order, these results suggest that TGase activity is controlled by ATP in vivo.     

The physiology of the smooth muscle: an interdisciplinary review—part II

Summary In the guinea pig taenia coli, when glycogen is depleted by repeating Ca-induced contracture in excess K solution containing no glucose, the tension cannot be maintained. The decrease in tension is accompanied by reduction of high energy phosphate compounds and oxygen consumption. When substrate is readmitted to the glycogendepleted preparation in the presence of 2.4 mM Ca and 20 mM K, the first response is hyperpolarization of the membrane and relaxation, and this is followed by depolarization and development of contracture. The latter response is blocked by verapamil, suggesting that energy supply increases the Ca conductance of the plasma membrane. The early response is considered to be due to activation of electrogenic Ca pump, since this is not affected by ouabain as well as removal of Na and K. ATP produced by substrate readmission is probably preferentially utilized for Ca pump activation to reduce the intracellular. Ca. The recovery of tension is likely to be brought about by ATP supply not only to the contractile machinery but also to the plasma membrane to remove inactivation of Ca conductance. It is postulated that as the energy source is depleted, energy consumption is automatically limited by suppressing Ca influx, as a selfdefence mechanism. Since HB is as effective as glucose in the recovery of these processes, and also in the activation of electrogenic Na pump, the matabolic pathway of oxidative phosphorylation alone can support these functions without a contribution of the glycolytic pathway.     

Inhibition of rat liver transglutaminase by nucleotides

Summary Tissue-type transglutaminase (TGase) was purified from rat liver, and the effects of nucleotides on its activity were examined. The enzyme activity is inhibited by ATP in a concentration-dependent way, with complete inhibition by 3 mM ATP. Partially-purified TGase from human brain was inhibited by ATP in a manner similar to that observed with the rat liver enzyme. This suggests that the inhibition is a common phenomenon for tissue-type TGase in all species and tissues. The inhibition is reversible since full activity is restored by lowering the ATP concentration. CTP has a TGase-inhibitory potency equivalent to that of ATP, whereas GTP and UTP possess about 50% of the inhibitory activity of ATP. ADP inhibits TGase activity to the same extent as ATP, but AMP causes much less inhibition, and there is no inhibition by adenosine or adenine. The inhibition by ATP is insensitive to ionic strength and is non-competitive with the substrate putrescine. Since ATP levels in cells are of mM order, these results suggest that TGase activity is controlled by ATP in vivo.     

稳定表达绿色荧光蛋白的B16细胞株的建立及其生物学特性研究

目的建立稳定表达绿色荧光蛋白的B16黑色素瘤细胞株，并研究其生物学特性。方法转染增强型绿色荧光蛋白基因入B16细胞，经G418联合单克隆培养，筛选阳性细胞。激光共聚焦荧光显微镜观察EGFP阳性细胞形态；流式细胞仪检测细胞内EGFP基因的蛋白表达；RT-PCR检测EGFP基因的mRNA表达；皮下接种及MTF法评价细胞在体内外的生长特性。结果流式细胞仪检测表达绿色荧光蛋白的细胞阳性率为99．81％，RT-PCR检测到EGFP基因的mRNA阳性表达。生长曲线显示其体外增殖趋势与B16相似，EGFP—B16体内成瘤速度较B16慢（P〈0．05）。结论稳定表达绿色荧光蛋白的EGFP—B16细胞株成功建立，可作为作为进一步研究恶性黑色素瘤的材料。     

Possibility of metabolic control of membrane excitation

In the guinea pig taenia coli, when glycogen is depleted by repeating Ca-induced contracture in excess K solution containing no glucose, the tension cannot be maintained. The decrease in tension is accompanied by reduction of high energy phosphate compounds and oxygen consumption. When substrate is readmitted to the glycogen-depleted preparation in the presence of 2.4 mM Ca and 20 mM K, the first response is hyperpolarization of the membrane and relaxation, and this is followed by depolarization and development of contracture. The latter response is blocked by verapamil, suggesting that energy supply increases the Ca conductance of the plasma membrane. The early response is considered to be due to activation of electrogenic Ca pump, since this is not affected by ouabain as well as removal of Na and K. ATP produced by substrate readmission is probably preferentially utilized for Ca pump activation to reduce the intracellular Ca. The recovery of tension is likely to be brought about by ATP supply not only to the contractile machinery but also to the plasma membrane to remove inactivation of Ca conductance. It is postulated that as the energy source is depleted, energy consumption is automatically limited by suppressing Ca influx, as a self-defence mechanism. Since beta HB is as effective as glucose in the recovery of these processes, and also in the activation of electrogenic Na pump, the metabolic pathway of oxidative phosphorylation alone can support these functions without a contribution of the glycolytic pathway.     

Lichthemmung der Oxydation von Krebscyclus-Säuren durch Blumenkohl-Mitochondrien

Summary The oxidation of succinic-, malic-, and -ketoglutaric acid by cauliflower mitochondria is slowed down by strong visible light. The cytochrome-c-oxidase activity is also inhibited by light and the pigment is completely reduced in time. It is concluded that the Krebscycle is inhibited by light.     

Phosphorylation and activation of the atypical kinase p53-related protein kinase (PRPK) by Akt/PKB

p53-related protein kinase (PRPK), the human homologue of yeast Bud32, belonging to a small subfamily of atypical protein kinases, is inactive unless it is previously incubated with cell lysates. Here we show that such an activation of PRPK is mediated by another kinase, Akt/PKB, which phosphorylates PRPK at Ser250. We show that recombinant PRPK is phosphorylated in vitro by Akt and its phospho-form is recognized by a Ser250-phospho-specific antibody; that cell co-transfection with Akt along with wild-type PRPK, but not with its Ser250Ala mutant, results in increased PRPK phosphorylation; and that the phosphorylation of p53 at Ser15, the only known substrate of PRPK, is markedly increased by co-transfection of Akt with wild-type PRPK, but not PRPK dead mutant, and is abrogated by cell treatment with the Akt pathway inhibitor LY294002. Our data disclose an unanticipated mechanism by which PRPK can be activated and provide a functional link between this enigmatic kinase and the Akt signaling pathway.     

Über die durch Licht bedingte «Missweisung» bei den Tänzen im Bienenstock

Summary Bees are able to indicate direction to their hive comrades by means of a waggling dance of 2 kinds: in the horizontal plane with regard to the sun they point directly towards the goal by a waggling walk using the same angle to the sun as they took in their flight. Inside the dark hive in the vertical honey-comb, they transpose the angle between goal and sun to the field of gravity, whereby the sun's direction is shown by a waggling walk upwards, and the angle to the right or left of the sun's position is given by a dance-direction in the corresponding angle to the right or left of the zenith.If a piece of blue sky is made visible in an observation hive to the bees which are dancing in orientation by gravity, they recognise the position of the sun by this polarisation sample, and the effort to orientate themselves directly by the sun (as in the horizontal plane) comes into conflict with the orientation by gravity. The result is a dance direction which corresponds remarkably well with the halving of the angle between what the dance direction should have been by gravity and what it should have been by light orientation (Figure 1). This is also true when the bee is orientating itself by polarised sky light over its back, while the sun is at the other side of the honey-comb under its front (Figure 2), a situation which does not occur during flight but which is important for its dance in the swarm. The bees receiving the information compensate the deviation of the angle determined by light, and fly to the right goal.As the sun itself, as well as the piece of blue sky, was made visible to the dancers, its influence dominated and they orientated themselves by its light (Figure 3).     

Dinemorphan N-demethylation by mouse liver microsomes

Dinemorphan, an antitussive drug, is N-demethylated in vitro by mouse liver microsomes with biphasic kinetics showing two apparent Km and Vmax. Moreover, dinemorphan N-demethylation is inhibited by CO, SKF-525A, metyrapone and it is specifically catalyzed by a phenobarbital-inducible form of cytochrome P-450.     

New technique for movement analysis: Application to biological systems

Summary A new system for motion analysis is described. Modulation of light intensity by an organism's movement over an opaque and transparent checkerboard grid is monitored by a photocell. The photocell's output is proportional to the organism's amplitude and frequency of movement. This output is analyzed by a continuous interval dot display and spectral analysis. The system was tested by analyzing the activity of 4 Drosophila strains which are known to differ in their activity. General applicability of the system is discussed.This research was supported by a grant from the United States-Israel Binational. Science Foundation (BSF), Jerusalem, Israel.     

Lysis of Rous sarcoma virus in the presence of anti-chicken fibroblast serum and complement]

Rous sarcoma virus produced by Chick embryo fibroblasts is inactivated by an antiserum prepared against uninfected fibroblasts in the presence, but not in the absence, of complement. This inactivation which demonstrates the presence of one or more antigens of the surface of the producer cell on the viral envelope, is due to virolysis. This is demonstrated by the release of the viral internal proteins and by the fact that the viral RNA becomes entirely degradable by RNase.     

Sur le rôle des groupes thiol dans la coagulation du plasma sanguin

Summary Coagulation of hen's blood plasma is inhibited by iodacetamid, dichloroethyl sulfide and sulfone, maleic acid; it is accelerated by chloropicrin. The inhibitory action is probably due to the formation of substitution products by reactions between the inhibiting agent and the -SH groups of the fibrinogen. The coagulating action is probably due to the formation of -S-S-bridges between molecules.     

Nitric oxide synthase reduces nitrite to NO under anoxia

Cultured bEND.3 endothelial cells show a marked increase in NO production when subjected to anoxia, even though the normal arginine pathway of NO formation is blocked due to absence of oxygen. The rate of anoxic NO production exceeds basal unstimulated NO synthesis in normoxic cells. The anoxic release of NO is mediated by endothelial nitric oxide synthase (eNOS), can be abolished by inhibitors of NOS and is accompanied by consumption of intracellular nitrite. The anoxic NO release is unaffected by the xanthine oxidase inhibitor oxypurinol. The phenomenon is attributed to anoxic reduction of intracellular nitrite by eNOS, and its magnitude and duration suggests that the nitrite reductase activity of eNOS is relevant for fast NO delivery in hypoxic vascular tissues. Received 20 August 2006; received after revision 21 September 2006; accepted 8 November 2006     

Regulatory properties of the citrate synthase from Rhodospirillum rubrum.

Citrate synthase, purified 600-fold from Rhodospirillum rubrum, is activated by KCl and inhibited by ATP and NADH; the effect of the latter inhibitor is completely counteracted by AMP and partially counteracted by KCl.     

Microtubule inhibitors block the morphological changes induced in Drosophila blood cells by a parasitoid wasp factor

The shape change of Drosophila melanogaster blood cells (lamellocytes) from discoidal to bipolar that is caused by a factor from the female parasitoid Leptopilina heterotoma is blocked by the tubulin inhibitors vinblastine and vincristine in vitro. The actin inhibitor, cytochalasin B, causes arborization of Drosophila lamellocytes and acts synergistically with the wasp factor to alter lamellocyte morphology. Lamellocyte aborization induced by cytochalasin B is blocked by simultaneous treatment with vinblastine. These observations indicate that the changes in lamellocyte shape induced by both the wasp factor and cytochalasin B require microtubule assembly.     

Correlations between the neurobiology of colour vision and the psycholinguistics of colour naming.

Neurobiological experiments demonstrate that colour sensation is perceived by the brain by processes which, in principle, follow the opponent colour pairs scheme proposed by Hering in 1874. Tests on colour naming in various European, Asian and Central American languages have shown that the opponent scheme is also reflected in psycholinguistics. The linguistic evolution of colour terms proposed by Berlin and Kay (1969) is correlated directly with the ontogenetic development of language in children as elucidated by Jakobson (1941). Colour vision is therefore a suitable field for interdisciplinary investigations of brain processes and linguistics.     

Behavioural and electrophysiological evaluation of oviposition attractants forCulex quinquefasciatus say (Diptera: Culicidae)

The attraction of gravidCulex quinquefasciatus by the oviposition pheromone,erythro-6-acetoxy-5-hexadecanolide, and by polluted water is described. Both materials increase oviposition and when combined the effect is additive. The oviposition behaviour is reflected by the antennal sensitivity to these compounds.     

一种基于自体集层次聚类的否定选择算法

否定选择算法是用于产生人工免疫检测器的重要算法,然而传统的否定选择过程需要将随机生成的候选检测器与全部自体数据进行匹配以排除识别了自体的无效检测器,该匹配过程导致检测器的生成效率过低,极大地限制了免疫算法的应用.为此,文中提出了一种基于自体集层次聚类的否定选择算法CB-RNSA.算法首先对自体数据进行层次聚类预处理,然后用聚类中心取代自体数据点与候选检测器进行匹配,以减少距离计算代价.在生成检测器的过程中,候选检测器被限定在非自体空间的低覆盖率区域内,以降低检测器冗余.对检测器的非自体空间覆盖率进行了概率分析,给出了中止生成检测器的条件,该条件较传统的预设检测器数量的中止条件更为合理.理论分析表明CB-RNSA的时间复杂度与自体集规模无关,从而解决了经典的否定选择算法的时间复杂度随自体数量呈指数增长这一难题,极大地提高了大自体样本空间下的检测器生成效率.对比实验结果表明:在相同的实验数据集与期望覆盖率下,CB-RNSA的检测率比经典的RNSA与V-detector算法分别提高了12.3%与7.4%,误警率分别降低了8.5%与4.9%,产生检测器的时间代价分别降低了67.6%和75.7%.     

Adipocyte extracellular matrix composition, dynamics and role in obesity

The central role of the adipose tissue in lipid metabolism places specific demands on the cell structure of adipocytes. The protein composition and dynamics of the extracellular matrix (ECM) is of crucial importance for the functioning of those cells. Adipogenesis is a bi-phasic process in which the ECM develops from a fibrillar to a laminar structure as cells move from the commitment phase to the growth phase characterized by storage of vast amounts of triglycerides. Mature adipocytes appear to spend a lot of energy on the maintenance of the ECM. ECM remodeling is mediated by a balanced complement of constructive and destructive enzymes together with their enhancers and inhibitors. ECM remodeling is an energy costing process regulated by insulin, by the energy metabolism, and by mechanical forces. In the obese, overgrowth of adipocytes may lead to instability of the ECM, possibly mediated by hypoxia.     

Haemagglutinating activity of modeccin

Summary Modeccin, the toxin ofAdenia digitata, agglutinates erythrocytes from several mammalian species. The haemagglutinating activity is enhanced by neuraminidase and is inhibited by galactose and by galactose-containing sugars.Acknowledgment. This research was supported by grants from the Consiglio Nazionale delle Ricerche, Rome, and by the Pallotti's legacy for cancer research.