Effects of corpora cardiaca extract,furosemide and ion substitution on sodium and chloride flux in perfused Malpighian tubules ofLocusta

Treatment with the co-transport inhibitor, furosemide decreased³⁶Cl^– flux across perfused Malpighian tubules ofLocusta. However, exclusion of³⁶Cl^– from the bathing medium had not effect on²²Na⁺ flux whereas substitution of bathing medium Na⁺ by K⁺ increased³⁶Cl^– flux. Diuretic extract of corpora cardiaca increased²²Na⁺ (by 106%) and³⁶Cl^– (by 335%) fluxes differentially.     

Invertebrate neuropeptides resembling vasotocin and some analogues: Synthesis and pharmacological properties

Summary The solid phase synthesis of three invertebrate vasopressin-oxytocin homologs: AVP-like factor, F₁ ¹, ([Leu², Thr⁴] AVT)² isolated from subesophageal and thoracic ganglia ofLocusta migratoria ³, Arg-conopressin-S⁴. ([Ile², Arg⁴] AVT), Lys-conopressin-G⁴ ([Phe², Arg⁴] LVT), both isolated from the venom of fish-hunting marine snails of the genusConus and six of their analogues is reported. These analogues are: [Arg⁴] AVT, [Ile²] AVT, [Leu²] AVT, [Phe², Arg⁴] AVT, [Arg⁴] LVT and [Ile², Arg⁴] LVT. All peptides were tested for antidiuretic and vasopressor activities.     

Announcement of the Editor-in-Chief

Announcement.  As decided in July 2005 we continue to publish once a year (in July) the names of the authors and the titles of the two most read (by Internet) Research Papers and Reviews published in Cell. Mol. Life Sci. the previous year. Thus we have the pleasure to provide you with the results of 2005. Research Articles  

Über Peptidsynthesen,XLIII. Darstellung einiger Oxytocin-Analoga

Summary Four new analogues of oxytocin, i.e. Phe²-Ser⁴-Ileu⁸-oxytocin, Phe³-Ser⁴-Ileu⁸-oxytocin, Gly⁴-Ileu⁸-oxytocin and Phe²-Val³-oxytocin have been synthesized and their biological activities compared with those of oxytocin (Syntocinon®).

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XLII. Mitt.:E. Schröder, Justus Liebigs Annln Chem., im Druck.     

Bicarbonate and chloride transport across rat ileal basolateral membrane

The mechanisms of HCO ₃ ^– and Cl^– transport across basolateral membranes from rat ileum were investigated in isolated vesicles by means of uptake experiments. Neither Cl^–/HCO ₃ ^– exchanger nor Na⁺–(HCO ₃ ^– )_n cotransport seem to be present in ileal basolateral membranes. Moreover Cl^– uptake is unaffected bycis Na⁺ and/or K⁺ gradients, indicating the absence of Na⁺–Cl^–, K⁺–Cl^– and Na⁺–K⁺–2Cl^– symport activity. An electrically conductive pathway seems to be responsible for both HCO ₃ ^– and Cl^– fluxes. Evidence is also given for the presence of a Na⁺/H⁺ exchanger at the basolateral pole of ileal enterocytes.     

Cancer spheres from gastric cancer patients provide an ideal model system for cancer stem cell research

Cancer stem cells have been hypothesized to drive the growth and metastasis of tumors. Because they need to be targeted for cancer treatment, they have been isolated from many solid cancers. However, cancer stem cells from primary human gastric cancer tissues have not been isolated as yet. For the isolation, we used two cell surface markers: the epithelial cell adhesion molecule (EpCAM) and CD44. When analyzed by flow cytometry, the EpCAM⁺/CD44⁺ population accounts for 4.5% of tumor cells. EpCAM⁺/CD44⁺ gastric cancer cells formed tumors in immunocompromised mice; however, EpCAM^?/CD44^?, EpCAM⁺/CD44^? and EpCAM^?/CD44⁺ cells failed to do so. Xenografts of EpCAM⁺/CD44⁺ gastric cancer cells maintained a differentiated phenotype and reproduced the morphological and phenotypical heterogeneity of the original gastric tumor tissues. The tumorigenic subpopulation was serially passaged for several generations without significant phenotypic alterations. Moreover, EpCAM⁺/CD44⁺, but not EpCAM^?/CD44^?, EpCAM⁺/CD44^? or EpCAM^?/CD44⁺ cells grew exponentially in vitro as cancer spheres in serum-free medium, maintaining the tumorigenicity. Interestingly, a single cancer stem cell generated a cancer sphere that contained various differentiated cells, supporting multi-potency and self-renewal of a cancer stem cell. EpCAM⁺/CD44⁺ cells had greater resistance to anti-cancer drugs than other subpopulation cells. The above in vivo and in vitro results suggest that cancer stem cells, which are enriched in the EpCAM⁺/CD44⁺ subpopulation of gastric cancer cells, provide an ideal model system for cancer stem cell research.     

Differences in membrane ion transport between hepatocytes from the periportal and the pericentral areas of the liver lobule

We studied the Na⁺/K⁺ pump, Na⁺/K⁺ ATPase activity, and oxygen consumption (QO₂) in hepatocytes isolated from the periportal (PH) and pericentral (CH) regions of the liver lobule, to provide an insight into the functional properties of these cells. Na⁺/K⁺ pump activity was determined using⁸⁶Rb⁺ (a functional analog of K⁺) and ouabain, a specific inhibitor of this transport system. Our results indicate the the Na⁺/K⁺, pump and Na⁺/K⁺ ATPase activity are significantly lower in CH than in PH, although basal ouabain-sensitive (OS) QO₂ was negligible in both of these cell preparations. However, OSQO₂ was significantly lower in CH than in PH when the Na⁺/K⁺ pump was activated using the ionophore nystatin in a Na⁺-containing medium. These results indicate that the differences in membrane ion transport exist between hepatocytes from different locations of the liver lobule.     

Na+,K+-ATPase as a docking station: protein–protein complexes of the Na+,K+-ATPase

The Na⁺,K⁺-ATPase, or sodium pump, is well known for its role in ion transport across the plasma membrane of animal cells. It carries out the transport of Na⁺ ions out of the cell and of K⁺ ions into the cell and thus maintains electrolyte and fluid balance. In addition to the fundamental ion-pumping function of the Na⁺,K⁺-ATPase, recent work has suggested additional roles for Na⁺,K⁺-ATPase in signal transduction and biomembrane structure. Several signaling pathways have been found to involve Na⁺,K⁺-ATPase, which serves as a docking station for a fast-growing number of protein interaction partners. In this review, we focus on Na⁺,K⁺-ATPase as a signal transducer, but also briefly discuss other Na⁺,K⁺-ATPase protein–protein interactions, providing a comprehensive overview of the diverse signaling functions ascribed to this well-known enzyme.     

On the synthesis of serine and homoserine samples asymmetrically labelled with tritium and deuterium in the hydroxymethylene group

Summary (1 R) [1-³H,²H₁] 3-Phenylpropanol, the key intermediate in the synthesis of (4 R) [4-³H,²H₁] D, L-homoserine and of the (4 S)-isomer, is obtained from (1 S) [1-²H₁] 3-phenylpropanol and (1 RS) [1-³H] ethanol upon incubation with yeast alcohol dehydrogenase and NAD⁺; under similar conditions 2-phenylethanol undergoes very small exchange with [1-²H₂] ethanol.     

Beeinflussung der ATPase aminspeichernder Granula von Nebennierenmark und Milznerven durch Thallium

Summary ATPases of the amine storing granules from bovine adrenal medulla and splenic nerves are inhibited by Tl⁺⁺⁺ 3×10^–5 and 5×10^–6 M, respectively. Tl⁺ up to 10^–3 M is ineffective. By T1⁺⁺⁺ in concentrations of 10^–4 M or more, proteins are precipitated, so that the enzyme inhibiton by these concentrations is unspecific. If T1⁺ is oxidized to Tl⁺⁺⁺ in the organism, the inhibition of granular ATPase may be responsible for the alterations of the catecholamine metabolism observed in thallium intoxication.     

Sopra un caso particolare della superficie F 4 (3) di M. Noether

Summary The Noetherian surfaceF ₄ ⁽³⁾ , which is represented on a plane by a linear ³ system ofC ₉(A ₁ ³ A ₂ ³ A ₃ ³ A ₄ ³ A ₅ ³ A ₆ ³ A ₇ ³ A ₈ ³ A ₉ ² A ₁₀), possesses generally only one linear pencil of elliptic cubics. IfA _i (i=1, 2, , 9) are the basis points of aHalphen pencil ofC ₉,A ₁₀ is infinitely near toA ₉, and in this caseF ₄ ⁽³⁾ is a not trivial example of such a surface with two pencils of elliptic cubics.     

Reactivity of some transition metals on nuclear protein biosynthesis in rat liver

Zusammenfassung An enzymatisch aktiven Leberzellkernen aus Ratten wurde die Reaktivität von Hg²⁺, Mn²⁺, Fe³⁺ und Cr³⁺ auf die nukleare Proteinbiosynthese geprüft. Hg²⁺-Ionen hemmen den¹⁴C-Aminosäure-Einbau vollständig, während Mn²⁺ und Fe³⁺ eine schwache Stimulierung verursachen. Die¹⁴C-Einbaurate ist nach Cr³⁺-Zusätzen stark erhöht. Es konnte gezeigt werden, dass die Cr³⁺-Stimulierung nicht durch Adsorption zu erklären ist.     

Na+ mechanism of δ-opioid receptor induced protection from anoxic K+ leakage in the cortex

Activation of δ-opioid receptors (DOR) attenuates anoxic K⁺ leakage and protects cortical neurons from anoxic insults by inhibiting Na⁺ influx. It is unknown, however, which pathway(s) that mediates the Na⁺ influx is the target of DOR signal. In the present work, we found that, in the cortex, (1) DOR protection was largely dependent on the inhibition of anoxic Na⁺ influxes mediated by voltage-gated Na⁺ channels; (2) DOR activation inhibited Na⁺ influx mediated by ionotropic glutamate N-methyl-D-aspartate (NMDA) receptors, but not that by non-NMDA receptors, although both played a role in anoxic K⁺ derangement; and (3) DOR activation had little effect on Na⁺/Ca²⁺ exchanger-based response to anoxia. We conclude that DOR activation attenuates anoxic K⁺ derangement by restricting Na⁺ influx mediated by Na⁺ channels and NMDA receptors, and that non-NMDA receptors and Na⁺/Ca²⁺ exchangers, although involved in anoxic K⁺ derangement in certain degrees, are less likely the targets of DOR signal. Received 26 November 2008; received after revision 26 December 2008; accepted 13 January 2009     

Die Abhängigkeit der Ca++-Aufnahme isolierter Mitochondrien des Herzmuskels von der Na+-und K+-Konzentration als mögliche Ursache der inotropen Digitaliswirkung

Summary In isolated mitochondria of heart muscle from rabbits and oxen there is, under suitable conditions, an accumulation of Ca⁺⁺, which is significantly enhanced by elevating the K⁺/Na⁺ quotient of the incubation medium. K-strophanthine (10^–5–10^–7) does not influence the accumulation of Ca⁺⁺ by the mitochondria of heart muscle. Therefore the intracellular increase in exchangeable Ca⁺⁺ observed after digitalis-glycosides could be explained by a decrease of the intracellular K⁺/Na⁺ quotient, which is caused by inhibition of the membrane ATPase and diminishes the capacity for Ca⁺⁺ accumulation in mitochondria.     

Blockade by zinc ions of K+-induced contraction and calcium in guinea pigTaenia coli

Preincubation with 0.3 mM Zn²⁺ markedly inhibited both the tonic response and Ca²⁺ binding at low affinity sites induced by K⁺ (60 mM), with smaller effects on the phasic response and the high affinity Ca²⁺ sites, inTaenia coli. However, when the muscle was kept in Zn²⁺-containing medium following the first stimulation with the K⁺, the phasic response and the high affinity Ca²⁺ sites were more severely inhibited during the second stimulation with K⁺. This probably indicates that Zn²⁺ reduced the tonic tension response to K⁺ mainly by inhibiting Ca²⁺ influx at the cell membranes ofTaenia coli. However, when Zn²⁺ is continuously present, Ca²⁺ is not supplied at the storage sites and is not available for the phasic response to a second stimulation with K⁺.     

Influence of ethanol on calcium,inositol phospholipids and intracellular signalling mechanisms

Summary Studies have implicated Ca⁺⁺ in the actions of ethanol at many biochemical levels. Calcium as a major intracellular messenger in the central nervous system is involved in many processes, including protein phosphorylation enzyme activation and secretion of hormones and neurotransmitters. The control of intracellular calcium, therefore, represents a major step by which neuronal cells regulate their activities. The present review focuses on three primary areas which influence intracellular calcium levels; voltage-dependent Ca⁺⁺ channels, receptor-mediated inositol phospholipid hydrolysis, and Ca⁺⁺/Mg⁺⁺-ATPase, the high affinity membrane Ca⁺⁺ pump.Current research suggests that a subtype of the voltage-dependent Ca⁺⁺ channel, the dihydropyridine-sensitive Ca⁺⁺ channel, is uniquely sensitive to acute and chronic ethanol treatment. Acute exposure inhibits, while chronic ethanol exposure increases⁴⁵Ca⁺⁺-influx and [³H]dihydropyridine receptor binding sites. In addition, acute and chronic exposure to ethanol inhibits, then increases Ca⁺⁺/Mg⁺⁺-ATPase activity in neuronal membranes. Changes in Ca⁺⁺ channel and Ca⁺⁺/Mg⁺⁺-ATPase activity following chronic ethanol may occur as an adaptation process to increase Ca⁺⁺ availability for intracellular processes. Since receptor-dependent inositol phospholipid hydrolysis is enhanced after chronic ethanol treatment, subsequent activation of protein kinase-C may also be involved in the adaptation process and may indicate increased coupling for receptor-dependent changes in Ca⁺⁺/Mg⁺⁺-ATPase activity.The increased sensitivity of three Ca⁺⁺-dependent processes suggest that adaptation to chronic ethanol exposure may involve coupling of one or more of these processes to receptor-mediated events.     

Extracellular Mg2+ regulates intracellular Mg2+ and its subcellular compartmentation in fission yeast, Schizosaccharomyces pombe

Effects of extracellular magnesium ions ([Mg²⁺]_o ) on intracellular free Mg²⁺ ([Mg²⁺]_i ) and its subcellular distribution in single fission yeast cells, Schizosaccharomyces pombe, were studied with digital-imaging microscopy and an Mg²⁺ fluorescent probe (mag-fura-2). Using 0.44 mM [Mg²⁺]_o , [Mg²⁺]_i in yeast cells was 0.91±0.08 mM. Elevation of [Mg²⁺]_o to 1.97 mM induced rapid (within 5 min) increments in [Mg²⁺]_i (2.18±0.11 mM). Lowering [Mg²⁺]_o to 0.06 mM, however, exerted no significant effects on [Mg²⁺]_i (0.93±0.14 mM), at least for periods of up to 30 min. Irrespective of the [Mg²⁺]_o used, the subcellular distribution of [Mg²⁺]_i remained hetero geneous, i.e. where the sub-plasma membrane region >cytoplasm >nucleus. [Mg²⁺] in all three subcellular compartments increased significantly, two- to threefold, concomitant with [Mg²⁺]_i when placed in 1.97 mM [Mg²⁺]_o . We conclude that [Mg²⁺]_i in fission yeast is maintained at a physiologic level when [Mg²⁺]_o is low, but intracellular free Mg²⁺ rapidly rises when [Mg²⁺]_o is elevated. Like most eukaryotic cells, yeast may have a Mg²⁺ transport system(s) which functions to maintain gradients of Mg²⁺ from the outside to inside the cell and among its subcellular compartments. Received 18 April 1996; received after revision 4 July 1996; accepted 26 July 1996     

Changes induced by the acute administration of iodide on secretion of iodinated components by the rat thyroid gland perfused in situ

Résumé Nous avons étudié l'effet de l'iodure 127 (10^–3 et 10^–4 M) sur la sécrétion de T₄ ^*, T₃ ^* et –I^* par la thyroïde de rat, prémarquée à l'iode 131 et perfusée in situ avec du sang non radioactif contenant 12 mU de TSH/ml. Il n'a pas été noté de changement significatif dans la sécrétion de T₄ ^* ou T₃ ^* jusqu'à 45 min après le début de l'administration d'iodure 127, qu'il y ait ou non du méthimazole (10^–3 M) dans le sang perfusé. Il y avait par contre une augmentation de la sécrétion d'-I^* pendant la perfusion de 10^–3 M d'ïodure, indiquant une décharge d'-I^* pendant la perfusion de 10^–3 M d'ïodure, indiquant une décharge d'-I^* de provenance intrathyroïdienne. Cet effet est similaire à celui noté auparavant pendant le perfusion de 10^–3 M de ClO ₄ ^– .     

Knockout of the Bcmo1 gene results in an inflammatory response in female lung, which is suppressed by dietary beta-carotene

Beta-carotene 15,15′-monooxygenase 1 knockout (Bcmo1 ^?/?) mice accumulate beta-carotene (BC) similarly to humans, whereas wild-type (Bcmo1 ^+/+) mice efficiently cleave BC. Bcmo1 ^?/? mice are therefore suitable to investigate BC-induced alterations in gene expression in lung, assessed by microarray analysis. Bcmo1 ^?/? mice receiving control diet had increased expression of inflammatory genes as compared to BC-supplemented Bcmo1 ^?/? mice and Bcmo1 ^+/+ mice that received either control or BC-supplemented diets. Differential gene expression in Bcmo1 ^?/? mice was confirmed by real-time quantitative PCR. Histochemical analysis indeed showed an increase in inflammatory cells in lungs of control Bcmo1 ^?/? mice. Supported by metabolite and gene-expression data, we hypothesize that the increased inflammatory response is due to an altered BC metabolism, resulting in an increased vitamin A requirement in Bcmo1 ^?/? mice. This suggests that effects of BC may depend on inter-individual variations in BC-metabolizing enzymes, such as the frequently occurring human polymorphisms in BCMO1.     

Untersuchungen über den Einfluss verschiedener Kationen auf die ATP-Spaltung durch die Zytoplasmafraktion vom Plexus chorioideus des Rindes

Summary The cytoplasma fraction of the bovine choroid plexus epithelial cells was found to contain a considerable ATPase activity. The influence of Na⁺, K⁺, Li⁺, Rb⁺, Cs⁺, Co⁺⁺, Mn⁺⁺, Zn⁺⁺ and Fe⁺⁺⁺ on the activity of the Mg⁺⁺-dependent enzyme has been studied. The monovalent cations do not influence the enzymic activity, whereas the effect of the bi- and trivalent cations is characterized by an inhibition of the ATPase.