Influence of the composition of the fusion medium on the yield of electrofused yeast hybrids

Abstract Electrofusion between cells of yeast strains with different genetic markers in isotonic sorbitol solutions leads to high yields of hybrids when 0.1 mM Ca²⁺ and 0.5 mM Mg²⁺ salts are aded. On average, 1000–2000 hybrids are obtained when electrofusion is performed (in a helical chamber) compared to a yield of about 40–120 in the absence of these bivalent cations. A further increase in yield can be achieved by the addition of 1 mg/ml albumin, which results in up to 4000 hybrids per experimental run. The entire fusion process leads to very reproducible results in the presence of these substances.     

The Legal Regulation of Floats and Gliders—In Quest of a New Regime?

This article analyzes the legal status of unmanned instruments (particularly, floats and gliders) for observation purposes in the ocean environment. These new kinds of instruments are being deployed by the thousands into the oceans, not the least as part of the Argo Project of the International Oceanographic Commission. Their uncontrolled drifting has raised legal questions, especially when such instruments enter waters subject to the jurisdiction of foreign states. The authors argue that the current international legal framework is insufficient to address the pertinent issues, and that a new legal regime is needed.     

Mechanisms and development of self–other distinction in dyads and groups

This opinion piece offers a commentary on the four papers that address the theme of the development of self and other understanding with a view to highlighting the important contribution of developmental research to understanding of mechanisms of social cognition. We discuss potential mechanisms linking self–other distinction and empathy, implications for grouping motor, affective and cognitive domains under a single mechanism, applications of these accounts for joint action and finally consider self–other distinction in group versus dyadic settings.     

Energetics and intermediates of the assembly of Protein OmpA into the outer membrane of Escherichia coli

OmpA is a major protein of the outer membrane of Escherichia coli. It is made as a larger precursor, pro-OmpA, which requires a membrane potential for processing. We now show that pro-OmpA accumulates in the cytoplasm of cells treated with carbonyl cyanide m-chlorophenylhydrazone, an uncouple which lowers the membrane potential. Upon restoration of the potential, this pro-OmpA is secreted, processed, and assembled into the outer membrane. Pro-OmpA made in vitro is also recovered with the postribosomal supernatant. It is efficiently processed to OmpA by liposomes which have bacterial leader peptidase that is exclusively internally oriented. These experiments show that: (i) the insertion of pro-OmpA into the plasma membrane is not coupled to its synthesis; (ii) insertion is promoted by the transmembrane electrochemical potential; (iii) pro-OmpA can cross a bilayer spontaneously; and (iv) pro-OmpA is processed by the same leader peptidase which converts M13 procoat to coat.     

Co‐occurrence patterns of trees along macro‐climatic gradients and their potential influence on the present and future distribution of Fagus sylvatica L.

Aim During recent and future climate change, shifts in large‐scale species ranges are expected due to the hypothesized major role of climatic factors in regulating species distributions. The stress‐gradient hypothesis suggests that biotic interactions may act as major constraints on species distributions under more favourable growing conditions, while climatic constraints may dominate under unfavourable conditions. We tested this hypothesis for one focal tree species having three major competitors using broad‐scale environmental data. We evaluated the variation of species co‐occurrence patterns in climate space and estimated the influence of these patterns on the distribution of the focal species for current and projected future climates. Location Europe. Methods We used ICP Forest Level 1 data as well as climatic, topographic and edaphic variables. First, correlations between the relative abundance of European beech (Fagus sylvatica) and three major competitor species (Picea abies, Pinus sylvestris and Quercus robur) were analysed in environmental space, and then projected to geographic space. Second, a sensitivity analysis was performed using generalized additive models (GAM) to evaluate where and how much the predicted F. sylvatica distribution varied under current and future climates if potential competitor species were included or excluded. We evaluated if these areas coincide with current species co‐occurrence patterns. Results Correlation analyses supported the stress‐gradient hypothesis: towards favourable growing conditions of F. sylvatica, its abundance was strongly linked to the abundance of its competitors, while this link weakened towards unfavourable growing conditions, with stronger correlations in the south and at low elevations than in the north and at high elevations. The sensitivity analysis showed a potential spatial segregation of species with changing climate and a pronounced shift of zones where co‐occurrence patterns may play a major role. Main conclusions Our results demonstrate the importance of species co‐occurrence patterns for calibrating improved species distribution models for use in projections of climate effects. The correlation approach is able to localize European areas where inclusion of biotic predictors is effective. The climate‐induced spatial segregation of the major tree species could have ecological and economic consequences.     

Zur h?henverbreitung der v?gel

Ohne Zusammenfassung     

Resistivity of Red Blood Cells Against High-Intensity, Short-Duration Electric Field Pulses Induced by Chelating Agents

The interaction of human red blood cells (RBCs) with diethylenetriamine-pentaacetic acid (DTPA) or its Gd-complex (Magnevist, a widely used clinical magnetic resonance contrast agent containing free DTPA ligands) led to the following, obviously interrelated phenomena. (i) Both compounds protected erythrocytes against electrohemolysis in isotonic solutions caused by a high-intensity DC electric field pulse. (ii) The inhibition of electrohemolysis was observed only when cells were electropulsed in low-conductivity solutions. (iii) The uptake of Gd-DTPA by electropulsed RBCs was relatively low. (iv) (Gd-) DTPA reduced markedly deformability of erythrocytes, as revealed by the electrodeformation experiments using high-frequency electric fields. Taken together, the results indicate that (Gd-) DTPA produce stiffer erythrocytes that are more resistant to electric field exposure. The observed effects of the chelating agents on the mechanical properties and the electropermeabilization of RBCs must have an origin in molecular changes of the bilayer or membrane-coupled cytoskeleton, which, in turn, appear to result from an alteration of the ionic equilibrium (e.g., Ca²⁺ sequestration) in the vicinity of the cell membrane. Received: 19 January 1999/Revised: 1 April 1999     

Über das Invertierungsvermögen der Speicheldrüsen und des Mitteldarmes von Bienen verschiedenen Alters

Zusammenfassung Die gewonnenen Ergebnisse zeigen mit völliger Klarheit, daß die Schlunddrüse ein stark ausgesprochenes Invertierungsvermögen besitzt, das sich jedoch in hohem Grade, je nach dem Alter der Bienen ändert. Im Gegensatz zum Mitteldarm, der Invertase auszuscheiden beginnt von dem Augenblicke des Ausschlüpfens der jungen Biene, bedarf die Schlunddrüse eines gewissen Zeitraumes zu ihrer Entwicklung. Die von uns angestellten Versuche können indessen die Frage noch nicht endgültig entscheiden, in welchem Moment die Schlunddrüse zu funktionieren beginnt. Wir haben die Sekretionstätigkeit der Drüse an verhältnismäßig alten Bienen — von 10 Tagen an — beobachtet und die maximalen Ziffern der Zuckerinversion bei 30tägigen Bienen ermittelt. Hieraus lassen sich noch keine verallgemeinernden Schlüsse ziehen. Unsere Versuche begannen im August und dauerten bis Mitte September, fanden also am Ende der Saison statt. Das Versuchsvolk war schwach und die Zahl der jungen Bienen und der Brut gering (auf einem Rahmen). Als Beleg dazu genügt der Hinweis, daß die Bienen ihre Orientierungsausflüge im Alter von 18 Tagen unternahmen. Alle diese Faktoren könnten zweifellos das Tempo der Entwicklung des Organismus der Biene — und der Sehlunddrüse im besonderen — beeinflussen. Es ist sehr wahrscheinlich, daß eine Untersuchung über die Veränderung des Invertierungsvermögens der Schiunddrüsen von Bienen, die zu verschiedenen Zeiten der Frühlings- und Sommersaison ausgeschlüpft waren, auch eine verschiedene Schnelligkeit der Entwicklung der Drüse ergeben würde.Was die Brustdrüse anbetrifft, so geben unsere Versuche kein genügendes Material zur Beurteilung, welche Rolle sie im Kohlehydratstoffwechsel der Biene spielt. Nur eines steht fest, daß sie an der Invertierung des Zuckers in keinem Falle beteiligt ist.Der Mitteldarm scheint auch mit dem Alter der Biene sein Invertierungsvermögen zu steigern, wenn auch nicht in solchem Umfange als die Schlunddrüse.Die von uns gewonnenen Resultate erlauben, einige Schlüsse über mehrere Fragen der Bienenbiologie zu ziehen, wie z. B. die Frage über den Anteil der Schlunddrüse an der Futtersaftbereitung usw. Doch müssen solche spezielle Fragen an einer anderen Stelle ausführlich besprochen werden.Zum Schlusse sprechen wir Dr. Resnitschenko sowie Dr. Alpatov für die freundliche Hilfe bei der Ausführung der Arbeit unseren Dank aus.