Data files of figures 1, 2 and 3

We investigated optimal conditions for characterization of bioactivity of lytic compound(s) excreted by Alexandrium tamarense based on a cell-bioassay system. Allelochemical response of the cryptophyte Rhodomonas salina indicated the presence oflytic compound(s) in a reliable and reproducible way and allows for quantification of this lytic effect. The parameters tested were the incubation time of putatively lytic extracts or fractions with the target organism R. salina, different techniques for cell harvest from A. tamarense cultures and the optimal harvest time. A three hour incubation time was found to be optimal to yield a rapid response while accurately estimating effective concentration (ECso) values. Harvest of A. tamarense cultures by filtration resulted in loss of lytic activity in most cases and centrifugation was most efficient in terms of recovery of lytic activity. Maximum yield of extracellular lytic activity of A. tamarense cultures was achieved in the stationary phase. Such optimized bioassay guided fractionation techniques are a valuable asset in the isolation and eventual stmctural elucidation of the unknown lytic substances.

Files of figures 2, 3 and table

Certain allelochemicals of the marine dinoflagellate Alexandrium tamarense cause lysis of a broad spectrum of target protist cells but the lytic mechanism is poorly defined. We first hypothesized that membrane sterols serve as molecular targets of these lytic compounds, and that differences in sterol composition among donor and target cells may cause insensitivity of Alexandrium and sensitivity of targets to lytic compounds. We investigated Ca2+ influx after application of lytic fractions to a model cell line PC12 derived from a pheochromocytoma of the rat adrenal medulla to establish how the lytic compounds affect ion flux associated with lysis of target membranes. The lytic compounds increased permeability of the cell membrane for Ca2+ ions even during blockade of Ca2+ channels with cadmium. Results of a liposome assay suggested that the lytic compounds did not lyse such target membranes non-specifically by means of detergent-like activity. Analysis of sterol composition of isolates of A. tamarense and of five target protistan species showed that both lytic and non-lytic A. tamarense strains contain cholesterol and dinosterol as major sterols, whereas none of the other tested species contain dinosterol. Adding sterols and phosphatidylcholine to a lysis bioassay with the cryptophyte Rhodomonas salina for evaluation of competitive binding indicated that the lytic compounds possessed apparent high affinity for free sterols and phosphatidylcholine. Lysis of protistan target cells was dose-dependently reduced by adding various sterols or phosphatidylcholine. For three tested sterols, the lytic compounds showed highest affinity towards cholesterol followed by ergosterol and brassicasterol. Cholesterol comprised a higher percentage of total sterols in plasma membrane fractions of A. tamarense than in corresponding whole cell fractions. We conclude therefore that although the molecular targets of the lytic compounds are likely to involve sterol components of membranes, A. tamarense must have a complex self-protective mechanism that still needs to be addressed.

Files related to figures 1 to 13

Members of the marine dinoflagellate genus Alexandrium are known to exude allelochemicals, unrelated to well-known neurotoxins (PSP-toxins, spirolides), with negative effects on other phytoplankton and marine grazers. Physico/chemical characterization of extracellular lytic compounds of A. tamarense, quantified by Rhodomonas salina bioassay, showed that the lytic activity, and hence presumably the compounds were stable over wide ranges of temperatures and pH and were refractory to bacterial degradation. Two distinct lytic fractions were collected by reversed-phase solid-phase extraction. The more hydrophilic fraction accounted for about 2% of the whole lytic activity of the A. tamarense culture supernatant, while the less hydrophilic one accounted for about 98% of activity. Although temporal stability of the compounds is high, substantial losses were evident during purification. Lytic activity was best removed from aqueous phase with chloroform-methanol (3:1). A "pseudo-loss" of lytic activity in undisturbed and low-concentrated samples and high activity of an emulsion between aqueous and n-hexane phase after liquid-liquid partition are strong evidence for the presence of amphipathic compounds. Lytic activity in the early fraction of gel permeation chromatography and lack of activity after 5 kD ultrafiltration indicate that the lytic agents form large aggregates or macromolecular complexes.

(Figures S2a-c) Phase diagrams of the upper crust, the lower crust and the upper mantle based on averaged compositions of the oceanic crust and mantle

The age of the subducting Nazca Plate off Chile increases northwards from 0 Ma at the Chile Triple Junction (46°S) to 37 Ma at the latitude of Valparaíso (32°S). Age-related variations in the thermal state of the subducting plate impact on (a) the water influx to the subduction zone, as well as on (b) the volumes of water that are released under the continental forearc or, alternatively, carried beyond the arc. Southern Central Chile is an ideal setting to study this effect, because other factors for the subduction zone water budget appear constant. We determine the water influx by calculating the crustal water uptake and by modeling the upper mantle serpentinization at the outer rise of the Chile Trench. The water release under forearc and arc is determined by coupling FEM thermal models of the subducting plate with stability fields of water-releasing mineral reactions for upper and lower crust and hydrated mantle. Results show that both the influx of water stored in, and the outflux of water released from upper crust, lower crust and mantle vary drastically over segment boundaries. In particular, the oldest and coldest segments carry roughly twice as much water into the subduction zone as the youngest and hottest segments, but their release flux to the forearc is only about one fourth of the latter. This high variability over a subduction zone of < 1500 km length shows that it is insufficient to consider subduction zones as uniform entities in global estimates of subduction zone fluxes. This article is protected by copyright. All rights reserved.

Recueil d'ouvrages curieux de mathematique et de mecanique, ou description du cabinet de monsieur Grollier de Serviere [Texto impreso] : avec des figures en taille douce

Sign.: a4, e4, i6, A-O4

Histoire naturelle des reptiles : avec figures dessinées d'apres nature / par C. S. Sonnini et P. A. Latreille.

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Histoire naturelle des reptiles : avec figures dessinées d'apres nature / par C. S. Sonnini et P. A. Latreille.

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Histoire naturelle des reptiles : avec figures dessinées d'apres nature / par C. S. Sonnini et P. A. Latreille.

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The nature-printed British sea-weeds : a history, accompanied by figures and dissections of the Algae of the British Isles / By William Grosart Johnstone and Alexander Croall. Nature printed by Henry Bradbury.

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