An improved method to isolate lichen algae by gel filtration

Photobiont cells of the lichen Evernia prunastri have completely been separated from their fungal partner by filtration through a bed of Sepharose 2B. Both mannitol and ribitol have been quantified by gas-liquid chromatography in the different steps of the isolation procedure. Absence of mannitol, which is exclusively produced by the mycobiont, has been used as the best probe to monitor isolation.     

Immunoelectrophoretic approach to the metabolic regulation of glutamine synthetase in Rhodopseudomonas capsulata E1F1: role of glutamine

Anti-glutamine synthetase serum was raised in rabbits by injecting purified glutamine synthetase (GS) of the phototrophic bacterium Rhodopseudomonas capsulata E1F1. The antibodies were purified to monospecificity by immunoaffinity chromatography in GS-sepharose gel. These anti-GS antibodies were used to measure the antigen levels in crude extracts from bacteria, grown phototrophically with dinitrogen, nitrate, nitrite, ammonia, glutamate, glutamine or alanine as nitrogen sources. The amount of GS detected by rocket immunoelectrophoresis was proportional to Mn²⁺-dependent transferase activity measured in the crude extracts. Addition of GS inhibitor l-methionine-d,l-sulfoximine (MSX) to the actively growing cells promoted increased antigen levels, that were not found in the presence of glutamine or chloramphenicol. The ammonia-induced decrease in GS relative levels was reverted by MSX. GS levels remained constant when phototrophically growing cells were kept in the dark.Abbreviations GS glutamine synthetase - MOPS 2-(N-morpholine) propane sulfonate - MSX l-methionine-d,l-sulfoximine     

Estimation of the mechanical parameters of the human respiratory system

A numerical algorithm has been developed for the estimation of the mechanical parameters of the human respiratory system. In order to estimate the pulmonary resistance and the dynamic pulmonary elastance, the transpulmonary pressure and the airflow at the mouth or nose are expanded in Chebyshev series. The nonlinear mathematical lung model and a set of measurements for airflow and pressure are then handled by the numerical technique. The lung model includes a component to account for turbulent flow in the larynx and trachea. This contribution presents an alternative method for lung parameter estimation and differs from most existing methods in that it does not need measurements for the tidal volume. It therefore eliminates the use of a body plethysmograph. The method may also find potential application to various other parameter identification problems.     

The dopaminergic innervation of the goldfish pituitary

Summary The dopaminergic innervation of the goldfish pituitary gland was studied by immunocytochemistry at the electron-microscope level using highly specific antibodies against dopamine coupled to bovine serum albumin with glutaraldehyde. A satisfactory preservation of the tissue was achieved after immersion in 5% glutaraldehyde in phosphate buffer containing sodium metabisulfite to prevent oxidation of the endogenous dopamine. The immunocyto-chemical procedure was performed on Vibratome sections using the preembedding method. Immunoreactivity was restricted to part of the neurosecretory type-B fibers (diameter of the secretory vesicles lower than 100 nm) in which it was found to occupy the whole cytoplasm. Labeled fibers were observed within the neurohypophysis in the different parts of the gland and in the adenohypophyseal tissue where immunoreactive profiles were detected in close apposition to the different cell types. These data are in agreement with previous results obtained by means of radioautography and further support a role for dopamine in the neuroendocrine regulation of pituitary functions in teleosts.     

Desangeloylshairidin,X-ray structure determination of a sesquiterpene lactone from Guillonea scabra

The structure and absolute configuration of desangeloylshairidin, a guaianolide isolated from Guillonea scabra, have been established by X-ray diffraction analysis. No conformational change was observed in its seven-membered ring between the crystal and deuterochloroform solution states.     

Calorimetric measurement of free energy utilization by Nitrosomonas and Nitrobacter

Summary Calorimetric estimates of the utilization efficiency of the free-energy derived from substrate oxidation by cell suspensions of two nitrifying bacteria, Nitrosomonas and Nitrobacter, provided two ranges of values: 11 to 27% and 15 to 51%, respectively. About 15 to 30% of the utilized free-energy is used for driving endergonic reactions other than CO₂ fixation, probably the synthesis of polyphosphates.The molar heat of substrate oxidation does not seem to be influenced by the age of cells harvested during growth or by the length of the incubation period during which cells have been kept in a buffer suspension in a starved condition. The loss of respiratory activity measured either by oxygen uptake or heat evolution in the presence of the specific substrate, nitrite or ammonium, decreases according to kinetics which are influenced by the aerobiosis of the suspension. The viability of the starved cells decreases in a way which is similar to that of the respiratory activity. It seemed impossible to obtain cells which had lost their viability but kept the ability to oxidize their substrate.Two inhibitors of the respiratory chain, quinacrine and cyanide, are without effect on the molar heat of substrate oxidation and consequently on the free-energy utilization efficiency. 2.4 dinitrophenol did decrease the rate of heat evolution during substrate oxidation at concentrations at which the rate of oxygen uptake was not depressed, with the consequences that free-energy efficiency was apparently increased.     

L'excursion en Provence (Sud-Est de la France) de la Societe Internationale de Phytosociologie

Conclusions La Provence apparaît, en définitive, comme un territoire des plus variés aux divers points de vue climatique, édaphique, biotique.C'est une région d'autant plus favorable aux études phytosociologiques que la variété des milieux s'y double d'une brutalité souvent étonnante des contrastes, conduisant à la juxtaposition parfois extrêmement précise d'associations végétales très différentes, dont les limites peuvent alors être cartographiquement figurées avec une très suffisante exactitude.Cette vigueur des contrastes est surtout marquée en Provence calcaire où les divers termes des séries évolutives dérivant du Quercetum ilicis climacique ou y conduisant, se juxtaposent souvent sans transition de quelque importance.Les contrastes sont cependant moins accusés en Provence cristalline et dans la Basse-Vallée du Rhône.En Crau comme en Camargue, les associations végétales se juxtaposent souvent en ceintures parallèles ou concentriques — elles s'individualisent alors facilement —, souvent en mosaïques irrégulières, au grè des variations locales des deux facteurs déterminants: l'eau en Crau, le sel en Camargue.En Provence cristalline, les Maures étant surtout faites de roches se délitant facilement (grès, micaschistes et phyllades) sous un climat chaud et humide, le sol se reconstitue facilement après sa destruction partielle par l'érosion; les contrastes sont moins tranchés; on observe des transitions nombreuses entre les divers termes des séries évolutives, dont les limites ne peuvent être fixées qu'approximativement. Nous y avons notamment abordé le problème de la forêt de chêne-liège dont la signification est loin d'être connue, non seulement en Provence mais sur toute l'étendue de son aire, en Afrique du Nord, en Espagne, au Portugal, en France méridionale, en Italie et dans les Iles de la Méditerranée occidentale.En provence cristalline, il est certain que l'homme en a considérablement assuré l'extension, directement pour l'utilisation du liège, indirectement par la coupe et le feu, et, en bien des points de la Méditerranée occidentale, le chêne-liège se présente comme une essence cultivée.Par ailleurs rien ne permet de mettre en doute son indigénat. Mais, dans les conditions actuelles du milieu en Provence cristalline:le chêne vert est possible partout;dans les conditions naturelles de la concurrence vitale le chêne vert l'emporte sur le chêne-liège.Si, donc, la forêt originelle a comporté le chêne-liège, ce ne peut être qu'à l'état d'essence secondaire et seulement en quelques points où une faible couverture forestière lui permettait de cohabiter avec le chêne vert.Les documents cartographiques au I/20 000^e tendent à montrer, d'aileurs que, dans les conditions naturelles de reconstitution de la forêt ou dans le peuplement naturel des sols neufs (Isthme de Giens), c'est la chênaie de chêne vert, non la chênaie de chêne-liège qui s'installe.Les grandes unités phytosociologiques de la Provence paraissent être maintenant convenablement définies floristiquement; il convient donc d'en poursuivre l'étude écologique, dont certains aspects seulement les plus apparents—mais peut-être pas toujours les plus actifs-, ont été abordés.Notons enfin la démonstration faite, au cours de l'excursion, de la parfaite application possible des méthodes phytosociologiques à l'étude des groupements marins et la nécessité, particulièrement apparente en milieu marin, de considérer tout autant les animaux que les végétaux. L'avenir est bien aux biocénoses dont les grands traits ont été mis en évidence, quant à la côte de la presqu'île de Giens, par Roger Molinier.Reçu par la rédaction le 20. XII. 1958.avec la collaboration de ROGER MOLINIER pour la partie marine et G. TALLON pour la Camargue.     

Implication of Pectic Components in Cell Surface Interactions between Tomato Root Cells and Fusarium oxysporum f. sp. radicis-lycopersici: A Cytochemical Study by Means of a Lectin with Polygalacturonic Acid-Binding Specificity

Aplysia gonad lectin, a polygalacturonic acid-binding lectin isolated from the sea mollusc Aplysia depilans, was complexed to colloidal gold and used for localizing polygalacturonic-acid-containing molecules in tomato root tissues infected with Fusarium oxysporum f. sp. radicis-lycopersici (FORL). Colonization of host tissues by FORL was associated with striking wall modifications including disruption and even loss of middle lamellae. According to the labeling pattern observed in host wall areas adjacent to fungal penetration channels, it is likely that FORL pectolytic enzymes act through localized wall degradation. The release of polygalacturonic acid-rich wall fragments and the accumulation of polygalacturonic acid-containing molecules in some altered phloem cells were frequently observed and considered to be specific host reactions to fungal attack. The heavy deposition of such molecules at strategic sites such as wall oppositions and intercellular spaces provides support to their implication in the plant defense system. The possible interrelation between polygalacturonic acid-containing molecules and other polymers such as lignin and phenolic compounds remains to be investigated further. The role of these molecules in host-pathogen interactions is discussed in relation to plant defense.     

The tryptophan residues of aspartate transcarbamylase: site-directed mutagenesis and time-resolved fluorescence spectroscopy.

Aspartate transcarbamylase (EC 2.1.3.2) contains two tryptophan residues in position 209 and 284 of the catalytic chains (c) and no such chromophore in the regulatory chains (r). Thus, as a dodecamer [(c3)2(r2)3] the native enzyme molecule contains 12 tryptophan residues. The present study of the regulatory conformational changes in this enzyme is based on the fluorescence properties of these intrinsic probes. Site-directed mutagenesis was used in order to differentiate the respective contributions of the two tryptophans to the fluorescence properties of the enzyme and to identify the mobility of their environment in the course of the different regulatory processes. Each of these tryptophan residues gives two independent fluorescence decays, suggesting that the catalytic subunit exists in two slightly different conformational states. The binding of the substrate analog N-phosphonacetyl-L-aspartate promotes the same fluorescence signal whether or not the catalytic subunits are associated with the regulatory subunits, suggesting that the substrate-induced conformational change of the catalytic subunit is the essential trigger for the quaternary structure transition involved in cooperativity. The binding of the substrate analog affects mostly the environment of tryptophan 284, while the binding of the activator ATP affects mostly the environment of tryptophan 209, confirming that this activator acts through a mechanism different from that involved in homotropic cooperativity.