Etude de quelques propriétés de la phosphofructokinase érythrocytaire de l'homme normal

Human erythrocyte phosphofructokinase was purified 150 fold by DEAE cellulose adsorption and ammonium sulfate precipitation.At pH 7,5 the enzyme exhibits allosteric kinetics with respect to ATP, fructose 6 phosphate, and Mg²⁺.ATP at high concentration acted as an inhibitor and ADP, 5′AMP, 3′,5′, AMP, acted as activators. Both effectors seemed to decrease the homotropic interactions beetween the fructose 6 phosphate molecules.The activators increased the affinity of phosphofructokinase for the substrate (F6P), the inhibitor decreased it.These ligands had no effect on the maximum velocity of the reaction except in the case of ADP.Interactions between the substrates and the effector ligands on the enzyme were considered in terms of the Monod - Changeux - Wyman model for allosteric proteins.With GTP and ITP, no inhibition was observed. At saturing concentration of GTP, ATP still inhibited phosphofructokinase.Both 3′5′ AMP and fructose 6 phosphate increased the concentration of ATP required to produce an inhibition of 50 %.Citrate, like ATP, inhibited phosphofructokinase by binding most likely at the same allosteric site. Erythrocyte phosphofructokinase is inhibited by 2–3 DPG.The study of the relation log V max = f (pH) suggested, that the active center contains at least one imidazole and one sulfhydryl group.     

Préparation et forme sérique de la protéine réactive C de lapin

The preparation of rabbit C-reactive protein (CRP) involves a single step affinity chromatography. This preparation takes advantage of the calcium-dependent affinity of CRP for an agarose gel bearing 2-aminoethanol dihydrogen-phosphate as a ligand. A prior chromatography on agarose gel without the ligand allows the uptake of the serum amyloid P-component (SAP).The CRP prepared according to this method is able to form precipitating complexes in agarose with rabbit lipoproteins. The specificity of these interactions is studied. It is demonstrated that CRP-High Density Lipoproteins (HDL) association produces a second precipitation arc when the pure CRP is revealed by a specific antiserum in agarose. Moreover, CRP in the serum is shown to be in the bound form only, and the binding involves Low Density Lipoproteins (LDL) exclusively.     

Aspartate transcarbamylase de la coquille Saint-Jacques Pecten maximus 1. (Mollusque lamellibranche): Méthode de dosage et variations de l'activité dans le manteau et la gonade

The search of an index for the instantaneous estimate of the in situ growth rate of marine animals led us to attempt to measure the specific activity of aspartate transcarbamylase (ATC). The experiments to test the value of the index were carried out on the scallop Pecten maximus L. p]The first step was to find the optimum conditions for enzyme activity measurement. At 35 °C. the scallop ATC shows an optimum pH of 9 and a K_m of 4.6 × 10^?3 M for aspartate and of 8.0 × 10^?4 M for carbamyl phosphate. The different types of inhibition by the substrates high concentrations and the products suggest an ordered sequential mechanism for the reaction. The decrease in enzyme activity due to metallic ions (Cu²⁺ and Zn²⁺) and to parahydroxymercuribenzoate is compatible with the presence of a sulphydryl group in the active site. p]The variations in ATC levels within the gonad and within the mantle of the scallop were measured and compared with the processes of the sexual maturation and somatic growth in a natural population. For the two tissues, a correlation between the ATC specific activity and the relative growth rate is demonstrated.     

Régulation de la synthèse du DNA dans la glande séricigène de Bombyx mori: II. Etude de l''activité DNA-polymérasique présente dans la fraction soluble brute

In the silkglands of Bombyx mori larvae, DNA synthesis suddenly stops in the middle of the fifth larval instar, when cellular growth is over and cell activity is turned on to massive silk synthesis. Of much interest is to determine what factors are implied in the quantitative and temporal control of DNA synthesis in this tissue.Previous work on thymidine metabolism in the intact organ lead us to look for several enzymatic activities in the soluble fraction of the cells during this period.The results presented here concern the kinases of the thymidylic pathway. As measured in vitro from crude cellular extracts, thymidine-kinase is present at a lower level than thymidylate-kinase but both activities vary in connection with the pattern of in situ DNA synthesis, reaching their highest levels on the fourth day of the instar and decreasing as DNA synthesis definitively stops in the glands.     

Influence de la température sur les acides aminés libres de Palaemon serratus et P. squilla

Free amino acids concentrations were determined in the abdominal muscle of Palaemon serratus and P. squilla submitted to several temperatures. Only serine, alanine and proline show significant differences between the two species. Temperature variations mainly affect glycine, but do not act in a similar manner on all other amino acids. Differences in the influence of temperature upon the total free amino acids of P. serratus and P. squilla can be related with their migratory behaviour.     

Applications biologiques de la spectroscopie Raman de résonance et de la spectroscopie micro-Raman de résonance

Advantages are given to illustrate the possibilities of resonance Raman spectroscopy in the studies of biological systems.A relation giving the size of porphinato-core of metalloporphyrins from resonance Raman frequency is proposed. It can be applied to heme, even in lived state.By illuminating the sample through a microscope, resonance Raman effect can be collected from 1 (μm)² of sample. Spectra of a vegetal-cell wall (pimento) and of a red corpuscle (hemoglobin) are shown as examples.     

Groupement et ordination appliqués à l''étude de la répartition de l''épifaune benthique de l''estuaire maritime et du Golfe du Saint-Laurent

Forty-seven navigation buoys were used to collect quantitative samples of epibenthic fauna in the Lower Estuary and Gulf of St. Lawrence. Stations, as described by their sessile species and biomass composition, were grouped using multivariate analysis. A Gower similarity index, modified to take into account the importance of each species in the ecosystem, was used to compare stations. Results obtained by cluster and principal coordinate analyses showed comparable relations between stations. North Shore and Gaspé Coast stations were separated from those of the Lower North Shore, the Mingan Island sector and the Lower Estuary. Stations from the Lower North Shore were also separated from those of the Mingan Island region and of the Lower Estuary, but at a higher level of similarity. All animal assemblages consisted of the following sessile species: Obelia longissima (Pallas), Hiatella arctica (L.), Mytilus edulis L., Balanus balanoides (L.) and B. crenatus (Bruguière). These species formed over 95% of biomass of all sessile species. Differences between the various station assemblages were mainly due to changes in the relative abundance of these species. The geographical gradient of summer surface water temperature best explains the observed clusters. In the Lower Estuary, the spatio-temporal variations of physico-chemical factors and the surface circulation may influence the number of species and their abundance. The circulation systems of the Lower Estuary and near the Mingan Islands might account for the similarity between their assemblages. The same dominant species, Obelia longissima, characterizes the fauna of these two regions.     

Etude de la fertilité des eaux marines au moyen de tests biologiques effectués avec des cultures d'algues. II. Limitation nutritionnelle et viabilité de l'inoculum

Ten cultures of phytoplankters, including four strains of Skeletonema costatum from different origins, were used to improve some aspects of the bioassay technology. Special attention was paid to the preliminary nutrient limitation of the inocula. When the cells are maintained in nutrient starvation, their carbon, nitrogen, carbohydrates, proteins and above all chlorophyll a contents decrease. The minimum appears after a range of 2–7 days, according to species. The survival of these limited cells and their capacity to give rise to active growing cultures when sub-cultured are different with species, but efficiency in the inocula they provide usually can occur only until the minimum content in cellular components appears. Starved cells and enriched cultures of Chaetoceros lauderi and Skeletonema costatum were used to inoculate several samples of sea water, in order to test the effect of the starvation on the experimental results. It appears that the starved cells increase the sensitivity of the method, but they are more susceptible to substances limiting their growth. In the opinion of the authors the best way would be to use both starved and enriched cells as inocula, but, when this is impossible, cautiously starved cells should be used with unpolluted sea waters.     

Distribution subcellulaire des protéine-kinases stimulables par l'AMP cyclique et des protéines réceptrices de l'AMP cyclique dans la thyroïde de porc

The distribution of cyclic AMP-dependent protein kinase activity in porcine thyroid glands has been studied. Enzyme activity catalyzing phosphorylation of exogenous substrate (protamine) from ATP, and cyclic AMP binding were determined in parallel in subcellular fractions purified by differential centrifugation and flotation on sucrose density layers. Both activities were found in all the studied fractions; they were quantitatively the highest in the cytosol but particles showed the highest specific activities.Latent protein-kinase activity was unmasked by action of detergents on microsomes (× 5–10 fold) and solubilized (85 to 99 p. cent of the initial total activity). Cyclic AMP binding capacity was also recovered in detergent-treated microsomal extracts in spite of reduced cyclic AMP binding in the presence of detergent.Protein kinase activity and cyclic AMP-binding proteins were less represented in purified nuclei than in microsomes. Again both activities were unmasked by detergent.Preparations highly enriched in Golgi membranes were compared to rough microsomal preparations. Higher protein kinase activity was detected in rough microsomes as compared to Golgi membranes, whereas the reverse was true for cyclic AMP binding. Both activities were equalized after detergent treatment. Since unmasking of protein kinase activity was the highest in Golgi membranes, this fraction contains more enzyme activity and cyclic AMP binding capacity than rough microsomes.The localization of endogeneous protein substrates of cyclic AMP-dependent protein kinases was investigated using purified soluble protein kinase subcellular fractions. The better endogeneous substrates seemed to be localized in the rough microsomal and in the nuclear fractions.     

Etude préliminaire des effets de la mise en eau du Réservoir de LG-2 (Territoire de la Baie James,Québec) sur le seston grossier et le zooplancton des rivières et des lacs inondés

Preliminary Study of the Effects of Impoundment of LG-2 Reservoir (James Bay Territory, Quebec) on the Net Seston and the Zooplankton of Impounded Rivers and Lakes The effects of impoundment on the biomass of net seston and zooplankton in lakes and rivers of Northern Quebec were investigated from 1978 to 1980, before, during and after the completion of LG2 Reservoir on the River La Grande (53° 54′ N, 76° 78′ W). In lotic stations, a 41–77% decrease in net seston was observed due to sedimentation of mineral particles (36–80 mg/m³ in 1978 to 10–21 mg/m³ in 1979). The ratio of organic to total seston increased from an average of 0.18 before the impoundment to 0.65 after. The flooding phase brought about a trophic upsurge; the biomass of the zooplankton increased by one to two orders of magnitude at the various sites; the greatest increases occurred in the central stations near the dam, the smallest in the upper stations near the inflow. In lake stations, no significant difference between years could be detected; the mean zooplankton biomass for all three years ranged from 6.59 mg/m³ to 34.34 mg/m³. Spatial variations between lakes were however significant. Results are compared with those in other natural lakes in Canada and reservoirs in the United States. Comparative examination of phytoplankton biomass before and after impoundment of LG2 Reservoir suggests that bacterioplankton and allochthonous organic material are key elements in the pelagic food chain after impoundment.     

Interets du dosage de l'ensemble des nucleotides adenyliques pour l'estiimation de la biomasse des populations phytoplanctoniques lacustres (Lac Pavin, France)

Three methods for phytoplanktonic standing crop estimationshave been used during ecological successions in a meso-oligotrophiclake (Le Pavin, France). In this study, it appears that cellcounting and pigment concentrations overestimate the biomassof pioneer population, by taking into account a high numberof dead cells. It seems preferable to estimate biomass fromATP concentrations, to measure living phytoplanktonic standingcrop and to calculate activity coefficients. However, continuousrearrangements exist between ATP, ADP and AMP in the pool ofadenine nucleotides. These rearrangements, which are revealedby energy charge variations, indicate changes in intracellularATP concentrations. Consequently it is desirable to integrateenergy charge variations in standing crop estimates from ATPconcentrations.     

Application paléoécologique de la méthode d''analyse factorielle en composantes principales: Interprétation des microfaunes de foraminifères planctoniques quaternaires en Méditerranée. III. Les séquences paléoclimatiques. Conclusions générales

The statistical analysis (Q mode) of the microfauna of 135 bottom and core samples from the Mediterranean provides evidence of the predominating local conditions in each of the two basins.

Temperature, which plays a primary role in the distribution of foraminiferal species, only takes second place when one considers the regrouping of the layers.

Once the temperature factor is identified, its variations provide a precise climatic curve with the advantage of not having to make any subjective interpretation of the assemblages.

Résumé

L'analyse en mode Q des microfaunes de 135 échantillons de Méditerrannée (niveaux de surface et carottages) met en évidence la prédominance des conditions locales propres à chacun des deux bassins.

La température, qui joue un rôle primordial dans la répartition des espèces de Foraminifères, n'intervient qu'en second lieu lorsqu'on considère les regroupements des niveaux.

Une fois le facteur thermique identifié, ses variations fournissent une courbe climatique précise qui présente l'avantage de ne nécessiter aucune interprétation subjective des assemblages.     

Role des corpora allata et cardiaca sur l'action cardioaccélératrice de la lumière et de la température chez Locusta migratoria

A rise in temperature as well as the passage from dark to light increases the cardiac beat of adult Locusta migratoria. Continuous exposure to light considerably decreases the sensibility to temperature variations. Neither the corpora allata nor the corpora cardiaca have any influence in this case.     

Controle de la chaine de biosynthese de la threonine chezE. coli

Resume Le travail décrit dans cet article s'appuie sur la théorie du contrôle du métabolisme et a pour but la mesure des coefficients de contrôle des différentes étapes sur le flux de production de thréonine. Le coefficient de contrôle d'une étape sur un flux mesure quantitativement la réponse du flux aux variations de l'étape. Cette notion est donc particulièrement importante aussi bien dans les situations pathologiques (diminution de l'activité d'une étape) qu'en biotechnologies où au contraire les étapes sont amplifiées.La mesure des coefficients de contrôle des étapes d'une chaîne métabolique permet donc de connaître celle(s) dont l'amplification doit entraîner une augmentation concomitante du flux.Nous avons appliqué ces concepts à l'étude de la voie de biosynthèse de la thréonine à partir de l'aspartate.La voie de la biosynthèse de la thréonine à partir de l'aspartate est constituée de cinq étapes catalysées par cinq activités enzymatiques: l'aspartokinase (AK), l'aspartate semi-aldéhyde déshydrogénase (ASA-DH), l'homosérine déshydrogénase (HDH), l'homosérine kinase (HK) et la thréonine synthase (TS).La mesure du coefficient de contrôle de la première étape (AK, insensible à la rétro-inhibition par la thréonine dans la souche étudiée) a montré qu'elle était faiblement contrôlante. L'étude a révélé la présence d'une inhibition jusqu'alors inconnue de l'homosérine kinase par la lysine.Un début de modélisation de cette chaîne de biosynthèse permet d'expliquer les résultats expérimentaux.

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This paper deals with the application of the metabolic control theory, especially the measurement of control coefficients, to the threonine pathway inE. coli. The control coefficient of a step on a metabolic flux quantitatively assesses the flux response to the step variations. This concept is particularly relevant both in pathological situations (decrease in the activity of an enzymatic step in the metabolism) and in biotechnologies, where, on the contrary steps are amplified.Measurement of the control coefficients of the steps of a metabolic network makes it possible to know those whose amplification should lead to a simultaneous increase in the fluxes.We have applied these concepts to threonine biosynthesis from aspartate inE. coli. The threonine pathway starting from aspartate involves five steps catalyzed by five enzyme activities: aspartokinase (AK), aspartate-semialdehyde-dehydrogenase (ASA-DH), homoserine dehydrogenase (HDH), homoserine kinase (HK) and hreonine synthetase activity (TS).Measurement of the control coefficient of the first step (AK, insensitive to threonine inhibition in the studied strain) has shown that it controls threonine production weakly. Our study has revealed a hitherto unknown inhibition of homoserine kinase activity by lysine.Mathematical modeling of this metabolic pathway has been undertaken to better understand our experimental results.

     

Évolution temporelle de la diversité génétique de Chaerophyllum bulbosum : conséquences sur la gestion des ressources génétiques

To increase the germplasm necessary for varietal improvement of tuberous-rooted chervil, a food apiaceae of increasing importance, two successive surveys of wild populations were carried out in Germany, in the Rhine and the Weser River basins. These mainly riparian populations are likely to be shaped by changes in hydrographic networks that characterize their habitat. Molecular studies have shown a strong structuration between wild populations (G_ST ∼ 32 %), but did not reveal any structuring effect of the hydrographic network on diversity or any global phenomenon of genetic erosion. A discussion about the strategy for maintaining the diversity of this species on a long-term period is proposed.     

Évolution horaire au cours d'une journée normale de la photosynthèse, de la transpiration, de la respiration foliaire et racinaire et de la nutrition N.P.K. chez Zea mays

Hourly development during a normal day of photosynthesis, transpiration, leaf and root respiration, and of N.P.K nutrition in Zea mays. Metabolism of Zea mays L. cv. INRA F₇×F₂ can be measured hourly with the “C₂3A system”, under favourable and constant growth conditions. The photosynthesis is especially stable and is submitted only to a development linked with the leaf surface growth. During the vegetative stage the leaf surface increases regularly both in the day and in the night. The water loss does not change during the diurnal period and remains important during the night if humidity is less than 100%. The leaf respiration is nearly stable. The root respiration, measured with O₂ and CO₂, fluctuates according to a typical rhythm with two maxima. Day and night mean rates were about the same. The respiratory quotient is about one during the vegetative stage. There is no decrease in the rate of phosphate absorption during the night, and a very small decrease in the rate of nitrate absorption. Ammonium is totally consumed in the first hours after renewal of the nutrient solution. At the same time the potassium consumption is decreased, and then presents a maximum followed by a night reduction. Under our conditions, all of the observations allowed us to assume the presence of an adequate reserve of assimilates and suggest the existence of a precise regulation process, which can ensure an even day night functioning of the plant metabolism, but which does not preclude the presence of internal rhythms, as indicated by the oscillation of the root respiration.     

Activité oxydasique de l''oxydoréductase de la xanthine, spécifique de la classe des mammifères

In every species other than mammals xanthine oxidoreductase behaves as a dehydrogenase, never as an oxidase. In three mammalian species, the enzyme acts intracellularly as a dehydrogenase, but its class-specific ambivalence allows its extracellular conversion into an oxidase.

Résumé

En dehors de la classe des Mammifères, l'oxydoréductase de la xanthine ne se comporte jamais comme une oxydase. Chez les Mammifères, une ambivalence de la molécule permet l'expression d'une activité déshydrogénasique (NAD) dans la cellule, et d'une activité oxydasique en dehors de la cellule.