Chemical and conformational study of the interactions involved in mycotoxin complexation with beta-D-glucans.

In a previous paper we reported that beta-D-glucans isolated from Saccharomyces cerevisiae could adsorb zearalenone, reduce its bioavailability in the digestive tract, and protect animals against its adverse effects. We have now investigated, in vitro, the kinetics of the interaction between other mycotoxins and beta-D-glucans from several sources at three pH values found along the digestive tract (3.0, 6.

Influence of pH on complexing of model beta-d-glucans with zearalenone.

Previous studies have shown that isolated beta-(1,3 and 1,6)-D-glucans and related alkali-extracted fractions from the cell wall of Saccharomyces cerevisiae are able to complex with zearalenone in vitro (affinity up to 50%) and thus may reduce the bioavailability of toxins in the digestive tract. The complexation mechanisms involve cooperative interaction between the two chemical entities that can be computed by Hill's model. Various linear or branched soluble or insoluble beta-D-glucans were evaluated to elucidate their roles in the adsorption mechanisms under three pH conditions (3.

Comprehensive conformational study of key interactions involved in zearalenone complexation with beta-D-glucans.

The beta-D-glucans from the cell wall of Saccharomyces cerevisiae have shown in vitro affinity for zearalenone. For this reason, their utilization as dietary adsorbent, to reduce the bioavailability of zearalenone, is of practical interest. Our study used powerful devices to elucidate the spatial conformation and molecular sites of interaction between ZEN and beta-D-glucans.

Adsorption of Zearalenone by beta-D-glucans in the Saccharomyces cerevisiae cell wall.

Cell walls of yeasts and bacteria are able to complex with mycotoxins and limit their bioavailability in the digestive tract when these yeasts and bacteria are given as feed additives to animals. To identify the component(s) of the yeast cell wall and the chemical interaction(s) involved in complex formation with zearalenone, four strains of Saccharomyces cerevisiae differing in their cell wall glucan and mannan content were tested. Laboratory strains wt292, fks1, and mnn9 were compared with industrial S.

Alkali extraction of beta-d-glucans from Saccharomyces cerevisiae cell wall and study of their adsorptive properties toward zearalenone.

The isolated cell wall of Saccharomyces cerevisiae has some capacity to adsorb zearalenone (affinity near 30%) and reduce the bioavailability of toxins in the digestive tract. The adsorption process was quantified in vitro, and the data obtained when plotted with Hill's equation indicated a cooperative process. The model showed that the adsorption capacity was related to the yeast cell wall composition.

Unusual structure of the dimeric 4-bromocalcimycin-Zn2+ complex.

The X-ray structure of [Zn(4-bromocalcimycin)2 x H2O] complex shows two highly different conformations of the ligand in the dimeric association, unusual in this ionophore family.

In vitro activity of the polyether ionophorous antibiotic monensin against the cyst form of Toxoplasma gondii.

Toxoplasma gondii. The experiments were conducted in vitro using 2 methods; cysts produced either in mice or in cell culture were exposed to monensin in vitro, and the infectivity of the parasites was then assessed in vivo or in vitro. The data obtained from these 2 systems of evaluation showed that monensin inhibits the infectivity and the viability of the bradyzoites.

Determination of absolute configurations of amines and amino acids using nonchiral derivatizing agents (NCDA) and deuterium NMR.

Enantiomeric analysis and empirical determination of the absolute configuration of amines and amino acids can be easily performed using acetyl-d(3) chloride as a nonchiral derivatizing agent (deuterium probe) and deuterium NMR in a chiral solvent (Courtieu's method). In the case of amino acids, derivatization to amido esters, performed with methanol-d(4) and acetyl-d(3) chloride, gives a double opportunity for enantiomeric analysis.

Ionophore-Phospholipid Interactions in Langmuir Films in Relation to Ionophore Selectivity toward Plasmodium-Infected Erythrocytes.

Carboxylic true ionophores were previously demonstrated to have efficient antimalarial activity against the human parasite Plasmodium falciparum, with a 50% inhibitory concentration around nM and generally high selectivity as compared to their toxic effects against mammalian cell lines. The decreased molecular packing of the erythrocyte membrane outer leaflet after malarial infection could explain the preferential ionophore interaction with infected erythrocytes. Monolayer penetration experiments using different phospholipid films showed strong incorporation of true carboxylic ionophores, from classes 1 (nigericin) and 2 (lasalocid), up to a surface pressure close to film collapse.

Cationomycin and monensin partition between serum proteins and erythrocyte membrane: consequences for Na+ and K+ transport and antimalarial activities.

The ionophore properties of cationomycin and monensin were studied on human erythrocytes by measuring Na+ influx by 23Na NMR and concomitant K+ efflux by potentiometry in the presence of increasing amounts of serum. Both ion currents (Na+ or K+) decreased linearly with the reciprocal of serum amount. The serum effects on ion currents were stronger with cationomycin than with monensin.

Ionophore properties of cationomycin in large unilamellar vesicles studied by 23Na- and 39K-NMR.

Cationomycin, isolated from Actinomadura azurea belongs to a large family of carboxylic polyether antibiotics, transporting monovalent cations through membranes by a mobile carrier mechanism, leading globally to an H+, M+ exchange. In this report the cation transporting properties of cationomycin were characterized in large unilamellar vesicles (LUVs) by 23Na- and 39K-NMR. Kinetic studies showed that cationomycin transported potassium more rapidly than sodium, and the more stable complex was formed with potassium at the water/membrane interface.

Characterization of the potent in vitro and in vivo antimalarial activities of ionophore compounds.

Large-scale in vitro screening of different types of ionophores previously pinpointed nine compounds that were very active and selective in vitro against Plasmodium falciparum; their in vitro and in vivo antimalarial effects were further studied. Addition of the ionophores to synchronized P. falciparum suspensions revealed that all P.

Differential in vitro activities of ionophore compounds against Plasmodium falciparum and mammalian cells.

Twenty-two ionophore compounds were screened for their antimalarial activities. They consisted of true ionophores (mobile carriers) and channel-forming quasi-ionophores with different ionic specificities. Eleven of the compounds were found to be extremely efficient inhibitors of Plasmodium falciparum growth in vitro, with 50% inhibitory concentrations of less than 10 ng/ml.

Ionophore properties of monensin derivatives studied on human erythrocytes by 23Na NMR and K+ and H+ potentiometry: relationship with antimicrobial and antimalarial activities.

Eight derivatives of monensin with a modified C25-C26 moiety were synthesized. Their ionophore properties were studied on human erythrocytes by measuring Na+ influx with 23Na NMR and concomitant K+ and H+ efflux by potentiometry. Modification of OH-26 led to inversion of selectivity of transport in favor of K+/Na+ in comparison with monensin.

Na+ and K+ transport by 4-chlorophenylurethane-monensin in Enterococcus hirae de-energized and energized cells studied by 23Na-NMR and K+ atomic absorption.

Na+ and K+ movements induced by 4-chlorophenylurethane-monensin, which presents an inverted ion selectivity (K+ > Na+) in model systems compared with monensin, were followed on Enterococcus hirae cells by 23Na-NMR and K+ atomic absorption. For de-energized cells, the urethane derivative is much more selective for K+ than monensin, but only at low concentrations (10(-3)-10(-4) mM). For higher concentrations, as previously shown for monensin, the sodium and potassium movements are driven by the ion gradients present.

Conditions modulating the ionic selectivity of transport by monensin examined on Enterococcus hirae (Streptococcus faecalis) by 23Na-NMR and K+ atomic absorption.

Factors likely to modulate the ionic selectivity of monensin were examined on Enterococcus hirae (Streptococcus faecalis) in two states previously characterized: the resting (de-energized) cell and the active (energized) cell. Internal and external Na+ were followed by corresponding 23Na-NMR resonances K+ concentrations were measured by atomic absorption. For a given cellular population of de-energized cells, the apparent transport rates and the final cationic concentrations reached at the steady state were decreasing with the ionophore dose.

A radioautographic study of [3H]-A 23187 (calcimycin) in components of the brain. Distribution in different cell types of the diencephalo-mesencephalic roof.

The distribution of the ionophore [3H]-A 23187 was examined by means of light and electron microscopy in elements of the central nervous system located in the diencephalo-mesencephalic roof. A 23187 is not evenly distributed in the components studied (ependyma, secretory ependyma of the subcommissural organ and neurons of the mesencephalon). At the cellular level, A 23187 appears preferentially associated with the cytoplasmic membrane as well as with the internal membranous system.

Phosphate-dependent sodium transport in S. faecalis investigated by 23Na and 31P NMR.

Na+ movements in S. faecalis were studied by 23Na NMR. They proved to be dependent on phosphate concentration in the buffer during the de-energization step.

Study by NMR of the mode of action of monensin on Streptococcus faecalis de-energized and energized cells.

Streptococcus faecalis was used as a bacterial model for studying the mode of action of monensin by NMR investigations. Experiments were carried out in two states, characterized by several complementary methods: (i) the resting (de-energized) cell which was considered as an inert biological membrane, on which cationic transport induced by the ionophore alone can be investigated; (ii) the active (energized) cell where the ionophore-sensitive response of the living organism, particularly the cation pumps and the glycolysis, is probed. Studies of resting cells were performed, with changing external ionic concentrations, in the presence of monensin, which is preferentially a sodium carrier.

In vitro study of the effect of different ionophore antibiotics and of certain derivatives on rumen fermentation and on protein nitrogen degradation.

An in vitro study was carried out to evaluate the effect of different ionophore antibiotics and some of their derivatives on rumen fermentation and on the degradation of peanut meal nitrogen. The increase in the production of propionic acid at the expense of acetic acid, observed with lonomycin, nigericin, cationomycin and lysocellin, was identical to that noted with monensin. The decrease in methanogenesis observed in the presence of monensin was also found with cationomycin and lysocellin.

Study of a delta-hydroxy ketone-hemiketal equilibrium by 2D-NMR exchange spectroscopy for the antibiotic grisorixin: involvement in cationic transport through membranes.

Study of a delta-hydroxyketone-hemiketal equilibrium in the polyether antibiotic grisorixin was performed with 2D-NMR spectroscopy. The efficiency of 13C chemical exchange spectroscopy for the assignment of 1H and 13C resonances, in the 2 forms, was shown, making possible a conformational investigation of both forms. This equilibrium was observed for grisorixin in solvents of varying polarity, such as CD2Cl2, CDCl3, CD3CN, or CD3OD, but not in C6D12 or C6D6.

Microbial conversion of nigericin in three successive steps, by Sebekia benihana.

A strain of Sebekia benihana NRRL 11111 was found to transform nigericin in three successive steps, giving three compounds which were isolated. Their structure were determined by IR, 1H and 13C NMR, and fast atom bombardment mass spectra. The first compound resulted from the reduction of the terminal hemiketal ring it was then transformed into the two other compounds as a result of the oxidation of methyl (C-33) into a CH2OH and COOH group respectively.

[In vitro study of various ionophore antibiotics and some of their derivatives. II. Characterization of the ionophore properties of the compounds in a model system for Na+ and K+ ions].

Transport of Na+ and K+ by different carboxylic polyether antibiotics and some of their derivatives was measured in a triphasic water/chloroform/water model system. Monensin, nigericin, narasin and lasalocid proved to be efficient K+ carriers, while grisorixin, alborixin and X 14547A gave lower ionic fluxes. Furthermore, the structural modifications of nigericin, grisorixin and lasalocid reduced the ionophore properties of the corresponding natural metabolite.

Semi-synthesis of A23187 (calcimycin) analogs. IV. Cation carrier properties in mitochondria of analogs with modified benzoxazole rings. Antimicrobial activity.

The transporting abilities in the mitochondrial membrane for Ca++ and Mg++ of ten semi-synthetic analogs A23187 (calcimycin) and X14885A are compared. Analogs classified as efficient divalent cation carrier retained the calcimycin antimicrobial activity against three Gram-positive strains tested.