Interactions of surfactants with living cells. Induction of apoptosis by detergents containing a beta-lactam moiety.

The toxicity of new surfactants containing a beta-lactam ring has been established by studying their interaction with a hybridoma cell line. An hour of contact is sufficient to generate an apoptotic signal after two days of culture. Under the experimental conditions chosen for the experiments, surfactants have been divided into three categories: i) biocompatible and non-apogenic; ii) surfactants triggering an apoptotic signal without inducing cell necrosis; iii) surfactants triggering an apoptotic signal at low concentrations and destroying the cells by necrosis at higher concentrations. The necrosis inducing surfactants also had haemolytic properties. These properties were related to the values of the hydrophilic-lipophilic balance of the molecules.

Possible effects of counterions on biological activities of anionic surfactants.

The aggressiveness in terms of apogenic and necrotic activities of lysine-derived anionic surfactants towards a mammalian cell line (U937) were studied. We used N(alpha)N(epsilon)-dioctanoyl lysine as the model surfactant with lysine, tris, Na(+) or Li(+) as counterions. The aggressiveness of the different surfactants was assessed as the concentrations leading to apoptosis or necrosis after the cells were incubated in the presence of surfactants and then cultivated in their absence. Used in the same conditions, acetates associated with the same cations, had no effects on the cells. Our results show that the aggressiveness of the surfactants depended on the nature of the counterions: it was high when surfactants were associated with small counterions, and low with large counterions.

Derivatives of glutamic acid as new surfactants.

Starting from glutamic acid, different types of surfactants have been synthesised by using original trimodular strategies. Monosubstituted zwitterionic amides of glutamic acid obtained with excellent yields show good surface activity. The grafting of a second hydrophobic side-chain leads to bicatenar cationic surfactants or to disubstituted nonionic cyclic compounds. In order to reduce the hydrophobic character of the bicatenar surfactants, a second synthetic method has been developed, allowing the introduction of a polar sugar group into these molecules. The surfactant properties of several of the products have been determined by physico-chemical methods such as surface tension measurements and compression isotherm studies by means of a Langmuir balance.

Preparation and antibiotic activity of monobactam analogues of nocardicins.

A series of diverse beta-lactam analogues of nocardicins with interesting antimicrobial properties were prepared. Coupling of glucosamine to these compounds improved their water solubility. Aminoacid derivatives produced a stereoinduction on the quaternary enantiotopic carbon of the starting compound 1. Evaluation of their antimicrobial activity showed that the introduction of alpha-amninoacids to monobactams increased their activity. The importance of asymmetric carbon is exemplified by the higher antibiotic activity of L-alpha-aminoacids than the D-series. No significant difference was observed between fluorinated and non-fluorinated monobactams.

Synthesis of perfluoroalkylated beta-alanine and some peptide derivatives: an access to original surfactants.

The reaction of amines of sodium azide with 3-perfluoroalkyl-3-fluoroprop-2-enoate, followed by hydrogenation, affords perfluoroalkylated beta-alanine analogues in very good yields. These compounds can be linked via an amide bond to produce peptide analogues such as carnosine or carcinine derivatives, which could have surfactive and complexing properties.

Erythrocyte hemolysis and shape changes induced by new lysine-derivate surfactants.

The effects of new synthetic lysine-derived anionic surfactants on human and rat erythrocytes were studied. The surfactants were salts of Nalpha,Nepsilon-dioctanoyl lysine with different counterions: lysine (77KK), tris (trishydroxymethyl amminomethane) (77KT), sodium (77KS), and lithium (77KL). 77KK and 77KT showed a biphasic hemolytic behavior in the erythrocytes. The surfactants 77KS and 77KL showed concentration-dependent hemolysis with a CH50 of about 3.4 and 2.6 mmol/l, respectively. 77KK and 77KT induced protection against hypotonic hemolysis in rat erythrocytes at the concentration which showed the least hemolytic activity under isotonic conditions. With human erythrocytes, 77KT did not show biphasic behavior in isotonic medium, but under hypotonic conditions biphasic behavior was present. Changes in shape of the erythrocyte, from discocytic to stomatocytic were observed after incubation with the anionic surfactants studied. Such shape changes occurred progressively over time, with total alteration in shape occuring after about 20 min of incubation.

Preliminary studies of the toxic effects of non-ionic surfactants derived from lysine.

The toxic effects of new synthetic monodisperse non-ionic long-chain N alpha, N epsilon-diacyl lysine polyoxyethylene glycol amide compounds with a structural resemblance to natural lecithin phospholipids were studied by the haemolytic method and the test of the chorioallantoic membrane of the hen's egg (HET-CAM). The following compounds were tested: symmetrical N alpha,N epsilon-diacyl lysine homologues (N alpha,N epsilon-dihexanoyl, N alpha,N epsilon-dioctanoyl and N alpha,N epsilon-didecanoyl lysine) with one methyl ether polyoxyethylene glycol chain of different oxyethylene units (dioxyethylene glycol, tetraoxyethylene glycol and hexaoxyethylene glycol) as headgroup; symmetrical N alpha,N epsilon-diacyl lysine homologues with two methyl ether dioxyethylene glycol chains and the asymmetrical N alpha-butanoyl, N epsilon-dodecyl lysine with two hydrophilic methyl ether dioxyethylene glycol chains as headgroup. A commercial (polydisperse) oleoyl polyoxyethylene glycol diethanolamide with an average of eight units of ethylene oxide was used as control. All the synthesized tested compounds appeared to be less haemolytic and less irritant than the control. The synthesized products were studied with regard to their hydrophobic and hydrophilic chains in order to evaluate the influence of their structure on their haemolytic and irritative action. The results of this study show that the acyl chain distribution of these compounds greatly influence toxic effects: the asymmetrical compound N alpha-butanoyl,N epsilon-dodecyl lysine-bis[methyl ether diethylene glycol]amide was found to be the most haemolytic and irritating compound. Among the symmetrical homologues, the shortest-chain compounds N alpha,N epsilon-dihexanoyl lysine methyl ether polyoxyethylene glycol amides present the least haemolytic and irritating activity, independently of the number and length of the hydrophilic methyl ether polyoxyethylene glycol chains. Taking into account their surface activity properties and their less haemolytic and irritant action, the compound N alpha,N epsilon-dioctanoyl lysine-bis[methyl ether diethylene glycol]amide would be the most suitable for practical purposes.

Transport of alkali-metal picrates through liquid membranes: Coupled action of w/o microemulsion droplets and lipophilic crown-ether carriers.

The transport of potassium and sodium picrates is measured by a liquid membrane technique in the presence of two mobile carriers: water-in-oil microemulsion droplets and lipophilic crown-ethers. The fluxes are compared for two cases: (i) the crown-ether (dicyclohexano-18-crown-6 is assumed to be independent of the microemulsion droplet and (ii the crown-ether (dodecano- 18-crown-6) is assumed to be anchored in the droplet. A lower flux is observed when the microemulsion droplet and the crown-ether diffuse as a single entity. These results suggest that larger synergistic effects are obtained when the extractant can diffuse independently of the microemulsion droplet.

[Conditions for the use of specific antibodies for immunohistochemical visualization of serotonin and melatonin in the pineal gland of sheep]. / Conditions d'utilisation d'anticorps spécifiques pour la visualisation immunohistochimique de la sérotonine et de la mélatonine dans la glande pinéale du mouton.

Having prepared antisera to serotonin and to melatonin, the authors were able to show that, in the pineal gland, the behaviour of these two antisera in immunohistochemical studies differs. The antiserum raised against 5HT, actually bound to the molecule formed by condensation of formaldehyde on 5HT and therefore could be used to reveal 5HT in tissue fixed with formaldehyde. On the other hand, the anti-melatonin antiserum bound to melatonin, and could therefore be used to reveal its presence in fresh tissue.

[Induction of reaginic hypersensitivity to tartrazine in the rabbit immunization by ingestion of the covalent conjugate tartrazine-human serum albumin (author's transl)]. / Induction d'une hypersensibilité réaginique à la tartrazine chez le lapin: immunisation par voie digestive par le conjugué covalent tartrazine-séralbumine humaine.

Sensitization to simple chemical substances acting as haptens is not well known in food allergy. For that purpose, a study was made in rabbit using tartrazine alone with adjuvant, or mixed with human serum albumin (HSA), or conjugated by covalent BOP bonds with HSA. Hypersensitivity was evaluated by rabbit basophil degranulation test which would offer more specificity through cell membrane IgE-antibody reactions, and thus, more accuracy than classical passive cutaneous anaphylaxis. The data showed that sensitization could not be induced by ingestion of tartrazine with adjuvant or mixed with a protein. But covalent conjugate T-HSA mixed with adjuvant sensitized 8 of the 9 rabbits studied. It seems that preceding irritation of intestinal mucous membrane by acetylsalicylic acid enables one to obtain a more long lasting sensitization.