Structural organisation of cationic dioctadecyldimethyammonium bromide monolayers in presence of hyaluronic acid.

Langmuir monolayers of dioctadecyldimethyammonium bromide and its interaction with the natural mucopolysaccharide hyaluronic acid are studied using thermodynamic methods and X-ray diffraction at grazing incidence. The 2D crystalline lattice parameters of different phases are determined. The monolayer compressibility, the linear crystalline compressibility components and the thermoelastic expansion coefficient are evaluated. The biopolymer stabilises the monolayer structural properties, increases the collapse pressure and the correlation length of the 2D crystalline domains. The results show that this lipid has a potential for developing of stabilised drug delivery systems of anionic biopolymers like hyaluronic acid, oligomers and genes.

Interactions of lipid monolayers with the natural biopolymer hyaluronic acid.

The interaction of the natural mucopolysaccharide hyaluronic acid with different lipids, present in the natural membranes, was studied at the lipid/water interface using thermodynamic methods and X-ray diffraction. The results show that this biopolymer modifies the properties and the structure of the lipid monolayer. The two-dimensional crystalline lattice and domain structure of the charged octadecylamine monolayer are strongly disturbed by the hyaluronic acid, the monolayer compressibility increases and the monolayer collapse pressure drops down. In addition, the presence of charged lipid interfaces influences the structural organisation of the hyaluronic acid at the membrane/water interfaces. The impacts of these results on the structural organisation at the membrane interface are discussed.

Changes in some nitrogenous compounds in the blood and tissues of freshwater pikeperch (Sander lucioperca) during salinity acclimation.

The effect of ambient salinity changes (0.9, 6 and 12 psu) on the levels of dissolved ammonia (DA), ninhydrin positive substances (NPS), trimethylamine (TMA) and trimethylamine oxide (TMAO) in the blood and tissue of medium-acclimated Sander lucioperca L. (also Stizostedion lucioperca) were investigated. In freshwater, blood and tissue total free amino acid levels (measured as NPS) were 3.62 mM and 60.61 mM, respectively. The NPS content increased significantly (P<0.05) in the tissue and blood on acclimation to 6 and 12 psu salinities. The mass-specific tissue TMAO concentration of pikeperch acclimated to normal freshwater was 0.413+/-0.084 micromol TMAO g(-1). Results reveal that TMAO levels are positively influenced by the external salinity medium where significant differences in mean levels occurred between the groups (P<0.05). The calculated p[NH(3)] and [NH(4)(+)] gradients reveal that the [NH(3)] gradient was consistently low (cf. the [NH(4)(+)] gradient). The gradient of p[NH(3)] decreased with the medium increased salinities. The results suggest that freshwater pikeperch may be able to resist salinity changes by manipulation of nitrogen metabolism. Free amino acids and TMAO are involved in mediating response to salinity exposure in freshwater pikeperch.

Genetic responses to metal contamination in two clams: Ruditapes decussatus and Ruditapes philippinarum.

Coastal ecosystems are subjected to a wide variety of disturbances, including those due to xenobiotics of agricultural and industrial origin. These pollutants as heavy metals can modify the genetic diversity of populations by favouring or counter-selecting certain alleles or genotypes by differential mortality. In the present study, two genetic markers (phosphoglucomutase and glucosephosphate isomerase) and a protein marker (metallothionein) were monitored in order to determine the impact of heavy metals in different clam populations. Analysis of the genetic structure of the clam populations examined reveals that those inhabiting environments contaminated by heavy metals exhibit a higher allelic diversity and possess alleles at PGM loci that could be selected by the presence of heavy metals. The evaluation of metallothionein levels using a specific polyclonal antibody developed in the Pacific oyster (Crassostrea gigas) demonstrated the existence of a relationship between metallothionein concentrations and the level of metal pollution for clam populations sampled from different sites. An inter-specific difference was also detected between Ruditapes decussatus and Ruditapes philippinarum living in sympatry at the same site, suggesting a differential response of these two species upon exposure to an identical heavy metal concentration.

Experimental evidence for an original two-dimensional phase structure: an antiparallel semifluorinated monolayer at the air-water interface.

We show the spontaneous formation of an antiparallel monolayer of diblock semifluorinated n-alkane molecules spread at the air-water interface. We used simultaneous measurements of surface pressure and surface potential versus molecular area and performed grazing x-ray reflectivity experiments to characterize the studied monolayer, which is obtained at almost zero surface pressure and precedes the formation of a bilayer at higher surface pressure. Its thickness, equal to 2.7 nm, was found to be independent of the molecular area. This behavior may be explained by van der Waals and electrostatic interactions.

Indium tellurium trioxide chloride, InTeO3Cl.

The new oxychloride InTeO(3)Cl was synthesized from a mixture of In(2)O(3), InCl(3) and TeO(2). Its structure has been determined from single-crystal X-ray diffraction data. The structure is composed of layers separated by a van der Waals gap. The layers consist of edge-sharing chains of [InO(4)Cl(2)] octahedra linked through [TeO(3)] trigonal pyramids. No free Cl atoms are located between the layers.

Growth of Sr6Rh5O15 single crystals from high-temperature solutions: structure determination using the traditional 3-D and the 4-D superspace group methods and magnetic measurements on oriented single crystals.

Single crystals of Sr6Rh5O15 were grown from a molten potassium carbonate flux. The structure was solved by both the traditional 3-D crystallographic approach and the 4-D superspace group approach using JANA2000. Both methods produced an equivalent structure determination, thereby confirming the 4-D superspace group approach as an effective structure solution method for 3-D commensurate composite structures. Sr6Rh5O15 corresponds to the n = 1, m = 1 member of the A3n+3mA'nB3m+nO9m+6n family of 2H hexagonal perovskite-related oxides. This compound is characterized by pseudo-one-dimensional polyhedral chains of four face-sharing RhO6 octahedra followed by one RhO6 trigonal prism. These chains in turn are separated by [Sr](infinity) chains. Magnetic measurements were carried out on oriented single crystals, and a very large magnetic anisotropy in the magnetic susceptibility was observed.

Observation of a large magnetic anisotropy in the new 2H-perovskite related oxide Ba8CoRh6O21: magnetic measurements on aligned single crystals.

Single crystals of Ba8CoRh6O21, grown out of a potassium carbonate flux, were characterized by single-crystal X-ray diffraction and magnetic measurements. X-ray data were collected in a superspace group approach and solved using the JANA2000 software package. Ba8CoRh6O21 represents the first example of a structurally characterized m = 5, n = 3 member of the A3n+3mA'nB3m+nO9m+6n family of 2H hexagonal perovskite related oxides and contains chains consisting of six consecutive RhO6 octahedra followed by one distorted CoO6 trigonal prism. Magnetic measurements were carried out on large aligned single crystals, and a very large magnetic anisotropy in the magnetic susceptibility, persisting up to room temperature, was observed.

Air-water interface-induced smectic bilayer

We show, using surface pressure versus molecular area isotherms measurements and x-ray reflectivity, that the long diblock semifluorinated n-hexaeicosane molecules, F(CF2)(8)-(CH2)18H, form a stable smectic bilayer phase, noted M1, with a total thickness of 3. 3 nm, at an apparent molecular area about 0.3 nm(2), though in the bulk the used molecules do not form smectic phases at any temperature. We discuss different molecular packing models according to our experimental data and deduce that molecules are antiparallel with fluorinated chains outwards and interleaved hydrocarbon chains inwards.

Asymmetrical ion-channel model inferred from two-dimensional crystallization of a peptide antibiotic.

The structural organization of ion channels formed in lipid membranes by amphiphilic alpha-helical peptides is deduced by applying direct structural methods to different lipid/alamethicin systems. Alamethicin represents a hydrophobic alpha-helical peptide antibiotic forming voltage-gated ion channels in lipid membranes. Here the first direct evidence for the existence of large-scale two-dimensional crystalline domains of alamethicin helices, oriented parallel to the air/water interface, is presented using synchrotron x-ray diffraction, fluorescence microscopy, and surface pressure/area isotherms. Proofs are obtained that the antibiotic peptide injected into the aqueous phase under phospholipid monolayers penetrates these monolayers, phase separates, and forms domains within the lipid environment, keeping the same, parallel orientation of the alpha-helices with respect to the phospholipid/water interface. A new asymmetrical, "lipid-covered ring" model of the voltage-gated ion channel of alamethicin is inferred from the structural results presented, and the mechanism of ion-channel formation is discussed.

Metallothionein-like protein: is It an efficient biomarker of metal contamination? A case study based on fish from the Tunisian coast.

The aim of this work was to assess the relative importance of natural fluctuations in metallothionein-like protein (MTLP) levels associated with the sexual status of fish versus fluctuations due to metal exposure. In order to see fluctuations due to metal exposure, comparisons were made on the same fish species Scorpaena porcus sampled in polluted and unpolluted sites. The hermaphrodite fish Serranus scriba and Scorpaena porcus, in which sexes are separate, were compared at the unpolluted site to see fluctuations caused by the sexual status. In both species, metals and the MTLP levels were determined in the gills and liver. In these organs, Cd, Cu, and Zn distributions were examined in different fractions: the insoluble fraction (IF) and the cytosol divided into thermolabile compounds (HDF) and the heat stable compounds including MTLP. MTLP levels were higher in the liver (3.09 mg/g in S. porcus, 1.59 mg/g in S. scriba) than in gills (0.13 mg/g in S. porcus, 0.40 mg/g in S. scriba). For Scorpaena porcus, metals and MTLP levels varied with sex, whereas in Serranus scriba, which is a hermaphrodite species, inherent variations were also observed. At the polluted site, MTLP, Cd, and Cu concentrations in the gills of S. porcus increased but the supplementary metals were not associated with the heat stable compounds including MTLP. At this site, hepatic MTLP bound more metals than at the unpolluted site, but its binding capacity was not sufficient to avoid the binding of metals to the insoluble and the heat denaturable fractions. In light of these results and in spite of its hermaphrodism, it is questionable whether to consider S. scriba as a good candidate for biomonitoring based on MTLP. S. porcus could be useful for this purpose only if the MTLP capacity in binding metals is not exceeded. The MTLP could be considered as a biomarker only if it is investigated in relatively unpolluted sites.

Median eminence dopamine and serotonin neuronal activity. Temporal relationship to preovulatory prolactin and luteinizing hormone surges.

Using a high-performance liquid chromatography (HPLC) system coupled with an electrochemical detector, the concentrations of dopamine (DA) and 5-hydroxytryptamine (5-HT) and their major specific metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindole-3-acetic acid (5-HIAA), respectively, were measured in the median eminence (ME) throughout the rat estrous cycle. The ME DA content remained fairly constant throughout the estrous cycle except on estrus when 17.00 h values were significantly lower than 10.00 h values (40% decrease, p less than 0.05). The ME 5-HT content determined at 10.00 h was higher on proestrus than on any other day of the cycle. The ME DOPAC concentrations did not differ between 10.00 and 17.00 h on diestrus I, diestrus II or estrus. On the contrary, there was an almost linear decline between 10.00 and 17.00 h on proestrus (36% decrease, p less than 0.05). The ME 5-HIAA content did not differ between 10.00 and 17.00 h on any day of the estrous cycle. Significant changes were recorded for the DOPAC/DA and 5-HIAA/5-HT ratio in the ME on proestrus. There was a progressive decrease, starting from 10.00 h in the DOPAC/DA ratio with minimal values (42% decrease, p less than 0.05) at 16.00 h followed by an increase from 16.00 to 19.00 h. On the other hand, the 5-HIAA/5-HT ratio increased between 10.00 and 17.00 h (97% increase, p less than 0.05) and subsequently declined until 19.00 h (67% decrease vs. 17.00 h, p less than 0.05).2+hese data show that a concomitant

Stimulation of estradiol-17 beta secretion by 7,12-dimethylbenz (a) anthracene during mammary tumor induction in Sprague-Dawley rats.

Testosterone, androstenedione, progesterone, 17-hydroxyprogesterone, estrone and estradiol-17 beta serum levels were measured at given times after dimethylbenz (a) anthracene (DMBA) treatment of a sensitive rat strain Sprague-Dawley (S-D) and a resistant strain Wistar (W). Tumors appeared with a 100% incidence around the 14th to 15th estrous cycle after DMBA treatment in in Sprague-Dawley rats. Hormonal determinations were made, during the 5th or 6th estrous cycle after DMBA treatment, in groups of 4-day cycling rats of both strains which were given DMBA or the carrier solution (sesame oil) when they were about 55-days old. In Sprague-Dawley female rats, DMBA treatment significantly stimulated estradiol-17 beta and estrone preovulatory surge on proestrous days. No such stimulation was found for any other steroid at any time of the estrous cycle. On the other hand, the resistant Wistar rats did not show any disturbed preovulatory or basal steroid hormone release after the carcinogen treatment. These results complete and explain previous findings concerning the hypothalamo-pituitary activity after DMBA treatment of S-D rats: an early and persistent alteration in the centers involved in the hormonal cyclicity of the hypothalamo-pituitary-ovarian axis must be a result of the DMBA treatment. This deregulation could probably account for the distant and selective production of tumors in the mammary gland induced by a single gastric administration of DMBA.

Binding studies of substance P anterior pituitary binding sites: changes in substance P binding sites during the rat estrous cycle.

Previous studies have shown that substance P (SP), an undecapeptide widely distributed in the gastrointestinal tract and in the peripheral and central nervous system, is a putative regulatory peptide involved in the control of reproductive function. Specifically, SP inhibited, at the anterior pituitary (AP) level, the stimulatory action of a physiological concentration (10(-8) M) of Gonadotropin Releasing Hormone (GnRH) on the release of the luteinizing hormone (LH). In the present work, we have demonstrated the presence of specific SP binding sites in the AP and related changes in the number of these sites to GnRH receptor number, hypothalamic SP and GnRH content and LH secretion during the rat estrous cycle. High affinity saturable SP binding sites (Kd, 1.5 approximately equal to 10 nM) were demonstrated in AP membranes using [3H]-SP or a novel analog, [125I]-(D-Tyr0, NorLeu11)SP. The binding affinity of SP fragments decreased with progressive removal of amino acid residues from N or C termini of the molecule. Other neuropeptides had low affinity for the SP binding sites. During the rat estrous cycle, SP and GnRH binding capacity of the anterior pituitary were inversely related. At the time of the proestrous LH surge, the AP binding capacity was low for GnRH but high for SP. The highest content of SP in the hypothalamus were recorded during the afternoon of proestrus when hypothalamic GnRH levels were lowest and the preovulatory surge occurred. These studies have established the presence of high affinity specific binding sites for SP in the AP which alter during the estrous cycle in a manner appropriate for mediating the direct inhibitory effects of SP on LH release in vitro.

Anterior pituitary dopamine receptors during the rat estrous cycle. A detailed analysis of proestrus changes.

The binding of [3H]-spiperone to partially purified rat anterior pituitary plasma membranes was quantified throughout the estrous cycle in relation with the serum prolactin (PRL) levels. Receptor affinity remained unchanged throughout the cycle (Kd:0.08-0.16 nM). The number of receptors was constantly high from diestrus I 10.30 h to proestrus 10.30 h, as long as serum PRL remained low. Between 10.30 and 17.30 h on proestrus, there was a rapid and marked decrease in receptor numbers (Bmax, from 180 +/- 50 to 48 +/- 10 fmol/mg protein: means +/- SEM of three independent determinations), which coincided with the preovulatory PRL surge. Subsequently, [3H]-spiperone binding gradually increased (from 48 +/- 10 to 110 +/- 21 fmol/mg protein, at 21.30 h), while PRL returned to basal levels. On the afternoon of estrus, the number of dopamine receptors was also negatively correlated with the increase in serum PRL. These results show that, while receptor affinity is constant, the number of dopamine binding sites changes significantly and rapidly on the afternoon of proestrus. A rapid decrease in receptor content is temporally correlated with the onset of the preovulatory PRL surge. Therefore, the number of [3H]-spiperone binding sites may be regulated during the estrous cycle. In addition, the proestrus decrease in the number of these receptors might be a decisive component in the initiation or maintenance of the preovulatory PRL surge in the rat.

[Inhibition of the preovulatory secretion of gonadotropins by prolactin in rats seems to depend on the strain used]. / L'inhibition de la sécrétion préovulatoire des gonadotropines par la prolactine chez la Ratte semble dépendre de la souche utilisée.

Intraperitoneal injection of Rat Prolactin to cyclic female Rats results in preovulatory changes which depend on the strain of animals. Specifically, preovulatory surges of luteinizing hormone and follicle stimulating hormone were suppressed in Sprague-Dawley Rats while delayed but enhanced preovulatory surges of both gonadotropins were observed in Wistar strain.

Oxidation of methionine residues in lutropin.

Bovine lutropin and its subunits were submitted to oxidation by sodium periodate or chloramine T. Methionine residues were easily oxidized but partial destruction of fucose was observed. After oxidation treatment most of the lutropin exhibits the same elution volume in gel filtration as the native hormone. Sucrose gradient sedimentation or gel filtration experiments show however that the oxidation of isolated subunits is accompanied by aggregation of conformational changes even in the case of porcine beta subunit which contains only one methionine residue, unit and intact alpha subunit exhibit very low residual activity.