N. M. Sanina

Thermotropic behavior of major phospholipids from marine invertebrates: changes with warm-acclimation and seasonal acclimatization.

The crystal-liquid crystal-isotropic melt phase transitions of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) from muscle tissue of five species (actinia Metridium senile fimbriatum, mussel Crenomytilus grayanus, sea-urchin Strongylocentrotus intermedius, starfish Distolasterias nipon and the ascidian Halocynthia aurantium) of marine invertebrates, collected in winter at 0 degrees C and then acclimated to 18.5 degrees C for 5 days, were studied by differential scanning calorimetry and polarising microscopy. To elevate temperature from 0 to 18.5 degrees C, we used the rate of 4.5 degrees C/h. Although phase transitions of both phospholipids from animals collected in summer occurred already at temperatures below -1.7 oC (minimal temperature of seawater in winter), compensatory mechanisms resulted in a decrease by 29-43 oC in the phase transition temperature of PE in winter. Thermotropic behavior of PCs changed in various trends. However, the total heat of their phase transitions always decreased in winter compared with summer. For all species, except the mussel, the time of warm-acclimation was insufficient to adjust the thermotropic behavior of either phospholipid. Nevertheless, the unsaturation index decreased to achieve summer values, due primarily to decreased proportions of eicosapentaenate and docosahexaenate. The accumulation of arachidonate, during warm-acclimation, might be connected to the signalling properties of n-6 eicosanoids. Absence of effective homeoviscous mechanisms suggests that most of the studied marine invertebrates have very limited capacity to survive an acute temperature elevation, e.g. at the appearance of thermal currents.

The Effect of Seasonal Shifts in Temperature on the Lipid Composition of Marine Macrophytes

The comparative study of lipid composition was carried out in four species of marine algae, Ahnfeltia tobuchiensis, Laminaria japonica, Sargassum pallidum, and Ulva fenestrata, as well as a higher plant grass wrack (Zostera marina). Plants were collected in the Japan Sea in spring at 2.9 and 5.5°C and in summer at 23°C. The main lipid components of membranes were determined, and the general patterns of the ratio of phospholipids (PL), glycolipids (GL), betaine (BL), and neutral (NL) lipids were discerned. The relative content of NL in all species (except A. tobuchiensis) was higher in summer. The level of triacylglycerols was as high as 18–37%. The content of individual classes of PL and GL varied between the spring and summer samples, the relative content of PL being higher in spring. In most species, the ratio of PL to GL decreased in summer. The content of free sterols did not depend on the season. The molar ratios of phosphatidylcholine and diacylglycerol-o-(hydroxymethyl)-(N,N,N-trimethyl)homoserine to free sterols varied from 0.9 to 1.7. The seasonal changes of lipid composition were apparently related to macrophyte adaptation to water temperature and to biology of their development.

Tubular immunostimulating complex based on cucumarioside A2-2 and monogalactosyldiacylglycerol from marine macrophytes

BackgroundThere is an urgent need to develop safe and effective adjuvants for the new generation of subunit vaccines. We developed the tubular immunostimulating complex (TI-complex) as a new nanoparticulate antigen delivery system. The morphology and composition of TI-complexes principally differ from the known vesicular immunostimulating complexes (ISCOMs). However, methodology for the preparation of TI-complexes has suffered a number of shortcomings. The aim of the present work was to obtain an antigen carrier consisting of triterpene glycosides from Cucumaria japonica, cholesterol, and monogalactosyldiacylglycerol from marine macrophytes with reproducible properties and high adjuvant activity.ResultsThe cucumarioside A2-2 - cholesterol - MGalDG ratio of 6:2:4 (by weight) was found to provide the most effective formation of TI-complexes and the minimum hemolytic activity in vitro. Tubules of TI-complexes have an outer diameter of about 16 nm, an inner diameter of 6 nm, and a length of 500 nm. A significant dilution by the buffer gradually destroyed the tubular nanoparticles. The TI-complex was able to increase the immunogenicity of the protein antigens from Yersinia pseudotuberculosis by three to four times.ConclusionsWe propose an optimized methodology for the preparation of homogeneous TI-complexes containing only tubular particles, which would achieve reproducible immunization results. We suggest that the elaborated TI-complexes apply as a universal delivery system for different subunit antigens within anti-infectious vaccines and enhance their economic efficacy and safety.

Seasonal changes in thermotropic behavior of phosphatidylcholine and phosphatidylethanolamine in different organs of the ascidian Halocynthia aurantium

Differential scanning calorimetry and polarising microscopy were used to investigate the crystal-liquid crystal-isotropic melt phase transitions of phosphatidylcholine (PC), and phosphatidylethanolamine (PE), isolated from muscles, gill pouches, gonads and digestive glands of Halocynthia aurantium, collected in summer and winter. We also analyzed the fatty chain composition of these phospholipids. In summer, the crystalline to liquid crystalline phase transitions of PC and PE from different organs were more co-operative than in winter. Their peak maximum temperatures were close and temperature ranges overlapped for summer samples. Peak maximum temperatures of winter samples decreased sharply, by 18-27 degrees C for PC and by 10-44 degrees C for PE, respectively, depending on the organ. Total heat changes of transitions also decreased. Thermograms were completely located at temperatures below -1.7 degrees C (minimal temperature of seawater in winter). In contrast to summer samples, peak maximum temperatures for PC and PE in winter differed significantly, (by 14-30 degrees C depending on organ), while the temperature ranges of their transitions still showed considerable overlap. Simultaneously, the temperature ranges of the liquid crystalline to isotropic phase transitions decreased. The main reason for changes in thermotropic behavior of phospholipids seems to be the decrease of saturated/unsaturated ratios. The existence of stable and thermoadaptative labile phospholipid pools in the membrane structure is proposed. The relationship of these transitions to low- and high-temperature adaptation is discussed.

The thermotropic behavior and major molecular species composition of the phospholipids of echinoderms

This study examines the molecular species composition and heat-induced crystalline-liquid crystalline phase transitions of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) from the muscle tissues of six species of echinoderms that were collected during the summer: the starfishes Distolasterias nipon and Asterias amurensis, the sea urchin Strongylocentrotus intermedius, and the holothurians Eupentacta fraudatrix, Cucumaria frondosa japonica, and Apostichopus japonicus. Phospholipids (PLs) were in the liquid crystalline state, which is optimal for the functioning of the cell membranes. The use of data on the molecular species composition of PLs for the interpretation of their thermotropic behavior indicated that homeoviscous adaptation is achieved by various rearrangements in the composition of the aliphatic groups of PLs. The phase transitions of PC and PE of echinoderms (except holothurians) were symbatic. The presence of a high-temperature peak on the PC thermograms of C. frondosa japonica and A. japonicus is attributable to the melting of the phospholipid domain, which is composed of molecular species with saturated aliphatic groups. Such domains are responsible for a significant shift in the temperature ranges of the phase transitions of phospholipids of holothurians and sea urchin towards temperatures above 0°C.

On the possibility of lipid-induced regulation of conformation and immunogenicity of influenza a virus H1/N1 hemagglutinin as antigen of TI-complexes

The tubular immunostimulating complex (TI-complex) consisting of cucumarioside A2-2, cholesterol and monogalactosyldiacylglycerol (MGDG) from marine macrophytes is the perspective antigen delivery system for subunit vaccines. MGDG is a lipid matrix for the protein antigen incorporated in the TI-complex. The aim of the present work was to study the influence of MGDGs from different macrophytes on conformation and immunogenicity of the secreted recombinant uncleaved hemagglutinin monomer (HA0S) of influenza A virus H1/N1. Differential scanning calorimetry, fluorescence spectroscopy and circular dichroism showed a dependence of the conformational changes of HA0S on the microviscosity of MGDG. The most viscous MGDG from Zostera marina induced the strongest rearrangements in protein conformation. Immunization of mice with HA0S within TI-complexes comprising different MGDGs resulted in an approximately 2-fold increase of the levels of anti-HA0S antibodies and granulocyte-macrophage colony-stimulating factor (GM-CSF) compared with those induced by HA0S alone. TI-complexes based on MGDG from Z. marina stimulated the maximal production of GM-CSF. However, humoral immune response (anti-HA0S antibodies), unlike cell-mediated immune response (GM-CSF), did not depend on the physicochemical properties of MGDGs. It is assumed that this is due to the different localization and conformational lipid sensitivity of the HA0S regions, which are responsible for these types of immune responses.

Phase transitions of phospholipids as a criterion for assessing the capacity for thermal adaptation in fish

This study examines the crystal-liquid crystal phase transitions of the major phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), from muscle tissue of marine fish living at temperatures of 0–4.1°C (the Pacific cod Gadus macrocephalus, banded Irish lord Hemilepidotus gilberti, Pacific halibut Hippoglossus hippoglossus, black edged sculpin Gymnocanthus herzensteini, dark colored flounder Pleuronectes obscurus, and plain sculpin Myoxocephalus jaok), as well as of fish living at 14 and 18°C (Pacific redfin Tribolodon brandti). The PC and PE phase-transition thermograms of all the investigated species displayed specific profiles. The largest share of the thermogram area at temperatures higher than those of the habitat was found for the PC (28–40%) and PE (47–82%) of the black-edged sculpin, dark-colored flounder, and the plain sculpin, which have reduced physiological activity at low temperatures. In the Pacific cod, banded Irish lord, and the Pacific redfin, this parameter was much lower, 0–18% (PC) and 0–39% (PE). The thermotropic behavior PC and PE was symbate in all fish, except for the cod and the plain sculpin. The transition enthalpy of PC in all the investigated species was 2.8 times higher than that of PE. To interpret the varied PC and PE thermogram profiles of fish with similar fatty-acid compositions, the data on the composition of the molecular species of these phospholipids appeared to be the most informative. This study suggests that each fish species has its own strategy for thermal adaptation, which is realized through a certain set of phospholipid molecular species.

The effects of triterpene glycosides and phospholipids from marine invertebrates in the composition of tubular immunostimulating complexes on the immunogenicity of human serum albumin

To study the adjuvant activity and to optimize tubular immunostimulating complexes (TI complexes) with respect to the soluble antigen, an investigation was performed on cucumariosides and their fractions, i.e., individual triterpene glycosides that were obtained from the holothurian Cucumaria japonica, as well as on phospholipids from the sea urchin Strongylocentrotus intermedius and the starfish Distolasterias nippon included in the composition of TI complexes in order to assess their effects on the immunogenicity of human serum albumin (HSA). Generally, TI complexes showed a moderate adjuvant activity with respect to HSA; this activity can be adjusted by substitution of a cucumarioside and/or a polar lipid in the composition of the TI complex. TI complexes based on monogalactosyldyacylglycerol (MGDG) from the sea alga Ulva lactuca and cucumariosides A2-2 and A4-4 maximally stimulated anti-HSA antibody production in mice. Substitution of MGDG for phosphatidylcholine (PC) from the starfish D. nippon with an increased (compared with other investigated phospholipids) ratio of n-3/n-6 polyunsaturated fatty acids did not result in a change in the immunostimulating activity of the TI complex. However, the use of other phospholipids led to a decrease in the adjuvant activity of the TI complex, compared with that of the TI complex based on PC from D. nippon and cucumarioside A2-2. The greatest fluctuations in the contents of cytokines IL-6, IL-12, IFN-γ and GM-CSF depending on the composition of the TI complexes indicate the possibility of regulation of the T-cell immune response. The values of hematological parameters were normal.

Phospholipids of the organs and tissues of echinoderms and tunicates from peter the great bay (Sea of Japan)

The phospholipid compositions of organs and tissues were determined in representatives of two phyla of marine invertebrates: Echinodermata [Asteroidea: Aphelasterias japonica (Bell, 1881), Evasterias echinosoma Fisher, 1926, Distolasterias nipon (Doderlein, 1902), Asterias amurensis Lutken, 1871; Echinoidea: Strongylocentrotus intermedius (A. Agassiz, 1863); Holothurioidea: Cucumaria frondosa japonica (Semper, 1868), Eupentacta fraudatrix (Djakonov et Baranova, 1958), Apostichopus japonicus (Selenka, 1867)], and Tunicata [Ascidia: Halocynthia aurantium (Pallas, 1787), H. roretzi (Drasche, 1884), and Styela clava (Herdman, 1881)]. The specificity of phospholipid distribution was shown to be related to the taxonomic position of marine invertebrates and the functional properties of their organs and tissues. Ceramide aminoethylphosphonate was found only in the digestive organs of all holothurians and the starfishes D. nipon and A. amurensis, suggesting its exogenous origin. Phosphatidylglycerol was found in all organs and tissues of the holothurians C. frondosa japonica and E. fraudatrix, as well as in the gastrointestinal tract of ascidians; its origin is unclear.

Individual variations of the phospholipid compositions of the organs of arthropods, echinoderms, and tunicates from peter the Great Bay of the Sea of Japan

Individual variations in the phospholipid compositions of the organs of marine invertebrates were investigated in the representatives of three phyla: Arthropoda [decapods: Paralithodes camtschaticus (Tilesius, 1815) and Erimacrus isenbeckii (Brandt, 1848)]; Echinodermata [starfishes: Distolasterias nipon (Doderlein, 1902) and Evasterias echinosoma Fisher, 1926]; and Tunicata [ascidians: Halocynthia aurantium (Pallas, 1787), H. roretzi (Drasche, 1884), and Styela clava Herdman, 1881]. The specificity of individual phospholipid variation was shown to be related to the systematic status and functional properties of the organs of marine invertebrates. Phosphatidylcholine and phosphatidylethanolamine were the least variable. Low variation of phospholipid composition was found in the liver, legs, and gills of decapods; in the liver of starfishes; and in the gonads and skin-muscular and gill sacs of ascidians. The contents of ceramide aminoethylphosphonate (CAEP) and phosphatidylglycerol (PG) showed the highest variation. These phospholipids were found but not in all individuals or organs studied. After 7 days of starvation, CAEP completely disappeared from the organs of the animals; thus, we suggest its exogenous origin.