Autophagy and Symbiosis: Membranes, ER, and Speculations.

The interaction of plants and soil bacteria rhizobia leads to the formation of root nodule symbiosis. The intracellular form of rhizobia, the symbiosomes, are able to perform the nitrogen fixation by converting atmospheric dinitrogen into ammonia, which is available for plants. The symbiosis involves the resource sharing between two partners, but this exchange does not include equivalence, which can lead to resource scarcity and stress responses of one of the partners. In this review, we analyze the possible involvement of the autophagy pathway in the process of the maintenance of the nitrogen-fixing bacteria intracellular colony and the changes in the endomembrane system of the host cell. According to in silico expression analysis, ATG genes of all groups were expressed in the root nodule, and the expression was developmental zone dependent. The analysis of expression of genes involved in the response to carbon or nitrogen deficiency has shown a suboptimal access to sugars and nitrogen in the nodule tissue. The upregulation of several ER stress genes was also detected. Hence, the root nodule cells are under heavy bacterial infection, carbon deprivation, and insufficient nitrogen supply, making nodule cells prone to autophagy. We speculate that the membrane formation around the intracellular rhizobia may be quite similar to the phagophore formation, and the induction of autophagy and ER stress are essential to the success of this process.

Key structural factors and intermolecular interactions underlying the formation, functional properties and behaviour in the gastrointestinal tract in vitro of the liposomal form of nutraceuticals coated with whey proteins and chitosan.

The aim of this study was to gain a better understanding of the key structural factors and intermolecular interactions underlying the formation, functionality, and in vitro gastrointestinal behaviour of the liposomal form of nutraceuticals coated with whey proteins (WPI) and chitosan (CHIT). Phosphatidylcholine (PC) liposomes were used to encapsulate a combination of hydrophobic and hydrophilic nutraceuticals. The hydrophobic constituents were long-chain (LC) n-3 PUFAs (DHA and EPA) from fish oil (FO), vitamin D3, and clove essential oil (CEO), while the hydrophilic component was γ-aminobutyric acid (GABA). A combination of physicochemical methods was used to achieve this goal, including electron paramagnetic resonance spectroscopy (EPRS), laser light scattering in dynamic, static, and electrophoretic modes, transmission electron microscopy, spectrophotometry and tensiometry. The efficiency of encapsulating the nutraceuticals in PC liposomes simultaneously was as follows: 100 ± 1% for both FO triglycerides and CEO, 82 ± 2% for vitamin D3, and 50 ± 1% for GABA. According to EPRS data, encapsulating LC PUFA reduced microviscosity at a depth of 20 Å in the PC bilayer. The co-encapsulation of other nutraceuticals in PC liposomes at selected concentrations did not alter this effect. The upper part (8 Å) of PC liposome bilayers showed an increase in rigidity parameter S, indicating the presence of D3, CEO, and partially GABA. The liposome layer-by-layer encapsulation efficiency (EE%) was achieved by using WPI to form the binary complex [WPI-(PC-FO-D3-GABA-CEO)] (EE = 50% at pH 7.0 and I = 0.001 M), followed by coating with chitosan to form the ternary complex [WPI-(PC-FO-D3-GABA-CEO)]-CHIT (EE = 80% at pH 5.1 and I = 0.001 M). The biopolymer-coated liposomes displayed high water solubility owing to their submicron sizes, thermodynamic affinity for the aqueous medium, and 20 mV ζ-potential values. The chitosan shell regulated the release of liposomes from the ternary complex during in vitro gastrointestinal digestion. In the stomach, the hydrolysis of chitosan by pepsin resulted in a 40% release of liposomes. In the small intestine, chitosan was separated from the WPI-liposome core, facilitatig its hydrolysis and resulting in a 60% release of liposomes. The bioavailability of nutraceuticals encapsulated in PC liposomes in the small intestine may be enhanced by the interactions of both non-hydrolysed and hydrolysed liposomes with bile salts and mucin.

The relationship between the structure and functionality of essential PUFA delivery systems based on sodium caseinate with phosphatidylcholine liposomes without and with a plant antioxidant: an in vitro and in vivo study.

The aim of this work was to establish the main relationship between the structure and functionality of supramolecular complexes formed by sodium caseinate (SC) with phosphatidylcholine (PC) liposomes filled with fish oil (FO) to an equal mass ratio of n-3 to n-6 polyunsaturated fatty acids (PUFA) in the absence and presence of one of the most effective plant antioxidants, namely the essential oil of clove buds (EOC). The functionality of the supramolecular complexes (SC-PC-FO and SC-PC-FO-EOC) was considered from the point of view of the possibility of their use as effective delivery systems for long-chain n-3 PUFAs (eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids from FO). The laser light scattering method was used in the static, dynamic and electrophoretic modes to characterize the structure and thermodynamic parameters of the supramolecular complexes in an aqueous medium. It was found that the SC-PC-FO and SC-PC-FO-EOC complex particles had the following similar properties: nanosize; a spherical shape; 100% solubility in an aqueous medium (pH 7.0, ionic strength = 0.001 M); a high encapsulating ability of SC (up to 70%) in relation to the studied liposomes; and a high protective ability relative to lipid autooxidation (up to 96% on the 20th day of storage at room temperature in light). In addition, a sequential transformation of both the structural and thermodynamic parameters has been observed for the complex particles under in vitro simulated gastrointestinal (GI) conditions in accordance with the INFOGEST protocol. A greater release of the encapsulated lipids from the enzymatically hydrolyzed complex particles was observed at the small intestine stage compared to their release at the gastric stage. These data were in good agreement with those on the assessment of the bioavailability of the target PUFAs in in vivo experiments based on the chronic intake of aqueous solutions of the complexes (both SC-PC-FO and SC-PC-FO-EOC) by experimental mice for 92 days. Liver lipid profiles of the mice, obtained by gas-liquid chromatography, showed the following: (i) an almost twofold increase in the DHA content as compared with that of the control; (ii) an almost threefold decrease in the mass ratio of arachidonic acid (AA) (C20:4 n-6) to DHA (C22:6 n-3) compared to that of the control due to both a significant decrease in the AA content and a simultaneous pronounced increase in the DHA content; and (iii) an almost twofold decrease in the mass ratio of the total amounts of n-6 to n-3 PUFAs compared to that of the control.

Sequential transformation of the structural and thermodynamic parameters of the complex particles, combining covalent conjugate (sodium caseinate + maltodextrin) with polyunsaturated lipids stabilized by a plant antioxidant, in the simulated gastro-intestinal conditions in vitro.

The present work is focused on the structural transformation of the complexes, formed between covalent conjugate (sodium caseinate + maltodextrin) and an equimass mixture of the polyunsaturated lipids (PULs): (soy phosphatidylcholine + triglycerides of flaxseed oil) stabilized by a plant antioxidant (an essential oil of clove buds), in the simulated conditions of the gastrointestinal tract. The conjugate was used here as a food-grade delivery vehicle for the PULs. The release of these PULs at each stage of the simulated digestion was estimated.

Biopolymer nanovehicles for essential polyunsaturated fatty acids: Structure-functionality relationships.

Design of stimuli-sensitive (i.e., smart) nano-sized delivery systems for nutraceuticals, having both a nutritional and pharmaceutical value, is very important for the formulation of novel functional food. Omega-3 and omega-6 polyunsaturated fatty acids (PUFAs) are among the most needed nutraceuticals for the maintenance of good health. It is medically proven that in order to get the best effect on the human health the weight ratio of ω-6/ω-3 PUFAs should be within the range between 1/1 and 5/1. Thus, our work was focused on the molecular design of the delivery systems based on the nano-sized complexes formed between covalent conjugate (sodium caseinate+maltodextrin (a dextrose equivalent=2)) and the combinations of polyunsaturated lipids, which are mutually complementary in the ω-6 and ω-3 PUFAs content: α-linolenic (ALA)+linoleic (LA) acids; liposomes of soy phosphatidylcholine (PC)+ALA; and micelles of soy lysophosphatidylcholine (LPC)+ALA. For such complex particles the high extent (>95%) of encapsulation of these all combinations of lipids by the conjugate was found along with both the high protection of the lipids against oxidation and their high solubility in an aqueous medium. To gain a better insight into such functionality of the complex particles a number of their structural (the weight-averaged molar weight, Mw; the radius of gyration, RG; the hydrodynamic radius, Rh; the architecture; the volume; the density; the ζ-potential; the microviscosity of both the bilayers of PC liposomes and LPC micelles), and thermodynamic (the osmotic second virial coefficient, A2, reflecting the nature and intensity of both the complex-complex and complex-solvent pair interactions) parameters were measured by a combination of such basic physico-chemical methods as static and dynamic multiangle laser light scattering, particle electrophoresis, atomic-force microscopy and electron spin resonance spectroscopy.

Thermodynamic analysis of the impact of the surfactant-protein interactions on the molecular parameters and surface behavior of food proteins.

This paper reports on the thermodynamics of the interactions between surfactants (anionic, CITREM, SSL; nonionic, PGE; zwitterionic, phospholipids) and food proteins (sodium caseinate, legumin) depending on the chemical structure and molecular state (individual molecules, micelles) of the surfactants and the molecular parameters (conformation, molar mass, charge) of the proteins under changes of pH in the range from 7.2 to 5.0 and temperature from 293 to 323 K. The marked effect of the protein-surfactant interactions on the molecular parameters (the weight-average molar mass, the gyration and hydrodynamic radii) and the thermodynamic affinity of the proteins for an aqueous medium were determined by a combination of static and dynamic laser light scattering. Thermodynamically justified schematic sketches of the molecular mechanisms of the complex formation between like-charged proteins and surfactants have been proposed. In response to the complex formation between the proteins and the surfactants, the more stable and fine foams have been detected generally.

Thermodynamic and functional properties of legumin (11S globulin from Vicia faba) in the presence of small-molecule surfactants: effect of temperature and pH.

We report on the effect of a set of water-dispersible small-molecule surfactants (the main and the longest-hydrocarbon components of which are a citric acid ester of monostearate, a sodium salt of stearol-lactoyl lactic acid, and a polyglycerol ester of stearic acid) on molecular, thermodynamic, and functional properties of the major storage protein of broad beans (Vicia faba) legumin in different molecular states (native, heated, and acid-denatured). The interaction between legumin and the surfactants has been characterized by a combination of thermodynamic methods, namely, mixing calorimetry and multiangle laser static and dynamic light scattering. It was found that hydrogen bonds, electrostatic interactions, and hydrophobic contacts provided a basis for the interactions between the surfactants and both the native and the denatured protein in aqueous medium. Intensive association of the protein molecules in a bulk aqueous medium in the presence of the surfactants was revealed by static and dynamic laser light scattering. In consequence of this, both the surface activity and the gel-forming ability of legumin increased markedly, which has been shown by tensiometry, estimation of protein foaming capacity, and steady-state viscometry. A likely molecular mechanism underlying the effects of small-molecule surfactants on legumin structure-forming properties at the interface and in a bulk aqueous medium is discussed.

Analysis of Light Scattering Data on the Calcium Ion Sensitivity of Caseinate Solution Thermodynamics: Relationship to Emulsion Flocculation.

We describe the quantitative interrelation between the thermodynamic parameters of caseinate submicelles in the presence of calcium ions (0-14 mM) in aqueous medium and the capacity of the protein to induce depletion flocculation in oil-in-water emulsions at pH 7.0 and ionic strength 0.05 mol dm(-3). Measurements have been made by static and dynamic multiangle laser light scattering of the weight-average molecular weight, the radius of gyration, the hydrodynamic radius, and the second virial coefficient of caseinate submicelles in aqueous solution. Successive thermodynamic approximations with and without consideration of correlations between caseinate submicelles have been used to calculate the osmotic pressure in caseinate aqueous solutions and the free energy of the depletion interaction between droplets in oil-in-water emulsions stabilized by caseinate. Numerical results from both thermodynamic approximations are in reasonably good agreement with experiment, predicting a pronounced decrease in the strength of the depletion attraction at concentrations of Ca(2+) in the range 4-8 mM (with a minimum value at 8 mM). This correlates well with the great enhancement of stability of these emulsions with respect to flocculation in comparison with systems having no added ionic calcium and emulsions with lower (2 mM) or higher (10 mM) Ca(2+) contents. Nevertheless, the allowance for interactive correlations between caseinate submicelles seems to lead to a better prediction of emulsion flocculation on a qualitative level over the whole range of Ca(2+) concentrations studied (2-14 mM). The calculated pronounced decrease in depletion interaction strength is attributable to marked changes in weight-average molecular weight and mean size of aggregates, and to more positive values of the second virial coefficient of caseinate submicelles with increasing Ca(2+) content. Finally, we discuss the part played by the electrical charge on the protein in determining the overall strength of the flocculation-inducing attractive interactions between droplets. Copyright 2001 Academic Press.