Epigenetic regulation in obesity.

The availability to the DNA strand and the activity of the transcription machinery is crucial for the cell to use the information in the DNA. The epigenetic mechanisms DNA methylation, modification of histone tails, other chromatin-modifying processes and interference by small RNAs regulate the cell-type-specific DNA expression. Epigenetic marks can be more or less plastic perpetuating responses to various molecular signals and environmental stimuli, but in addition apparently stochastic epigenetic marks have been found. There is substantial evidence from animal and man demonstrating that both transient and more long-term epigenetic mechanisms have a role in the regulation of the molecular events governing adipogenesis and glucose homeostasis. Intrauterine exposure such as poor maternal nutrition has consistently been demonstrated to contribute to a particular epigenotype and thereby developmental metabolic priming of the exposed offspring in animal and man. Epigenetic modifications can be passed not only from one cell generation to the next, but metabolic disease-related epigenotypes have been proposed to also be transmitted germ-line. Future more comprehensive knowledge on epigenetic regulation will complement genome sequence data for the understanding of the complex etiology of obesity and related disorder.

The Richmond Agitation-Sedation Scale: translation and reliability testing in a Swedish intensive care unit.

BACKGROUND: Awareness about adequate sedation in mechanically ventilated patients has increased in recent years. The use of a sedation scale to continually evaluate the patient's response to sedation may promote earlier extubation and may subsequently have a positive effect on the length of stay in the intensive care unit (ICU). The Richmond Agitation-Sedation Scale (RASS) provides 10 well-defined levels divided into two different segments, including criteria for levels of sedation and agitation. Previous studies of the RASS have shown it to have strong reliability and validity. The aim of this study was to translate the RASS into Swedish and to test the inter-rater reliability of the scale in a Swedish ICU. METHODS: A translation of the RASS from English into Swedish was carried out, including back-translation, critical review and pilot testing. The inter-rater reliability testing was conducted in a general ICU at a university hospital in the south of Sweden, including 15 patients mechanically ventilated and sedated. Forty in-pair assessments using the Swedish version of the RASS were performed and the inter-rater reliability was tested using weighted kappa statistics (linear weighting). RESULT: The translation of the RASS was successful and the Swedish version was found to be satisfactory and applicable in the ICU. When tested for inter-rater reliability, the weighed kappa value was 0.86. CONCLUSION: This study indicates that the Swedish version of the RASS is applicable with good inter-rater reliability, suggesting that the RASS can be useful for sedation assessment of patients mechanically ventilated in Swedish general ICUs.

Mixed micelles of fluorocarbon and hydrocarbon surfactants. A small angle neutron scattering study.

Mixtures of the partly fluorinated cationic surfactant HFDePC (N-(1, 1,2,2-tetrahydroperfluorodecanyl)-pyridinium chloride and deuterated headgroup) with C16TAC, hexadecyl-trimethylammonium chloride, have been investigated using small angle neutron scattering with contrast matching. Earlier results from this system suggested that a demixing occurred, into two coexisting populations of micelles, hydrocarbon-rich and fluorocarbon-rich, respectively. The present results could be explained by one type of mixed micelles with an inhomogeneous distribution of fluorinated and hydrogenated surfactants within the micelles although a demixing cannot be definitely excluded.

Mixed micelles and other structures in the solubilization of bilayer lipid membranes by surfactants.

The solubilization of lipid bilayers by surfactants is accompanied by morphological changes of the bilayer and the emergence of mixed micelles. From a phase equilibrium perspective, the lipid/surfactant/water system is in a two-phase area during the solubilization: a phase containing mixed micelles is in equilibrium with bilayer structures of the lamellar phase. In some cases three phases are present, the single micelle phase replaced by a concentrated and a dilute solution phase. In the case of non-ionic surfactants, the lipid bilayers reach saturation when mixed micelles, often flexible rod-like or thread-like, start to form in the aqueous solution, at a constant chemical potential of the surfactant. The composition of the bilayers also remains fixed during the dissolution. The phase behavior encountered with many charged surfactants is different. The lamellar phase becomes destabilized at a certain content of surfactant in the membrane, and then disintegrates, forming mixed micelles, or a hexagonal phase, or an intermediate phase. Defective bilayer intermediates, such as perforated vesicles, have been found in several systems, mainly with charged surfactants. The perforated membranes, in some systems, go over into thread-like micelles via lace-like structures, often without a clear two-phase region. Intermediates in the form of disks, either micelles or bilayer fragments, have been observed in several cases. Most noteworthy are the planar and circular disks found in systems containing a large fraction of cholesterol in the bilayer. Bile salts are a special class of surfactants that seem to break down the bilayer at low additions. Originally, disk-like mixed micelles were conjectured, with polar membrane lipids building the disk, and the bile salts covering the hydrophobic rim. Later work has shown that flexible cylinders are the dominant intermediates also in these systems, even if the disk-like structures have been re-established as transients in the transformation from mixed micelles to vesicles.

Phase Behavior and Aggregate Structure in Aqueous Mixtures of Sodium Cholate and Glycerol Monooleate.

The phase behavior of the glycerol monooleate (GMO)-sodium cholate-water (or 0.9 wt% NaCl) system has been examined in the solvent-rich part, using small-angle X-ray scattering and conventional methods. Addition of cholate up to 7% of the total amphiphile swells the cubic phase of the binary GMO-water system so that it takes up almost 70% of water in the salt-free case and 55% in salt. With more bile salt the lamellar phase also appears highly swollen (up to 85% in water, 75% in brine). In the salt solution a small isotropic L3-phase region replaces the lamellar phase at a solvent content of about 79%. The lamellar phase can accept only about 0.2 cholate molecule per GMO, in both water and brine, and a phase with globular micelles (L1) follows and dominates the diagram. No threadlike micelles appear in this system. Investigation of the particle structures with cryo-transmission electron microscopy (TEM) in dilute systems (99% solvent) show globular micelles and coexisting vesicles and globular micelles. In the presence of salt, dilution of the L3 phase results in dispersed globular particles with an irregular internal morphology that suggests they are a dispersed L3 phase. These particles coexist with faceted particles having an inner structure giving a hexagonal pattern in projection, suggested to derive from the cubic phase. The cubic phase in the salt-free systems did not give dispersions stable enough for cryo-TEM examination. Copyright 1999 Academic Press.

A Comparison between Hydrophobically End-Capped Poly(ethylene oxide) with Ether and Urethane Bonds

Two models of HEUR (hydrophobically modified ethylene oxide urethane) associative polymers (AP) are compared. The two polymers are polyethyleneoxides simply end-capped with dodecyl groups, one through an ether bond H25C12-O-(CH2CH2O)304-C12H25 (AP14, Mw = 13,700) and the other through an urethane bond H25C12-NHCOO-(CH2CH2O)304-CONH-C12H25 (AP14NCO, Mw is about the same as for AP14). The results indicate that this subtle difference in polymer architecture dramatically influences the initial association and the clouding of the polymer in water, so that AP14NCO starts to aggregate earlier but has a higher clouding temperature. However, at more elevated polymer contents (above 2-3 wt%) the differences in solution behavior and transport dynamics between the two polymers seems less significant. These conclusions are drawn through fluorescence, dynamic light scattering, NMR self-diffusion, turbidimetry, and viscosity measurements. Copyright 1997 Academic Press. Copyright 1997Academic Press

Cryo-TEM of Polyelectrolyte-Micelle Complexes

Cryo-TEM was carried out on samples containing polyelectrolyte-micelle complexes, formed by combining poly(diallyldimethylammonium chloride) (PDADMAC), a strong cationic polyelectrolye, with oppositely charged mixed micelles of sodium dodecyl sulfate (SDS) and nonionic Triton X-100 (TX100), in 0.40 M NaCl. Complexation appears to involve the formation of micelle-rich regions, presumably within the domains of polymer chains, without any particular organization or restructuring of the micelles. At polymer concentrations (CP)

Complexes between cationic liposomes and DNA visualized by cryo-TEM.

The association structures formed by cationic liposomes and DNA-plasmids have been successfully employed as gene carriers in transfection assays. In the present study such complexes was studied by cryo-TEM (cryo-transmission electron microscopy). Cationic liposomes made up by DOPE (dioleoylphosphatidylethanolamine) and various amounts of three different cationic surfactants were investigated. The cryo-TEM analysis suggests that an excess of lipid in terms of charge, leads to entrapment of the DNA molecules between the lamellas in clusters of aggregated multilamellar structures. With increasing amounts of DNA free or loosely bound plasmids were found in the vicinity of the complexes. The importance of the choice of surfactant, as reported from many transfection assays, was not reflected in changes of the type of DNA-vesicle association. A tendency towards polymorphism of the lipid mixtures is reported and its possible implications are discussed.

Two-dimensional diffusion of amphiphiles in phospholipid monolayers at the air-water interface.

Steady-state and time-resolved fluorescence spectroscopy has been used to examine lateral diffusion in dipalmitoyl-L-alpha-phosphatidylcholine (DPPC) and dimyristoyl-L-alpha-phosphatidylcholine (DMPC) monolayers at the air-water interface, by studying the fluorescence quenching of a pyrene-labeled phospholipid (pyrene-DPPE) by two amphiphilic quenchers. Steady-state fluorescence measurements revealed pyrene-DPPE to be homogeneously distributed in the DMPC lipid matrix for all measured surface pressures and only in the liquid-expanded (LE) phase of the DPPC monolayer. Time-resolved fluorescence decays for pyrene-DPPE in DMPC and DPPC (LE phase) in the absence of quencher were best described by a single-exponential function, also suggesting a homogeneous distribution of pyrene-DPPE within the monolayer films. Addition of quencher to the monolayer film produced nonexponential decay behavior, which is adequately described by the continuum theory of diffusion-controlled quenching in a two-dimensional environment. Steady-state fluorescence measurements yielded lateral diffusion coefficients significantly larger than those obtained from time-resolved data. The difference in these values was ascribed to the influence of static quenching in the case of the steady-state measurements. The lateral diffusion coefficients obtained in the DMPC monolayers were found to decrease with increasing surface pressure, reflecting a decrease in monolayer fluidity with compression.

Cationic surfactant/bile salt interaction studied by fluorescence quenching.

Fluorescence quenching measurements were performed on aqueous solutions of the cationic surfactant cetyltrimethylammonium halide (CTAX) and two bile salts, sodium cholate (NaC) and sodium deoxycholate (NaDC), to study the state of aggregation in the mixtures. Pyrene was used as a photoluminiscence probe in the study, and dimethylbenzophenone (DMBP) as the quencher. Analysis of time-resolved decay data with and without quencher using a simple kinetic model gave information of the different aggregation characteristics in the above two cases. Mixed micelles of CTAX/NaC were small and spherical at all compositions, while those of CTAX/NaDC tended to grow from spherical micelles to larger rod-like mixed aggregates at equimolar and close-to-equimolar concentrations. In the latter case more complex kinetics ensues and the fluorescence decays were treated using a generatized model for diffusion-controlled quenching along one dimension for infinitely long rod-like micelles. The mutual diffusion coefficient for the probe-quencherpair was determined.

Diffusion-influenced fluorescence quenching dynamics in one to three dimensions.

The fluorescence decay curves obtained from diffusion-influenced quenching in various spatial dimensions are discussed. The two-dimensional quenching has, because of intractable fitting functions, previously been dealt with only in the completely diffusion-controlled case (corresponding to the Smoluchowski boundary condition). In this paper, an approximation for the two-dimensional (2D)-quenching behavior with the Collins-Kimball boundary condition is presented. The nonlinear least-squares method has been used to analyze simulated decay data. The consequences the choice of an incorrect model has on the final results as well as the possibility to discriminate between different dimensionalities are investigated. Also, some inherent properties of the fitting functions are studied.