Lipids, curvature stress, and the action of lipid prodrugs: free fatty acids and lysolipid enhancement of drug transport across liposomal membranes

Molecular shape and its impact on bilayer curvature stress are powerful concepts for describing the effects of lipids and fatty acids on fundamental membrane properties, such as passive permeability and derived properties like drug transport across liposomal membranes. We illustrate these relationships by studying the effects of fatty acids and lysolipids on the permeation of a potent anti-cancer drug, doxorubicin, across the bilayer of a liposome in which the drug is encapsulated. Using a simple fluorescence assay, we have systematically studied the passive permeation of doxorubicin across liposomal membranes in different lipid phases: the solid-ordered phase (DPPC bilayers), the liquid-disordered phase (POPC lipid bilayers), and the liquid-ordered phase induced by high levels of cholesterol (DOPC + cholesterol lipid bilayers). The effect of different free fatty acids (FA) and lysolipids (LL), separately and in combination, on permeability was assessed to elucidate the possible mechanism of phospholipase A₂-triggered release in cancer tissue of liposomal doxorubicin formulations. In all cases, FAs applied separately lead to significant enhancement of permeability, most pronounced in liquid-disordered bilayers and less pronounced in solid and solid-ordered bilayers. LLs applied separately had only a marginal effect on permeability. FA and LL applied in combination lead to a synergistic enhancement of permeability in solid bilayers, whereas in liquid-disordered bilayers, the combined effect suppressed the otherwise strong permeability enhancement due to the FAs.     

Night-Migratory Songbirds Possess a Magnetic Compass in Both Eyes

Previous studies on European robins, Erithacus rubecula, and Australian silvereyes, Zosterops lateralis, had suggested that magnetic compass information is being processed only in the right eye and left brain hemisphere of migratory birds. However, recently it was demonstrated that both garden warblers, Sylvia borin, and European robins have a magnetic compass in both eyes. These results raise the question if the strong lateralization effect observed in earlier experiments might have arisen from artifacts or from differences in experimental conditions rather than reflecting a true all-or-none lateralization of the magnetic compass in European robins. Here we show that (1) European robins having only their left eye open can orient in their seasonally appropriate direction both during autumn and spring, i.e. there are no strong lateralization differences between the outward journey and the way home, that (2) their directional choices are based on the standard inclination compass as they are turned 180° when the inclination is reversed, and that (3) the capability to use the magnetic compass does not depend on monocular learning or intraocular transfer as it is already present in the first tests of the birds with only one eye open.     

MHC molecules protect T cell epitopes against proteolytic destruction.

There is a subtle duality in the role of proteolytic enzymes in Ag processing. They are required to fragment protein Ag ingested by APC. However, prolonged exposure to proteolytic enzymes may lead to a complete degradation of the Ag, leaving nothing for the T cell system to recognize. What ensures that some of the Ag is salvaged? Using a cell-free system we demonstrate that an Ag fragment, once bound to a MHC class II molecule, is effectively protected against proteolytic destruction by cathepsin B and pronase E. The bound fragment, however, can be modified by aminopeptidase N. We suggest that MHC class II molecules play an important regulatory role in the physiologic processing of Ag.     

Genomic analysis of the relationship between gene expression variation and DNA polymorphism in Drosophila simulans

Background

Understanding how DNA sequence polymorphism relates to variation in gene expression is essential to connecting genotypic differences with phenotypic differences among individuals. Addressing this question requires linking population genomic data with gene expression variation.

Results

Using whole genome expression data and recent light shotgun genome sequencing of six Drosophila simulans genotypes, we assessed the relationship between expression variation in males and females and nucleotide polymorphism across thousands of loci. By examining sequence polymorphism in gene features, such as untranslated regions and introns, we find that genes showing greater variation in gene expression between genotypes also have higher levels of sequence polymorphism in many gene features. Accordingly, X-linked genes, which have lower sequence polymorphism levels than autosomal genes, also show less expression variation than autosomal genes. We also find that sex-specifically expressed genes show higher local levels of polymorphism and divergence than both sex-biased and unbiased genes, and that they appear to have simpler regulatory regions.

Conclusion

The gene-feature-based analyses and the X-to-autosome comparisons suggest that sequence polymorphism in cis-acting elements is an important determinant of expression variation. However, this relationship varies among the different categories of sex-biased expression, and trans factors might contribute more to male-specific gene expression than cis effects. Our analysis of sex-specific gene expression also shows that female-specific genes have been overlooked in analyses that only point to male-biased genes as having unusual patterns of evolution and that studies of sexually dimorphic traits need to recognize that the relationship between genetic and expression variation at these traits is different from the genome as a whole.     

Hitch-hiking parasite: a dark horse may be the real rider

Many parasites engaged in complex life cycles manipulate their hosts in a way that facilitates transmission between hosts. Recently, a new category of parasites (hitch-hikers) has been identified that seem to exploit the manipulating effort of other parasites with similar life cycle by preferentially infecting hosts already manipulated. Thomas et al. (Evolution 51 (1997) 1316) showed that the digenean trematodes Microphallus papillorobustus (the manipulator) and Maritrema subdolum (the hitch-hiker) were positively associated in field samples of gammarid amphipods (the intermediate host), and that the behaviour of Maritrema subdolum rendered it more likely to infect manipulated amphipods than those uninfected by M. papillorobustus. Here I provide experimental evidence demonstrating that M. subdolum is unlikely to be a hitch-hiker in the mentioned system, whereas the lucky candidate rather is the closely related but little known species, Microphallidae sp. no. 15 (Parassitologia 22 (1980) 1). As opposed to the latter species, Maritrema subdolum does not express the appropriate cercarial behaviour for hitch-hiking.     

Orientation and conformation of a lipase at an interface studied by molecular dynamics simulations

Electron density profiles calculated from molecular dynamics trajectories are used to deduce the orientation and conformation of Thermomyces lanuginosa lipase and a mutant adsorbed at an air-water interface. It is demonstrated that the profiles display distinct fine structures, which uniquely characterize enzyme orientation and conformation. The density profiles are, on the nanosecond timescale, determined by the average enzyme conformation. We outline a computational scheme that from a single molecular dynamics trajectory allows for extraction of electron density profiles referring to different orientations of the lipase relative to an implicit interface. Profiles calculated for the inactive and active conformations of the lipase are compared with experimental electron density profiles measured by x-ray reflectivity for the lipase adsorbed at an air-water interface. The experimental profiles contain less fine structural information than the calculated profiles because the resolution of the experiment is limited by the intrinsic surface roughness of water. Least squares fits of the calculated profiles to the experimental profiles provide areas per adsorbed enzyme and suggest that Thermomyces lanuginosa lipase adsorbs to the air-water interface in a semiopen conformation with the lid oriented away from the interface.     

The Hydrobia ulvae-Maritrema subdolum association: influence of temperature,salinity, light,water-pressure and secondary host exudates on cercarial emergence and longevity

The effects of environmental factors and exudates from the amphipod Corophium volutator on the emergence of Maritrema subdolum cercariae (Digenea: Microphallidae) from the snail Hydrobia ulvae were investigated in the laboratory. Increasing the temperature (15 to 25 degrees C) caused an overall 11-fold increase in emergence rate under varying salinities (24 to 36 per thousand). The effect of salinity depended on the experimental temperature. Emergence increased with increasing salinity at higher temperatures, but decreased with increasing salinity at 15 degrees C.Whereas the different levels of salinity had no effect, increasing the temperature significantly reduced the life span of cercariae. In comparison with complete darkness, light caused a two-fold increase in emergence, whereas an increment of the water pressure from 1.0 to 1.3 ATM (corresponding to 0 and 3 m of depth) left the shedding rate unaffected. Unidentified exudates from the second intermediate host, C. volutator, significantly depressed the cercarial emergence rate. The main transmission window of M. subdolum seems to occur during low water in tidal pools where light levels are high and solar radiation rapidly elevates the water temperature, as well as salinity through evaporation. The consequence of such a transmission strategy is discussed in relation to the impact of M. subdolum on the population dynamics of the second intermediate host.     

The Hydrobia ulvae-Maritrema subdolum association: cercarial emergence controlled by host activity

The release of Maritrema subdolum cercariae (Digenea: Microphallidae) from the marine mud snail Hydrobia ulvae is significantly affected by temperature, salinity, light and exudates from the second intermediate amphipod host. Based on (i) previously published data on temperature-salinity dependent H. ulvae activity, (ii) new experimental data on H. ulvae activity in light and darkness as well as in the presence and absence of host exudates, and (iii) the cercarial emergence rate from free moving snails and snails prevented from crawling, the present analysis indicates that emergence of M. subdolum larvae is regulated mainly by host activity as the ultimate factor for release. The adaptive significance of such an emergence strategy is emphasized.     

Freeze/thaw effects on lipid-bilayer vesicles investigated by differential scanning calorimetry

Differential scanning calorimetry (DSC) has been used to study the effects of repeated freezing and thawing on dipalmitoylphosphatidylcholine (DPPC) vesicles. Aqueous suspensions of both multilamellar vesicles (MLVs) and large unilamellar vesicles (LUVs) were cycled between -37 and 8 degrees C, and for each thawing event, the enthalpy of ice-melting was measured. In the case of MLVs, the enthalpy increased each time the vesicles were thawed until a steady state was attained. In contrast, the enthalpies measured for LUV suspensions were independent of the number of previous thawing events. It was concluded that MLVs in terms of freezing characteristics contain two pools of water, namely bulk water and interlamellar water. Interlamellar water does not freeze under the conditions employed in the present study, and the MLVs therefore experience freeze-induced dehydration, which is the reason for the observed increase in ice-melting enthalpy. Furthermore, the thermodynamic results suggest that the osmotic stress resulting from the freeze-induced dehydration changes the lamellarity of the MLVs.