Advanced drug delivery systems that target the vascular endothelium

Targeted drug delivery to endothelial cells lining the vascular lumen will provide effective, precise and safe therapeutic interventions for treatment of diverse disease conditions. Rational design of such drug delivery systems (DDS) includes the following intertwined tasks: 1) selection of proper target determinants on endothelial surfaces, such as cell adhesion molecules, ectopeptidases, or caveolar antigens; 2) production of affinity ligands useful for targeting, such as affinity peptides, antibodies, or their fragments; 3) selection and adopting of suitable delivery vehicles (such as liposomes or polymer nanocarriers); and 4) formulation of DDS with optimal targeting and therapeutic features. Important therapeutic features of DDS include: 1) sufficient targeting effectiveness, circulation time, and safety (i.e., lack of systemic and local adverse effects); 2) precise subcellular localization of drugs targeted to endothelial cells; and 3) adequate amplitude, kinetics, and duration of effects. This review utilizes examples of DDS-mediated interventions in vascular inflammation, oxidative stress, and thrombosis and analyzes them in an attempt to create design parameters that best regulate the pharmacological and therapeutic features of DDS that target endothelial cells.     

Interhemisphere differences of extrastriate cortical activation during attention and selection of lateralized visual stimuli in humans

The cortical activation was estimated by event-related potentials (ERPs) methods during selection tasks of lateralized visual stimuli in right and left hemi-fields requiring different forms of attention: 1. Attention of a stimuli form, 2. Attention of a stimuli position, 3. Combined attention of form and position. ERPs were recorded in 15 young healthy adults in 6 leads: P3, P4, T3, T4, T5, T6 and endogenous ERPs components: CNV (contingent negative variation), N1, P3 and complex [N1--P3]. The differences between ERPs at contra- and ipsilateral stimuli in the right and left hemispheres were considered as indices of asymmetry. The asymmetry was revealed in right hemisphere in all kinds of attention forms. The level (amplitude) of right-side asymmetry was depended on the level of attention: The significant relation between the right-side asymmetry and subjects' reaction time was also revealed. It is proposed that such an asymmetry is the evidence of better spatial differentiation of visual stimuli in right hemisphere in humans.     

Early-glycation of apolipoprotein E: effect on its binding to LDL receptor,scavenger receptor A and heparan sulfates

Glycation is responsible for disruption of lipoprotein functions leading to the development of atherosclerosis in diabetes. The effects of apolipoprotein E (apoE) glycation were investigated with respect to its interaction with receptors. The interaction of apoE with the low density lipoprotein receptor (LDL-R) and scavenger receptor A (SR-A) was measured by competition experiments performed using, respectively, on a human fibroblast cell line ¹²⁵I-LDL, and on a murine macrophage cell line (J774) ¹²⁵I-acetylated LDL, and unlabeled apoE/phospholipid complexes. Glycated apoE binding to heparin and heparan sulfates (HS) was assessed by surface plasmon resonance (SPR) technology. Site-directed mutagenesis was then performed on Lys-75, the major glycation site of the protein. The prepared mutant protein proved to be useful as a tool to study the role of Lys-75 in apoE glycation. The findings showed that, although glycation has no effect on apoE binding either to the LDL-R or to SR-A, it impairs its binding to immobilized heparin and HS. The glycation of Lys-75 was found to be proceed rapidly and contributed significantly to total protein glycation. We propose that, in the case of diabetes, glycation may lead to the atherogenicity of apoE-containing lipoproteins disturbing their uptake via the HS proteoglycan pathway.     

Basal ganglia and behavior

A comparative analysis of the most well known hypotheses of basal ganglia participation in organisation and control of behavioural functions, is presented.     

Apolipoprotein E activates Akt pathway in neuro-2a in an isoform-specific manner

Apolipoprotein E (apoE) is a ligand for members of the low density lipoprotein (LDL) receptor family, receptors highly expressed in neurons. A study of one of the mechanisms by which apoE might affect neuronal cell metabolism is reported herein. ApoE can induce Akt/protein kinase B phosphorylation in Neuro-2a via two different pathways. Both pathways are mediated by phosphatidylinositol 3-kinase and cAMP-dependent protein kinase. The first pathway is stimulated by apoE3 and E4, but not by E2, after a 1-h incubation. The process requires the binding of apoE to the heparan sulfate proteoglycan/LDL receptor-related protein complex. The second pathway is activated after a 2-h incubation of the cells, in another isoform-dependent manner (E2 = E3 dbl greater-than sign E4) and is mediated by calcium. Our results suggest that apoE might affect cell metabolism and survival in neurons in an isoform-specific manner by inducing novel signaling pathways.     

Activation of the human cortex during visual attention and selection

Cortical activation in visual discrimination tasks was estimated by measurement of the CNV (contingent negative variation) and N1-P3 components of visual ERPs in frontal, parietal, occipital and temporal leads recorded in 18 young healthy adults. In all investigated tasks, the maximal values of CNV and ERPa were observed in parietal regions. The estimation of cortical readiness state (CNV) is quite a useful procedure in the attention tasks because amplitude and stability of ERPs depend on preceding cortical excitability. The prevalence of parietal activation in visual attention tasks may be considered as the dominance of occipito-parietal way (stream) in human visual attention system.     

Supermolecular organization of apolipoprotein E in solution]

The aggregation behaviour and stability in solution of apolipoprotein E (apoE) isolated from human blood plasma very low density lipoproteins were investigated. The equilibrium denaturation of fluorescein-labeled apoE by guanidine-hydrochloride determined by anisotropy and overall intensity of fluorescence, shift of the emission spectrum maximum and gel-chromatographic behaviour was characterized by reversibility, biphasity, apoE concentration dependence and the existence of native structure of the apoE monomer. The contribution of the long-living component to the kinetic dependence of fluorescence anisotropy in the presence of the 6 M denaturant increased with an increase in apoE concentration. The data obtained fit into the following scheme: oligomer (upon aging of the preparation) in equilibrium tetramer in equilibrium native monomer in equilibrium denaturated monomer. The presence in the tetrameric structure of apoE of two domains is postulated; one of those is formed by lipid-binding fragments during aggregation of individual molecules of apoE. Monoclonal antibody 3D12F11 (subclass IgG1) showed a high affinity for the apoE (Kd = 3.5 +/- 0.5 nM) without any effect on the apoprotein binding to heparin-Sepharose and apoE-induced destruction of dipalmitoylphosphatidylcholine liposomes. It is concluded that the 3D12F11 epitope is localized outside heparin- and lipid-binding sites of the apoprotein molecule.     

Features of the organization of biopotentials of a cerebral cortex and visceral status at the person at neurotic depression

Researches are spent on 20 patients with various clinical variants of neurotic depression. The regional organization of bioelectric activity of a brain by a method cross-correlation and coherent analysises was studied. Vegetovisceral status was studied by auricular criotest (measuring of cold sensibility of auricular points). It is shown, that the clinical picture of neurotic depression finds reflectance in frame of regional organization EEG. Regional organization EEG is modified depending on a degree of manifestation of the most neurotic depression and concomitant syndromes of alarm and an asthenia. In bunch with a depressive syndrome without concomitant asthenic and disturbing exhibitings the maximum changes are taped in the right frontotemporal range--left posttemporal ranges. In bunch where along with depression the alarm--depression cross-correlation and coherent communications frontotemporal ranges of both hemispheres was taped, strongly pronounced rising cross-correlation attitudes in right occipital ranges. In bunch where depressive and disturbing syndromes were combined with the expressed asthenic exhibitings, depression cross-correlation and coherent communications in frontotemporal ranges of both hemispheres was observed. The clinic of neurotic infringements finds reflection in a specific picture of variations of the spatial organization of electric activity of a brain and in variations of parameters of the vegetovisceral status. Realization of negative emotional conditions at the person is accompanied by variations visceral functions. Thus variations in the central brain structures cover zones of representation of emotional reactions and the zones connected with cortical representation visceral functions. The minimal central regulation, even insignificant central variations can cause vegetovisceral dysfunctions.