5,7-Dihydroxy-3,4,6-trimethoxyflavone inhibits intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 via the Akt and nuclear factor-κB-dependent pathway, leading to suppression of adhesion of monocytes and eosinophils to bronchial epithelial cells.

5,7-Dihydroxy-3',4',6'-trimethoxyflavone (eupatilin), the active pharmacological ingredient from Artemisia asiatica Nakai (Asteraceae), is reported to have a variety of anti-inflammatory properties in intestinal epithelial cells. However, little information is known about the molecular mechanism of eupatilin-induced attenuation of bronchial epithelial inflammation. This study investigates the role of eupatilin in the adhesion of inflammatory cells such as monocytes and eosinophils to bronchial epithelial cells. Stimulation of a human bronchial epithelial cell line (BEAS-2B) with tumour necrosis factor-α (TNF-α) increased the expression of surface adhesion molecules, including intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1), in which eupatilin significantly inhibited the expression of those adhesion molecules in a dose-dependent manner. Eupatilin suppressed the TNF-α-induced activation of IκBα and nuclear factor-κB (NF-κB) signals in BEAS-2B cells. The IκB kinase (IKK) activation was also significantly reduced in eupatilin-pre-treated BEAS-2B and primary normal human bronchial epithelial (NHBE) cells. However, eupatilin did not influence AP-1 activity in TNF-α-stimulated cells. Suppression of NF-κB signalling induced by eupatilin resulted in the inhibition of the expression of adhesion molecules and the adhesion of monocytes and eosinophils to BEAS-2B cells. Furthermore, eupatilin suppressed the phosphorylation of Akt in TNF-α-stimulated BEAS-2B and NHBE cells, leading to down-regulation of NF-κB activation and adhesion molecule expression and finally to suppression of the inflammatory cell adhesion to epithelial cells. These results suggest that eupatilin can inhibit the adhesion of inflammatory cells to bronchial epithelial cells via a signalling pathway, including activation of Akt and NF-κB, as well as expression of adhesion molecules.

Pharmacokinetics of oral amlodipine orotate in vagotomized dogs.

It was reported that gastric motility was delayed and gastric acid secretion was reduced in vagotomized dogs which mimics a low gastric acidity in humans. A delay in gastric motility causes long residence of amlodipine in the stomach. More unionized fractions of amlodipine could exist in less acidic conditions of gastrointestinal fluids, since amlodipine is a weak basic drug with pKa of 8.7. Hence, gastrointestinal absorption of amlodipine is expected to be enhanced and the time to reach a peak plasma concentration of amlodipine (Tmax) is faster in vagotomized dogs. This was proven after oral administration of an amlodipine orotate tablet at a dose of 5 mg as amlodipine in vagotomized dogs. For example, in vagotomized dogs, the total area under the plasma concentration-time curve from time zero to the last measured time, 48 h, in plasma (AUC(0-48 h)) was significantly greater (725 versus 348 ng h/ml) and Tmax was significantly shorter (1.50 versus 5.00 h) than those in dogs without vagotomy.

Multiplexed analysis of polymorphisms in the HLA gene complex using bead array chips.

A novel custom bead array technology is introduced, and it is applied to multiplexed analysis of highly polymorphic human leukocyte antigen (HLA) genetic loci. Our technology combines the construction of probe libraries on color-encoded microparticles (beads) with semiconductor chip processing to produce custom-designed high-density bead array chips in large quantities. Using this novel assay format, two modes of parallel molecular typing of the HLA complex were implemented, namely direct hybridization, illustrated here for class II HLA-DR, and a novel format of on-chip polymerase-mediated primer elongation, illustrated here for class I HLA-A, HLA-B, and HLA-DR using patient and reference cell-line DNA samples. Hybridization-mediated multiplexed analysis of polymorphism method was validated with 142 samples, resulting in 100% concordance with sequence-specific oligonucleotide typing results and a concomitant average of 40% less allele ambiguity. In addition, elongation and hybridization reactions were combined to identify multiple polymorphisms on the same phase of DNA for allele identification.

Domain shapes and patterns: the phenomenology of modulated phases.

A wide variety of two- and three-dimensional physical-chemical systems display domain patterns in equilibrium. The phenomenology of these patterns, and of the shapes of their constituent domains, is reviewed here from a point of view that interprets these patterns as a manifestation of modulated phases. These phases are stabilized by competing interactions and are characterized by periodic spatial variations of the pertinent order parameter, the corresponding modulation period generally displaying a dependence on temperature and other external fields. This simple picture provides a unifying framework to account for striking and substantial similarities revealed in the prevalent "stripe" and "bubble" morphologies as well as in commonly observed, characteristic domain-shape instabilities. Several areas of particular current interest are discussed.

Scale transformation of magnetic "bubble" arrays: coupling of topological disorder and polydispersity.

The evolution of disorder in response to period adaptation in a hexagonal magnetic bubble array is shown to arise from the proliferation of dislocations and to proceed by means of intermediate states of steadily decreasing hexatic order to an amorphous final state. Remarkably, each dislocation core imposes a size adjustment on bubbles decorating its constituent pair of five- and sevenfold coordinated sites. Topological disorder thus induces intrinsic polydispersity and converts the initially unimodal size distribution into a trimodal one. This intimate interplay between geometry and topology provides an explicit mechanism by which structural disorder arises as a result of frustration.

Morphology and local structure in labyrinthine stripe domain phase.

Analysis of globally disordered, nonequilibrium "labyrinthine" stripe-domain patterns in thin ferrimagnetic garnet films has revealed a well-defined local structure containing an oblong polygonal plaquette as the fundamental motif. Two types of labyrinths were found: one having topological defects, the other composed of a single, unbranched, meandering line. These patterns emerge when the labyrinthine state is approached either by heating at constant magnetic field or by demagnetizing from saturation at constant temperature. Size and aspect ratios of the oblong polygonal plaquettes prove to be independent of the choice of these two mutually orthogonal trajectories within the phase diagram, which is surprising in view of the different mechanisms and concomitant topological constraints governing the evolution of disorder. The significance of this unique local structure is discussed in the general context of defectmediated melting of two-dimensional stripe phases.

Lateral diffusion of specific antibodies bound to lipid monolayers on alkylated substrates.

We have measured the lateral mobility of fluoresceinated monoclonal IgG antibodies bound specifically to a spin label lipid hapten in phospholipid monolayers supported on alkylated silicon oxide surfaces. Dimyristoyl phosphatidylcholine and dipalmitoyl phosphatidylcholine monolayers containing 5 mol% of the lipid hapten were transferred by conventional Langmuir-Blodgett techniques onto substrates alkylated with hydrocarbon chains containing 10, 16, and 18 carbon atoms. We show that the diffusion of the bound antibodies depends on their lateral density, the composition of the lipid monolayer, and the nature of lipid coupling to hydrocarbon chains on the alkylated substrate. Antibody diffusion coefficients at low antibody densities are within a factor of 2 of those displayed by the lipid hapten in the absence of the bound antibody. High antibody densities result in reduced antibody mobility, but the lateral diffusion of unbound lipids is unaffected.

Superoxide enhances photobleaching during cellular immune attack against fluorescent lipid monolayer membranes.

Lipid hapten-containing monolayer membranes with bound, anti-hapten antibody molecules serve as model immunological target membranes. Targets with bound-IgG trigger guinea pig macrophages to (a) adhere, (b) spread, (c) release lysosomal enzymes, and (d) increase cyanide-insensitive oxygen consumption. When the target membranes are derivatized with fluorescein, there is a 2-3-fold enhancement in the rate of fluorescein photobleaching in regions of cell-monolayer contact. This effect is due to release of O2- from macrophages, as shown by inhibition with superoxide dismutase and by the fact that enhanced photobleaching is not observed with cells of the RAW264 macrophage line, which undergo responses (a)-(d), but do not release O2- extracellularly. The O2- dependent photobleaching reaction appears to be relatively specific for fluorescein, as it did not occur with two other fluorophores, 4-nitrobenz-2-oxa-1,3-diazole and tetramethyl-rhodamine. Because stimulated neutrophils release large quantities of O2-, the photobleaching of fluorescein-labeled target membranes in response to neutrophils was examined. Monolayer membranes with specifically bound IgG caused neutrophils to adhere and become markedly motile during incubation at 37 degrees C. Like macrophages, neutrophils induced O2- -dependent photobleaching of fluorescein-labeled IgG in regions of cell-monolayer contact. In addition, neutrophils gave rise to a slower, nonphotochemical loss of fluorescence in the same contact regions. The latter effect is apparently due to cleavage of target-bound fluorescent IgG by proteolytic enzymes secreted by the neutrophils in response to the target surface.

X-ray diffraction by phospholipid monolayers on single-crystal silicon substrates.

Monolayers of dipalmitoyl phosphatidylcholine have been transferred from an air-water interface to single-crystal silicon wafers previously alkylated with octadecyltrichlorosilane. By using synchrotron radiation it has been found possible to observe diffraction by single-crystal regions of these monolayers with negligible background scattering from the solid substrate. In one series of experiments, diffraction signals at Bragg spacings of 0.4247 +/- 0.0002 nm and 0.4253 +/- 0.0002 nm were observed. The supported phospholipid monolayer crystals show remarkably high in-plane order: the positional coherence length is at least 500 nm and the orientational order is better than 0.01 degrees . Preliminary temperature scans were carried out. The data reveal the existence of a phase transition at about 65 degrees C.