Cellulose dimethylphenylcarbamate-bonded carbon-clad zirconia for chiral separation in high performance liquid chromatography.

Porous zirconia particles are very robust material and have received considerable attention as a stationary phase support for HPLC. We prepared cellulose dimethylphenylcarbamate-bonded carbon-clad zirconia (CDMPCCZ) as a chiral stationary phase (CSP) for separation of enantiomers of a set of 14 racemic compounds in normal phase (NP) and reversed-phase (RP) liquid chromatography. Retention and enantioselectivity on CDMPCCZ were compared to those on CDMPC-coated zirconia (CDMPCZ) to see how the change in immobilization method of the chiral selector affects the retention and chiral selectivity. In NPLC, retention was longer and the number of resolved racemates was smaller on CDMPCCZ than on CDMPCZ. However, chiral selectivity factors for some resolved racemates were better on CDMPCCZ than on CDMPCZ. The longer retention on CDMPCCZ is likely due to strong, non-chiral discriminating interactions with the carbon layer on CDMPCZ. In RPLC only two racemates were resolved on CDMPCCZ, but retention times were shorter than, and resolutions were comparable to, those in NPLC, indicating a potential for improving chromatographic performance of the CDMPCCZ column in RPLC with optimized column preparation and separation conditions.     

Validation of a method for predicting the precision, limit of detection and range of quantitation in competitive ELISA.

The mathematical model for predicting the precision, limit of detection (LOD) and range of quantitation (ROQ) in a competitive enzyme-linked immunosorbent assay (ELISA) proposed by Hayashi et al. (Anal. Chem., 2004, 76, 1295) was validated. The model describes the relative standard deviation (RSD) of concentration estimates by the RSDs of pipetting volumes of analyte, enzyme-conjugated antigen, antibody and substrate solutions, and the standard deviation (SD) of inherent absorbances between the wells in an ELISA plate. For 6 kinds of direct competitive ELISA kits, the LOD and ROQ predicted by the model agreed well with those obtained by experiments with real samples. It was also confirmed that the model is applicable to the prediction of uncertainty that depends on the pipetting error of the viscous antiserum solution. The model was demonstrated to be useful for estimating the LOD and ROQ of competitive ELISA.     

Mesomorphic properties of orthopalladated complexes of laterally substituted azobenzene derivatives, 2-alkoxycarbonyl-4-(4-ethoxyphenylazo)phenyl 4-methoxybenzoates

Homologous series of 2-alkoxycarbonyl-4-(4-ethoxyphenylazo)phenyl 4-methoxybenzoates (L) and their orthopalladated μ-chloro dimers (Pd₂Cl₂L₂) have been synthesized and their thermotropic phase transition behaviour has been characterized. The ligands, L, are three-ring nematogens with their central rings substituted in the 2-position by n-alkyl chains of varied length. The thermal stability of the nematic phases decreases rapidly with increasing side chain length and then levels off as the series is ascended. In every case the mesomorphic phase appears only over a small temperature range (less than 20° C). The orthopalladated species also proved to be nematogenic. Melting and clearing points rise on the average by 87° C and 126° C, respectively, and thus the temperature ranges of the nematic phases are enlarged considerably upon metallation. In relation to the observed stabilization of the nematic phases, particularly of higher homologues, a lath-like molecular structure, in which the side chains of the ligands fill up the clefts of the core part, has been deduced for a Pd2Cl2L2 homologue from its solution ¹H-NMR spectrum.     

Illuminating Molecular Processes in Living Cells

Lately, scientists have explored approaches to developing fluorescent and/or bioluminescent indicators to pinpoint cellular processes in single living cells. These analytical methods have become a key technology for visualizing and detecting what was otherwise unseen in live cells. The target signaling included second messengers, protein phosphorylations, protein–protein interactions, and protein localizations.     

Electrical conductance of solutions of lithium nitrate in 1,3-diaminopropane

Electrical conductances and densities of the LiNO₃-1,3-diaminopropane system have been measured as a function of composition and temperature. The results suggest that the solvent molecules may act as bridges between Li⁺ ions at higher concentration of LiNO₃.     

High-quality hydroxyapatite coating on biocompatible materials by laser-assisted laser ablation method

A new method for high-quality hydroxyapatite (HAp) coating is developed, the laser-assisted laser ablation method. In this method, two lasers are used. One is used for ablation of a HAp target. The other, the assist laser, is used to irradiate a Ti substrate surface. The effects of the assist laser irradiation are to anneal the HAp coating and to improve its adhesive strength to the Ti substrate. The quality of the HAp coating depends on the delay time of the assist-laser irradiation. HAp coatings obtained at a delay time of 10 microseconds or more are more amorphous. It was confirmed that the amorphous part of a coating dissolves in simulated body fluid, while the crystalline part does not. The value of the Ca/P ratio in a coating obtained at a delay time of a few microseconds is close to the stoichiometric value. PACS 81.16.Mk     

Keratinized epithelial transport of beta-blocking agents. I. Relationship between physicochemical properties of drugs and the flux across rat skin and hamster cheek pouch.

The maximum fluxes (Jmax) of beta-blockers through keratinized membranes were determined in vitro and compared with their physiochemical parameters such as lipophilicity (log k'0) and melting point (mp). Rat abdominal skin and hamster cheek pouch mucosa were used as the model membranes. Propranolol, metoprolol, timolol, pindolol, nadolol and agenolol were used as beta-blockers with a variety of physicochemical characters. Linear relations of Jmax with either log k'0 or mp were observed both in intact rat skin and in intact hamster cheek pouch, suggesting that the lipophilicity and thermodynamic activity of a drug in the crystal state primarily affect the drug's permeation through these membranes. However, the slope, dJmax/d(log k'0), for cheek pouch mucosa was greater than that for rat skin, corresponding to the lack of appendigeal shunt pathways in cheek pouch. Penetration studies using the delipidized membranes and the isolated stratum corneum sheet of hamster cheek pouch mucosa clarified that the primary rate-limiting barrier function might exist in the lipid layer of the stratum corneum. Jmax values for the tape-stripped and delipidized skins correlated with both the solubilities of drugs in the vehicle and with the mp, suggesting the polar porous characteristics of both model membranes. However, a theoretical approach confirmed that the contribution of an intracellular or aqueous pore route in the intact membrane to the permeation of drugs with positive lipophilic indexes is negligible.     

Simultaneous Measurement of Droplet Size and Three-Components of Velocity in Spray

The combination of the optical diagnostics and image processing technique has produced various methods of practical measurement in engineering. The measurement of size and velocity of droplets in fuel spray is one typical example. In general, the spray is a complicated and three-dimensional two-phase flow; therefore multi-dimensional field measurement of the spray field is indispensable. In the present paper, a technique of simultaneous measurement of droplet size and three-components of velocity is proposed and applied to spray issuing from the swirl nozzle as a practical application. The focused image of glare points is captured by a stereoscopic arrangement, and droplet size and the three-components of velocity are evaluated from the doublet image. Fundamental optical arrangements to capture the image of the glare points with a stereo view are established, and the feasibility and the accuracy of the technique are confirmed. © 2004 The Optical Society of Japan     

Genetically Encoded Bioluminescent Indicator for ERK2 Dimer in Living Cells

In this study, a genetically encoded bioluminescent indicator for ERK2 dimer was developed with the split Renilla luciferase complementation method, in which the formation of ERK2 dimer induces a spontaneous emission of bioluminescence in living cells. In response to extracellular stimuli, such as epidermal growth factor (EGF) or 17β‐estradiol (E2), extracellular signal‐regulated kinase 2 (ERK2) is phosphorylated by its upstream kinase MEK, and also phosphorylates its substrates in various regions of the cell, including the nucleus. Phosphorylated ERK2 is led to form its dimer, thereby transporting itself into the nucleus. We demonstrated with the indicator that stimulation with EGF or E2 induces the formation of ERK2 dimer in living MCF‐7 cells. The dynamics of this dimer formation was examined and discussed.