Dentin and pulp sense cold stimulus.

Dentin hypersensitivity is a common symptom, and recent convergent evidences have reported transient receptor potential (TRP) channels in odontoblasts act as mechanical and thermal molecular sensor, which detect stimulation applied on the exposed dentin surface, to drive multiple odontoblastic cellular functions, such as sensory transduction and/or dentin formation. In the present study, we confirmed expression of TRP melastatin subfamily member-8 (TRPM8) channels in primary cultured cells derived from human dental pulp cells (HPCs) and mouse odontoblast-lineage cells (OLCs) as well as in dentin matrix protein-1 (DMP-1) and dentin sialoprotein (DSP) positive acutely isolated rat odontoblasts from dental pulp tissue slice culture by immunohistochemical analyses. In addition, we detected TRPM8 channel expression on HPCs and OLCs by RT-PCR and Western blotting analyses. These results indicated that both odontoblasts and dental pulp cells express TRPM8 channels in rat, mouse and human, and therefore we hypothesize they may contribute as cold sensor in tooth.

Effects of postnatal dexamethasone treatment on mRNA expression profiles of genes related to alveolar development in an emphysema model in mice.

Emphysema can be induced in animals by postnatal treatment with dexamethasone (Dex) and such models have been widely used for various research. However, it is not clear what are the effects of Dex on assembly of alveolar elastic fibers in the emphysema model in mice. This study compared the expression profile of genes related to alveolar development between Dex treated and control mice during the treatment from postnatal day 3 (P3) to P14 with a 2-day break. From morphological observation of lung sections on P42, we confirmed the induction of emphysema in the treated mice. The mRNA expression level of fibrillin-1, which consists of microfibrils as a scaffold to form elastic fibers, and fibulin-5, which is a key protein reinforcing the fibers, reached maximum on P7 in control mice. However, in the Dex group, expression levels both types of mRNA were much lower with no clear expression peak. On the other hand, mRNA expression of tropoelastin, the main component in elastic fibers, reached maximum on P5 in the Dex group, which was 9 days earlier than in the control group. At this time, the amount of microfibrils might not be enough for tropoelastin to be deposited completely in Dex treated mice. This imbalance in the expression of tropoelastin and microfibril might interfere with the efficient formation of elastic fibers.

Hyperosmotic stress induces cell death in an odontoblast-lineage cell line.

INTRODUCTION: Osmotic stress is one of the stimulations related to dental pain caused by caries or dentin hypersensitivity. The mechanism of osmotic-induced dental pain is not completely understood. The purpose of this study was to examine the responses of odontoblasts under sucrose-induced hyperosmotic stress. METHODS: We used an odontoblast-lineage cell (OLC) line in our experiments. OLCs were stimulated with sucrose to produce hyperosmotic stress. The expressions of dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP 1) were detected by using reverse transcriptase polymerase chain reaction assay. The cell viability of OLCs was detected by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium assay. The responses accompanied with cell death were detected by using 4-6-diamidino-2-phenylindole staining, Western blotting of caspase-3, and annexin V assay. The expression of mitogen-activated protein kinases (MAPKs) was detected by using Western blot analysis. RESULTS: DSPP and DMP 1 were not affected by hyperosmotic stress in OLCs. Cell viability decreased over 700 mOsm for 3 hours of cell culture. The shapes of cells and nuclei became irregular and vacuolar under hyperosmotic stress. The expression of cleaved caspase-3 was increased after treatment with hyperosmotic stress. Some propidium iodide-positive cells were detected in flow cytometry analysis. Phosphorylation of 3 MAPKs was induced by hyperosmotic stress. Inhibitors of 3 MAPKs inhibited the hyperosmotic stress-induced decline in cell viability at 500 and 700 mOsm. CONCLUSIONS: Hyperosmotic stress induces cell death of OLCs with sucrose through a MAPK pathway.

Enantioseparation using urea- and imide-bearing chitosan phenylcarbamate derivatives as chiral stationary phases for high-performance liquid chromatography.

Completely deacetylated chitosan was prepared by the treatment of commercial chitosan with 50% aqueous NaOH, and then derivatized into several new chitosan phenylcarbamate derivatives having a urea and an imide moiety at the 2-position of the glucosamine ring by the reaction with isocyanate and phthalic anhydride/isocyanate, respectively. The chitosan derivatives were coated on macroporous silica gel and evaluated as chiral stationary phases (CSPs) for high-performance liquid chromatography. The chiral recognition ability of the chitosan derivative was improved using the completely deacetylated chitosan. Among the novel chitosan derivatives, the 3,5-dimethyl-, 3,5-dichloro-, and 3,4-dichlorophenylcarbamate derivatives were found to possess relatively high chiral resolution abilities. The CSPs based on the chitosan phenylcarbamate-urea and -imide derivatives were stable in the presence of chloroform and ethyl acetate as a component of the eluents, and some racemates were better resolved by such eluents. The dichlorophenylcarbamate-imide derivatives showed a high chiral recognition for metal acetylacetonate complexes. The enantiomerization of Al(acac)3 was performed on the chitosan 3,5-dichlorophenylcarbamate-imide derivative CSP and the resulting chromatogram showed a 26% (+)-isomer enrichment.

Syntheses and properties of enantiomerically pure higher (n > or = 7) [n-2]triangulanedimethanols and sigma-[n]helicenes.

(P)-(+)-Hexaspiro[2.0.0.0. 0.0.2.1.1.1.1.1]pentadecane [(P)-17] as well as (M)-(-)- and (P)-(+)-octaspiro[2.0.0.0.0.0.0.0.2.1.1.1.1.1.1.1]nonadecanes [(M)- and (P)-25]-enantiomerically pure unbranched [7]- and [9]triangulanes-have been prepared starting from racemic THP-protected (methylenecyclopropyl)methanol 6. The relative configurations of all important intermediates as well as the absolute configurations of the key intermediates were established by X-ray crystal structure analyses. This new convergent approach to enantiomerically pure linear [n]triangulanes for n=7, 9 was also tested in two variants towards [15]triangulane. Some of the most prominent and unexpected features of the newly prepared compounds are the remarkable modes of self-assembly of the diols (P)-14, (E)-(3S,3'S,4S,4'S,5R,5'R)-21, (P)-(+)-22, and (E)-31 in the solid state through frameworks of intermolecular hydrogen bonds leading to, depending on the respective structure, nanotube- [(P)-14, (P)-(+)-22, and (E)-31], honeycomb-like structures [(E)-(3S,3'S,4S,4'S,5R,5'R)-21] or a supramolecular double helix [(P)-(+)- and (M)-(-)-22]. Liquid crystalline properties of the esters and ethers of the diols (P)-14, (P)-, and (M)-22 have also been tested. Although all of these [n]triangulanes have no chromophore which would lead to significant absorptions above 200 nm, they exhibit surprisingly high specific rotations even at 589 nm with [alpha](20)(D)=+672.9 (c=0.814 in CHCl(3)) for (P)-(+)-17, +909.9 (c=0.96 in CHCl(3)) for (P)-(+)-25, -890.5 (c=1.01 in CHCl(3)) for (M)-(-)-25, and -1302.5 (c=0.36 in CHCl(3)) for (M)-(-)-39, and the specific rotations increase drastically on going to shorter wavelengths. This outstanding rotatory power is in line with their rather rigid helical arrangement of sigma bonds, and accordingly these helically shaped unbranched [n]triangulanes may be termed "sigma-[n]helicenes", as they represent the sigma-bond analogues of the aromatic pi-[n]helicenes. Density functional theory (DFT) computations at the B3 LYP/6-31+G(d,p) level of theory for the geometry optimization and time-dependent DFT for determining optical rotations with a triplet-zeta basis set (B3 LYP/TZVP) reproduce the optical rotatory dispersions (ORD) very well for the lower members (n=4, 5) of the sigma-[n]helicenes. For the higher ones (n=7, 9, 15) the computed specific rotations turn out increasingly larger than the experimental values. The remarkable increase of the specific rotation with an increasing number of three-membered rings is proportional neither to the molecular weight nor to the number of cyclopropane rings in these sigma-[n]helicenes.