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Histamine receptors are densely expressed in the mesencephalic trigeminal nucleus (MesV) and trigeminal motor nucleus. However, little is known about the functional roles of neuronal histamine in controlling oral-motor activity. Thus, using the whole-cell recording technique in brainstem slice preparations from Wistar rats aged between postnatal days 7 and 13, we investigated the effects of histamine on the MesV neurons innervating the masseter muscle spindles and masseter motoneurons (MMNs) that form a reflex arc for the jaw-closing reflex. Bath application of histamine (100 μM) induced membrane depolarization in both MesV neurons and MMNs in the presence of tetrodotoxin, whereas histamine decreased and increased the input resistance in MesV neurons and MMNs, respectively. The effects of histamine on MesV neurons and MMNs were mimicked by an H1 receptor agonist, 2-pyridylethylamine (100 μM). The effects of an H2 receptor agonist, dimaprit (100 μM), on MesV neurons were inconsistent, whereas MMNs were depolarized without changes in the input resistance. An H3 receptor agonist, immethridine (100 μM), also depolarized both MesV neurons and MMNs without changing the input resistance. Histamine reduced the peak amplitude of postsynaptic currents (PSCs) in MMNs evoked by stimulation of the trigeminal motor nerve (5N), which was mimicked by 2-pyridylethylamine but not by dimaprit or immethridine. Moreover, 2-pyridylethylamine increased the failure rate of PSCs evoked by minimal stimulation and the paired-pulse ratio. These results suggest that histaminergic inputs to MesV neurons through H1 receptors are involved in the suppression of the jaw-closing reflex although histamine depolarizes MesV neurons and/or MMNs.

Although it is known that osteoclasts are multinucleated cells that are responsible for bone resorption, the mechanism by which their size is regulated is unclear. We previously reported that an actin-rich superstructure, termed the zipper-like structure, specifically appears during the fusion of large osteoclast-like cells (OCLs). Actin cytoskeleton reorganization in osteoclasts is regulated by a signaling network that includes the macrophage colony-stimulating factor (M-CSF) receptor, a proto-oncogene, Src, and small GTPases. Here, we examined the role of actin reorganization in the multinucleation of OCLs differentiated from RAW 264.7 cells using various pharmacological agents. Jasplakinolide, which stabilizes actin stress fibers, induced the development of small OCLs, and the Src inhibitor SU6656 and the dynamin inhibitor dynasore impaired the maintenance of the podosome belt and the zipper-like structure. These inhibitors decreased the formation of large OCLs but increased the number of small OCLs. M-CSF is known to stimulate osteoclast fusion. M-CSF signaling via Src up-regulated Rac1 activity but down-regulated Rho activity. Rac1 and Rho localized to the center of the zipper-like structure. Rho activator II promoted the formation of small OCLs, whereas the Rho inhibitor Y27632 promoted the generation of large OCLs. These results suggest that the status of the actin cytoskeleton signaling network determines the size of OCLs during cell fusion.

To evaluate the ease of manipulation and durability of 11 commercially available silicone-based resilient denture liners, extrusion force, hardness, weight change, and bond strength were determined. Extrusion force from the cartridge of each material ranged from 0.25 to 1.26 MPa at an extrusion rate of 1 cm/min. Durometer hardness, after set materials were stored in distilled water at 37 degrees C for one day, ranged from A5.9 to A47.7, and after four weeks their values increased by 4.0 to 275%. Bond strength ranged from 1.01 to 2.88 MPa after set materials were stored in distilled water at 37 degrees C for one day, but decreased to 0.59 to 1.99 MPa after 10,000 thermal cycles. These results suggested that except for one material, the rest of the evaluated materials exhibited good handling properties--for example, mixing and spreading of material can be done easily. However, some materials exhibited inadequate durability for clinical service, because hardness increased during storage and/or bond strength decreased after thermal cycling.

The aim of this study was to investigate the setting characteristics of seven silicone-based resilient denture liners during setting. Maximum extrusion force--at the flow rate of 1 cm/min--ranged from 0.183 to 1.26 MPa. Working time and setting time ranged from 2.0 to 5.0 minutes and 1.9 to 6.8 minutes respectively. Consistency as described in JIS T6519-2000 was 32.4 to 46.5 mm. As normal load of 1 kgf was applied to a specimen held between two glass plates after mixing, the time at which film thickness reached up to 1 mm was 4.0 to 13.0 minutes at 23 degrees C and 2.2 to 5.5 minutes at 37 degrees C. These results suggested that the reaction rate of materials used in this study differed from one brand to another. Against this backdrop of differing reaction rates amongst brands, it is therefore vital that a denture liner material's setting characteristics be carefully considered whenever it is used in clinical dentistry to prevent any undesirable or unexpected results and outcome.

Fusarium oxysporum, a soilborne phytopathogenic fungus, produces endo polygalacturonase (PG) which has been suggested to be related to disease development. A polyclonal antibody (PAb) APG1 was prepared against purified PG from F. oxysporum f. sp. lycopersici race 2. By direct tissue-blotted immunobinding assay (DT-IBA) with the antibody, PG production by the pathogen in the stem of the host plant was confirmed. Degenerate primers were designed based on partial amino acid residue information for the PG protein from the fungus: part of the PG-encoding gene was obtained by PCR. Using TAIL-PCR, the complete gene encoding PG was cloned and sequenced. The structural gene comprises 1318 bp coding for 371 amino acids with a putative signal peptide of 22 amino acids, and the open reading frame is interrupted by four introns of 47, 51, 50, and 54bp. The deduced amino acid sequence of the mature protein showed 82.9, 29.0, 27.6, 14.2, and 26.9% homology with those of F. moniliforme PG, Cochliobolus carbonum PGNI, Aspergillus niger PG, A. oryzae PG, and Sclerotinia sclerotiorum PGI, respectively.

Fructanase enzymes hydrolyze the β-2,6 and β-2,1 linkages of levan and inulin fructans, respectively. We analyzed the influence of fructan on the growth of Prevotella intermedia. The growth of P. intermedia was enhanced by addition of inulin, implying that P. intermedia could also use inulin. Based on this finding, we identified and analyzed the genes encoding a putative fructanase (FruA), sugar transporter (FruB), and fructokinase (FruK) in the genome of strain ATCC25611. Transcript analysis by RT-PCR showed that the fruABK genes were co-transcribed as a single mRNA and semi-quantitative analysis confirmed that the fruA gene was induced in response to fructose and inulin. Recombinant FruA and FruK were purified and characterized biochemically. FruA strongly hydrolyzed inulin, with slight degradation of levan via an exo-type mechanism, revealing that FruA is an exo-β-d-fructanase. FruK converted fructose to fructose-6-phosphate in the presence of ATP, confirming that FruK is an ATP-dependent fructokinase. These results suggest that P. intermedia can utilize fructan as a carbon source for growth, and that the fructanase, sugar transporter, and fructokinase proteins we identified are involved in this fructan utilization.

Dental materials that fluoresce affect the reading of the laser fluorescence device DIAGNOdent. Tetracycline hydrochloride (TCH) shows fluorescence and is retained in teeth. The aim of this study was to evaluate the effects of TCH on the DIAGNOdent reading. Filter-paper discs that contained various amounts of TCH were prepared (0.16–10 mg per disc). One-day-old newborn rats were subcutaneously injected with TCH for 29 days, and their mandibles were then removed. The DIAGNOdent values (D-V) of the discs and first molars of the rats were measured before and after they were subjected to ultraviolet irradiation (UV). The D-V of discs containing TCH increased depending on the amount of TCH. The D-Vs of discs with lower amounts of TCH (0.16–1.25 mg) were approximately 10–15, and these values increased to 30–40 under UV. In addition, the D-Vs of molars after UV were twofold greater than those before UV. These results suggest that TCH might affect the readings obtained by DIAGNOdent.

The xenoestrogenic substance bisphenol-A is widely used as a synthetic precursor of resin monomers, such as bisphenol-A diglycidyl methacrylate. Reports describing the release of bisphenol-A from polymerized resin into saliva have aroused considerable concern regarding exposure to xenoestrogen by dental treatment. The purpose of the present study was to demonstrate a reliable methodology of detecting the trace amounts of bisphenol-A in dental materials. Bisphenol-A was separable from bisphenol-A diglycidyl methacrylate, which is often employed as the principal dimethacrylate monomer, by selective extraction with a Sep-Pak C18 cartridge. Using this extraction method in combination with a gas-chromatography mass-spectrometry, we have obtained evidence that all unpolymerized materials used in this study were contaminated with bisphenol-A. Quantitative analysis using a deuterium-labeled compound as an internal standard revealed bisphenol-A contents in commercial dental materials ranging from < 1 microgram/g material to about 20 micrograms/g material. The polymerized dental materials released up to 91.4 ng bisphenol-A/g material into phosphate buffered saline during 24-h incubation. These results indicate that bisphenol-A can be released from dental materials, however the leachable amount would be less than 1/1000 of the reported dose (2 micrograms/kg body weight/day) required for xenoestrogenisity in vivo.

Qualitative and quantitative analyses were conducted on four kinds of resin-modified glass ionomer (RMGI) cement liquids, LC, LC II, LC III (hereinafter referred to as LCs) and VM, using HPLC and laser Raman spectroscopic methods. HPLC revealed that among the RMGI liquids LCs contain 31-32% HEMA (2-Hydroxyethyl methacrylate), and VM contains 18% of the same. The composition of RMGI cement liquids varied significantly between manufacturers. In Raman spectroscopic analyses, the spectra of liquids of various ratios of polyacrylic acid and HEMA were measured, and calculations were made on the peak intensity ratios of C=C stretch vibration to C=O stretch vibration, common in both HEMA and polyacrylic acid. The composition ratio of polycarboxylic acid to HEMA of commercial glass ionomer cements was assessed by the regression curve generated by a combination of peak intensity ratios and composition ratios. In addition, Raman spectroscopy was able to identity the differences in form of the methacryloyloxy group.