Catalytic Dealkylative Synthesis of Cyclic Carbamates and Ureas via Hydrogen Atom Transfer and Radical-Polar Crossover.

Guided by the transition-metal hydrogen atom transfer and radical-polar crossover concepts, we developed a functional-group-tolerant and scalable method for the synthesis of cyclic carbamates and ureas, which are found in the structures of bioactive compounds. This method provides not only a common five-membered ring but also six-to-eight-membered ring products. The reaction proceeds through the intramolecular displacement of an alkylcobalt(IV) intermediate and dealkylation by 2,4,6-collidine; the activation energies of these steps were calculated by DFT.

Inhibition of RANKL-dependent cellular fusion in pre-osteoclasts by amiloride and a NHE10-specific monoclonal antibody.

The functions of Na(+) /H(+) exchangers (NHEs) during osteoclastic differentiation were investigated using the NHE inhibitor amiloride and a monoclonal antibody (MAb). Compared with sRANKL-stimulated control cells, amiloride decreased the number of large TRAP-positive osteoclast cells (OCs) with ≥10 nuclei and increased the number of small TRAP-positive OCs with ≤10 nuclei during sRANKL-dependent osteoclastic differentiation of RAW264.7 cells. NHE10 mRNA expression and OC differentiation markers were increased by sRANKL stimulation in dose- and time-dependent manners. NHEs 1-9 mRNA expression was not increased by sRANKL stimulation. Similar to amiloride, a rat anti-mouse NHE10 MAb (clone 6B11) decreased the number of large TRAP-positive OCs, but increased the number of small TRAP-positive OCs. These findings suggested that inhibition of NHEs by amiloride or an anti-NHE10 MAb prevented sRANKL-promoted cellular fusion. The anti-NHE10 MAb has the potential for use as an effective inhibitor of bone resorption for targeted bone disease therapy.

Low-loss high-speed silicon IQ modulator for QPSK/DQPSK in C and L bands.

A low-loss high-speed silicon in-phase (I) quadrature (Q) modulator is designed, fabricated and characterized. The fabricated IQ modulator has a low passive optical loss of 9 dB in C and L bands. Using the modulator, differential quadrature phase-shift keying (DQPSK) transmission at 44.6 Gb/s with differential detection is confirmed with an optical signal-to-noise ratio (OSNR) of 16.3 dB for a bit error rate (BER) of 10(-3) and a dispersion tolerance of -96 to 107 ps/nm. Moreover, in digital coherent detection, quadrature phase-shift keying (QPSK) up to 64 Gb/s are achieved with an OSNR of 11.6-11.8 dB for a BER of 10(-2) at 1530, 1550, and 1610 nm.

The usefulness of an autologous blood clot combined with gelatin for regeneration of periodontal tissue.

Several growth factors have been used in tissue regeneration therapy. Here we describe the use of an autologous blood clot combined with gelatin for teeth affected by severe periodontitis and vertical root fracture treated using three oral surgical procedures: periodontal flap surgery, intentional tooth replantation (IR), and tooth autotransplantation. Treatment with a blood clot improved the condition of the periodontal tissue, including reduction of pocket depth. Radiographical images demonstrated no evidence of ankylosis, and revealed the presence of alveolar bone regeneration. Our successful clinical outcome suggests that use of an autologous blood clot combined with gelatin is clinically effective for regeneration of lost periodontal tissue.

11-Gb/s 80-km transmission performance of zero-chirp silicon Mach-Zehnder modulator.

The 11-Gbps 80-km transmission performance of a zero-chirp silicon Mach-Zehnder modulator has been characterized. The zero-chirp characteristic of the silicon modulator is confirmed in the constellation measurement, and gives high tolerance both for positive and negative chromatic dispersion. A low-dispersion-penalty transmission up to 80 km using the 11-Gbps non return-to-zero on-off-keying format is confirmed via bit-error-rate measurements with a performance comparable to that of a commercial lithium-niobate modulator. The dispersion tolerance at 2-dB power penalty for a bit-error-rate of 10(-3) is more than ± 950 ps/nm. Further, 22.3-Gbps binary phase-shift-keying is demonstrated, and the back-to-back power penalty with respect to the lithium-niobate modulator is less than 0.5 dB.

The usefulness of three-dimensional imaging for prognostication in cases of intentional tooth replantation.

This paper describes the utility of three-dimensional (3D) images obtained with cone beam computed tomography (CBCT) for prediction of successful clinical outcome in two cases of intentional tooth replantation (IR). IR was performed for teeth affected by vertical root fracture and root perforation with local application of blood clot and oxy-tetracycline antibiotic. High-resolution 3D images demonstrated no evidence of ankylosis, but did reveal the presence of alveolar bone regeneration, suggesting a good long-term prognosis. Our observations in these cases suggested that local application of the above two materials might help to induce the regeneration of lost periodontal tissues in IR.

Fabrication of solid and hollow carbonate apatite microspheres as bone substitutes using calcite microspheres as a precursor.

Spherical carbonate apatite (CO3Ap) microspheres approximately 1 mm in diameter were fabricated by granulation of calcium hydroxide around a core followed by carbonation and phosphatization through dissolution-precipitation reaction. CO3Ap microspheres with high uniformity could not be achieved without using a core. Solid CO3Ap microspheres were obtained using a calcite core whereas hollow CO3Ap microspheres were obtained using a NaCl core. The obtained microsphere was identified as B-type CO3Ap by Fourier transform infrared analysis and the carbonate content was approximately 7-8 wt% regardless of the type of core used for sample preparation. The mechanical strength of both the solid and hollow CO3Ap microspheres was sufficient for practical use as a bone substitute.

Structural requirements essential for elastin coacervation: favorable spatial arrangements of valine ridges on the three-dimensional structure of elastin-derived polypeptide (VPGVG)n.

The elastin precursor tropoelastin possesses a number of polymeric peptides with repeating 3-9 mer sequences. One of these is the pentapeptide Val-Pro-Gly-Val-Gly (VPGVG) present in almost all animal species, and its polymer (VPGVG)n coacervates just as does tropoelastin. In the present study, in order to explore the structural requirements essential for coacervation, (VPGVG)n and its shortened repeat analogs (VPGV)n, (VPG)n, and (PGVG)n were synthesized and their structural properties were investigated. In our turbidity measurements, (VPGVG)n demonstrated complete reversible coacervation in agreement with previous findings. The Gly(5) -deleted polymer (VPGV)n also achieved self-association, though the onset of self-association occurred at a lower temperature. However, the dissociation of (VPGV)n upon temperature lowering was found to occur in a three-step process; the Val(i) (4) -Val(i+1) (1) structure arising in the VPGV polypeptide appeared to perturb the dissociation. No self-association was observed for (VPG)n or (PGVG)n repeats. Spectroscopic measurements by CD, FT-IR, and (1) H-NMR showed that the (VPGV)n and (VPG)n both assumed ordered structures similar to that of (VPGVG)n. These results demonstrated that VPGVG is a structural element essential to achieving the ß-spiral structure required for self-association followed by coacervation, probably due to the ideal spatial arrangement of the hydrophobic Val residues.

Blocking of sodium and potassium ion-dependent adenosine triphosphatase-α1 with ouabain and vanadate suppresses cell-cell fusion during RANKL-mediated osteoclastogenesis.

To examine the possible enrolment of Na(+)/K(+)-ATPase during osteoclast differentiation, Na(+)/K(+)-ATPase inhibitors, including ouabain and vanadate, were used in this study. These inhibitors significantly inhibited cell-cell fusion of RAW264.7 cells and bone marrow cells induced by RANKL. Interestingly, in response to RANKL-stimulation, ouabain and vanadate decreased the number of large TRAP+ osteoclasts in the culture of RAW264.7 cells, as well as bone marrow cells. In contrast, the number of small TRAP+ osteoclasts either increased in RAW264.7 cells or were otherwise less affected in bone marrow cells than large TRAP+ osteoclasts. Large TRAP+ osteoclasts are defined as having ≥ 10 nuclei/cell and having more potency in bone resorption than small multinuclear osteoclasts with <9 nuclei/cell. Na(+)/K(+)-ATPase α1 and ß2 mRNAs were detected in sRANKL-stimulated RAW264.7 cells. Moreover, real-time quantitative PCR showed that ouabain and vanadate suppressed the RANKL-dependent induction of the osteoclast fusion-promotion molecule DC-STAMP at the mRNA level. Finally, and importantly, RNAi-mediated suppression of Na(+)/K(+)-ATPase α1 resulted in a diminished number of large TRAP+ osteoclasts in the sRANKL-stimulated RAW264.7 cells, along with the decreased level of DC-STAMP mRNA expression. These findings strongly suggest that blockage of the Na(+)/K(+)-ATPase α1 subunit by ouabain or vanadate caused the inhibition of RANKL-induced cell-cell fusion, resulting in the generation of large osteoclasts through suppression of DC-STAMP expression. Thus, in addition to its known function of sodium and potassium ion exchange during bone resorption by mature osteoclasts, this study has revealed a novel molecular role of the Na(+)/K(+)-ATPase α1 subunit in osteoclastogenesis.

Antimicrobial and antifungal effects of tissue conditioners containing a photocatalyst.

This study examined the antimicrobial/antifungal ability of a tissue conditioner containing a photocatalyst for Escherichia coli, Streptococcus mutans, Staphylococcus aureus and Candida albicans. The photocatalyst was mixed with tissue conditioners powders at concentrations of 0, 10, 15, and 20 wt%. Tissue conditioners powders containing a photocatalyst were mixed with liquid to make test specimens. Test specimens inoculated by each microorganism were irradiated by ultraviolet light for 0-, 2- and 4 hours. The antimicrobial/antifungal effects were evaluated by the CFU technique. The CFU values of each microorganism for tissue conditioners containing a photocatalyst showed significant decrease following UV-irradiation. The improvement in antimicrobial/antifungal effects was concomitant with the increase of the mixing ratio and the irradiation time. Therefore, the results indicated that tissue conditioners containing a photocatalyst might have photocatalytic ability.

Involvement of PRIP, phospholipase C-related, but catalytically inactive protein, in bone formation.

PRIP (phospholipase C-related, but catalytically inactive protein) is a novel protein isolated in this laboratory. PRIP-deficient mice showed increased serum gonadotropins, but decreased gonadal steroid hormones. This imbalance was similar to that for the cause of bone disease, such as osteoporosis. In the present study, therefore, we analyzed mutant mice with special reference to the bone property. We first performed three-dimensional analysis of the femur of female mice. The bone mineral density and trabecular bone volume were higher in mutant mice. We further performed histomorphometrical assay of bone formation parameters: bone formation rate, mineral apposition rate, osteoid thickness, and osteoblast number were up-regulated in the mutant, indicating that increased bone mass is caused by the enhancement of bone formation ability. We then cultured primary cells isolated from calvaria prepared from both genotypes. In mutant mice, osteoblast differentiation, as assessed by alkaline phosphatase activity and the expression of osteoblast differentiation marker genes, was enhanced. Moreover, we analyzed the phosphorylation of Smad1/5/8 in response to bone morphogenetic protein, with longer phosphorylation in the mutant. These results indicate that PRIP is implicated in the negative regulation of bone formation.

Protogenin, a new member of the immunoglobulin superfamily, is implicated in the development of the mouse lower first molar.

BACKGROUND: Protogenin (Prtg) has been identified as a gene which is highly expressed in the mouse mandible at embryonic day 10.5 (E10.5) by a cDNA subtraction method between mandibles at E10.5 and E12.0. Prtg is a new member of the deleted in colorectal carcinoma (DCC) family, which is composed of DCC, Neogenin, Punc and Nope. Although these members play an important role in the development of the embryonic central nervous system, recent research has also shed on the non-neuronal organization. However, very little is known regarding the fetal requirement of the non-neuronal organization for Prtg and how this may be associated with the tooth germ development. This study examined the functional implications of Prtg in the developing tooth germ of the mouse lower first molar. RESULTS: Ptrg is preferentially expressed in the early stage of organogenesis. Prtg mRNA and protein were widely expressed in the mesenchymal cells in the mandible at E10.5. The oral epithelial cells were also positive for Prtg. The expression intensity of Prtg after E12.0 was markedly reduced in the mesenchymal cells of the mandible, and was restricted to the area where the tooth bud was likely to be formed. Signals were also observed in the epithelial cells of the tooth germ. Weak signals were observed in the inner enamel epithelial cells at E16.0 and E18.0. An inhibition assay using a hemagglutinating virus of Japan-liposome containing Prtg antisense-phosphorothioated-oligodeoxynucleotide (AS-S-ODN) in cultured mandibles at E10.5 showed a significant growth inhibition in the tooth germ. The relationship between Prtg and the odontogenesis-related genes was examined in mouse E10.5 mandible, and we verified that the Bmp-4 expression had significantly been decreased in the mouse E10.5 mandible 24 hr after treatment with Prtg AS-S-ODN. CONCLUSION: These results indicated that the Prtg might be related to the initial morphogenesis of the tooth germ leading to the differentiation of the inner enamel epithelial cells in the mouse lower first molar. A better understanding of the Prtg function might thus play a critical role in revealing a precious mechanism in tooth germ development.

Repeated administration of methamphetamine blocked cholecystokinin-octapeptide injection-induced c-fos mRNA expression without change in capsaicin-induced junD mRNA expression in rat cerebellum.

In the cerebellum, there are numerous cholecystokinin (CCK-8)-containing fibers. Since systemic CCK-8 injection-induced anxiety (psychological stress) activates the locus coeruleus cells that send mossy fiber inputs to the cerebellum, we examined whether systemic CCK-8 injections activate the rat and mouse cerebellum. First, injections of CCK-8 were found to induce c-fos mRNA expression in a vague patchy pattern that is different from single methamphetamine-induced Zebrin band-like c-fos mRNA expression, suggesting that the CCK-8 activating mossy fibers induce gene expression differently from the dopamine-containing mossy fibers in the ventral tegmental area. Second, since CCK-8 facilitates neural activity of dopamine in the midbrain, we examined whether repeated methamphetamine administration that induced behavioral sensitization had similar effects on the cerebellar CCK system. Repeated administration of methamphetamine suppressed the CCK-8-induced c-fos mRNA expression in the rat cerebellum. Third, capsaicin injections (physical stress) into a hind limb of the rat increased junD mRNA expression with no effect on c-fos mRNA expression, and repeated methamphetamine injections had no effect on the capsaicin-induced expression of junD mRNA. Fourth, either single injection of methamphetamine or CCK-8 to mice increased c-fos mRNA expression in the locus coeruleus, and so noradrenalin, but not dopamine, might interact with CCK-8-activating system. However, we considered the possibility unlikely. Thus, we conclude that repeated methamphetamine administration though dopamine selectively inhibits the c-fos mRNA expression after CCK-8 injection in the cerebellum.

Fabrication of carbonate apatite block based on internal dissolution-precipitation reaction of dicalcium phosphate and calcium carbonate.

In this study, we investigated a novel method for fabrication of carbonate apatite block without ionic movement between precursor and solution by using precursor that includes all constituent ions of carbonate apatite. A powder mixture prepared from dicalcium phosphate anhydrous and calcite at appropriate Ca/P ratios (1.5, 1.67, and 1.8) was used as starting material. For preparation of specimens, the slurry made from the powder mixture and distilled water was packed in a split stainless steel mold and heat - treated, ranging from 60 degrees C to 100 degrees C up to 48 hours at 100% humidity. It appeared that carbonate apatite could be obtained above 70 degrees C and monophasic carbonate apatite could be obtained from the powder mixture at Ca/P ratio of 1.67. Carbonate content of the specimen was about 5-7%. Diametral tensile strength of the carbonate apatite blocks slightly decreased with increasing treatment temperature. The decrease in diametral tensile strength is thought to be related to the crystal size of the carbonate apatite formed.

Involvement of phospholipase C-related inactive protein in the mouse reproductive system through the regulation of gonadotropin levels.

Phospholipase C-related but catalytically inactive protein (comprising PRIP-1 and PRIP-2 [officially designated PLCL1 and PLCL2]) was first identified in our laboratory, but the biological functions have remained elusive. Therefore, we generated Plcl1 and Plcl2 double-knockout mice (Plcl1(tm1Mh); Plcl2(tm1Tta)) to gain insight into the biological function. Double-knockout mice apparently grew normally and became fertile; however, during animal maintenance, we noticed that mutant couples exhibited decreased litter events and litter size, indicating dysfunction of the reproductive system. Cross-mating experiments to discriminate whether males or females were defective indicated that the cause appeared to be on the female side. Mutant female mice had an apparently smaller uterus by gross anatomical observation and had more estrous days during the cycles. Levels of serum luteinizing hormone (LH) and follicle-stimulating hormone were measured for 5-6 consecutive days and were significantly higher in the mutant, which was also confirmed by examining the secretion of LH from the explant culture of anterior pituitary glands of wild-type and double-knockout mice. These results suggest that through gonadotropin secretion, PRIP plays an important role in female reproduction.

Development of tissue conditioner capable of binding with anti-microbial protein lactoferrin.

PURPOSE: This study focused on the anti-microbial protein human lactoferrin (hLF) commonly found in saliva, and tried to develop biocompatible dental materials that have higher anti-microbial effects. METHODS: A lyophilized cation exchange resin was added to tissue conditioner at 4 wt% and 8 wt%. The amount of hLF binding to the tissue conditioner and their anti-microbial effect against Candida albicans was investigated. Then their mechanical properties and cytotoxicity were examined. RESULTS: Tissue conditioner containing cation exchange resin was bound with hLF and had an anti-microbial effect against C. albicans. In addition, their physical properties were sufficient and they were harmless to human fibroblasts. CONCLUSION: The clinical application of cation exchange resin for tissue conditioner can be effective for the prevention and treatment of denture stomatitis and systemic opportunistic infections since it is thought that these materials will increase the local concentration of anti-microbial protein in saliva at the lesion site.

Self-cleaning ability of a photocatalyst-containing denture base material.

This study examined the ability of a photocatalyst mixed in a denture base resin to decompose organic substances which adhered to the denture base resin surface. The photocatalyst was mixed with denture base resin at concentrations of 0, 5, 10, and 15% (w/w). Decomposition test, bending test, and surface roughness measurement were performed at 1, 7, 30, 90, and 180 days after polymerization. Decomposition ability was evaluated based on the residual amount of methylene blue (MB) dissolved in ethanol after UV irradiation for 12 hours. As the mixing ratio increased, the amount of MB in the solution decreased. Meanwhile, no changes in the amount of MB in the immersion solution were observed in the photocatalyst-free resin specimen. Therefore, the results indicated that a denture base resin containing a photocatalyst might have a photocatalytic ability.

Reversal of the expression pattern of Aldolase C mRNA in Purkinje cells and Ube 1x mRNA in Golgi cells by a dopamine D1 receptor agonist injections in the methamphetamine sensitized-rat cerebellum.

The cerebellum has a parasagittal modular structure, in which Zebrin (Aldolase) positive and negative bands expressed in Purkinje cell layers alternate, and is involved in amphetamine psychosis. Administration of SKF38393, a D1 receptor agonist, reversed the behavioral sensitization of methamphetamine. In the vermis, there were the binding sites of SKF38393. In methamphetamine-sensitized rats the expression of the Aldolase mRNA positive bands move laterally in the rat vermis. We provide here the evidence that the D1 agonist injections also reversed the expression pattern of both the Aldolase mRNA in Purkinje cells and Ube (ubiquitin activating enzyme) 1x mRNA in Golgi interneurons of the sensitized rats. Thus the reverse changes in gene expression pattern in the vermis may be involved in the mechanisms of the behavioral plasticity and suggests the new treatment of drug abuse.

[Planning guidelines for prosthodontic treatment].

In recent years "practice guidelines" based on EBM techniques have even been attracting attention at a societal level, and guidelines modeled after the procedure for preparing practice guideline (described at http://www.niph.go.jp/glgl-4.3rev.htm) have begun to be drafted and made public. With the aim of ensuring the quality and presenting the basic concepts of prosthodontic therapy, the Japan Prosthodontic Society, which bears a great obligation and responsibility toward society and the Japanese public, has decided to undertake the formulation of guidelines related to prosthodontic therapy, and decided to first undertake the formulation of "Practice guideline for denture relining and rebasing", and to prepare a guideline model. We tried to prepare the guidelines according to the "Procedure for preparing practice guidelines", but because of the scientific uniqueness of prosthodontic treatment and dentistry, research to elucidate the basis of treatment has been insufficient, and we ultimately reconfirmed the current state of affairs in which it is difficult to perform. We therefore prepared the guidelines based on the limited evidence obtained in a search of the scientific literature and on the consensus of experts. The Japan Prosthodontic Society has investigated and prepared a Society guideline "model" to the extent possible at the present time, and it has prepared "Guidelines for adhesion bridge" and "Practice guidelines for denture prosthodontics" based on it. Nevertheless, the fact of the matter is that we are faced with numerous problems, and we think that in the future new bases and clinical knowledge will be accumulated by promoting scientific clinical research, and that the guidelines should be revised regularly based on them.

Effect of molding pressure on fabrication of low-crystalline calcite block.

We have reported that low-crystalline porous calcite block, which is useful as a bone substitute or a source material to prepare apatite-type bone fillers could be fabricated by exposing calcium hydroxide compact to carbon dioxide gas saturated with water vapor. In the present study, we investigated the effect of molding pressure on the transformation of calcium hydroxide into calcite and the mechanical strength of the carbonated compact. Transformation into calcite was almost completed within 72 h, however, a small amount of Ca(OH)(2) still remained unreacted at higher molding pressure because of incomplete penetration of CO(2) gas into the interparticle space due to dense packing of Ca(OH)(2) particles. On the other hand, high molding pressure resulted in an increase in diametral tensile strength (DTS) of the calcite compact formed. Critical porosity of the calcite block was calculated as approximately 68%.