Application of Synephrine to Grape Increases Anthocyanin via Production of Hydrogen Peroxide, Not Phytohormones.

Global warming has caused such problems as the poor coloration of grape skin and the decreased production of high-quality berries. We investigated the effect of synephrine (Syn) on anthocyanin accumulation. Anthocyanin accumulation in cultured grape cells treated with Syn at concentrations of 1 mM or higher showed no significant difference, indicating that the accumulation was concentration-independent. On the other hand, anthocyanin accumulation was dependent on the compound used for treatment. The sugar/acid ratio of the juice from berries treated with Syn did not differ from the control. The expression of anthocyanin-biosynthesis-related genes, but not phytohormones, was increased by the treatment with Syn at 24 h or later. The Syn treatment of cultured cells increased SOD3 expression and hydrogen peroxide (H2O2) production from 3 to 24 h after treatment. Subsequently, the expression of CAT and APX6 encoding H2O2-scavenging enzymes was also increased. Treatment of cultured cells with Syn and H2O2 increased the expression of the H2O2-responsive gene Chit4 and the anthocyanin-biosynthesis-related genes mybA1 and UFGT 4 days after the treatment and increased anthocyanin accumulation 7 days after the treatment. On the other hand, the treatment of berries with Syn and H2O2 increased anthocyanin accumulation after 9 days. These results suggest that Syn increases anthocyanin accumulation through H2O2 production without changing phytohormone biosynthesis. Syn is expected to improve grape skin coloration and contribute to high-quality berry production.

Effects of the percentage of air/water in spray on the efficiency of tooth ablation with erbium, chromium: yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation.

We aimed at examining the effects of a percentage of air/water in spray on the cutting efficiency of Er,Cr:YSGG laser for enamel and dentin. The intensity and frequency of irradiation were 3.0 W and 20 Hz for the enamel surface and 2.0 W and 20 Hz for the dentin surface, respectively. Flattened surfaces of enamel and dentin were irradiated at nine points for approximately 1 s under various percentages of air/water in spray using Er,Cr:YSGG laser. A high-speed video microscope was used to record each laser irradiation on the tooth surface. A slow video image was used to count the number of water micro-explosions yielded on the tooth surface during laser irradiation. A surface roughness tester was used to measure the depth of the dimple prepared with laser irradiation on each specimen. Each individual depth of dimple was divided by the number of water micro-explosions (pulse). This allowed for the calculation of the cutting depth per pulse. Following laser irradiation, several representative specimens were observed using an SEM. Two-way ANOVA was used as the statistical analysis. This revealed that there was no significant effect of the percentage of air/water in spray on the cutting depth for enamel surface (p > 0.05). On the contrary, a significant effect was observed in air-ratio for dentin cutting (p < 0.05). Both enamel and dentin were characterized by the presence of rough surfaces, as shown by the SEM images of the dimples. The percentage of air/water in spray was not significantly effective in laser cutting for enamel. Air-percentage was significantly effective in laser cutting for dentin.

Molecular basis of a shattering resistance boosting global dissemination of soybean.

Pod dehiscence (shattering) is essential for the propagation of wild plant species bearing seeds in pods but is a major cause of yield loss in legume and crucifer crops. Although natural genetic variation in pod dehiscence has been, and will be, useful for plant breeding, little is known about the molecular genetic basis of shattering resistance in crops. Therefore, we performed map-based cloning to unveil a major quantitative trait locus (QTL) controlling pod dehiscence in soybean. Fine mapping and complementation testing revealed that the QTL encodes a dirigent-like protein, designated as Pdh1. The gene for the shattering-resistant genotype, pdh1, was defective, having a premature stop codon. The functional gene, Pdh1, was highly expressed in the lignin-rich inner sclerenchyma of pod walls, especially at the stage of initiation in lignin deposition. Comparisons of near-isogenic lines indicated that Pdh1 promotes pod dehiscence by increasing the torsion of dried pod walls, which serves as a driving force for pod dehiscence under low humidity. A survey of soybean germplasm revealed that pdh1 was frequently detected in landraces from semiarid regions and has been extensively used for breeding in North America, the world's leading soybean producer. These findings point to a new mechanism for pod dehiscence involving the dirigent protein family and suggest that pdh1 has played a crucial role in the global expansion of soybean cultivation. Furthermore, the orthologs of pdh1, or genes with the same role, will possibly be useful for crop improvement.

Shear bond strengths of tooth coating materials including the experimental materials contained various amounts of multi-ion releasing fillers and their effects for preventing dentin demineralization.

We examined shear bond strengths (SBSs) of various tooth-coating-materials including the experimental materials to dentin and demineralization resistance of a fractured adhesive surface after the SBS testing. Three resin-type tooth-coating-materials (BC, PRG Barrier Coat; HC, Hybrid Coat II; and SF, Shield force plus) and two glass-ionomer-type tooth-coating-materials (CV, Clinpro XT Varnish; and FJ, Fuji VII) were selected. The experimental PRG Barrier Coat containing 0, 17, and 33 wt% S-PRG filler (BC0, BC17, and BC33, respectively) were developed. Each tooth-coating-material was applied to flattened dentin surfaces of extracted human teeth for SBS testing. After storing in water for 32 days with 4000 thermal cycling, the specimens were subjected to the SBS test. Specimens after SBS testing were subjected to a pH cycling test, and then, demineralization depths were measured using a polarized-light microscope. ANOVA and Tukey's HSD test were used for statistical analysis. The SBS value of FJ and CV was significantly lower than those of other materials except for BC (p < 0.01). The lesion depth of FJ was significantly shallower than those of other materials (p < 0.01); that of CV was significantly shallower than those of BC, HC, SF, and the control; and those of BC0 and BC17 were significantly shallower than that of the control (p < 0.05). The resin-type tooth-coating-materials demonstrated significantly higher SBS for dentin than the glass-ionomer-type tooth-coating-materials; however, they were inferior to the glass ionomer-type tooth-coating-materials in regards to the acid resistance of the fractured adhesion surface.

Effect of adhesive system application for cavities prepared with erbium, chromium: yttrium scandium gallium garnet laser on rat dental pulp tissue.

We examined the effects of adhesive systems under study applied for a laser-cut cavity using an Er,Cr:YSGG laser on rat dental pulp at 24 h and 14 days postoperatively. Group 1, laser-cut cavities were treated with a self-etching-primer and bonding agent; group 2, pretreated with a phosphoric-acid, and then treated with a self-etching-primer and bonding agent; group 3, pretreated with a phosphoric-acid and sodium-hypochlorite, and then treated with a self-etching-primer and bonding agent; and group 4, treated with an all-in-one adhesive. A flowable resin composite was used as filling material for each cavity treated with each group. A glass-ionomer-cement was used as a control. The following items were evaluated: pulp-tissue-disorganization (PTD), inflammatory-cell-infiltration (ICI), tertiary-dentin-formation (TDF), and bacterial-penetration (BP). The results were statistically analyzed using the Kruskal-Wallis test and Mann-Whitney U test. No significant differences were observed among the experimental groups for all parameters after 24 h and 14 days (P > 0.05). The majority of the specimens showed PTD with edema formation after 24 h; however, all the specimens demonstrated pulpal healing with TDF after 14 days. On the parameter of TDF, all groups showed significant differences between the two postoperative periods (P < 0.01). On the parameter of ICI, a significant difference was found between the two postoperative periods in group 4 (P < 0.05). No specimens showed BP. The pretreatment on the cavity prepared with the laser using phosphoric-acid or sodium-hypochlorite did not affect the dental pulp healing of rat tooth.

Effects of refractive errors on visual evoked magnetic fields.

BACKGROUND: The latency and amplitude of visual evoked cortical responses are known to be affected by refractive states, suggesting that they may be used as an objective index of refractive errors. In order to establish an easy and reliable method for this purpose, we herein examined the effects of refractive errors on visual evoked magnetic fields (VEFs). METHODS: Binocular VEFs following the presentation of a simple grating of 0.16 cd/m(2) in the lower visual field were recorded in 12 healthy volunteers and compared among four refractive states: 0D, +1D, +2D, and +4D, by using plus lenses. RESULTS: The low-luminance visual stimulus evoked a main MEG response at approximately 120 ms (M100) that reversed its polarity between the upper and lower visual field stimulations and originated from the occipital midline area. When refractive errors were induced by plus lenses, the latency of M100 increased, while its amplitude decreased with an increase in power of the lens. Differences from the control condition (+0D) were significant for all three lenses examined. The results of dipole analyses showed that evoked fields for the control (+0D) condition were explainable by one dipole in the primary visual cortex (V1), while other sources, presumably in V3 or V6, slightly contributed to shape M100 for the +2D or +4D condition. CONCLUSIONS: The present results showed that the latency and amplitude of M100 are both useful indicators for assessing refractive states. The contribution of neural sources other than V1 to M100 was modest under the 0D and +1D conditions. By considering the nature of the activity of M100 including its high sensitivity to a spatial frequency and lower visual field dominance, a simple low-luminance grating stimulus at an optimal spatial frequency in the lower visual field appears appropriate for obtaining data on high S/N ratios and reducing the load on subjects.

Effect of adrenalectomy on expression and induction of UDP-glucuronosyltransferase 1A6 and 1A7 in rats.

Uridine 5'-diphosphate (UDP)-glucuronosyltransferase 1A (UGT1A), which catalyzes major phase II reactions, is regulated by endogenous and exogenous factors via nuclear receptors such as the aryl hydrocarbon receptor (AhR). Glucocorticoid, one of the adrenocortical hormones, regulates AhR and UGT1A expression. We examined the effects of adrenalectomy on the expression and induction of UGT1A via AhR in the rat liver and small intestine. Rats were adrenalectomized bilaterally (ADX) or sham-operated (SHAM) and received intraperitoneal treatment with ß-naphthoflavone (BNF) for 4 d. Hepatic UGT1A6 and UGT1A7 mRNA levels were altered by ADX (0.1-fold and 1.6-fold, respectively). BNF treatment increased UGT1A6 and UGT1A7 mRNA expression and the intrinsic clearance of acetaminophen (APAP) glucuronidation, which is primarily catalyzed by UGT1A6 and UGT1A7, in both SHAM and ADX rats. Therefore, ADX rats maintained a functional AhR signaling pathway in the presence of BNF, expressed UGT1A6 and UGT1A7 mRNA, and showed APAP glucuronidation, namely induction by BNF via AhR was not abolished. Our results indicate that adrenal-dependent factors such as glucocorticoids are partially involved in the basal regulation of UGT1A6 and UGT1A7 transcription.

Identification and association study with lung cancer for novel insertion polymorphisms of human endogenous retrovirus.

Sequences of human endogenous retroviruses (HERVs) are members of the long terminal repeat (LTR) retrotransposon family. Although the expression of HERV has long been a topic of investigation, HERV-insertion polymorphisms are not well known, and a genetic association between HERV-insertion polymorphisms and cancer has never been reported. To identify novel HERV loci in the genome from cancer tissues, we carried out the inverse PCR method targeting a conserved LTR region of HML-2, which is the most recently acquired HERV group. Novel two insertions, HML-2_sLTR(1p13.2) and HML-2_sLTR(19q12), were identified as insertionally polymorphic solo LTRs. Furthermore, a significant prevalence of HML-2_sLTR(1p13.2) homozygosity was detected in female never-smoking patients aged 60 years and over who had lung adenocarcinoma [versus the other genotyping; odds ratio (OR): 1.97; 95% confidence interval (CI): 1.01-3.81]. In another cohort consisting of female never-smoking patients with lung adenocarcinoma, a prevalence of HML-2_sLTR(1p13.2) homozygosity tended to be high in patients aged 60 years and over (versus the other genotyping; OR: 2.03; 95% CI: 0.96-4.29), whereas a low prevalence of HML-2_sLTR(1p13.2) homozygosity was detected in patients <60 years old (versus the other genotyping; OR: 0.31; 95% CI: 0.11-0.94). Our results suggest that HML-2_sLTR(1p13.2) is involved with the susceptibility to lung adenocarcinoma in female never-smokers in an age-dependent manner and that other HERV polymorphisms related to human diseases might remain to be identified in the human genome.

Lipid peroxidation-induced DNA adducts in human gastric mucosa.

DNA adducts are a major cause of DNA mutation and DNA mutation-related diseases, but the simultaneous identification of multiple DNA adducts has been a challenge for a decade. An adductome approach using consecutive liquid chromatography and double mass spectrometry after micrococcal nuclease treatment has paved the way to demonstrations of numerous DNA adducts in a single experiment and is expected to contribute to the comprehensive understanding of overall environmental and endogenous exposures to possible mutagens in individuals. In this report, we applied an adductome approach to gastric mucosa samples taken at the time of a gastrectomy for gastric cancer in Lujiang, China, and in Hamamatsu, Japan. Seven lipid peroxidation-related DNA adducts [1,N6-etheno-2'-deoxyadenosine, butanone-etheno-2'-deoxycytidine (BεdC), butanone-etheno-2'-deoxy-5-methylcytidine, butanone-etheno-2'-deoxyadenosine (BεdA), heptanone-etheno-2'-deoxycytidine, heptanone-etheno-2'-deoxyadenosine (HεdA) and heptanone-etheno- 2'-deoxyguanosine] were identified in a total of 22 gastric mucosa samples. The levels of these adducts ranged from 0 to 30,000 per 10(9) bases. Although the presence of Helicobacter pylori DNA in the mucosa was not related to these adducts level, the levels of BεdC, BεdA and HεdA were higher in the Japanese gastric mucosa samples. The profiles of these 7 adduct levels among the 21 cases were capable of discriminating between the possible origins (China or Japan) of the gastric mucosa samples. Our report is the first demonstration of lipid peroxidation-related DNA adducts in the human stomach, and these observations warrant further investigation in the context of the significance of DNA adducts in human gastric carcinogenesis.

Effects of spatial frequency on visual evoked magnetic fields.

Psychophysical and visual evoked potential (VEP) studies have shown that spatial frequency of a visual stimulus affects contrast sensitivity and VEPs in humans. However, it is not clear whether and how the effect of spatial frequency varies among cortical areas. Considering that all visual inputs to the retina could be expressed as a sum of sinusoidal gratings of different spatial frequencies, the effect of spatial frequency must be clarified to separate the brain activity specific to each visual stimulus. In order to examine the effect of spatial frequency on different cortical areas, the present study compared cortical responses to sinusoidal gratings of seven different spatial frequencies using magnetoencephalography (MEG). MEG waveforms of twelve healthy adults in response to sinusoidal gratings of 0.3-18.1 cycles per degree were subjected to a multi-dipole analysis. As a result, the effect of spatial frequency was significant on the first peak latency and amplitude of the source activity around V1 and V2 but not on the source activity around V3 and V6, indicating that the effect of spatial frequency varies across different visual areas in the human brain. Our results also suggest that the responses in V1 and V2 that have a peak around 90 ms and that of V6 peaking around 120 ms should be separated to investigate the stimulus-specific cortical response, particularly when examining effects of spatial frequency on the response latency.

The Effect of Laser Irradiation to Surfaces of Computer-Aided Design/Computer-Aided Fabrication Resin Blocks Coated with a Silane Coupling Agent on Bond Strength between the Resin Blocks and Composite Resin.

The aim of this study was to investigate the effect of laser irradiation to computer-aided design/computer-aided fabrication (CAD/CAM) resin blocks coated with a silane coupling agent on the bond strength between resin blocks and composite resin. The CAD/CAM resin blocks used in this study were Cerasmart 300 (GC) and Vita Enamic (Vita); they were cut into plates and then subjected to a series of treatments. After processing with a silane coupling agent, treatment with a semiconductor laser was performed at 3.0, 5.0, and 7.0 W, followed by bonding procedures. The control group included those exposed to silane and bonded without laser application. After bonding, a mold with a simulated cavity was formed on the specimen and filled with flowable composite resin, and they were stored for 24 h or stressed by thermal cycling for subsequent testing that assessed the shear bond strength (n = 10). The results revealed that the bond strength was significantly enhanced by laser irradiation after applying a silane coupling agent (p < 0.03), whereas significant increase was not detected between the materials (p > 0.05). Particularly, 7 W laser irradiation had a significant increase on the bond strength between the composite resin and Cerasmart block after thermal cycling (p = 0.009). The SBS of the composite resin to CAD/CAM resin blocks was significantly enhanced by laser irradiation after silane coupling agent application.

The CRKL gene encoding an adaptor protein is amplified, overexpressed, and a possible therapeutic target in gastric cancer.

BACKGROUND: Genomic DNA amplification is a genetic factor involved in cancer, and some oncogenes, such as ERBB2, are highly amplified in gastric cancer. We searched for the possible amplification of other genes in gastric cancer. METHODS AND RESULTS: A genome-wide single nucleotide polymorphism microarray analysis was performed using three cell lines of differentiated gastric cancers, and 22 genes (including ERBB2) in five highly amplified chromosome regions (with a copy number of more than 6) were identified. Particular attention was paid to the CRKL gene, the product of which is an adaptor protein containing Src homology 2 and 3 (SH2/SH3) domains. An extremely high CRKL copy number was confirmed in the MKN74 gastric cancer cell line using fluorescence in situ hybridization (FISH), and a high level of CRKL expression was also observed in the cells. The RNA-interference-mediated knockdown of CRKL in MKN74 disclosed the ability of CRKL to upregulate gastric cell proliferation. An immunohistochemical analysis revealed that CRKL protein was overexpressed in 24.4% (88/360) of the primary gastric cancers that were analyzed. The CRKL copy number was also examined in 360 primary gastric cancers using a FISH analysis, and CRKL amplification was found to be associated with CRKL overexpression. Finally, we showed that MKN74 cells with CRKL amplification were responsive to the dual Src/BCR-ABL kinase inhibitor BMS354825, likely via the inhibition of CRKL phosphorylation, and that the proliferation of MKN74 cells was suppressed by treatment with a CRKL-targeting peptide. CONCLUSION: These results suggested that CRKL protein is overexpressed in a subset of gastric cancers and is associated with CRKL amplification in gastric cancer. Furthermore, our results suggested that CRKL protein has the ability to regulate gastric cell proliferation and has the potential to serve as a molecular therapy target for gastric cancer.

Reduced expression of MUTYH with suppressive activity against mutations caused by 8-hydroxyguanine is a novel predictor of a poor prognosis in human gastric cancer.

The MUTYH gene encodes a DNA glycosylase that can initiate the excision repair of adenine mispaired with 8-hydroxyguanine (8OHG) and is responsible for a susceptibility to multiple colorectal adenomas and carcinomas. To determine whether the MUTYH gene is involved in gastric carcinogenesis, we first examined the expression level of MUTYH in gastric cancer. The reduced expression of MUTYH mRNA transcript was detected in both gastric cancer cell lines and primary gastric cancers using qRT-PCR analysis. Immunohistochemical analysis also showed a significant reduction in MUTYH protein expression in gastric cancer, compared with non-cancerous gastric epithelium (immunohistochemical score, 175.5 ± 43.0 versus 281.5 ± 24.8; p < 0.0001). Among the gastric cancers, the MUTYH expression level was significantly associated with the histopathology (p < 0.0001) and the pT stage (p < 0.001). The outcome of patients with gastric cancer exhibiting low MUTYH expression was significantly worse than the outcome of patients with gastric cancer exhibiting high MUTYH expression (p = 0.0007, log-rank test) and a multivariate analysis revealed that reduced MUTYH expression was an independent predictor of a poor survival outcome among the gastric cancer patients (hazard ratio, 1.865; 95% confidence interval, 1.028-3.529; p = 0.0401). We next compared the functional effects of MUTYH on gastric cancer cells, based on their MUTYH expression levels. MUTYH-over-expressing stable clones of the gastric cancer cell line AGS showed: (a) higher DNA cleavage activity towards adenine:8OHG mispair-containing substrates; (b) higher suppressive activity against mutations caused by 8OHG in a supF forward mutation assay; and (c) higher suppressive activity for cellular proliferation than empty vector-transfected AGS clones. These results suggested that MUTYH is a suppressor of mutations caused by 8OHG in gastric cells and that its reduced expression is associated with a poor prognosis in gastric cancer.

Two new approaches to improve the analysis of BALB/c 3T3 cell transformation assay data.

Validation activities of the BALB/c 3T3 cell transformation assay (CTA) - a test method used for the assessment of the carcinogenic potential of compounds - have revealed the need for statistical analysis tailored to specific features of BALB/c 3T3 CTA data. Whereas a standard statistical approach for the Syrian hamster embryo (SHE) CTA was considered sufficient, an international expert group was gathered by the European Centre for the Validation of Alternative Methods (ECVAM) to review commonly applied statistical approaches for BALB/c 3T3 CTA. As it was concluded that none of the commonly applied approaches is entirely appropriate, two novel statistical approaches were found to be recommended for the evaluation of BALB/c 3T3 CTA data accounting for possible non-monotone concentration-response relationship and variance heterogeneity: a negative binomial generalised linear model with William's-type downturn-protected trend tests and a normalisation of the data by a specific transformation allowing for application of a general linear model that estimates effects assuming a normal distribution with William's-type protected tests. Both approaches are described in this article and their performance and the quality of the results they generate is demonstrated using exemplary data. Our work confirmed that both approaches are suitable for the statistical analysis of BALB/c 3T3 CTA data and that each of them is superior to commonly used methods. Furthermore, a procedure dichotomising data into negatives and positives is proposed which allows re-testing in cases where inconclusive data are encountered. The scripts of the statistical evaluation programs written in R - a freely available statistical software - are appended including exemplary outputs (Appendix A).

Detection of kinase amplifications in gastric cancer archives using fluorescence in situ hybridization.

To test the feasibility of using bacterial artificial chromosomes (BAC) containing kinases for pathological diagnosis using fluorescence in situ hybridization (FISH), 10 BAC probes containing a gene amplified in 5% or more of a pilot cohort were selected from a previous survey using arbitrarily selected BAC clones harboring 100 kinases. In this report, we describe the prevalence and association with the clinicopathological profile of these selected 10 BAC probes in 365 gastric cancer tissues. FISH analyses using these 10 BAC probes containing loci encoding EGFR, ERBB2(HER2), EPHB3, PIK3CA, MET, PTK7, ACK1, STK15, SRC, and HCK showed detectable amplifications in paraffin-embedded tissue in 2.83% to 13.6% of the gastric cancer tissues. Considerable numbers of the cases showed the co-amplification of two or more of the probes that were tested. BAC probes located within a genome neighborhood, such as PIK3CA, EPHB3, and ACK1 at 3q26-29 or HCK, SRC, and STK15 at 20q11-13.1, were often co-amplified in the same cases, but non-random co-amplifications of genes at distant genomic loci were also observed. These findings provide basic information regarding the creation of a strategy for personalizing gastric cancer therapy, especially when using multiple kinase inhibitors.

Mapping and use of QTLs controlling pod dehiscence in soybean.

While the cultivated soybean, Glycine max (L.) Merr., is more recalcitrant to pod dehiscence (shattering-resistant) than wild soybean, Glycine soja Sieb. & Zucc., there is also significant genetic variation in shattering resistance among cultivated soybean cultivars. To reveal the genetic basis and develop DNA markers for pod dehiscence, several research groups have conducted quantitative trait locus (QTL) analysis using segregated populations derived from crosses between G. max accessions or between a G. max and G. soja accession. In the populations of G. max, a major QTL was repeatedly identified near SSR marker Sat_366 on linkage group J (chromosome 16). Minor QTLs were also detected in several studies, although less commonality was found for the magnitudes of effect and location. In G. max × G. soja populations, only QTLs with a relatively small effect were detected. The major QTL found in G. max was further fine-mapped, leading to the development of specific markers for the shattering resistance allele at this locus. The markers were used in a breeding program, resulting in the production of near-isogenic lines with shattering resistance and genetic backgrounds of Japanese elite cultivars. The markers and lines developed will hopefully contribute to the rapid production of a variety of shattering-resistant soybean cultivars.

Conditioning effects of cavities prepared with an Er,Cr:YSGG laser and an air-turbine.

The purpose of this study was to examine, morphologically and histochemically, five types of conditioning effects on cavities prepared with an Er,Cr:YSGG laser and an air-turbine. Cavities were prepared using a Waterlase(®) MD turbo handpiece (W) and an air-turbine (AT) on human extracted molars. The cavity conditionings used were non-conditioned (G1), K-etchant Gel (G2), K-etchant Gel + AD Gel (G3), Clearfil SE Bond primer (G4) and Clearfil S(3) Bond (G5). On naked eye observations, enamel of G1, G2 and G3 in the W cavities and etched enamel of G2 and G3 in the AT cavities were observed as rough and dull in appearance. G4 and G5 in W and AT cavities were observed as shiny surfaces. On SEM observations, no smeared layer was observed in W cavities, while a smeared layer and bur-scratches were observed in AT cavities. In W cavities, rough surfaces were observed on enamel. That is, cracks and minute rough surfaces were observed. In contrast, equally etched scale-shaped enamel rods were observed in AT cavities. Widely opened dentinal tubules and protruding peritubular matrices of dentin were observed in W cavities. A few remaining smeared plugs could be observed at the AT cavities. On LM observations, 13-16 µm layers of the dentin in G1, G2, G4 and G5 of W cavities were stained red in color by the Azan staining method, while redness was not observed in G3. No groups were stained red in AT cavities. It was considered that layers stained red in color were thermal degeneration layers of dentin induced by W. Namely 30 s etching of 40% phosphoric acid gel followed by 90 s treatment of 10% NaClO gel should be recommended for use when combined with an Er,Cr:YSGG laser for cavity preparation.

Dynactin is involved in a checkpoint to monitor cell wall synthesis in Saccharomyces cerevisiae.

Checkpoint controls ensure the completion of cell cycle events with high fidelity in the correct order. Here we show the existence of a novel checkpoint that ensures coupling of cell wall synthesis and mitosis. In response to a defect in cell wall synthesis, S. cerevisiae cells arrest the cell-cycle before spindle pole body separation. This arrest results from the regulation of the M-phase cyclin Clb2p at the transcriptional level through the transcription factor Fkh2p. Components of the dynactin complex are required to achieve the G2 arrest whilst keeping cells highly viable. Thus, the dynactin complex has a function in a checkpoint that monitors cell wall synthesis.

Effect of CO2 laser irradiation on wound healing of exposed rat pulp.

This study examined the effects of direct pulp capping treatment using super-pulsed CO2 laser preirradiation on the wound healing process of exposed rat pulp on days 1, 3, 7, 14, and 28 postoperatively. Group 1 was irradiated with a CO2 laser and directly capped with a self-etching adhesive system. The laser was operated in super-pulse mode (pulse duration, 200 µs; interval, 5800 µs; 0.003 J/pulse). The irradiation conditions were a power output of 0.5 W, an irradiation time of 3 s, and repeat mode (10 ms of irradiation at 10-ms intervals for a total beam exposure time of 1.5 s), defocused beam diameter of 0.74 mm (approximately 20 mm from the exposed pulp surface), energy density of 0.698 J/cm² per pulse, total applied energy of 0.75 J, and an activated air-cooling system. Group 2 was capped with the self-etching adhesive system. Group 3 was capped with commercially available calcium hydroxide, and the self-etching adhesive system was applied to the cavity. The following parameters were evaluated: pulp tissue disorganization, inflammatory cell infiltration, reparative dentin formation, and bacterial penetration. The results were statistically analyzed using the Kruskal-Wallis test for differences among the groups at each observation period (P < 0.05). There were no significant differences among the experimental groups in any parameters at any postoperative period (P > 0.05). CO2 laser irradiation was effective in arresting hemorrhaging but showed a tendency to delay reparative dentin formation compared with the application of calcium hydroxide.

Direct pulp capping effect with experimentally developed adhesive resin systems containing reparative dentin-promoting agents on rat pulp: mixed amounts of additives and their effect on wound healing.

This study examined the wound-healing process of exposed rat pulp when treated with experimental adhesive resin systems. The experimental direct pulp capping adhesive resin systems were composed of primer-I, primer-II and an experimental bonding agent. Primer-I was Clearfil SE Bond (CSE) primer containing 1.0 or 5.0 wt% CaCl(2), and primer-II was CSE primer containing 0.1, 1.0 or 5.0 wt% compound of equal mole of pA and pB with synthetic peptides derived from dentin matrix protein 1 (DMP1). Primer-I containing 1.0 and 5.0 wt% CaCl(2) was assigned to the experimental groups 1-3 and 4-6, respectively. Primer-II containing 0.1, 1.0 or 5.0 wt% compound of pA and pB was assigned to the experimental groups 1 and 4, 2 and 5, and 3 and 6, respectively. In all experimental groups, CSE bond containing 10 wt% hydroxyapatite powder was used as the experimental bonding agent. The positive control teeth were capped with calcium hydroxide preparation (Dycal), and the negative control teeth were capped with CSE. The specimens were alternately stained with Mayer's H&E and the enhanced polymer one-step staining methods. Experimental groups 1, 4, 5 and 6 showed a higher level of reparative dentin formation compared to the negative control 14 days postoperatively. At 28 days postoperatively, all experimental groups showed the formation of extensive reparative dentin, and experimental groups 4, 5 and 6 demonstrated similar dentin bridge formation as that of the positive control. How quickly reparative dentin formation occurs may depend on the concentration of CaCl(2) and the compound of pA and pB in the experimental primer.