A myosin from a higher plant has structural similarities to class V myosins

In plant cells, myosin is believed to be the molecular motor responsible for actin-based motility processes such as cytoplasmic streaming and directed vesicle transport. In an effort to characterize plant myosin, a cDNA encoding a myosin heavy chain was isolated from Arabidopsis thaliana. The predicted product of the MYA1 gene is 173 kDa and is structurally similar to the class V myosins. It is composed of the highly-conserved NH2-terminal "head" domain, a putative calmodulin-binding "neck" domain an alpha-helical coiled-coil domain, and a COOH-terminal domain. Northern blot analysis shows that the Arabidopsis MYA1 gene is expressed in all the major plant tissues (flower, leaf, root, and stem). We suggest that the MYA1 myosin may be involved in a general intracellular transport process in plant cells.

Marginal leakage in Class V cavities pretreated with different laser energy densities

The purpose of the present study was to evaluate in vitro the degree of marginal leakage in Class V cavities involving the cementoenamel junction. Cavities were 4 rum wide and 2 mm deep. The specimens received dentin pretreatment (37% phosphoric acid) followed by the Single Bond (3M) adhesive system application. The 40 specimens were then divided into four groups: Group I (control); Group 2 (Nd:YAG laser at 120 mJ/pulse, frequency of 10 Hz, power of 1.2 W); Group 3 (Nd:YAG laser at 140 mJ/pulse, frequency of 10 Hz, power of 1.4 W); Group 4 (Nd:YAG laser at 160 mJ/pulse, frequency of 10 Hz, power of 1.6 W). The cavities were restored with Z100 composite resin (3M) and light cured at 300-600 mW/cm(2) light intensity. Specimens were thermocycled to 500 cycles from 2-50 degrees C. After that, they were dried and sealed with nail varnish, respecting 1 mm around the restorations, and immersed in 0.5% methylene blue solution for 4 h. After this period, the teeth were rinsed, dried, sectioned, and analyzed in a stereoscopic loupe. The highest leakage scores were considered for each specimen. The results were statistically analyzed by the analysis of variance (ANOVA) Kruskal-Wallis test to the 5% level. For both the enamel and cementum, there was a decrease in marginal leakage with the application of laser energy; no significant differences were observed for Groups 2, 3, and 4. The results also showed a smaller tendency to marginal leakage on the cementum than on the enamel.

Study of microleakage at class V cavities prepared by Er : YAG laser using rewetting surface treatment

Objective: This study was conducted to analyze microleakage in Class V cavity preparation, using rewetting (or not) just after burr or Er:YAG laser preparation of enamel and dentin walls in permanent teeth. Background Data: Several studies reported microleakage around composite restorations when cavity preparation was done or treated by Er:YAG laser. As the hybridized laser is removed when this laser is used to cut dental hard tissue, there is a need for new materials or techniques to minimize gaps and microleakage. Results: Primer solution showed significant effect in enamel and dentin, at the level of 5%, when Er:YAG laser was used as a cutting tool. Using primer solution after phosphoric acid in preparations with the laser, microleakage was similar in degree to when cavities were prepared with the burr. Conclusion: Re-wetting surface just after Er:YAG irradiation and chemical treatment with phosphoric acid using HEMA aqueous solution seems to improve the quality of bioattachment between the adhesive system and enamel/dentin, showing similarities between restoration behaviors independently of the cutting tool, whether burr or laser.

Microleakage of hydrophilic adhesive systems in Class V composite restorations

Purpose: To investigate the microleakage of four hydrophilic adhesive systems: one multiple-bottles (Scotchbond Multi-Purpose Plus); two one-bottle (Single Bond, Stae); and one self-etching (Etch & Prime 3.0). Materials and Methods: 120 bovine incisor teeth were divided into four groups (n = 30) and Class V cavities were prepared at the cemento-enamel junction. The cavities were restored with the adhesive systems and with Z100 composite. The teeth were thermocycled 1,000 times between 5 +/- 2 degreesC and 55 +/- 2 degreesC with a dwell time of 1 min, and then placed in a 2% methylene blue dye (pH 7.0) for 4 hrs, washed and sectioned vertically through the center of the restorations. The qualitative evaluation was made by three examiners who distributed pre-established scores (0-4) for each tooth using a stereomicroscope at x30 magnification. Results: In enamel margins little microleakage was observed and the Kruskal-Wallis analysis did not show differences. In dentin margins the KruskaI-Wallis and multiple comparison analyses were applied: microleakage was significantly greater with Stae (median 3) and Scotchbond MP Plus (median 4). Single Bond (median 1) and Etch & Prime 3.0 (median 2) showed the best results in dentin margins, and the statistical analysis did not demonstrate differences in microleakage among these groups.

Influence of artificial aging in marginal adaptation of mixed class V cavities

The aim of this study was to investigate whether the artificial aging by thermal cycling had influenced the marginal adaptation of class V restorations with/without chlorhexidine application in the bond process. Twelve intact human third molars were used. Class V cavity preparations were performed on the buccal surface and the teeth received 35% phosphoric acid-etching procedure (Ultradent Products Inc., South Jordan, Utah, USA). Subsequently, the samples were divided in two groups: Untreated acid-etched dentin and chlorhexidine application as an adjunct in the bond process. The adhesive Single Bond 2 (3M ESPE, St. Paul, MN, USA) was used after 2% chlorhexidine application, and the restorations were performed with FiltekTM Z350 XT (3M ESPE) composite resin. The specimens were submitted to artificial aging by thermal cycling with 3,000 cycles. Analyzes were performed on scanning electron microscopy using replicas of marginal adaptation in percentage of continuous margin before and after the artificial aging. The data were analyzed by paired test and the results showed statistically significant differences in the percentage of continuous margin with/without chlorhexidine treatment before and after thermal cycling. This study concluded that the artificial aging by thermal cycling influenced the marginal adaptation of mixed class V composite restorations.

Influence of light intensity and curing cycle on microleakage of Class V composite resin restorations

O objetivo deste estudo foi determinar o efeito da polimerização gradual, mediante a utilização de aparelhos de Quartzo-Tungustênio-Halógena (QTH) e Arco de Plasma de Xenônio (PAC), no selamento marginal de restaurações classe V em resina composta com margens localizadas em dentina. Setenta e cinco incisivos bovinos receberam preparos de cavidades classe V, na raiz, com o intuito de situar as margens cavitárias em dentina. Os dentes foram divididos em cinco grupos de acordo com o método de fotoativação. As cavidades, depois de condicionadas, foram tratadas com o sistema adesivo Single Bond (3M Dental) e restauradas com a resina composta Z100 (3M Dental) pela técnica incremental. A fotoativação foi realizada para cada grupo como descrito a seguir: Grupo I: PAC pelo método de fotoativação constante: 1600mW/cm² – 3s; Grupo II: PAC pelo método de fotoativação por passos (800mW/cm² – 2s, subindo automaticamente para 1600mW/cm² – 4s); Grupo III: QTH pelo método de fotoativação constante: 400 mW/cm² – 40s; Grupo IV: QTH pelo método de fotoativação em rampa: 100 a 600 mW/cm² – 15s, permanecendo a 600mW/cm² por mais 25s; Grupo V: QTH pelo método de fotoativação por pulso: 200 mW/cm² – 3s, tempo de espera de 3min.e a seguir 600mW/cm² – 30s. Os dentes foram armazenados em água destilada a 37ºC por 30 dias e então submetidos à ciclagem térmica, por 500 ciclos à 5 ºC e 55 ºC. Os ápices dos dentes foram selados com resina composta e os dentes foram cobertos com duas camadas de esmalte para unha, antes da sua imersão em fucsina básica a 0,5%. Os dentes foram seccionados e os cortes foram escaneados para avaliação da área infiltrada por corante por um programa de computador (Image Tools). Os cortes foram também visualizados com lupa para a determinação do grau de penetração do corante na interface dente-restauração por escores. Diferenças estatisticamente significantes foram observadas entre os grupos quanto ao grau e à área de penetração de corante (p < 0,05). Os grupos I e II apresentaram valores significantemente mais altos de infiltração e penetração do corante que os grupos III, IV e V. Em conclusão, o uso da fonte de PAC, no modo constante e por passos, resultou em valores significantemente maiores de infiltração marginal quando comparados com a intensidade de luz média emitida pelos aparelhos de QTH. Os métodos de fotoativação por pulso, rampa e continuo com a fonte de QTH resultaram num grau similar de microinfiltração.

Impact of Quantity of Resin, C-factor, and Geometry on Resin Composite Polymerization Shrinkage Stress in Class V Restorations

Objective: This study evaluated the effect of quantity of resin composite, C-factor, and geometry in Class V restorations on shrinkage stress after bulk fill insertion of resin using two-dimensional finite element analysis.Methods: An image of a buccolingual longitudinal plane in the middle of an upper first premolar and supporting tissues was used for modeling 10 groups: cylindrical cavity, erosion, and abfraction lesions with the same C-factor (1.57), a second cylindrical cavity and abfraction lesion with the same quantity of resin (QR) as the erosion lesion, and then all repeated with a bevel on the occlusal cavosurface angle. The 10 groups were imported into Ansys 13.0 for two-dimensional finite element analysis. The mesh was built with 30,000 triangle and square elements of 0.1 mm in length for all the models. All materials were considered isotropic, homogeneous, elastic, and linear, and the resin composite shrinkage was simulated by thermal analogy. The maximum principal (MPS) and von Mises stresses (VMS) were analyzed for comparing the behavior of the groups.Results: Different values of angles for the cavosurface margin in enamel and dentin were obtained for all groups and the higher the angle, the lower the stress concentration. When the groups with the same C-factor and QR were compared, the erosion shape cavity showed the highest MPS and VMS values, and abfraction shape, the lowest. A cavosurface bevel decreased the stress values on the occlusal margin. The geometry factor overcame the effects of C-factor and QR in some situations.Conclusion: Within the limitations of the current methodology, it is possible to conclude that the combination of all variables studied influences the stress, but the geometry is the most important factor to be considered by the operator.

Marginal adaptation of Class V restorations with current generation dentin-bonding agents: effect of different dentin surface treatments.

This study sought to assess the use of chlorhexidine with several excipients as a dentin surface treatment and its effect on marginal adaptation of class V restorations with current-generation dentin bonding agents. A total of 120 human third molars were selected and allocated into 12 groups, with standardized buccal class V restorations randomly divided into preconditioned dentin rinsed with: water; water + chlorhexidine; ethanol; or ethanol + chlorhexidine. After rinsing of dentin (previously conditioned with 35% phosphoric acid) with the test solutions, the Adper single bond 2, prime and bond 2.1, and Excite bonding systems were applied randomly. Restorations were performed with FiltekTM Z350 XT composite resin. The resulting specimens were subjected to thermal and mechanical load cycling. Quantitative analysis of marginal adaptation was performed on epoxy replicas by means of scanning electron microscopy. Results were assessed by means of the Kruskal-Wallis test (percentages of continuous margins) and Wilcoxon test (differences between percentages of continuous margins before and after thermal cycling and mechanical loading), at a significance level of p < 0.05. Outcomes in the chlorhexidine-treated groups were not superior to those obtained with other treatments.

Molecular basis for class V beta-tubulin effects on microtubule assembly and paclitaxel resistance.

Vertebrates produce at least seven distinct beta-tubulin isotypes that coassemble into all cellular microtubules. The functional differences among these tubulin isoforms are largely unknown, but recent studies indicate that tubulin composition can affect microtubule properties and cellular microtubule-dependent behavior. One of the isotypes whose incorporation causes the largest change in microtubule assembly is beta5-tubulin. Overexpression of this isotype can almost completely destroy the microtubule network, yet it appears to be required in smaller amounts for normal mitotic progression. Moderate levels of overexpression can also confer paclitaxel resistance. Experiments using chimeric constructs and site-directed mutagenesis now indicate that the hypervariable C-terminal region of beta5 plays no role in these phenotypes. Instead, we demonstrate that two residues found in beta5 (Ser-239 and Ser-365) are each sufficient to inhibit microtubule assembly and confer paclitaxel resistance when introduced into beta1-tubulin; yet the single mutation of residue Ser-239 in beta5 eliminates its ability to confer these phenotypes. Despite the high degree of conservation among beta-tubulin isotypes, mutations affecting residue 365 demonstrate that amino acid substitutions can be context sensitive; i.e. an amino acid change in one isotype will not necessarily produce the same phenotype when introduced into a different isotype. Modeling studies indicate that residue Cys-239 of beta1-tubulin is close to a highly conserved Cys-354 residue suggesting the possibility that disulfide formation could play a significant role in the stability of microtubules formed with beta1- but not with beta5-tubulin.

The Tail of a Yeast Class V Myosin, Myo2p, Functions as a Localization Domain

Myo2p is a yeast class V myosin that functions in membrane trafficking. To investigate the function of the carboxyl-terminal-tail domain of Myo2p, we have overexpressed this domain behind the regulatable GAL1 promoter (MYO2DN). Overexpression of the tail domain of Myo2p results in a dominant–negative phenotype that is phenotypically similar to a temperature-sensitive allele of myo2, myo2–66. The tail domain of Myo2p is sufficient for localization at low- expression levels and causes mislocalization of the endogenous Myo2p from sites of polarized cell growth. Subcellular fractionation of polarized, mechanically lysed yeast cells reveals that Myo2p is present predominantly in a 100,000 × g pellet. The Myo2p in this pellet is not solubilized by Mg++-ATP or Triton X-100, but is solubilized by high salt. Tail overexpression does not disrupt this fractionation pattern, nor do mutations in sec4, sec3, sec9, cdc42, or myo2. These results show that overexpression of the tail domain of Myo2p does not compete with the endogenous Myo2p for assembly into a pelletable structure, but does compete with the endogenous Myo2p for a factor that is necessary for localization to the bud tip.

A Gene Encoding a Hevein-Like Protein from Elderberry Fruits Is Homologous to PR-4 and Class V Chitinase Genes1

We isolated SN-HLPf (Sambucus nigra hevein-like fruit protein), a hevein-like chitin-binding protein, from mature elderberry fruits. Cloning of the corresponding gene demonstrated that SN-HLPf is synthesized as a chimeric precursor consisting of an N-terminal chitin-binding domain corresponding to the mature elderberry protein and an unrelated C-terminal domain. Sequence comparisons indicated that the N-terminal domain of this precursor has high sequence similarity with the N-terminal domain of class I PR-4 (pathogenesis-related) proteins, whereas the C terminus is most closely related to that of class V chitinases. On the basis of these sequence homologies the gene encoding SN-HLPf can be considered a hybrid between a PR-4 and a class V chitinase gene.

Class V injection wells and your drinking water.

Mode of access: Internet.