Efeito da termociclagem e da técnica de inserção na infiltração de restaurações em resina composta

The objective of this study was to evaluate in vitro the effect on leakage of two incremental filling techniques and two composite resins with different elastic modulus and similar polymerization shrinkage. Eighty Class V cavities (4x4x2mm) were prepared in bovine incisors and were randomly restored with Z-250 (Z) or Durafill VS (D) + Single Bond in axial (a) or oblique (o) increments. The restorations were divided into two groups: Not Aged - N (4-hour-storage in water at 37ºC) and Aged - A (1-week storage in water at 37ºC + 1000 x - 5º-55ºC / 1-min dwell time). The specimens were covered with 2 coats of nail varnish so that only the restoration margins were exposed to silver nitrate 50% (2h) and developed under fluorescent light (8h). After they were sectioned twice in buccal-lingual direction, the four exposed surfaces were digitized (Vidcap) and the silver nitrate penetration was measured (ImageLab) at the incisal and gingival walls. Data were analyzed by a 3-way ANOVA (Resin, Filling Technique and Aging) separately for incisal and gingival walls (alpha=0.05). Resin and Aging were statistically significant either for the incisal and the gingival walls. The microfill composite resin infiltrated more than the hybrid composite. The thermal cycling caused an overall increase in silver nitrate penetration. The filling technique affected leakage depending on the composite resin and aging regimen.O objetivo deste estudo foi avaliar o efeito de duas técnicas incrementais de inserção na infiltração de restaurações de resinas compostas com módulos de elasticidade distintos e contração de polimerização semelhante. Para isto, foram preparadas 80 cavidades Classe V (4x4x2 mm) em incisivos bovinos que foram restauradas de modo aleatório com Z-250 (Z) ou Durafill VS (D) + Single Bond em incrementos axiais (a) ou oblíquos (o). As restaurações foram divididas em dois grupos: Não Envelhecidas - N (4 h em água destilada a 37º C) e Envelhecidas - E (1 semana de armazenagem em água a 37º C + 1000 x - 5º-55ºC / 1 min de imersão). Os espécimes foram recobertos com duas camadas de esmalte cosmético de modo que apenas as margens das restaurações ficassem expostas ao nitrato de prata (2h) seguido de imersão em revelador (8h) sob luz fluorescente. Posteriormente, eles foram secionados duas vezes no sentido vestíbulo-lingual, as superfïcies expostas (4) foram digitalizadas (Vidcap) e a penetração do nitrato de prata medido (ImageLab) nas paredes incisal e gengival. Os dados foram analisados por análise de variância de três fatores (Resina, Técnica de Inserção e Envelhecimento) separadamente para as paredes incisal e gengival. Resina e Envelhecimento foram significativos. A resina composta de micropartículas mostrou maior infiltração que a híbrida. A técnica de inserção influenciou a infiltração marginal dependendo da resina e do emprego ou não de ciclos térmicos

Assessing Lanthanide‐Dependent Methanol Dehydrogenase Activity: The Assay Matters

Artificial dye-coupled assays have been widely adopted as a rapid and convenient method to assess the activity of methanol dehydrogenases (MDH). Lanthanide(Ln)-dependent XoxF-MDHs are able to incorporate different lanthanides (Lns) in their active site. Dye-coupled assays showed that the earlier Lns exhibit a higher enzyme activity than the late Lns. Despite widespread use, there are limitations: oftentimes a pH of 9 and activators are required for the assay. Moreover, Ln-MDH variants are not obtained by isolation from the cells grown with the respective Ln, but by incubation of an apo-MDH with the Ln. Herein, we report the cultivation of Ln-dependent methanotroph Methylacidiphilum fumariolicum SolV with nine different Lns, the isolation of the respective MDHs and the assessment of the enzyme activity using the dye-coupled assay. We compare these results with a protein-coupled assay using its physiological electron acceptor cytochrome cGJ (cyt cGJ). Depending on the assay, two distinct trends are observed among the Ln series. The specific enzyme activity of La-, Ce- and Pr-MDH, as measured by the protein-coupled assay, exceeds that measured by the dye-coupled assay. This suggests that early Lns also have a positive effect on the interaction between XoxF-MDH and its cyt cGJ thereby increasing functional efficiency

Tensile bond strength of adhesive systems: effects of primer and thermocycling

O objetivo desta pesquisa foi o de verificar o efeito do "primer" e dos ciclos térmicos na resistência da união entre adesivos multiuso e esmalte dental, sob ensaios de tração. Os seguintes sistemas adesivos foram aplicados, de acordo com as instruções dos fabricantes, na superfície vestibular (sem desgaste) de 96 pré-molares com ou sem a aplicação prévia do "primer": Scotchbond MP, OptiBond FL, Amalgambond Plus e OptiBond - "dual cure". Após a aplicação do sistema adesivo, foi confeccionado um cone de resina composta (Z100, 3M), e fotoativado dentro de um molde metálico. Metade do total de espécimes foi submetida a 3.000 ciclos térmicos (5-37ºC; 37-55ºC, 60 s de imersão); a outra metade permaneceu imersa em água a 37ºC pelo mesmo tempo dispensado no procedimento anterior. Os dados foram submetidos a uma análise de variância (p = 0,05) e nenhum efeito significante foi detectado, indicando que a resistência de união não foi afetada pelo sistema adesivo, pela aplicação do "primer" ou pelos ciclos térmicos.The objective of this study was to evaluate the effects of primer and thermocycling on the bond strength of multi-purpose adhesive systems applied to enamel, under tensile stress. The following bonding systems were applied, according to the manufacturers' instructions, on unground enamel buccal surfaces of 96 premolars, with or without the application of primer: Scotchbond MP, OptiBond FL, Amalgambond Plus and OptiBond (dual-cure). A composite resin (Z100, 3M) was applied and light-cured in a cast metal hollow cone, which was previously fixed to the enamel surfaces. Half of the sample was subjected to 3,000 thermocycles (5-37ºC; 37-55ºC, dwell time of 60 s), and the other half was stored in water at 37ºC for the same period. The data were treated by means of ANOVA and no significant effects were detected, which indicates that tensile bond strength was not affected by the adhesive systems, application of primer or thermocycling

Understanding the chemistry of the artificial electron acceptors PES, PMS, DCPIP and Wurster’s Blue in methanol dehydrogenase assays

Methanol dehydrogenases (MDH) have recently taken the spotlight with the discovery that a large portion of these enzymes in nature utilize lanthanides in their active sites. The kinetic parameters of these enzymes are determined with a spectrophotometric assay first described by Anthony and Zatman 55 years ago. This artificial assay uses alkylated phenazines, such as phenazine ethosulfate (PES) or phenazine methosulfate (PMS), as primary electron acceptors (EAs) and the electron transfer is further coupled to a dye. However, many groups have reported problems concerning the bleaching of the assay mixture in the absence of MDH and the reproducibility of those assays. Hence, the comparison of kinetic data among MDH enzymes of different species is often cumbersome. Using mass spectrometry, UV–Vis and electron paramagnetic resonance (EPR) spectroscopy, we show that the side reactions of the assay mixture are mainly due to the degradation of assay components. Light-induced demethylation (yielding formaldehyde and phenazine in the case of PMS) or oxidation of PES or PMS as well as a reaction with assay components (ammonia, cyanide) can occur. We suggest here a protocol to avoid these side reactions. Further, we describe a modified synthesis protocol for obtaining the alternative electron acceptor, Wurster’s blue (WB), which serves both as EA and dye. The investigation of two lanthanide-dependent methanol dehydrogenases from Methylorubrum extorquens AM1 and Methylacidiphilum fumariolicum SolV with WB, along with handling recommendations, is presented

Measurement of H₂O₂ within living drosophila during aging using a ratiometric mass spectrometry probe targeted to the mitochondrial matrix

Hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) is central to mitochondrial oxidative damage and redox signaling, but its roles are poorly understood due to the difficulty of measuring mitochondrial H<sub>2</sub>O<sub>2</sub> in vivo. Here we report a ratiometric mass spectrometry probe approach to assess mitochondrial matrix H<sub>2</sub>O<sub>2</sub> levels in vivo. The probe, MitoB, comprises a triphenylphosphonium (TPP) cation driving its accumulation within mitochondria, conjugated to an arylboronic acid that reacts with H<sub>2</sub>O<sub>2</sub> to form a phenol, MitoP. Quantifying the MitoP/MitoB ratio by liquid chromatography-tandem mass spectrometry enabled measurement of a weighted average of mitochondrial H<sub>2</sub>O<sub>2</sub> that predominantly reports on thoracic muscle mitochondria within living flies. There was an increase in mitochondrial H<sub>2</sub>O<sub>2</sub> with age in flies, which was not coordinately altered by interventions that modulated life span. Our findings provide approaches to investigate mitochondrial ROS in vivo and suggest that while an increase in overall mitochondrial H<sub>2</sub>O<sub>2</sub> correlates with aging, it may not be causative

Cumulative Load of Depressive Symptoms Is Associated With Cortisol Awakening Response in Very Old Age.

This study examined links of cumulative and present depressive symptoms with present cortisol diurnal profiles in oldest-old adults. Five waves of data from 50 older adults (M age = 89.05 years; 64% women) who participated in the Australian Longitudinal Study of Ageing were used to combine 15 years of longitudinal data with seven cortisol samples per day over a one-week period. Findings revealed that individuals with more past depressive symptoms showed a lower cortisol awakening response (CAR). Interestingly, present depressive symptoms were not associated with the CAR. These findings inform our understanding of distal health factors in very old age

Epidemiological Methods: About Time

Epidemiological studies often produce false positive results due to use of statistical approaches that either ignore or distort time. The three time-related issues of focus in this discussion are: (1) cross-sectional vs. cohort studies, (2) statistical significance vs. public health significance, and (3), how risk factors “work together” to impact public health significance. The issue of time should be central to all thinking in epidemiology research, affecting sampling, measurement, design, analysis and, perhaps most important, the interpretation of results that might influence clinical and public-health decision-making and subsequent clinical research