Cr+3 distribution in Al1 and Al2 Sites of alexandrite (BeAl2O4: Cr3+) induced by annealing, investigated by optical spectroscopy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Investigação das propriedades ópticas, morfológicas e elétricas da heterojunção SnO2: Ce3+/ GaAs

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Monitoramento e avaliação de processos erosivos marginais em reservatórios de usinas hidrelétricas por meio de varredura a laser

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Montagem e avaliação de um sistema de varredura a LASER embarcado em VANT

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeitos de lasers de alta potência na superfície de cerâmicas vítreas de dissilicato de lítio e sua resistência de união à dentina humana

Pós-graduação em Odontologia Restauradora - ICT

Montagem e avaliação de um sistema de varredura a LASER embarcado em VANT

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeitos de lasers de alta potência na superfície de cerâmicas vítreas de dissilicato de lítio e sua resistência de união à dentina humana

Pós-graduação em Odontologia Restauradora - ICT

Porcelain monolayers and porcelain/alumina bilayers reinforced by Al2O3/GdAlO3 fibers

Aim. This work tested the effect of the addition of Al2O3/GdAlO3 longitudinal fibers in different contents to veneering porcelain of two dental all ceramic systems. Methods: Fibers (0.5 mm diameter) obtained by the Laser Heated Pedestal Growth (LHPG) method were added to bar-shaped specimens made by veneer porcelain (monolayers) or both the veneer and the core ceramic (bilayers) of two all-ceramic systems: In-Ceram Alumina - glass infiltrated alumina composite (GIA) and In-Ceram 2000 AL Cubes - alumina polycrystal (AP) (VITA Zahnfabrik). The longitudinal fibers were added to veneering porcelain (VM7) in two different proportions: 10 or 17 vol%. The bars were divided into nine experimental conditions (n = 10) according to material used: VM7 porcelain monolayers, VM7/GIA, VM7/AP; and according to the amount of fibers within the porcelain layer: no fibers, 10 vol% or 17 vol%. After grinding and polishing the specimens were submitted to a three point bending test (crosshead speed = 0.5 mm/min) with porcelain positioned at tensile side. Data were analyzed by means of one-way ANOVA and a Tukey's test (alpha = 5%). Scanning electronic microscopy (SEM) was conducted for fractographic analysis. Results. Regarding the groups without fiber addition, VM7/AP showed the highest flexural strength (MPa), followed by VM7/GIA and VM7 monolayers. The addition of fibers led to a numerical increase in flexural strength for all groups. For VM7/GIA bilayers the addition of 17 vol% of fibers resulted in a significant 48% increase in the flexural strength compared to the control group. Fractographic analysis revealed that the crack initiation site was in porcelain at the tensile surface. Cracks also propagated between fibers before heading for the alumina core. Conclusions. The addition of 17 vol% of Al2O3/GdAlO3 longitudinal fibers to porcelain/glass infiltrated alumina bilayers significantly improved its flexural strength. 10 vol% or 17 vol% of fibers inclusion increased the flexural strength for all groups. (C) 2011 Elsevier Ltd. All rights reserved.

Microtensile bond strength of resin composite to dentin treated with Er:YAG laser of bleached teeth

The objective of this study was to evaluate the influence of Er:YAG laser (lambda = 2.94 mu m) on microtensile bond strength (mu TBS) and superficial morphology of bovine dentin bleached with 16% carbamide peroxide. Forty bovine teeth blocks (7 x 3 x 3 mm(3)) were randomly assigned to four groups: G1- bleaching and Er:YAG irradiation with energy density of 25.56 J/cm(2) (focused mode); G2 - bleaching; G3 - no-bleaching and Er:YAG irradiation (25.56 J/cm(2)); G4 - control, non-treated. G1 and G2 were bleached with 16% carbamide peroxide for 6 h during 21 days. Afterwards, all blocks were abraded with 320 to 600-grit abrasive papers to obtain flat standardized dentin surfaces. G1 and G3 were Er:YAG irradiated. Blocks were immediately restored with 4-mm-high composite resin (Adper Single Bond 2, Z-250-3 M/ESPE). After 24 h, the restored blocks (n = 9) were serially sectioned and trimmed to an hour-glass shape of approximately 1 mm(2) at the bonded interface area, and tested in tension in a universal testing machine (1 mm/ min). Failure mode was determined at a magnification of 100x using a stereomicroscope. One block of each group was selected for scanning electron microscope (SEM) analysis. mu TBS data was analyzed by two-way ANOVA and Tukey test (alpha = 0.05). Mean bond strengths (SD) in MPa were: G1- 32.7 (5.9)(A); G2- 31.1 (6.3)(A); G3- 25.2 (8.3)(B); G4- 36.7 (9.9).(A) Groups with different uppercase letters were significantly different from each other (p < .05). Enamel bleaching procedure did not affect mu TBS values for dentin adhesion. Er:YAG laser irradiation with 25.56 J/cm(2) prior to adhesive procedure of bleached teeth did not affect mu TBS at dentin and promoted a dentin surface with no smear layer and opened dentin tubules observed under SEM. On the other hand, Er:YAG laser irradiation prior to adhesive procedure of non-bleached surface impaired mu TBS compared to the control group.

Silicon(IV) phthalocyanine-loaded-nanoparticles for application in photodynamic process

The aim of this study was to evaluate the potential application of biodegradable nanoparticles containing a photosensitizer in photodynamic therapy. The poly (D,L lactic-co-glycolic acid) nanoparticles were studied by steady-state techniques, time-resolved fluorescence, and laser flash photolysis. The external morphology of the nanoparticles was established by scanning electron microscopy, and the biological activity was evaluated by in vitro cell culture by 3-(4,5 dimethylthiazol-2,5 biphenyl) tetrazolium bromide assay. The particles were spherical in shape exhibiting a 435 nm diameter with a low tendency to aggregate. The loading efficiency was 77%. The phthalocyanine-loaded-nanoparticles maintained their photophysical behavior after encapsulation. The cellular viability was determined, obtaining 70% of cellular death. All the performed physical-chemical, photophysical, and photobiological measurements indicated that the phthalocyanine-loaded-nanoparticles are a promising drug delivery system for photodynamic therapy and photoprocesses. (C) 2012 Laser Institute of America.

Effect of different root caries treatments on the sealing ability of conventional glass ionomer cement restorations

In this study we compared the microleakage of conventional glass ionomer cement (GIC) restorations following the use of different methods of root caries removal. In vitro root caries were induced in 75 human root dentin samples that were divided in five groups of 15 each according to the method used for caries removal: in group 1 spherical carbide burs at low speed were used, in group 2 a hand-held excavator was used, and in groups 3 to 5 an Er,Cr:YSGG laser was used at 2.25 W, 40.18 J/cm(2) (group 3), 2.50 W, 44.64 J/cm(2) (group 4) and 2.75 W, 49.11 J/cm(2) (group 5). The air/water cooling during irradiation was set to 55%/65% respectively. All cavities were filled with GIC. Five samples from each group were evaluated by scanning electron microscopy (SEM) and the other ten samples were thermocycled and submitted to a microleakage test. The data obtained were compared by ANOVA followed by Fisher's test (pa parts per thousand currency sign0.05). Group 4 showed the lowest microleakage index (56.65 6.30; p < 0.05). There were no significant differences among the other groups. On SEM images samples of groups 1 and 2 showed a more regular interface than the irradiated samples. Demineralized dentin below the restoration was observed, that was probably affected dentin. Group 4 showed the lowest microleakage values compared to the other experimental groups, so under the conditions of the present study the method that provided the lowest microleakage was the Er,Cr:YSGG laser with a power output of 2.5 W yielding an energy density of 44.64 J/cm(2).

Mast cells act as phagocytes against the periodontopathogen Aggregatibacter Actinomycetemcomitans

Background: Evidence to date shows that mast cells play a critical role in immune defenses against infectious agents, but there have been no reports about involvement of these cells in eliminating periodontopathogens. In this study, the phagocytic ability of mast cells against Aggregatibacter actinomycetemcomitans compared with macrophages is evaluated. Methods: In vitro phagocytic assays were conducted using murine mast cells and macrophages, incubated with A. actinomycetemcomitans, either opsonized or not, with different bacterial load ratios. After 1 hour, cells were stained with acridine orange and assessed by confocal laser-scanning electronmicroscopy. Results: Phagocytic ability of murine mast cells against A. actinomycetemcomitans was confirmed. In addition, the percentage of mast cells with internalized bacteria was higher in the absence of opsonization than in the presence of opsonization. Both cell types showed significant phagocytic activity against A. actinomycetemcomitans. However, the percentage of mast cells with non-opsonized bacteria was higher than that of macrophages with opsonized bacteria in one of the ratios (1:10). Conclusions: This is the first report about the participation of murine mast cells as phagocytes against A. actinomycetemcomitans, mainly in the absence of opsonization with human serum. Our results may indicate that mast cells act as professional phagocytes in the pathogenesis of biofilmassociated periodontal disease

Quantification of cellular action fibers alignment from confocal microscopic images.

The cellular rheology has recently undergone a rapid development with particular attention to the cytoskeleton mechanical properties and its main components - actin filaments, intermediate filaments, microtubules and crosslinked proteins. However it is not clear what are the cellular structural changes that directly affect the cell mechanical properties. Thus, in this work, we aimed to quantify the structural rearrangement of these fibers that may emerge in changes in the cell mechanics. We created an image analysis platform to study smooth muscle cells from different arteries: aorta, mammary, renal, carotid and coronary and processed respectively 31, 29, 31, 30 and 35 cell image obtained by confocal microscopy. The platform was developed in Matlab (MathWorks) and it uses the Sobel operator to determine the actin fiber image orientation of the cell, labeled with phalloidin. The Sobel operator is used as a filter capable of calculating the pixel brightness gradient, point to point, in the image. The operator uses vertical and horizontal convolution kernels to calculate the magnitude and the angle of the pixel intensity gradient. The image analysis followed the sequence: (1) opens a given cells image set to be processed; (2) sets a fix threshold to eliminate noise, based on Otsu's method; (3) detect the fiber edges in the image using the Sobel operator; and (4) quantify the actin fiber orientation. Our first result is the probability distribution II(Δθ) to find a given fiber angle deviation (Δθ) from the main cell fiber orientation θ0. The II(Δθ) follows an exponential decay II(Δθ) = Aexp(-αΔθ) regarding to its θ0. We defined and determined a misalignment index α of the fibers of each artery kind: coronary αCo = (1.72 ‘+ or =’ 0.36)rad POT -1; renal αRe = (1.43 + or - 0.64)rad POT -1; aorta αAo = (1.42 + or - 0.43)rad POT -1; mammary αMa = (1.12 + or - 0.50)rad POT -1; and carotid αCa = (1.01 + or - 0.39)rad POT -1. The α of coronary and carotid are statistically different (p < 0.05) among all analyzed cells. We discussed our results correlating the misalignment index data with the experimental cell mechanical properties obtained by using Optical Magnetic Twisting Cytometry with the same group of cells.

Bioimaging of animal embryogenesis: mathematical methods and computational algorithms

Some fundamental biological processes such as embryonic development have been preserved during evolution and are common to species belonging to different phylogenetic positions, but are nowadays largely unknown. The understanding of cell morphodynamics leading to the formation of organized spatial distribution of cells such as tissues and organs can be achieved through the reconstruction of cells shape and position during the development of a live animal embryo. We design in this work a chain of image processing methods to automatically segment and track cells nuclei and membranes during the development of a zebrafish embryo, which has been largely validates as model organism to understand vertebrate development, gene function and healingrepair mechanisms in vertebrates. The embryo is previously labeled through the ubiquitous expression of fluorescent proteins addressed to cells nuclei and membranes, and temporal sequences of volumetric images are acquired with laser scanning microscopy. Cells position is detected by processing nuclei images either through the generalized form of the Hough transform or identifying nuclei position with local maxima after a smoothing preprocessing step. Membranes and nuclei shapes are reconstructed by using PDEs based variational techniques such as the Subjective Surfaces and the Chan Vese method. Cells tracking is performed by combining informations previously detected on cells shape and position with biological regularization constraints. Our results are manually validated and reconstruct the formation of zebrafish brain at 7-8 somite stage with all the cells tracked starting from late sphere stage with less than 2% error for at least 6 hours. Our reconstruction opens the way to a systematic investigation of cellular behaviors, of clonal origin and clonal complexity of brain organs, as well as the contribution of cell proliferation modes and cell movements to the formation of local patterns and morphogenetic fields.

Influence of climate changes on cross allergies: possible involvement of pollen transglutaminase

Allergies are a complex of symptoms derived from altered IgE-mediated reactions of the immune system towards substances known as allergens. Allergic sensibilization can be of food or respiratory origin and, in particular, apple and hazelnut allergens have been identified in pollens or fruits. Allergic cross-reactivity can occur in a patient reacting to similar allergens from different origins, justifying the research in both systems as in Europe a greater number of people suffers from apple fruit allergy, but little evidence exists about pollen. Apple fruit allergies are due to four different classes of allergens (Mal d 1, 2, 3, 4), whose allergenicity is related both to genotype and tissue specificity; therefore I have investigated their presence also in pollen at different time of germination to clarify the apple pollen allergenic potential. I have observed that the same four classes of allergens found in fruit are expressed at different levels also in pollen, and their presence might support that the apple pollen can be considered allergenic as the fruit, deducing that apple allergy could also be indirectly caused by sensitization to pollen. Climate changes resulting from increases in temperature and air pollution influence pollen allergenicity, responsible for the dramatic raise in respiratory allergies (hay fever, bronchial asthma, conjunctivitis). Although the link between climate change and pollen allergenicity is proven, the underlying mechanism is little understood. Transglutaminases (TGases), a class of enzymes able to post-translationally modify proteins, are activated under stress and involved in some inflammatory responses, enhancing the activity of pro-inflammatory phospholipase A2, suggesting a role in allergies. Recently, a calcium-dependent TGase activity has been identified in the pollen cell wall, raising the possibility that pollen TGase may have a role in the modification of pollen allergens reported above, thus stabilizing them against proteases. This enzyme can be involved also in the transamidation of proteins present in the human mucosa interacting with surface pollen or, finally, the enzyme itself can represent an allergen, as suggested by studies on celiac desease. I have hypothesized that this pollen enzyme can be affected by climate changes and be involved in exhacerbating allergy response. The data presented in this thesis represent a scientific basis for future development of studies devoted to verify the hypothesis set out here. First, I have demonstrated the presence of an extracellular TGase on the surface of the grain observed either at the apical or the proximal parts of the pollen-tube by laser confocal microscopy (Iorio et al., 2008), that plays an essential role in apple pollen-tube growth, as suggested by the arrest of tube elongation by TGase inhibitors, such as EGTA or R281. Its involvement in pollen tube growth is mainly confirmed by the data of activity and gene expression, because TGase showed a peak between 15 min and 30 min of germination, when this process is well established, and an optimal pH around 6.5, which is close to that recorded for the germination medium. Moreover, data show that pollen TGase can be a glycoprotein as the glycosylation profile is linked both with the activation of the enzyme and with its localization at the pollen cell wall during germination, because from the data presented seems that the active form of TGase involved in pollen tube growth and pollen-stylar interaction is more exposed and more weakly bound to the cell wall. Interestingly, TGase interacts with fibronectin (FN), a putative SAMs or psECM component, inducing possibly intracellular signal transduction during the interaction between pollen-stylar occuring in the germination process, since a protein immunorecognised by anti-FN antibody is also present in pollen, in particular at the level of pollen grain cell wall in a punctuate pattern, but also along the shank of the pollen tube wall, in a similar pattern that recalls the signal obtained with the antibody anti TGase. FN represents a good substrate for the enzyme activity, better than DMC usually used as standard substrate for animal TGase. Thus, this pollen enzyme, necessary for its germination, is exposed on the pollen surface and consequently can easily interact with mucosal proteins, as it has been found germinated pollen in studies conducted on human mucus (Forlani, personal communication). I have obtained data that TGase activity increases in a very remarkable way when pollen is exposed to stressful conditions, such as climate changes and environmental pollution. I have used two different species of pollen, an aero allergenic (hazelnut, Corylus avellana) pollen, whose allergenicity is well documented, and an enthomophylus (apple, Malus domestica) pollen, which is not yet well characterized, to compare data on their mechanism of action in response to stressors. The two pollens have been exposed to climate changes (different temperatures, relative humidity (rH), acid rain at pH 5.6 and copper pollution (3.10 µg/l)) and showed an increase in pollen surface TGase activity that is not accompanied to an induced expression of TGase immunoreactive protein with AtPNG1p. Probably, climate change induce an alteration or damage to pollen cell wall that carries the pollen grains to release their content in the medium including TGase enzyme, that can be free to carry out its function as confirmed by the immunolocalisation and by the in situ TGase activity assay data; morphological examination indicated pollen damage, viability significantly reduced and in acid rain conditions an early germination of apple pollen, thus possibly enhancing the TGase exposure on pollen surface. Several pollen proteins were post-translationally modified, as well as mammalian sPLA2 especially with Corylus pollen, which results in its activation, potentially altering pollen allergenicity and inflammation. Pollen TGase activity mimicked the behaviour of gpl TGase and AtPNG1p in the stimulation of sPLA2, even if the regulatory mechanism seems different to gpl TGase, because pollen TGase favours an intermolecular cross-linking between various molecules of sPLA2, giving rise to high-molecular protein networks normally more stable. In general, pollens exhibited a significant endogenous phospholipase activity and it has been observed differences according to the allergenic (Corylus) or not-well characterized allergenic (Malus) attitude of the pollen. However, even if with a different intensity level in activation, pollen enzyme share the ability to activate the sPLA2, thus suggesting an important regulatory role for the activation of a key enzyme of the inflammatory response, among which my interest was addressed to pollen allergy. In conclusion, from all the data presented, mainly presence of allergens, presence of an extracellular TGase, increasing in its activity following exposure to environmental pollution and PLA2 activation, I can conclude that also Malus pollen can behave as potentially allergenic. The mechanisms described here that could affect the allergenicity of pollen, maybe could be the same occurring in fruit, paving the way for future studies in the identification of hyper- and hypo- allergenic cultivars, in preventing environmental stressor effects and, possibly, in the production of transgenic plants.