Influence of Different Professional Prophylactic Methods on Fluorescence Measurements for Detection of Occlusal Caries

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The Efficacy of Laser Fluorescence to Detect in Vitro Demineralization and Remineralization of Smooth Enamel Surfaces

Objective: The purpose of this study was to evaluate the efficacy of the laser fluorescence (LF) device in detecting in vitro demineralization and remineralization of smooth surface caries-like lesions. Background Data: The early detection of smooth surface caries-like lesions is important to provide proper management of carious lesions, and allows monitoring of them over time. Also, some authors suggest that LF could be useful in monitoring the caries pathological process. Materials and Methods: Seventy-eight blocks of bovine teeth were obtained, and then submitted to artificial caries lesion induction and to a pH-cycling process. Superficial microhardness (SMH) and laser fluorescence analysis were performed at baseline, after induction of caries-like lesions, and after the pH-cycling regimen to promote remineralization. Results: Friedman's and multiple comparison tests were performed for all variables. SMH analysis showed significant differences (p < 0.05) between baseline (286.77 +/- 1.49 Vickers hardness number [VHN] units), before (38.48 +/- 0.85 VHN), and after remineralization (131.93 +/- 2.63 VHN). Baseline values for LF were extremely low (2.71 +/- 0.05), and a statistically significant difference was observed only after remineralization (3.61 +/- 0.08), as demonstrated by the increase in LF values. Conclusion: The LF device did not show efficacy for monitoring in vitro demineralization and remineralization of smooth enamel surfaces.

Influence of the Light-Curing Unit, Storage Time and Shade of a Dental Composite Resin on the Fluorescence

The aim of this study was to determine the influence of three light-curing units, storage times and colors of the dental composite resin on the fluorescence. The specimens (diameter 10.0 +/- 0.1 mm, thickness 1.0 +/- 0.1 mm) were made using a stainless steel mold. The mold was filled with the microhybrid composite resin and a polyethylene film covered each side of the mold. After this, a glass slide was placed on the top of the mold. To standardize the top surface of the specimens a circular weight (1 kg) with an orifice to pass the light tip of the LCU was placed on the top surface and photo-activated during 40 s. Five specimens were made for each group. The groups were divided into 9 groups following the LCUs (one QTH and two LEDs), storage times (immediately after curing, 24 hours, 7 and 30 days) and colors (shades: A(2)E, A(2)D, and TC) of the composite resin. After photo-activation, the specimens were storage in artificial saliva during the storage times proposed to each group at 37 C and 100% humidity. The analysis of variance (ANOVA) and Tukey's post-hoc tests showed no significant difference between storage times (immediately, 24 hours and 30 days) (P > 0.05). The means of fluorescence had difference significant to color and light-curing unit used to all period of storage (P < 0.05). The colors had difference significant between them (shades: A2D < A2E < TC) (P < 0.05). The Ultraled (LED) and Ultralux (QTH) when used the TC shade showed higher than Radii (LED), however to A2E shade and A2D shade any difference were found (P > 0.05).

Diffusion analysis of one photosensitizer in bovine teeth using fluorescence optical imaging

Some photosensitizers (PSs) used for PACT (Antimicrobial Photodynamic Therapy) show an affinity for bacterial walls and can be photo-activated to cause the desired damage. However, on dentine bacterias may be less susceptible to PACT as a result of limited penetration of the PS. The aim of this study was to evaluate the diffusion of one PS based on hematoporphyrin on dentine structures. Twelve bovine incisors were used. Class III cavities (3 x 3 x 1 mm) were prepared on the mesial or distal surfaces using a diamond bur. Photogem (R) solution at 1 mg/mL (10 uL for each cavity) was used. The experimental Groups were divided according to thickness of dentine remaining and etched or no-etched before the PS application. The fluorescence excitation source was a VelScope (R) system. For image capture a scientific CCD color camera PixelFly (R) was coupled to VelScope. For image acquisition and processing, a computational routine was developed at Matlab (R). Fick's Law was used to obtain the average diffusion coefficient of PS. Differences were found between all Groups. The longitudinal temporal diffusion was influenced by the different times, thickness and acid etching.

Laurdan Spectrum Decomposition as a Tool for the Analysis of Surface Bilayer Structure and Polarity: a Study with DMPG, Peptides and Cholesterol

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

Prevention of experimental diabetes by Uncaria tomentosa extract: Th2 polarization, regulatory T cell preservation or both?

Ethnopharmacological relevance: Uncaria tomentosa (Willd.) DC (Rubiaceae) is a species native to the Amazon rainforest and surrounding tropical areas that is endowed with immunomodulatory properties and widely used around the world. In this study we investigated the immunomodulatory potential of Uncaria tomentosa (UT) aqueous-ethanol extract on the progression of immune-mediated diabetes.Materials and methods: C57BL/6 male mice were injected with MLDS (40 mg/kg) and orally treated with UT at 10-400 mg/kg during 21 days. Control groups received MLDS alone or the respective dilution vehicle. Pancreatic mononuclear infiltrate and beta-cell insulin content were analyzed by HE and immunohistochemical staining, respectively, and measured by digital morphometry. Lymphocyte immunophenotyping and cytokine production were determined by flow cytometry analysis.Results: Treating the animals with 50-400 mg/kg of UT caused a significant reduction in the glycemic levels, as well as in the incidence of diabetes. The morphometric analysis of insulitis revealed a clear protective effect. Animals treated with UT at 400 mg/kg presented a higher number of intact islets and a significant inhibition of destructive insulitis. Furthermore, a significant protection against the loss of insulin-secreting presented beta-cells was achieved, as observed by a careful immunohistochemical evaluation. The phenotypic analysis indicated that the groups treated with higher doses (100-400 mg/kg) presented CD4(+) and CD8(+) T-cell values similar to those observed in healthy animals. These same higher doses also increased the number of CD4(+)CD25(+)Foxp3(+) regulatory T-cells. Moreover, the extract modulated the production of Th1 and Th2, with increased levels of IL-4 and IL-5.Conclusions: The extract was effective to prevent the progression of immune-mediated diabetes by distinct pathways. (C) 2011 Elsevier B.V. All rights reserved.

Detection of numerical chromosome anomalies in interphase cells of benign and malignant thyroid lesions using fluorescence in situ hybridization

Benign and malignant thyroid tumors constitute a wide range of neoplasias showing recurrent chromosome abnormalities. In an attempt to characterize specific numerical chromosome abnormalities in thyroid tissues, We present here the findings from a study of archival samples depicted by 10 malignant tumors, 30 benign lesions, and 10 normal thyroid tissues. Fluorescence in situ hybridization was performed on noncultured samples using biotinylated centromere-specific probes for chromosomes 7, 10, and 17. Trisomy or tetrasomy 7 were present in 19 benign and in 7 malignant tumors. Trisomy 10 or 17 were observed in 18 adenomas or goiters and in 9 carcinomas, and monosomy 17 was seen in 2 carcinomas. Our findings suggest that such abnormalities are an in vivo phenomenon and may be important in the neoplastic proliferation of thyroid gland. (C) Elsevier B.V., 2000. All rights reserved.

The effect of acidic residues and amphipathicity on the lytic activities of mastoparan peptides studied by fluorescence and CD spectroscopy

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

Effect of Sonication on the Thermotropic Behavior of DODAB Vesicles Studied by Fluorescence Probe Solubilization

The effect of sonication on fluorescence probe solubilization in cationic vesicles of dioctadecyldimethylammonium bromide (DODAB) was investigated by steady-state fluorescence of pyrene (Py), trans-diphenylpolyenes-diphenylbutadiene (DPB), diphenylhexatriene (DPH), and their corresponding 4,4'-dialkyl derivatives 4B4A and 4H4A fluorescence probes. The data indicate that sonication affects the bilayer polarity, the melting temperature (T (m)), and the cooperativity of the melting process due to changes in vesicle morphology. The effect of temperature on the fluorescence intensity and yielding I broken vertical bar(f) and anisotropy < r > shows that the ionizable probes 4B4A and 4H4A are solubilized close to the vesicle interfaces, whereas the non-ionizable DPH and DPB are deeper in the bilayers. Py solubilization indicates that sonicated vesicles exhibit less densely packed bilayers.

A fluorescence probe study of gemini surfactants in aqueous solution: a comparison between n-2-n and n-6-n series of the alkanediyl-a,w-bis (dimethylalkylammonium bromides)

Two series of alkanediyl-a,w-bis (dimethylalkylammonium bromide (n-2-n and n-6-n; n=8, 10,12, and 16) have been synthesized and their micelles properties studied in aqueous solution using pyrene, pyrenecarboxaldehyde (PCA) and 1,8 anilinonaphtalene sulfonic acid sodium salt (ANS) as fluorescent probes. The micelles from these surfactants have been characterized on the basis of the information provided by micelle-solubilized fluorescent probes. The obtained results indicated that the surfactant concentration at which a marked decrease in l max parameter of pyrenecarboxaldehyde (PCA) occurs corresponds to the CMC determined by conductimetric measurements. Changes in the emission spectra of ANS and PCA observed in the submicellar range for both surfactants series (n-2-n and n-6-n) were interpreted as formation of pre-aggregates. It was found that the dimeric surfactants with long spacer (s= 6) form more hydrated aggregates when compared with those formed by the n-2-n and CnTAB surfactants series. This was attributed to a more difficult packing of n-6-n surfactant molecules to form micelles.

Aggregation behavior of hydrophobically modified dextran in aqueous solution: a fluorescence probe study

Dextrans (M-W = 11.000 and M-w = 40.000) have been modified with 4-hexyl benzoyl chloride and their aggregation behavior was studied in aqueous solution employing the fluorescent probes pyrene and 1,8 anilinonaphtalene sulfonic acid sodium salt (1,8 ANS). The photophysical studies showed that above a critical concentration the derivatives tend to form aggregates having different properties, which depend on both the degree of substitution (alpha) and the molecular weight of the sample. The parameter alpha has a marked effect on the critical aggregation concentrations (CAC) and aggregate proper-ties. Hydrophobic microenvironments can be detected for substituted dextrans having alpha values varying from 0.01 to 0.19. CAC values decreased by two orders and magnitude when the molecular weight increased from 11 to 40 kDa, leading to formation of more apolar aggregates and diminishing by about 30% the polarity of the microenviromnents. Pre-aggregation was evidenced by pyrene excimer emission and intermolecular interactions were responsible by the formation of aggregates leading to solution behaviour similar to that of common surfactants. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Simultaneous Determination of Phenanthrene and Benzo(a)pyrene in Water Samples by Synchronous Fluorescence Spectroscopy

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

Fluorescence spectroscopy for the detection of tongue carcinoma - validation in an animal model

The efficacy of fluorescence spectroscopy to detect squamous cell carcinoma is evaluated in an animal model following laser excitation at 442 and 532 nm. Lesions are chemically induced with a topical DMBA application at the left lateral tongue of Golden Syrian hamsters. The animals are investigated every 2 weeks after the 4th week of induction until a total of 26 weeks. The right lateral tongue of each animal is considered as a control site (normal contralateral tissue) and the induced lesions are analyzed as a set of points covering the entire clinically detectable area. Based on fluorescence spectral differences, four indices are determined to discriminate normal and carcinoma tissues, based on intraspectral analysis. The spectral data are also analyzed using a multivariate data analysis and the results are compared with histology as the diagnostic gold standard. The best result achieved is for blue excitation using the KNN (K-nearest neighbor, a interspectral analysis) algorithm with a sensitivity of 95.7% and a specificity of 91.6%. These high indices indicate that fluorescence spectroscopy may constitute a fast noninvasive auxiliary tool for diagnostic of cancer within the oral cavity. (C) 2008 Society of Photo-Optical Instrumentation Engineers.