Sistema de espectroscopia de fluorescência na avaliação de margens cirúrgicas de carcinoma de células escamonas da cavidade oral nos momentos in situ e ex vivo

Orientadores: Luiz Paulo Kowalski, Cristina KurachiTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: O tratamento mais empregado para os carcinomas de células escamosas da boca é a ressecção cirúrgica, sendo ou não acompanhada de radio e/ou quimioterapia. O tratamento é simplificado, diminuindo número de recidivas e aumentando a sobrevida, quando as lesões encontram-se em estágios iniciais, conforme localização anatômica e diagnóstico de margens cirúrgicas livres. Frente a isto, o desenvolvimento e aperfeiçoamento de técnicas para um diagnóstico precoce, assim como de uma acurada definição das margens cirúrgicas livres e correta delimitação da extensão do câncer boca, com o intuito de melhorar a qualidade de vida e a taxa de sobrevida desses pacientes, tornam-se de grande importância. A espectroscopia de fluorescência é uma ferramenta diagnóstica não invasiva que pode auxiliar na detecção do câncer em tempo real, com o potencial de fornecer sensibilidade e especificidade semelhantes ao diagnóstico clínico de profissionais experientes. É uma técnica relativamente simples, rápida e acurada que consiste em avaliar a composição bioquímica e a estrutura do tecido pelo espectro de fluorescência emitido por ele, após aplicação de um feixe de luz. Quando há progressão de um estado normal para um estado alterado, isso é refletido nas características espectrais da fluorescência dos tecidos, podendo ser correlacionada com o exame histopatológico destes tecidos. O objetivo do presente estudo consistiu em discriminar, na mucosa bucal, tecido sadio de neoplásico, por meio de espectroscopia de fluorescência avaliando as margens ressecadas cirurgicamente. As avaliações ocorreram nos pacientes do Departamento de Cirurgia de Cabeça e Pescoço e Otorrinolaringologia do AC Camargo Câncer Center. O estudo obteve aprovação dos Comitês de Ética em Pesquisa das instituições participantes. A amostra consistiu de 75 indivíduos nos quais se realizou a espectroscopia de fluorescência dos quais 45 pacientes eram portadores de carcinoma oral e 30 voluntários com mucosa oral clinicamente normal. 29 casos (64.4%) do sexo masculino e a média de idade foi de 61.3 anos. Foram realizadas biópsias e os resultados destas duas metodologias foram comparados, usando o diagnóstico histopatológico como padrão ouro, para identificar características espectrais de entre tecidos clinicamente não alterados das margens cirúrgicas da mucosa de voluntários. Os espectros foram classificados e comparados com a histopatologia para determinação da eficiência na discriminação diagnóstica empregando-se a fluorescência. A análise inicial foi qualitativa e após consistiu de processamentos matemáticos dos espectros com excitação nos comprimentos de 532 e 406nm. Observou-se a variabilidade entre os indivíduos, entre os sítios anatômicos, entre regiões da mesma lesão e entre tecido clinicamente normal de voluntários e tecido das margens cirúrgicas em momentos in situ. Foram observadas também grandes diferenças entre espectros in situ e ex vivo, em concordância com resultados de outros estudos. A acurácia da técnica variou em função do tipo de análise empregada, mas pode-se constatar o seu potencial de uso como instrumento auxiliar para avaliar margens cirúrgicas no câncer de bocaAbstract: The most widely used treatment for squamous cell carcinoma of the mouth is surgical resection, whether or not accompanied by radiation and/or chemotherapy. The treatment is simplified, reducing the number of recurrences and increasing survival when the lesions are in the early stages, according to anatomical site and diagnosis of disease-free surgical margins. The development and improvement of techniques for early diagnosis, as well as an accurate definition of disease-free surgical margins and correct delineation of the extent of the mouth cancer, is an important part of improving the quality of life and survival rate for these patients. Fluorescence spectroscopy is a noninvasive diagnostic tool that can aid in real-time cancer detection, with the potential to provide similar sensitivity and specificity to that of the clinical diagnoses of experienced professionals. It is a relatively simple, fast and accurate technique that assesses the biochemical composition and structure of the tissue by the fluorescence spectrum emitted after the application of a beam of light. When there is progression from a normal state to an altered state, this is reflected in the spectral characteristics of the fluorescence of the tissues, which may be correlated with the histopathological examination of these tissues. The aim of this study was to discriminate, in oral mucosa, healthy tissue through fluorescence spectroscopy by evaluating surgically resected margins. Assessments occurred in patients of the Head and Neck Surgery and Otorhinolaryngology Department of A.C. Camargo Cancer Center. The study was approved by the Research Ethics Committee of the institution. The sample consisted of 75 individuals who underwent fluorescence spectroscopy, of which there were 45 individuals with oral carcinoma and 30 healthy volunteers with normal oral mucosa. Twenty-nine cases (64.4%) were male and the mean age was 61.3 years. Biopsies were performed and the results of these two methods were compared using histopathology as the gold standard to identify spectral characteristics from clinically unchanged tissues to surgical margins of the mucosa of volunteers. The spectra were classified and compared with histopathology for determining the efficiency of diagnostic discrimination of employing fluorescence. The initial analysis was qualitative and consisted of following mathematical processing of the spectra with excitation in lengths of 532 and 406 nm. Variability was observed among individuals, anatomical sites, regions of the same lesion and clinically normal tissue of volunteers and the tissue of surgical margins in situ. Also observed were large differences between spectra in situ and ex vivo, in agreement with results of other studies. The accuracy of the technique varied according to the type of analysis used, but its potential use is noted as an aid to evaluated surgical margins in oral cancerDoutoradoEstomatologiaDoutora em Estomatopatologi

Fluorescence spectroscopy for detection of premalignant lesions and squamous cell carcinoma of the oral cavity : prospective study

Orientador: Luiz Paulo KowalskiDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: O câncer da boca é um problema de saúde pública, com grande incidência na população brasileira. O paciente afetado geralmente apresenta-se para o tratamento com a doença em estádios avançados, com conseqüente diminuição das taxas de sobrevida. Diante disso, torna-se imprescindível a detecção do câncer de boca em estádios precoces. A espectroscopia de fluorescência é uma ferramenta diagnóstica não invasiva que pode auxiliar na detecção do câncer em tempo real, com o potencial de fornecer sensibilidade e especificidade semelhantes ao diagnóstico clínico de profissionais experientes. É uma técnica relativamente simples, rápida e acurada que consiste em avaliar a composição bioquímica e a estrutura do tecido pelo espectro de fluorescência emitido por ele, após aplicação de uma fonte de luz. Quando há progressão de um estado normal para um estado alterado, isso é refletido nas características espectrais da fluorescência dos tecidos, podendo ser correlacionada com o exame histopatológico destes tecidos. O objetivo do presente estudo consistiu em discriminar, na mucosa bucal, tecido sadio de neoplásico, por meio de espectroscopia de fluorescência. As avaliações ocorreram nos pacientes do Departamento de Cirurgia de Cabeça e Pescoço e Otorrinolaringologia do Hospital A. C. Camargo e do Laboratório Especial de Laser em Odontologia da USP, São Paulo, Brasil. O estudo obteve aprovação dos Comitês de Ética em Pesquisa das instituições participantes. A amostra consistiu de 150 indivíduos nos quais se realizou a espectroscopia de fluorescência dos quais 55 pacientes eram portadores de carcinoma oral, 30 voluntários com mucosa oral clinicamente normal, 35 pacientes sem lesão oral, com história prévia de neoplasia, que foram submetidos à cirurgia ou à cirurgia e radioterapia, e 30 pacientes com lesão clinicamente detectável e potencialmente maligna. 81 casos (54%) do sexo masculino e a média de idade foi de 62,7 anos. Em relação aos fatores de risco, 76 (50,6%) eram fumantes e 31(20,7%) etilistas. Foram realizadas biópsias e os resultados destas duas metodologias foram comparados, usando o diagnóstico histopatológico como padrão ouro, para identificar características espectrais de cada tipo de lesão. Os espectros foram classificados e comparados com a histopatologia para determinação da eficiência na discriminação diagnóstica empregando-se a fluorescência. A análise consistiu de diversos tipos de processamentos matemáticos dos espectros, sendo as acurácias Bayes 0.513 e 0.618 e J48 0.829 e 0.887 nos comprimentos 532 e 406nm, respectivamente. Observou-se a variabilidade entre as doenças, entre os sítios anatômicos, entre os indivíduos e entre regiões da mesma lesão. Os valores de especificidade e sensibilidade da técnica variam em função do tipo de análise empregada, como na excitação em 406 nm o algoritmo J48+CFS mostrou valores de sensibilidade igual a 88,5% e de especificidade igual a 93,8%, podendo em alguns casos, pode-se constatar o seu potencial de uso como instrumento auxiliar do diagnóstico do câncer de bocaAbstract: Oral cancer is a public health problem with high incidence in the Brazilian population. The affected patient usually presents for clinical investigation, with advanced stage disease, the consequence of this delay is a reduction in survival rates. Given this, it's essential to detect oral cancer in early stages. Fluorescence spectroscopy is a non-invasive diagnostic tool that can aid in cancer detection in real time, with the potential to provide similar sensitivity and specificity for clinical diagnosis of experienced professionals. This technique is relatively simple, fast and accurate which evaluates the biochemical composition and structure of the tissue fluorescence spectrum emitted by it, after application of a light source. When there is a progression from normal state to an altered state, this is reflected in the spectral characteristics of fluorescence of tissues, which may be correlated with the histopathological examination of tissues. The aim of this study was to discriminate, in the oral mucosa, healthy tissue and tumor, through fluorescence spectroscopy. Assessments occurred in patients of the Department of Head and Neck Surgery and Otorhinolaryngology, Hospital A. C. Camargo and the Special Laboratory of Laser in Dentistry, USP, São Paulo, Brazil. The study was approved by the Ethics Research Committee of the participant institutions. The sample consisted of 150 individuals who were submitted to fluorescence spectroscopy, of whom 55 patients had oral carcinoma, 30 volunteers with normal oral mucosa, 35 patients without oral lesions, but with a previous history of malignancy, who underwent surgery or surgery and radiotherapy, and 30 patients with a clinically detectable potentially malignant lesions. 81 cases (54%) were male and the mean age was of 62.7 years. Regarding risk factors, 76 (50.6%) were smokers and 31 (20.7%) consumed alcohol. Biopsies were performed and the results of these two methods were compared using the histopathologic diagnosis as the gold standard for comparison to the spectral characteristics of each type of lesion. The spectra were classified and compared to histopathology for determining the efficiency in diagnostic discrimination employing fluorescence. The analysis consisted of various types of mathematical processing of spectra, and the accuracies Bayes 0,513 and 0,618 and J48 0,829 and 0,887 in lengths of 532 and 406nm, respectively. It can be observed variability among diseases, among the anatomical sites among individuals and among regions of the same injury. The specificity and sensitivity of the technique varies depending on the type of analysis employed as the excitation at 406 nm the algorithm J48 + CFS showed sensitivity of 88,5% and specificity of 93,8% and may in some cases, one can see its potential use as an aid to diagnosis of oral cancerMestradoPatologiaMestre em Estomatopatologi

Clinicopathologic analysis of 14 cases of odontogenic myxoma and review of the literature

Odontogenic myxoma is a rare benign neoplasm that originates from odontogenic ectomesenchyme. There is no standard of care and recurrences are frequent after conservative surgical procedures. A retrospective study conducted at a single cancer center, with analysis of medical records of all patients diagnosed with odontogenic myxoma from 1980 to 2010, along with a literature review. There were 14 patients with diagnosis of odontogenic myxoma (OM). Most patients were female (78.6%) and Caucasian (100%), with ages ranging from 7 to 51 years (21.6 ± 11.6 years). The time period between the first symptom and first consultation ranged from 0 to 60 months (19.4 ± 19.97 months). The most frequent complaints were increased local volume or failure to tooth eruption. The most common tumor site was the mandible (11 cases, 78.5%). About radiological findings, most lesions were multilocular (9 cases, 64.3%) and with imprecise limits (12 cases, 85.7%). Surgery was performed in all cases and curettage was the most applied technique (10 cases, 71.4%). Three patients underwent mandibulectomy and complex reconstructions including iliac crest microvascular flap. Three patients had postoperative complications and 4 had local recurrences of the tumor. The follow up time ranged from 12 to 216 months (112 ± 70.8 months). All patients are without clinical and radiographic evidence of disease. OM is a locally aggressive and rare tumor. There is no gold standard surgical management and the therapeutic decision should be individualized taking into account the characteristics and extension of the tumor

Analysis Of Surgical Margins In Oral Cancer Using In Situ Fluorescence Spectroscopy.

Oral cancer is a public health problem with high prevalence in the population. Local tumor control is best achieved by complete surgical resection with adequate margins. A disease-free surgical margin correlates with a lower rate of local recurrence and a higher rate of disease-free survival. Fluorescence spectroscopy is a noninvasive diagnostic tool that can aid in real-time cancer detection. The technique, which evaluates the biochemical composition and structure of tissue fluorescence, is relatively simple, fast and, accurate. This study aimed to compare oral squamous cell carcinoma lesions to surgical margins and the mucosa of healthy volunteers by fluorescence spectroscopy. The sample consisted of 56 individuals, 28 with oral squamous cell carcinoma and 28 healthy volunteers with normal oral mucosa. Thirty six cases (64.3%) were male and the mean age was 60.9 years old. The spectra were classified and compared to histopathology to determine fluorescence efficiency for diagnostic discrimination of tumors. In the analysis of the other cases we observed discrimination between normal mucosa, injury and margins. At two-year follow up, three individuals had local recurrence, and in two cases investigation fluorescence in the corresponding area showed qualitative differences in spectra between the recurrence area and the area without recurrence at the same anatomical site in the same patient. In situ analysis of oral mucosa showed the potential of fluorescence spectroscopy as a diagnostic tool that can aid in discrimination of altered mucosa and normal mucosa.50593-

Fluorescence spectroscopy for the detection of potentially malignant disorders and squamous cell carcinoma of the oral cavity

Oral cancer is a public health problem with relevant incidence in the world population. The affected patient usually presents advanced stage disease and the consequence of this delay is a reduction in survival rates. Given this, it is essential to detect oral cancer at early stages. Fluorescence spectroscopy is a non-invasive diagnostic tool that can improve cancer detection in real time. It is a fast and accurate technique, relatively simple, which evaluates the biochemical composition and structure using the tissue fluorescence spectrum as interrogation data. Several studies have positive data regarding the tools for differentiating between normal mucosa and cancer, but the difference between cancer and potentially malignant disorders is not clear. The aim of this study was to evaluate the efficacy of fluorescence spectroscopy in the discrimination of normal oral mucosa, oral cancer, and potentially malignant disorders. The fluorescence spectroscopy was evaluated in 115 individuals, of whom 55 patients presented oral squamous cell carcinoma, 30 volunteers showing normal oral mucosa, and 30 patients having potentially malignant disorders. The spectra were classified and compared to histopathology to evaluate the efficiency in diagnostic discrimination employing fluorescence. In order to classify the spectra, a decision tree algorithm (C4.5) was applied. Despite of the high variance observed in spectral data, the specificity and sensitivity obtained were 93.8% and 88.5%, respectively at 406 nm excitation. These results point to the potential use of fluorescence spectroscopy as an important tool for oral cancer diagnosis and potentially malignant disorders1128290CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP477439/2007-198/14270-8; 07/57126-5; 09/12938-8; 2008/57887-

DNA methylation-based depiction of the immune microenvironment and immune-associated long non-coding RNAs in oral cavity squamous cell carcinomas

Oral cavity squamous cell carcinoma (OSCC) is a complex and dynamic disease characterized by clinicopathological and molecular heterogeneity. Spatial and temporal heterogeneity of cell subpopulations has been associated with cancer progression and implicated in the prognosis and therapy response. Emerging evidence indicates that aberrant epigenetic profiles in OSCC may foster an immunosuppressive tumor microenvironment by modulating the expression of immune-related long non-coding RNAs (lncRNAs). DNA methylation analysis was performed in 46 matched OSCC and normal adjacent tissue samples using a genome-wide platform (Infinium HumanMethylation450 BeadChip). Reference-based computational deconvolution (MethylCIBERSORT) was applied to infer the immune cell composition of the bulk samples. The expression levels of genes encoding immune markers and differentially methylated lncRNAs were investigated using The Cancer Genome Atlas dataset. OSCC specimens presented distinct immune cell composition, including the enrichment of monocyte lineage cells, natural killer cells, cytotoxic T-lymphocytes, regulatory T-lymphocytes, and neutrophils. In contrast, B-lymphocytes, effector T-lymphocytes, and fibroblasts were diminished in tumor samples. The hypomethylation of three immune-associated lncRNAs (MEG3, MIR155HG, and WFDC21P) at individual CpG sites was confirmed by bisulfite-pyrosequencing. Also, the upregulation of a set of immune markers (FOXP3, GZMB, IL10, IL2RA, TGFB, IFNG, TDO2, IDO1, and HIF1A) was detected. The immune cell composition, immune markers alteration, and dysregulation of immune-associated lncRNAs reinforce the impact of the immune microenvironment in OSCC. These concurrent factors contribute to tumor heterogeneity, suggesting that epi-immunotherapy could be an efficient alternative to treat OSCC

The functional effect of soybean extract and isolated isoflavone on myocardial infarction and ventricular dysfunction The soybean extract on myocardial infarction

Made available in DSpace on 2019-09-12T16:53:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2012Pontifícia Universidade Católica do Paraná (PUCPR)Background: Myocardial infarction is a public health problem. Functional food is an alternative treatment for cardiovascular diseases. Objective: The objective was to analyze the functional and anatomopathological post-myocardial-infarction effects of soybean extract (SE) and isoflavone (IF). Methods: Myocardial infarction was induced in adult Wistar rats. After 5 days, an echocardiogram was performed to determine heart rate (HR), ejection fraction (EF), systolic volume (LVESV) and diastolic volume (LVEDV). Animals with ventricular dysfunction (EF<45%) were selected for study. The animals were divided into three groups: control (n=14), SE (n=15) and IF (n=12). The IF group received 120 mg/kg/day isolated IF, and the SE group received 12.52 g/day. After 30 days, a new echocardiogram was performed. A histological exam was carried out to determine the collagen. Activity of biochemical markers [arginase. lactate dehydrogenase (LDH) and malate dehydrogenasel was measured. Results: The animals of the control, IF and SE groups showed a reduction in EF after the infarction (P=.432, P=.017 and P=.320, respectively). An increase of LVESV and LVEDV was observed in all groups (P=.009, P=.001 and P=.140; and P=.003, P=.008 and P=.205, respectively). A reduction of HR was found in the SE group (P=.020). There was a greater activity of LDH in the SE group. A smaller quantity of mature collagen was found in the region proximal to the myocardial infarction in the SE group. Conclusion: A protective effect in the SE group was observed 30 days after the myocardial infarction. (C) 2012 Elsevier Inc. All rights reserved.[Miguez, Ana C.; Francisco, Julio C.; Barberato, Silvio H.; Simeoni, Rossana; Precoma, Dalton; do Amaral, Vivian Ferreira; Olandoski, Marcia; de Noronha, Lucia; Greca, Fernando H.; Faria-Neto, Jose Rocha; Guarita-Souza, Luiz C.] Pontifical Catholic Univ Parana PUCPR, Ctr Biol & Hlth Sci, Expt Lab, Curitiba, Parana, Brazil[Rodrigues, Edson] Universidade de Taubaté (Unitau), Unitau, SP, Brazil[Teixeira de Carvalho, Katherine Athayde] Pele Little Prince Inst Chid & Adolescent Hlth Re, Curitiba, Parana, Brazi