The Fagerström and AUDIT Tests as Probable Screening Tools in Oral Cancer and Their Correlation with CYP1A1, GSTM1, GSTP1, and GSTT1 Gene Expression.

BACKGROUND: Cancer is currently a major public health problem worldwide, with a marked increase of about 70% in the number of expected diagnosed cases over the next two decades. The amount of tobacco and alcohol consumed is calculated based on the subjective information provided by the user. Tobacco exposure can be assessed using the Fagerström Test for Cigarette Dependence (FTCD) and alcohol consumption by the Alcohol Use Disorder Identification Test (AUDIT). MATERIALS AND METHODS: Forty-eight subjects answered the Fagerström, and AUDIT tests and we studied them as likely screening tools for oral cancer and their correlation with the expression of CYP1A1, GSTM1, GSTP1, and GSTT1 genes by the RT-qPCR method. RESULTS: There were significant differences in the AUDIT score and CYP1A1 expression between cancer and control groups. Participants in advanced stages, whether due to tumor size or regional metastasis, showed significant differences in the duration of tobacco use, FTCD, AUDIT score, and CYP1A1 expression when compared to patients in early stages. Among subjects without cancer, we found a significant correlation between participant age and GSTP1 expression. Furthermore, the expression of GSTP1 was significantly correlated with the number of cigarettes smoked per day, duration of tobacco use, and FTCD. CONCLUSIONS: Questionnaires designed to evaluate the degree of tobacco and alcohol exposure and dependence combined with gene expression tests can be useful to assess the risk of developing oral cancer. Furthermore, raising the awareness of individuals regarding their degree of dependence and encouraging them to participate in cessation programs are important educational measures for the prevention of tobacco-related malignancies.

Influence of nicotine dependence, current smoking status and gender on CYP1A1, CYP1B1 and CYP2A6 expression in oral squamous cell carcinoma / Influência da dependência nicotínica, parâmetros de consumo de tabaco e gênero na expressão de CYP1A1, CYP1B1 e CYP2A6 em carcinoma de células escamosas oral

Objective: Tobacco smoke is composed of cancer-causing chemicals referred to as carcinogens. These carcinogens are metabolized by the enzymes of the cytochrome P450 (CYP) family. Our objective was to evaluate the correlation of tobacco consumption parameters with CYP1A1, CYP1B1 and CYP2A6 expression using qRT-PCR in samples of oral squamous cell carcinoma (OSCC). Material and Methods: The sample was divided into 2 groups: Cancer (36 subjects) and non-Cancer (12 subjects). The smokers' participants (36) were evaluated regarding their Nicotine dependence (ND) was assessed by the Fagerström test for cigarette dependence (FTCD). Questions regarding tobacco consumption like the number of cigarettes/day (CPD), duration of use, and pack-years were also evaluated. The Mann-Whitney and Spearman correlation tests were used at a significance level of 5%. Results: 48 participants were included, 32 men (66.7%), 36 smokers (75%) and 27 smokers with OSCC (56.3%). Samples of OSCC expressed more CYP1A1, CYP1B1, and CYP2A6. Especially, the CYP1B1 gene was significantly expressed in OSCC samples, regardless gender or tobacco use. No women expressed CYP2A6, as well as, non-smokers did not express the CYP1A1 and CYP2A6 genes. CYP1A1 gene was higher among men (P = 0.021). Conclusion: Lack of exposure to tobacco may justify the absence of CYP1A1 and CYP2A6 expression in non-smokers. The CYP1B1 gene was significantly expressed in the cancer presence despite gender or tobacco use. The assessment of ND and quantification of tobacco consumption are important instruments in monitoring smokers with benign oral lesions and, especially, in the presence of cancer.(AU)

Objetivo: A fumaça do tabaco é composta de substâncias químicas cancerígenas conhecidas como carcinógenos. Esses carcinógenos são metabolizados pelas enzimas da família do citocromo P450 (CYP). Nosso objetivo foi avaliar a correlação dos parâmetros do consumo de tabaco com a expressão de CYP1A1, CYP1B1 e CYP2A6 por qRT-PCR em amostras de carcinoma de células escamosas bucal (CCEB). Material e Métodos: A amostra foi dividida em 2 grupos: Câncer (36 indivíduos) e sem Câncer (12 indivíduos). Os participantes fumantes (36) foram avaliados quanto à dependência nicotínica (DN) pelo teste de Fagerström para dependência de cigarro (TFDC). Questões relacionadas ao consumo de tabaco como número de cigarros / dia (CPD), tempo de uso e anos-maço também foram avaliadas. Os testes de correlação de Mann-Whitney e Spearman foram utilizados com nível de significância de 5%. Resultados: foram incluídos 48 participantes, 32 homens (66,7%), 36 fumantes (75%) e 27 fumantes com CCEB (56,3%). Amostras de CCEB expressaram mais CYP1A1, CYP1B1 e CYP2A6. Especialmente, o gene CYP1B1 foi significativamente expresso em amostras de CCEB, apesar do sexo ou uso de tabaco. Nenhuma mulher expressou CYP2A6, assim como, não fumantes não expressaram os genes CYP1A1 e CYP2A6. O gene CYP1A1 foi maior entre os homens (P = 0,021). Conclusão: A falta de exposição pode justificar a ausência da expressão dos genes CYP1A1 e CYP2A6 entre não fumantes. O gene CYP1B1 foi significativamente expresso na presença de câncer, independentemente do sexo ou do uso de tabaco. A avaliação da DN e a quantificação do consumo de tabaco são importantes instrumentos no acompanhamento de fumantes com lesões bucais benignas e, principalmente, na presença de câncer (AU)

Expression of DNA repair genes in oral squamous cell carcinoma using reverse transcription-quantitative polymerase chain reaction.

OBJECTIVE: The aim of this study was to evaluate the expression of DNA repair genes in cases of oral squamous cell carcinoma (OSCC). STUDY DESIGN: Expression of the MLH1, MSH2, MLH3, ATM, MRE11A, XRCC1, and PMS2 genes was evaluated by reverse transcription-quantitative polymerase chain reaction in the OSCC group (32 patients) and the control group (15 patients). The groups were compared by using the Mann-Whitney test, with Bonferroni correction. Associations between gene expression levels and clinical data were explored by using Pearson's and Spearman's correlation coefficients, with P value less than .05 indicating a significant difference. RESULTS: The MLH1, MSH2, MLH3, ATM, MRE11A, XRCC1, and PMS2 genes were downregulated in the OSCC group compared with the control group, with significant values for MLH1 (P < .0001); MSH2 (P = .038); MLH3 (P < .0001); ATM (P < .0001); MRE11A (P < .0001); XRCC1 (P = .0004); and PMS2 (P = .008). Analysis of the correlation between gene expression and clinical data only revealed a significant negative correlation between age and expression of the PMS2 gene. CONCLUSIONS: Expression of the DNA repair genes MLH1, MSH2, MLH3, ATM, MRE11 AMRE11A, XRCC1, and PMS2 was reduced in OSCC.

Immediate placement of denture over undiagnosed ameloblastoma / Instalação de prótese total imediata sobre ameloblastoma não diagnosticado

Ameloblastoma is an odontogenic tumor that affects the bones of the jaw, often affecting posterior region of the mandible. It is a benign neoplasm and is frequently associated to an unerupted tooth. The purpose of this article is to report a case of failure in making an immediate full denture in a patient that had an ameloblastoma. A female patient aged 67 years complained of a hard swelling in the right mandible at the premolar region, that she noted after teeth extraction and confection of an immediate denture. Intraoral examination showed buccal and lingual cortical plate expansion and radiograph examination showed multilocular radiolucency with a well-defined margin. Aspiration was nonproductive and the provisional diagnosis was ameloblastoma. An incisional biopsy was performed and the histopathological report was conclusive of an acanthomatous ameloblastoma. The patient was sent for surgical excision of the lesion and after three years, the patient returned reporting that she was operated elsewhere in the past year and had an unsuccessful bone graft. She was sent to a prosthodontist to make a complete denture. In the present case the ameloblastoma was diagnosed only after the teeth extraction and immediate denture (AU)

O ameloblastoma é um tumor odontogênico que afeta os ossos maxilares, geralmente a região posterior de mandíbula. É uma neoplasia benigna frequentemente associada a um dente não irrompido. O objetivo deste artigo é relatar um caso de prótese total imediata confeccionada para uma paciente com ameloblastoma na região anterior da mandíbula. Paciente do sexo feminino, 67 anos, queixou-se de aumento de volume na região de pré-molares inferiores do lado direito após extração dentária e confecção de prótese total imediata. Ao exame clínico intrabucal foi observada expansão da cortical vestibular e lingual e o exame radiográfico revelou radiolucência multilocular com margem bem definida. A punção aspirativa não foi produtiva e o diagnóstico provisório foi de ameloblastoma. Foi realizada biópsia incisional e o laudo histopatológico foi conclusivo para ameloblastoma acantomatoso. A paciente foi encaminhada para ressecção cirúrgica da lesão e não retornou. Depois de três anos procurou atendimento e relatou que tinha sido submetida a cirurgia com colocação de enxerto ósseo para implante sem sucesso. O enxerto ósseo foi perdido e a paciente necessitava de nova prótese total. Ela foi encaminhada a um protesista. No presente caso, o diagnóstico de ameloblastoma foi feito somente após a instalação da prótese total imediata, quando a paciente notou aumento de volume (AU).

Quality of life in a smoking cessation program: lifestyle, anxiety, and depression / Qualidade de vida em um programa de cessação do tabagismo: estilo de vida, ansiedade e depressão

Objective: Tobacco dependence is quite variable among individuals because it includes many aspects such as psychological, emotional, behavioral, and cultural perspectives. This study aimed to evaluate the social and emotional issues of patients in a smoking cessation program through the generic and specific quality of life questionnaires. Material and Methods: Two generic questionnaires (SF36 and Fantastic Lifestyle) and two specific (SRQ-20 for depression and STAI s/t for levels of state and anxiety trait) were applied at the first meeting and after three months of treatment. Data were statistically analyzed by the Pearson correlation coefficient (significance level of 5%). Results: In total, 15 participants (10 men, 5 women), with mean age of 60 and 56, respectively. The mean of the smoking intake was 65.88±34.37 for men and 35.66±11.17 for women. SRQ20 and STAI s/t indicated that poor improvement in depression and anxiety was related to relapse; SF36 questionnaire indicated improvement in the domains of physical aspect limit, pain, social and emotional aspects; Fantastic Lifestyle indicated improvement according to its scale, being in a first moment "Regular" (53 points), and later, "Good" (55.67 points). Conclusion: In conclusion, smoking cessation treatment is correlated to the decrease in anxiety and depression levels and improvement of the general quality of life (AU)

Objetivo: A dependência do tabaco é bastante variável entre os indivíduos porque inclui muitos aspectos, como perspectivas psicológicas, emocionais, comportamentais e culturais. Este estudo teve como objetivo avaliar as questões sociais e emocionais de pacientes em um programa de cessação tabagágica por meio de questionários genéricos e específicos de qualidade de vida. Material e Métodos: Dois questionários genéricos (SF36 e Estilo de Vida Fantástico) e dois específicos (SRQ-20 para depressão e IDATE s/t para níveis de estado e traço de ansiedade) foram aplicados no primeiro encontro e após três meses de tratamento. Os dados foram analisados estatisticamente pelo coeficiente de correlação de Pearson (nível de significância de 5%). Resultados: No total, 15 participantes (10 homens, 5 mulheres), com média de idade de 60 e 56 anos, respectivamente. A média do consumo tabágico foi de 65,88 ± 34,37 para homens e 35,66 ± 11,17 para mulheres. SRQ20 e IDATE s/t indicaram que a fraca melhora na depressão e ansiedade estava relacionada à recaída; o questionário SF36 indicou melhora nos domínios limite do aspecto físico, dor, aspectos sociais e emocionais; Estilo de Vida Fantástico indicou melhora de acordo com sua escala, sendo em um primeiro momento "Regular" (53 pontos), e posteriormente, "Bom" (55.67 pontos). Conclusão: Em conclusão, o tratamento para cessação do tabagismo está correlacionado à diminuição dos níveis de ansiedade e depressão e à melhora da qualidade de vida geral (AU)

Tobacco influence in heavy metals levels in head and neck cancer cases.

Heavy metals intoxication is known to be risk factors for various diseases, including cancer. These metals may be presented in food and soil as well as in leaf and tobacco smoke. The aim of this study was to correlate the exposure to heavy metals stemming from tobacco and head and neck squamous cell carcinoma carcinogenesis. Analysis of lead, copper, manganese, arsenic, chromium, and cadmium by atomic absorption spectrophotometry was performed in whole blood samples from 91 patients: 68 smokers with oral cavity, pharynx, or laryngeal cancer; 8 non-smokers with oral or larynx cancer; and 15 non-cancer smokers with tobacco-related diseases (control group). No differences were found in metals quantifications, except a significant difference was observed (p = 0.0223) with higher mean in copper levels for non-smokers with cancer. The present study concluded, for the groups evaluated, it was not possible to prove the relationship between the studied metals in the development of the neoplasm. On the other hand, the results of copper demonstrated a correlation with smokers with cancer and lower levels of circulating copper.

Screening methylation of DNA repair genes in the oral mucosa of chronic smokers.

OBJECTIVE: The aim of this study was to evaluate the epigenetic changes in the process of oral carcinogenesis by screening the methylation of repair genes in chronic smokers. DESIGN: Two groups were formed: Group 1: 16 smokers with consumption of 20 cigarettes/day for at least 10 years; and Group 2: 10 non-smoking. Exfoliative cytology of the tongue was performed, and the extracted DNA was treated by enzymes. The PCR Array System performed methylation screening to evaluate 22 DNA repair genes, and the results were validated by RT-qPCR for each gene with methylation levels ≥10%. RESULTS: Highest percentages of methylation were observed for MLH3 and XRCC1 genes (11-20% methylation) and in one case for MRE11A and PMS2 (>50% methylation). Statistical analysis showed significant differences in the expression of the genes MRE11A (p = 0.0002), PMS2(p = 0.0068), XRCC1 (p = 0.0080) and MLH3 (0.0057) between the two groups. CONCLUSION: The effects of chronic smoking on oral mucosa led to the methylation of genes MRE11A PMS2, XRCC1 and MLH3, but resulted in a reduction of gene expression of MRE11A and PMS2, which showed ≥50% methylation. These results provide evidence that smoking cause methylation and reduced expression of repair genes.

Repair genes expression profile of MLH1, MSH2 and ATM in the normal oral mucosa of chronic smokers.

OBJECTIVE: The aim of this study was to evaluate the effect of chronic smoking on the expression profile of the repair genes MLH1, MSH2 and ATM in the normal oral mucosa of chronic smokers and never smokers. MATERIALS AND METHODS: The sample consisted of thirty exfoliative cytology smears per group obtained from Smokers and Never Smokers. Total RNA was extracted and expression of the MLH1, MSH2 and ATM genes were evaluated by quantitative real-time and immunocytochemistry. The gene and protein expression data were correlated to the clinical data. Gene expression was analyzed statistically using the Student t-test and Pearson's correlation coefficient, with p<0.05. RESULTS: MLH1, MSH2 and ATM genes were downregulated in the smoking group compared to the control with significant values for MLH1 (p=0.006), MSH2 (p=0.0001) and ATM (p=0.0001). Immunocytochemical staining for anti-MLH1, anti-MSH2 and anti-ATM was negative in Never Smokers; in Smokers it was rarely positive. No significant correlation was observed among the expression of MLH1, MSH2, ATM and age, number of cigarettes consumed per day, time of smoking during life, smoking history or levels of CO in expired air. CONCLUSION: The expression of genes and proteins related to DNA repair mechanism MLH1, MSH2 and ATM in the normal oral mucosa of chronic smokers was reduced.

How to improve the early diagnosis of oral cancer? / Como melhorar o diagnóstico precoce do câncer bucal?

Objective: Oral squamous cell carcinoma (OSCC) is one of the most common malignant neoplasms, with 300,000 new cases annually diagnosed in the world. Although it appears in the fifth position among men in Brazil, late diagnosis continues to be a leading cause of poor prognosis. We reviewed the literature to evaluate factors that contribute to diagnostic delay in oral cancer considering aspects related to the patient, health team and public health system. Material and Methods: The Medline (PubMed), Scopus, Web of Science and Embase databases were searched in the last 5 years using the following MeSH descriptors: carcinoma, squamous cell; mouth neoplasms; delayed diagnosis. The articles should contain in their objectives factors contributing to the delayed diagnosis of OSCC related to the patient, healthcare professionals and public health system in general. Results: One hundred twenty articles were retrieved and 27 on the delayed diagnosis of oral cancer were eligible. Conclusion: The literature showed that the delay in the diagnosis and treatment of patients with oral cancer could be minimized by providing health information, particularly about risk factors, by improving the training and retraining of physicians and dentists and by improving access to the health system. The communication technologies such as telemedicine, can play a key role in the early diagnosis of oral cancer. (AU)

Objetivo: o carcinoma de células escamosas de cavidade bucal (CCECB) é uma das neoplasias malignas mais comuns, com cerca de 300 mil novos casos diagnosticados anualmente no mundo. Embora esteja na quinta posição entre os mais frequentes em homens no Brasil, o diagnóstico tardio continua a ser a principal causa de mau prognóstico. Revisamos a literatura para avaliar fatores que possam contribuir para o atraso de diagnóstico no câncer bucal considerando aspectos relacionados ao paciente, equipe de saúde e o Sistema de Saúde Pública. Material e Métodos: Foram pesquisados os bancos de dados Medline (PubMed), Scopus, Web of Science e Embase nos últimos 5 anos utilizando os seguintes descritores MeSH: carcinoma de células escamosas, neoplasias de boca e atraso diagnóstico. Os artigos deveriam conter em seus objetivos fatores que possam ter contribuído para o diagnóstico tardio do CCECB relacionado ao paciente, aos profissionais de saúde e ao Sistema de Saúde Pública em geral. Resultados: Cento e vinte artigos foram recuperados, sendo elegíveis, vinte e sete relacionados ao diagnóstico tardio de câncer bucal. Conclusão: a literatura mostrou que o atraso no diagnóstico e tratamento de pacientes com câncer bucal poderia ser minimizado, fornecendo informações sobre saúde, particularmente sobre fatores de risco a população, melhorando o treinamento e reciclagem de médicos e dentistas e melhorando o acesso ao sistema de saúde. As tecnologias de comunicação, dentre elas a telemedicina, pode desempenhar um papel fundamental no diagnóstico precoce de câncer bucal. (AU)

Challenges to be overcome: Current situation involving smoking cessation in Dental setting / Desafios a serem superados: situação atual envolvendo cessação do tabagismo no cenário odontológico

Objetivo: O uso do tabaco é a principal causa de doenças evitáveis no mundo. A consulta odontológica representa uma oportunidade de abordar os fumantes no controle do uso do tabaco. Embora esta conclusão seja bem conhecida, a intervenção para cessação tabágica entre dentistas permanece menos explorada do que o esperado. Este estudo avaliou artigos publicados sobre intervenções entre dentistas e profissionais de saúde dentária na cessação do tabagismo e discute as barreiras a serem superadas para atingir esse objetivo. Material e Métodos: As bases de dados Medline (PubMed), Scopus, Web of Science, Cochrane Reviews, LILACS e Google Scholar foram pesquisadas por artigos publicados em inglês entre janeiro de 2010 e dezembro de 2016 usando os seguintes termos MeSH: tobacco use cessation, smoking cessation, tobacco use disorder and dentistry. A busca manual complementar para potenciais artigos foi feita verificando-se referências bibliográficas. Resultados: setenta e duas publicações foram elegíveis e os resultados foram divididos em oito categorias: 1) o papel dos dentistas no controle do uso do tabaco; 2) conhecimento sobre tratamento e intervenções sobre uso do tabaco; 3) características da abordagem do dentista aos fumantes; 4) barreiras e facilitadores; 5) intervenções futuras para cessação do tabagismo, 6) reembolso de intervenções de tabaco, 7) pesquisas e 8) revisões sistemáticas e diretrizes. Conclusão: Os dentistas desempenham um papel importante na prevenção e controle do tabagismo. O treinamento na cessação do uso do tabaco tem aumentado nas últimas décadas. Ensinar a lidar com o tabagismo em cursos de graduação deve ser uma prioridade para os próximos anos, de modo que uma nova geração de dentistas incorpore em suas atividades habituais uma abordagem natural e eficiente para os fumantes.(AU)

Objective: Tobacco use is the leading cause of preventable illness in the world. The dental office represents an opportunity to smokers approach tobacco use control. Despite this well-known statement tobacco use cessation intervention delivered by dentists remain less explored than expected. This study evaluated published articles about interventions delivered by dentists and dental healthcare professionals on smoking cessation and discusses the barriers to be overcome to achieve this goal. Material and Methods: The Medline (PubMed), Scopus, Web of Science, Cochrane Reviews, LILACS and Google Scholar databases were searched for articles published in English between January 2010 and December 2016 using the following MeSH terms keywords: tobacco use cessation, smoking cessation, tobacco use disorder and dentistry. Complementary hand searching was done checking bibliographic references for potentially articles. Results: Seventy-two publications were eligible and the results were divided into eight categories: 1) the role of dentists in tobacco use control; 2) knowledge about tobacco use treatment and interventions; 3) characteristics of the dentist's approach to smokers; 4) barriers and facilitators; 5) future interventions for smoking cessation, 6) reimbursement for tobacco interventions, 7) surveys and 8) systematic reviews or guidelines Conclusion: Dentists play an important role in the prevention and control of smoking. Training in tobacco use cessation has been increased in the past few decades. Teaching how to deal with smoking in undergraduate courses should be a priority for the next few years, so that a new generation of dentists will incorporate into their habitual activities a natural and efficient approach to smokers.(AU)

Comparison of RNA Extraction Methods for Molecular Analysis of Oral Cytology.

OBJECTIVE OF WORK: The aim of this study was to compare three methods of RNA extraction for molecular analysis of oral cytology to establish the best technique, considering its concentration and purity for molecular tests of oral lesions such as real-time reverse transcriptase reaction. MATERIAL AND METHODS: The sample included exfoliative cytology from the oral cavity mucosa of patients with no visible clinical changes, using Orcellex Rovers Brush®. The extraction of total RNA was performed using the following three techniques: 30 samples were extracted by Trizol® technique, 30 by the Direct-zolTM RNA Miniprep system and 30 by the RNeasy mini Kit. The absorbance was measured by spectrophotometer to estimate the purity. The estimated RNA concentration was obtained by multiplying the value of A260 (ng/mL) by 40. Statistical analysis of the obtained data was performed using GraphPad Prism 5.03 software with Student t, analysis of variance and Bonferroni tests, considering p ≤0.05. RESULTS: Trizol® group revealed higher average concentration, followed by Direct-zolTM and Rneasy group. It was observed that the RNA Direct-zolTM group had the highest purity, followed by RNeasy and Trizol® groups, allowing for the two ratios. CONCLUSION: Considering all aspects, concentration, purity and time spent in the procedures, the Direct-zolTM group showed the best results.