Is DNA ploidy related to smoking?

BACKGROUND: In the oral cavity, genomic instability is caused by long-term exposure to carcinogens. The aim of this study was to evaluate the relationship between smoking and DNA ploidy. METHODS: Cytological material was obtained from patients participating in the Outpatient Smoking Treatment Program of the Heart Institute (INCOR-HCFMUSP), and of the Discipline of Oral Medicine (ICT-UNESP). The inclusion criteria for all groups were the absence of a history of malignant tumors, absence of clinical signs of changes in the selected area, and alcohol consumption of less than 3 units per week. Group 1:30 smokers before smoking cessation treatment; Group 2:30 non-smokers; Group 3:30 ex-smokers abstinent for at least 1 year. Cytological smears were collected from the floor of the mouth and border of the tongue and stained by Feulgen. Aneuploidy was evaluated using the ACIS® III system. RESULTS: The Kruskal-Wallis test showed no statistically significant difference (P = .4383) between the groups studied. No association between tobacco consumption and aneuploidy was observed in group 1 (P = 1) or group 2 (P = .68; Fisher's exact test). CONCLUSION: Smoking was not associated with changes in DNA content or the incidence of aneuploidy in normal oral mucosa.

Effect of HIV infection in the micronuclei frequency on the oral mucosa.

BACKGROUND: The genotoxic impact of HIV infection on the oral cavity malignancies is unknown. The aim of this study was to evaluate the effect of HIV infection in micronucleus (MN) frequency on the oral mucosa of HIV+ patients and establish a relationship with early cytogenetic changes in oral carcinogenesis. METHODS: Thirty HIV+ individuals who are under highly active antiretroviral therapy (HAART) and 30 non-HIV patients were evaluated. Two smears were taken from the lateral border of the tongue and mouth floor and stained by Feulgen. The frequency of MN was examined in 3000 cells per subject under common microscopy. RESULTS: MN analysis showed no significant difference between groups by Mann-Whitney U-test for total MNs (P = 0.178). The presence of single MN was greater in control group with statistical significance (P = 0.009), while in HIV group, multiple MNs were exhibited in higher mean. CONCLUSIONS: HIV patients under HAART therapy and low viral load values showed higher frequency of multiple MNs, which, although not statistically significant, may be caused by the action of the Vpr gene, an accessory gene of HIV. These results corroborate the theory of HIV infection cytogenetic damage.

Effect of smoking cessation in saliva compounds by FTIR spectroscopy.

INTRODUCTION: Smoking is currently considered one of the biggest risk factors for the development of various diseases and early death. Fourier transform infrared (FTIR) spectroscopy is a valuable tool for analysis of biofluids such as saliva and is considered useful for diagnostic purposes. The aim of this study was to evaluate the effect of smoking cessation on saliva composition by FTIR spectroscopy. METHODS: We analyzed the saliva of participants in two groups: a smoker group made up of 10 chronic smokers and a former smoker group made up of 10 individuals who had stopped smoking. Members of both groups had similar smoking history. RESULTS: The results showed few differences in spectral intensity between the groups; however, spectral peaks were slightly increased in the group of smokers in the bands for DNA, indicating modification of its content or cell necrosis. They were also increased for the mannose-6-phosphatase molecule, which is expressed in prostate and breast carcinomas. In the former smoker group, the peak of thyociante was decreased and the band referring to collagen increased in intensity, which indicates a better tissue regeneration capacity. CONCLUSION: Considering these results and the fact that tobacco intake was similar between the groups, it can be concluded that there was recovery of tissue regeneration capacity with smoking cessation during the study period, although the effects found in smokers persisted in the bodies of those who had given up smoking.

Oral Adenosquamous Carcinoma Mimicking a Pyogenic Granuloma: a Challenging Diagnosis.

Adenosquamous carcinoma is an aggressive variant of squamous cell carcinoma. This report describes a case of adenosquamous carcinoma with clinical features of a benign lesion and discusses the differential diagnoses, especially regarding histopathological and immunohistochemical analyses. A 45-year-old male was referred to our outpatient clinic complaining about a rapid-growing enlargement in hard palate. Clinical examination revealed an erythematous and pedunculated nodule with lobulated non-ulcerated surface. Excisional biopsy was performed following clinical diagnosis of pyogenic granuloma. Histologically, the specimen consisted of areas characterizing both well-differentiated squamous cell carcinoma and true adenocarcinoma. After a broad list of immunohistochemical markers was evaluated (AE1/AE3, CEA, CK5, CK7, CK8/18, p53, p63 and Ki67), the diagnosis of adenosquamous carcinoma was rendered and the patient referred to complementary surgery. Adenosquamous carcinoma represents a challenge in diagnostic routine due to its rarity, diverse range of clinical presentations and histological features. Furthermore, classical clinical benign features may be present in malignant lesions; hence the submission of every surgical specimen to histological analysis is mandatory to provide the patient the adequate treatment.

Oral Adenosquamous Carcinoma Mimicking a Pyogenic Granuloma: a Challenging Diagnosis

Abstract Adenosquamous carcinoma is an aggressive variant of squamous cell carcinoma. This report describes a case of adenosquamous carcinoma with clinical features of a benign lesion and discusses the differential diagnoses, especially regarding histopathological and immunohistochemical analyses. A 45-year-old male was referred to our outpatient clinic complaining about a rapid-growing enlargement in hard palate. Clinical examination revealed an erythematous and pedunculated nodule with lobulated non-ulcerated surface. Excisional biopsy was performed following clinical diagnosis of pyogenic granuloma. Histologically, the specimen consisted of areas characterizing both well-differentiated squamous cell carcinoma and true adenocarcinoma. After a broad list of immunohistochemical markers was evaluated (AE1/AE3, CEA, CK5, CK7, CK8/18, p53, p63 and Ki67), the diagnosis of adenosquamous carcinoma was rendered and the patient referred to complementary surgery. Adenosquamous carcinoma represents a challenge in diagnostic routine due to its rarity, diverse range of clinical presentations and histological features. Furthermore, classical clinical benign features may be present in malignant lesions; hence the submission of every surgical specimen to histological analysis is mandatory to provide the patient the adequate treatment.

Resumo O carcinoma adenoescamoso é uma variante agressiva do carcinoma de células escamosas. Este relato descreve um caso de carcinoma adenoescamoso que apresenta características clínicas de uma lesão benigna e discute o diagnóstico diferencial, especialmente em relação à análise histopatológica e imuno-histoquímica. Um homem de 45 anos foi encaminhado ao nosso serviço queixando-se um aumento de volume de rápido crescimento no palato duro. O exame clínico revelou um nódulo eritematoso e pedunculado com superfície lobulada não ulcerada. Foi realizada biópsia excisional seguindo a hipótese clínica de granuloma piogênico. Histologicamente o fragmento consistia em áreas de carcinoma de células escamosas bem diferenciado e áreas de adenocarcinoma verdadeiro. Após a avaliação de uma ampla lista de marcadores imuno-histoquímicos (AE1/AE3, CEA, CK5, CK7, CK8/18, p53, p63 e Ki67), o diagnóstico de carcinoma adenoescamoso foi atribuído e o paciente foi encaminhado à cirurgia complementar. O carcinoma adenoescamoso representa um desafio na rotina de diagnóstico, devido à sua raridade, diversificada gama de apresentações clínicas e características histológicas. Além disso, características clínicas benignas podem estar presentes em lesões malignas, por isto a submissão de todas as peças cirúrgicas à análise histológica é obrigatória para fornecer ao paciente o tratamento adequado.

Immunocytochemistry associated with oral exfoliative cytology: methodological analysis..

OBJECTIVE: To evaluate different immunocytochemical protocol variations to find the most effective protocol for the analysis of involucrin, epidermal growth factor receptor (EGFR), and E-cadherin antibodies. Exfoliative cytology is a noninvasive method used to monitor and screen for early changes in the oral mucosa of patients exposed to carcinogens such as tobacco and alcohol. It has been postulated that its association with immunocytochemistry may improve the effectiveness of the screening process. STUDY DESIGN: Four graduate students from Porto Alegre in southern Brazil had oral smears collected from the border of the tongue using a cytobrush. The following variables were analyzed: cell membrane permeability, antigen retrieval method (microwave oven or water bath), antibody incubation time (overnight or 1 hour), detection system used (Envision or LSAB), and chromogen incubation time (10 seconds or 5 minutes). RESULTS: Best results were obtained with the following combinations: (1) for involucrin: water bath, 1-hour incubation for primary antibody, Envision, and chromogen incubation for 10 seconds; (2)for EGFR: microwave, overnight incubation, LSAB, and chromogen incubation for 5 minutes; and (3) for E-cadherin: water bath, over-night incubation, Envision, and chromogen incubation for 5 minutes. CONCLUSION: Our findings suggest that each antibody requires a specific immunocytochemical protocol to guarantee optimal results with oral smears.

Percepção dos pacientes em uso de enxaguatórios bucais: óleos essenciais e cloreto de cetilperidíneo / Perception of patients with use mouthwashes: essential oils and cetylpyridinium chloride

Objetivo: Avaliar a percepção dos pacientes quanto às sensações após o uso de enxagua­tórios à base de óleos essenciais e àlcool e à base de cloreto de cetilperidíneo. Métodos: Foram constituídos 2 grupos (n= 15) que realizaram 2 bochechos diários durante 3 semanas: G 1 - enxaguatório a base de óleos essenciais e álcool, em bochechos de 30 segundos; G2 - enxaguatório à base de cloreto de cetilperidínio, em bochechos de 60 segundos. Ao final de 3 semanas, os participantes responderam a um questionário sobre desconforto, alteração de paladar, sensibi­lidade dentária e ressecamento da mucosa oral. Resultados: O desconforto foi de 93,3% para G1 e de 26,6% em G2, sendo o ardor o mais relatado (86,6% em G1 e 26,6% em G2). Não foi relatada sensibilidade ou alteração dentária nos grupos avaliados. O ressecamento da mucosa ocorreu em 66,6% no G 1 e 33,3% no G2. A alteração de paladar ocorreu somente em 13,3% do G1 com diminuição da sensibilidade e gosto metálico. Conclusão: A utilização contínua de enxaguatórios com óleos essenciais e álcool causou maior desconforto quando comparado ao cloreto de cetilperidínio.

Objective: Evaluate the perceptions of patients about the effects of the use of mouthwash based on essential oils and alcohol -based and cetylpyridinium chloride. Methods: 2 groups ( n = 15) who underwent 2 mouthwash daily for 3 weeks were formed: G1 - a mouthwash based on essential oils and alcohol in mouthwash for 30 seconds; G2 - the base of mouthwash cetil­ peridínio chloride in mouthwashes 60 seconds. At the end of 3 weeks, participants answered a questionnaire on discomfort , change in taste, tooth sensitivity and dryness of the oral mucosa. Results: The discomfort was 93.3 % for G1 and 26.6% in G2, with the ardor as reported (86.6% in G1 and 26.6 % in G2) . It was reported tooth sensitivity or change in study groups. The dryness of the mucosa occurred in 66.6 % in G1 and 33.3% in G2. The taste change occurred in only 13.3% of G 1 with decreased sensitivity and metallic taste . Conclusion: The continuous use of mouthwashes with essential oils and alcohol caused greater discomfort when compared to cetilperidínio chloride.

Comparative study of oral mucosa micronuclei in smokers and alcoholic smokers.

OBJECTIVE: To compare the frequency of micronuclei and metanucleated anomalies in the oral mucosa of smokers, alcoholic smokers, and nonsmokers. STUDY DESIGN: Three groups were studied: group I, 15 smokers; group II, 16 alcoholic smokers; and group III, 20 nonsmokers. Three smears from the lateral left border of the tongue were processed for Feulgen staining. A minimum of 300 cells per participant were examined for the quantification of micronuclei and metanucleated anomalies. RESULTS: The Kruskal-Wallis test showed no significant difference in the frequency of micronuclei (p = 0.602) or karyorrhexis (p = 0.114) among the three groups, but there was a significant difference in the frequency of broken eggs, binucleated cells, and karyolysis (p = 0.001). Spearman's correlation indicated an influence of the number of cigarettes per day on micronuclei frequency. Tobacco caused significant alterations in the exfoliative cytology (broken eggs, binucleated cells, and karyolysis) of chronic smokers, but not in the frequency of micronuclei or karyorrhexis, despite the observation of a larger absolute number of micronuclei in group II. CONCLUSION: The action of genotoxic agents (tobacco and alcohol) causes alterations in the frequency of micronuclei and metanucleated anomalies.

Cytokeratin profile in exfoliative cytology of smokers.

OBJECTIVE: To compare the expression of cytokeratins (CKs) 6, 16, 19 and pan-cytokeratin (PAN) in oral mucosa cells between smokers and nonsmokers to determine the proliferative activity and expression indicative of a potential for malignant transformation. STUDY DESIGN: Smears were obtained from the left lateral border of the tongue with a cytobrush from 25 smokers and 20 nonsmokers seen at the clinics of São José dos Campos Dental School, São Paulo State University, São José dos Campos, São Paulo, Brazil, and processed for immunohistochemistry. Conventional microscopy was used for qualitative analysis. Proportions were compared statistically by the z-test and Fisher's exact test. RESULTS: The expression of CK6 (p = 0.002), CK16 (p = 0.003), CK19 (p = 0.0001) and PAN (p = 0.008) was higher in oral mucosa smears from smokers compared to nonsmokers. CONCLUSION: The expression of CK6 and CK16 demonstrated increased epithelial proliferation in the oral mucosa of smokers, and expression of CK19 indicated alterations in epithelial maturation. The expression of PAN indicates the need for the investigation of other types of CK in further studies.

Lateral periodontal cyst: a case report and literature review.

BACKGROUND: The lateral periodontal cyst is considered a developmental odontogenic cyst with unusual occurrence. In most cases it is preliminary diagnosed as a radiographic finding, presenting as well circumscribed or as a round or teardrop-shaped radiolucent area. Due to its location it can easily be misdiagnosed as a lesion of endodontic origin. Final diagnosis should be based on histopatological examination. The purpose of this paper is to report a classic case of lateral periodontal cyst located in the anterior region of mandible and to review the relevant literature which describes the clinical, radiological and histopathological features of lateral periodontal cysts. METHODS: A 50 years female patient complained of an asymptomatic gingival swelling in the region between the left mandibular lateral incisor and canine. Radiographic examination revealed a well circumscribed radiolucency with approximately 0.5 cm diameter with a radiopaque margin between the roots of the left mandibular lateral incisor and canine. The adjacent teeth had vital pulp. RESULTS: A total enucleation of the lesion was performed, and intraoperative examination showed a single lesion with no communication between the cyst's cavity and the oral environment. Histological examination revealed that the lesion was "lateral periodontal cyst of developmental origin". There was no recurrence or complications for 24 months follow-up. CONCLUSIONS: The lateral periodontal cyst can be considered in the differential diagnosis when a radioloucent lesion appears adjacent to the roots of vital teeth. The treatment of choice is surgical removal and subsequent histological evaluation to confirm the diagnosis. Relapses are infrequent.

Exfoliative cytology of the oral mucosa: comparison of two collection methods.

This study compared the sampling efficacy of a cytobrush and metal spatula for exfoliative cytology of the oral mucosa. Thirty students with no detectable oral alterations upon clinical examination were submitted to exfoliative cytology of the lateral border of the tongue, using a metal spatula on the left side and a cytobrush on the right side. The smears were stained using the Papanicolaou technique and evaluated for cellularity, cell type, cell distribution, homogeneity, and cellular distortion, as well as the presence of mucus, inflammatory infiltrate, and hemorrhage. A statistical test (Z-test) with a 95% confidence interval (CI) showed a significant difference between the metal spatula and cytobrush in terms of cellularity (p = 0.02) and homogeneity (p = 0.01). No difference between the two methods was observed regarding cell type (p = 0.4, Z-test) or cell distribution for the 95% confidence interval (p = 0.2, Fisher's test). Cell distortion and the presence of mucus were observed in five cases that used the metal spatula and in two cases that used the cytobrush. No hemorrhage or inflammatory infiltrate was detected in any of the slides. Based on the results of this study, the cytobrush produced qualitatively better smears in terms of cellularity and homogeneity compared to the metal spatula.

Cytogenetic damage of oral mucosa by consumption of alcohol, tobacco and illicit drugs.

BACKGROUND: The aim of the present study was to quantitatively evaluate micronuclei (MN) in the mucosa of users of alcohol, tobacco and illicit drugs in a Brazilian population. METHODS: Patients were divided: (i) experimental group - 24 patients users of alcohol, tobacco and illicit drugs from the Center of Psychosocial Care for Alcohol and other Drugs (CAPSad), São José dos Campos city, and (ii) control group: 24 patients attending the clinics of the São José dos Campos Dental School FOSJC-UNESP. Criterion for inclusion in the two groups was no visible clinical alteration in the oral mucosa. Exfoliative cytology specimens were obtained from the left side of the border of the tongue. Feulgen staining was used and 600 cells per subject were evaluated by light microscopy. RESULTS: The frequency of MN and micronucleated cells was analysed statistically using the Kruskal-Wallis and Mann-Whitney tests. The incidence of MN was 3.08 +/- 3.20 in the CAPSad group and 2.08 +/- 1.93 in the control group. The frequency of micronucleated cells was 2.38 +/- 2.57 in the CAPSad group and 1.42 +/- 1.25 in the control group. CONCLUSION: The results showed a higher frequency of MN and micronucleated cells in the CAPSad group, but the difference compared with the control group was not significant.

Oral exfoliative cytology for the diagnosis of paracoccidoidomycosis in a patient with human immunodeficiency virus: a case report.

BACKGROUND: Paracoccidioidomycosis is not the most common fungal disease in patients infected with human immunodeficiency virus (HIV), except for endemic regions in Latin America countries. CASE: A 33-year-old man with HIV presented with mulberry-like lesions on the palate. The diagnosis was made by exfoliative cytology and Papanicolaou staining. Microscopic analysis revealed fungal structures with birefringent walls and exosporulation conferring an airplane radial motor appearance, or even bowel-like or goblet-like forms compatible with Paracoccidioides brasiliensis. CONCLUSION: This process spares the immunosuppressed patient from undergoing invasive biopsy procedures.

Expressão da survivina em diferentes condições relacionadas à carcinogênese intra-bucal humana / Expression of survivin in different conditions related to human intra-oral carcinogenesis

A survivina é uma proteína inibidora da apoptose que desempenha papel de controle no ciclo celular e no mecanismo de carcinogênese. Este trabalho teve como proposição verificar a correlação clinico patológica da expressão da survivina nas diferentes condições relacionadas à carcinogênese intra-bucal humana, o que pode ser útil para destacar aspectos importantes das etapas da carcinogênese bucal. Foram constituídos três grupos, formados em parte por material citológico coletado de pacientes participantes do Programa Ambulatorial de Tratamento de Tabagismo do Instituto do Coração do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (INCOR-HCFMUSP); e por material que se encontra incluído em blocos de parafina no Laboratório de Patologia Bucal da Faculdade de Odontologia de São José dos Campos FOSJC –UNESP. O primeiro grupo foi constituído por material citológico obtido do bordo lateral lingual esquerdo e soalho bucal de 30 pacientes que fumavam mais de 20 cigarros/dia/10 anos e que não apresentavam histórico de neoplasia bucal maligna, nem sinais clínicos visíveis no local avaliado; o segundo grupo foi constituído por amostras teciduais de 21 pacientes com lesões brancas clinicamente classificadas como leucoplasias. O terceiro grupo foi formado por 42 amostras teciduais de pacientes com diagnóstico de carcinoma epidermóide bucal. Os pacientes que foram submetidos à citologia esfoliativa foram examinados através de anamnese, exame clínico extra e intra -bucal. A citologia esfoliativa foi realizada com cytobrush para obtenção de duas lâminas de cada local selecionado. Após a realização da imunoistoquímica com anticorpo primário anti-survivina as lâminas foram analisadas qualitativamente através da microscopia óptica. Uma lâmina de assoalho e uma de língua foram coradas e avaliadas pelo método de Papanicolaou. A análise estatística foi realizada pelo teste exato de Fisher, X 2 e Mann-Whitney. A survivina foi positiva...

Survivin is an inhibitor of apoptosis protein that plays a role in cell cycle control and the mechanism of carcinogenesis. The aim of the present work was to study the clinico pathological correlation of survivin expression in different conditions related to intra-oral carcinogenesis. This may be useful to highlight important aspects of oral carcinogenesis steps. Three groups were analyzed. They were formed in part by cytological material collected from patients of Heart Institute’s Patient Center and the Smoking Cessation Program of the University Hospital, University of São Paulo Medical School (INCOR-HCFMUSP) and material of Laboratory of Oral Pathology, São José dos Campos Dental School. The first group consisted of cytologic material obtained from the left side of the tongue and mouth floor of 30 patients who smoked more than 20 cigarrettes/day/10years and had no history of malignant oral neoplasm or clinical signs at the site evaluated; the second group consisted of tissue samples from 21 patients with white lesions clinically classified as leukoplakia. The third group consisted of 42 tissue samples from patients diagnosed with oral squamous cell carcinoma. Patients who underwent exfoliative cytology were examined by medical history, extra and intra-oral clinical examination. The exfoliative cytology was performed using cytobrush to obtain two smears of each selected location. After performing the immuno histochemistry for anti-survivin the slides were analyzed qualitatively by light microscopy. One smear of mouth floor and tongue was stained and evaluated by the method of Papanicolaou. Statistical analysis was performed by Fisher's exact test, Mann-Whitney and X2. Survivin was positive in 100% of cytological material from the smokers, 85.7% of oral leukoplakia and 83.3% of oral squamous cell carcinoma. Fisher's exact test showed no association between the expression of survivin and daily cigarette consumption and duration of smoking on the...

Evaluation of staining methods for cytologic diagnosis of oral lesions.

OBJECTIVE: To compare the efficacy ofPapanicolaou, hematoxylin-eosin (H-E), Leishman and periodic acid-Schiff (PAS) staining for cytologic diagnosis of oral lesions. STUDY DESIGN: Patients from the Discipline of Stomatology, São José dos Campos Dental School, from the wards of Hosapital Heliópolis and from the dentistry outpatient clinic of the University Hospital, University of São Paulo Medical School, with the following diseases, were selected: erythematous candidiasis (n=9), pseudomembranous candidiasis (n=10), squamous cell carcinoma (n=19), herpes simplex (n=8), paracoccidioidomycosis (n=8) and pemphigus vulgaris (n=1). RESULTS: The different staining methods were compared regarding the quality of definition of cytoplasmic and nuclear morphologic characteristics and the identification of bacteria, fungi, inflammatory cells and secretions. Papanicolaou and H-E staining were considered better methods. In cases of fungal infections, PAS staining is useful and should be applied as a complementary method. CONCLUSION: Within the limitations of this study, it can be concluded that the cytologic diagnosis of oral lesions along with different staining methods is a useful tool for oral diagnosis.

Calcifications in a clear cell mucoepidermoid carcinoma: a case report with histological and immunohistochemical findings.

The aim of this study was to report an unusual case of mucoepidermoid carcinoma (MEC) in a 39-year-old woman. The tumor showed a prominent population of clear and intermediate basal cells. Clear cells rarely predominate over other cell types. Such cases are called clear cell variant of MEC. The case also revealed a variable amount of calcified material in the tumor mass. Calcifications are rare in clear cell MEC. These structures were periodic acid-Schiff positive and diastase resistant, excluding glycogen origin. Immunohistochemistry was performed, and the epidermoid component was positive for cytokeratin (CK)7, CK13, CK14, and CK19. The mucous and clear cells presented mild staining for CK7. Cytokeratins 7, 13, and 19 stained luminal cells, and intermediate cells exhibited positivity for CK7, CK14, and vimentin. The origin of the calcifications is speculated to be the result of dystrophic calcification of the amorphous eosinophilic material secreted by intermediate basal cells.