Yes / Patients with adenomatous colonic polyps are at increased risk of developing further polyps suggesting field-wide alterations in cancer predisposition. The current study aimed to identify molecular alterations in the normal mucosa in the proximity of adenomatous polyps and to assess the modulating effect of butyrate, a chemopreventive compound produced by fermentation of dietary residues. A cross-sectional study was undertaken in patients with adenomatous polyps: biopsy samples were taken from the adenoma, and from macroscopically normal mucosa on the contralateral wall to the adenoma and from the mid-sigmoid colon. In normal subjects biopsies were taken from the mid-sigmoid colon. Biopsies were frozen for proteomic analysis or formalin-fixed for immunohistochemistry. Proteomic analysis was undertaken using iTRAQ workflows followed by bioinformatics analyses. A second dietary fibre intervention study arm used the same endpoints and sampling strategy at the beginning and end of a high-fibre intervention. Key findings were that keratins 8, 18 and 19 were reduced in expression level with progressive proximity to the lesion. Lesional tissue exhibited multiple K8 immunoreactive bands and overall reduced levels of keratin. Biopsies from normal subjects with low faecal butyrate also showed depressed keratin expression. Resection of the lesion and elevation of dietary fibre intake both appeared to restore keratin expression level. Changes in keratin expression associate with progression towards neoplasia, but remain modifiable risk factors. Dietary strategies may improve secondary chemoprevention. / Food Standards Agency (Ref N12017), EPSRC (EP/E036252/1), University of Sheffield

A presença de áreas displásicas em queilites actínicas constitui um fator preditivo importante para o câncer de lábio. Entretanto, a determinação histopatológica da presença e intensidade da displasia epitelial nestas lesões continua sendo um aspecto subjetivo para os patologistas. O objetivo do presente estudo foi verificar em 45 queilites actínicas e 20 carcinomas espinocelulares (CECs) de lábio tratados no Departamento de Otorrinolaringologia e Oftalmologia, da Faculdade de Medicina de Botucatu, UNESP, a expressão imuno-histoquímica da citoqueratina 10 (CK10) e citoqueratina 13 (CK13) na região labial e comparar a expressão dessas citoqueratinas com o índice de proliferação celular determinado pela imunomarcação com o Ki-67. A intensidade da displasia epitelial nas queilites actínicas foi determinada em coloração de Hematoxilina e Eosina com base nos critérios definidos pela Organização Mundial da Saúde. A associação entre a expressão da CK10 e da CK13 nas queilites actínicas com e sem displasia e nos CECs foi calculada pelo teste do qui-quadrado ou teste exato de Fisher, com nível de significância de 5%. Os resultados demonstraram uma positividade para CK13 no vermelhão do lábio e mucosa labial, com perda de expressão nas áreas displásicas das queilites actínicas e marcação heterogênea nos CECs de lábio. A imunomarcação da CK10 foi observada no vermelhão do lábio e epiderme, com ausência total de expressão nas áreas displásicas e nos CECs de lábio. Houve uma associação estatisticamente significante entre a expressão da CK10 nos CECs quando comparada às queilites actínicas com displasia epitelial (p<0,001) e sem displasia epitelial (p<0,001). Não houve diferença significativa (p=0,104) no índice de proliferação celular, determinado pela imunomarcação do Ki-67, nas queilites actínicas sem e com displasia epitelial e nos carcinomas espinocelulares de lábio. Assim como também não houve associação, estatisticamente significativa, entre a expressão das citoqueratinas 13 e 10 e os altos índices de proliferação celular, determinados pelo Ki-67, nas queilites actínicas sem e com displasia epitelial. Esses resultados sugerem que as citoqueratinas 13 e 10 participam do processo de malignização das queilites actínicas sendo a expressão alterada ou ausente destes biomarcadores indicativa de áreas displásicas ou carcinoma espinocelular invasivo na região de lábio inferior. / The presence of dysplastic areas in actinic cheilitis is an important predictive factor for lip cancer. However, the histopathologic evaluation of the presence and severity of epithelial dysplasia in these lesions remains subjective. The aim of this study was to evaluate the immunohistochemical expression of cytokeratin 10 (CK10) and cytokeratin 13 (CK13) in labial region and compare the expression of these cytokeratins with the cell proliferation index, which was determined by Ki-67 immunostaining. The sample was constituted by 45 cases of actinic cheilitis and 20 of squamous cell carcinomas (SCC) of the lip treated at the Department of Otorhinolaryngology and Ophthalmology of Botucatu School of Medicine, UNESP. The severity of epithelial dysplasia in actinic cheilitis was determined by hematoxylin and eosin staining based on World Health Organization criteria. The association between the expression of CK10 and CK13 in actinic cheilitis with and without dysplasia and SCC was calculated by Chi-square test or Fischer´s exact test, with significance level at 5%. The results showed positive correlation for CK13 in the lip vermilion and labial mucosa, with loss of expression in the dysplastic areas of actinic cheilitis, and heterogeneous positivity in SCC of the lip. There was a statistically significant association between the expression of CK10 in SCC and actinic cheilitis with epithelial dysplasia (p < 0.001) or without epithelial dysplasia (p < 0.001).There was not a statistically significant difference (p=0.104) for cell proliferation index in actinic cheilitis with and without dysplasia, and SCC. Furthermore, there was not a statistical association (p>0.05) between the expression of CK10 and CK13 and high cell proliferation index in actinic cheilitis with and without epithelial dysplasia. These results suggest that the cytokeratins 13 and 10 participate in the process of malignant transformation of actinic cheilitis, and absence or alteration of these biomarkers expression are indicative of dysplastic areas or invasive squamous cell carcinoma in the lower lip region.

Apesar da incidência relativamente baixa, as neoplasias de glândulas salivares (NGSs) constituem um grupo de doenças caracterizado pela notável heterogeneidade em sua apresentação clínica, histológica e comportamento biológico, abrigando um fator de extrema relevância a despeito de seu prognóstico: a ocorrência de metástases, considerada a principal intercorrência relacionada à mortalidade em pacientes oncológicos. Neste estudo, analisamos as características clínicas e histopatológicas de pacientes diagnosticados com metástases à distância de carcinomas de glândulas salivares tratados no A.C. Camargo Cancer Center, São Paulo, no período de 1970 a 2010. Investigamos alterações fenotípicas entre os tumores primários e tumores metastáticos de glândulas salivares, a partir do estudo da caracterização do padrão de expressão de citoqueratinas, procurando identificar a associação entre a expressão destes antígenos a fatores de risco, ou não, ao potencial metastático à distância e sua importância prognóstica. A casuística total compreendeu 43 pacientes diagnosticados com metástase à distância, com a média de idade de 42.5 anos sendo a glândula parótida o órgão primário mais acometido, e os pulmões o sítio metastático mais incidente. O tipo histológico mais comumente relacionado à metástase foi o carcinoma adenoide cístico. A análise imunoistoquímica da expressão de citoqueratinas fora realizada em 10 neoplasias primárias e 10 metastáticas, as quais revelaram um padrão fenotípico similar no tumor primário e no tumor metastático, apresentando a evidenciação majoritária de células luminais de estruturas ductais. A perda de expressão dos marcadores em áreas tumorais periféricas, adjacentes ao parênquima pulmonar, corrobora a predição de alteração do citoesqueleto das células neoplásicas, possivelmente promovendo uma maior capacidade de migração das células neoplásicas. / Salivary gland neoplasms consist of a notorious group of malignancies best known for its clinical features, biological behavior and histological heterogeneity. An important facto related to prognosis is distant metastasis (uncommon in salivary gland tumors). Cytokeratins (CK) are important differentiation markers frequently used for diagnosis processes. Its employment may be useful to identify metastatic disease. In this study, a retrospective evaluation was performed on patients undergoing treatment for metastatic salivary glands tumors between 1970 and 2010, from A.C. Camargo Cancer Center, São Paulo, Brazil. Demographic data and histopathological specimens were obtained from the medical records. Investigation of the expression of cytokeratins in primary and metastatic salivary gland neoplasms was performed and correlated with their phenotypical patterns. Forty three eligible patients were obtained from the database. Parotid gland represented the primary tumor site most frequently related to metastasis. Lung was the most common metastatic site. Adenoid cystic carcinoma was the histological type more frequently associated to metastatic disease. Immunohistochemistry analysis was performed in 10 primary salivary gland tumors and 10 metastatic salivary gland neoplasms. CK expression patterns were similar in both primary and metastatic neoplams, except for the tumor periphery in close contact with lung parenchyma. Cytokeratins were absent in the invasive front of metastatic lesions, and this may be related to an increased capacity of tumor cells migration and proliferation.

A presença de áreas displásicas em queilites actínicas constitui um fator preditivo importante para o câncer de lábio. Entretanto, a determinação histopatológica da presença e intensidade da displasia epitelial nestas lesões continua sendo um aspecto subjetivo para os patologistas. O objetivo do presente estudo foi verificar em 45 queilites actínicas e 20 carcinomas espinocelulares (CECs) de lábio tratados no Departamento de Otorrinolaringologia e Oftalmologia, da Faculdade de Medicina de Botucatu, UNESP, a expressão imuno-histoquímica da citoqueratina 10 (CK10) e citoqueratina 13 (CK13) na região labial e comparar a expressão dessas citoqueratinas com o índice de proliferação celular determinado pela imunomarcação com o Ki-67. A intensidade da displasia epitelial nas queilites actínicas foi determinada em coloração de Hematoxilina e Eosina com base nos critérios definidos pela Organização Mundial da Saúde. A associação entre a expressão da CK10 e da CK13 nas queilites actínicas com e sem displasia e nos CECs foi calculada pelo teste do qui-quadrado ou teste exato de Fisher, com nível de significância de 5%. Os resultados demonstraram uma positividade para CK13 no vermelhão do lábio e mucosa labial, com perda de expressão nas áreas displásicas das queilites actínicas e marcação heterogênea nos CECs de lábio. A imunomarcação da CK10 foi observada no vermelhão do lábio e epiderme, com ausência total de expressão nas áreas displásicas e nos CECs de lábio. Houve uma associação estatisticamente significante entre a expressão da CK10 nos CECs quando comparada às queilites actínicas com displasia epitelial (p<0,001) e sem displasia epitelial (p<0,001). Não houve diferença significativa (p=0,104) no índice de proliferação celular, determinado pela imunomarcação do Ki-67, nas queilites actínicas sem e com displasia epitelial e nos carcinomas espinocelulares de lábio. Assim como também não houve associação, estatisticamente significativa, entre a expressão das citoqueratinas 13 e 10 e os altos índices de proliferação celular, determinados pelo Ki-67, nas queilites actínicas sem e com displasia epitelial. Esses resultados sugerem que as citoqueratinas 13 e 10 participam do processo de malignização das queilites actínicas sendo a expressão alterada ou ausente destes biomarcadores indicativa de áreas displásicas ou carcinoma espinocelular invasivo na região de lábio inferior. / The presence of dysplastic areas in actinic cheilitis is an important predictive factor for lip cancer. However, the histopathologic evaluation of the presence and severity of epithelial dysplasia in these lesions remains subjective. The aim of this study was to evaluate the immunohistochemical expression of cytokeratin 10 (CK10) and cytokeratin 13 (CK13) in labial region and compare the expression of these cytokeratins with the cell proliferation index, which was determined by Ki-67 immunostaining. The sample was constituted by 45 cases of actinic cheilitis and 20 of squamous cell carcinomas (SCC) of the lip treated at the Department of Otorhinolaryngology and Ophthalmology of Botucatu School of Medicine, UNESP. The severity of epithelial dysplasia in actinic cheilitis was determined by hematoxylin and eosin staining based on World Health Organization criteria. The association between the expression of CK10 and CK13 in actinic cheilitis with and without dysplasia and SCC was calculated by Chi-square test or Fischer´s exact test, with significance level at 5%. The results showed positive correlation for CK13 in the lip vermilion and labial mucosa, with loss of expression in the dysplastic areas of actinic cheilitis, and heterogeneous positivity in SCC of the lip. There was a statistically significant association between the expression of CK10 in SCC and actinic cheilitis with epithelial dysplasia (p < 0.001) or without epithelial dysplasia (p < 0.001).There was not a statistically significant difference (p=0.104) for cell proliferation index in actinic cheilitis with and without dysplasia, and SCC. Furthermore, there was not a statistical association (p>0.05) between the expression of CK10 and CK13 and high cell proliferation index in actinic cheilitis with and without epithelial dysplasia. These results suggest that the cytokeratins 13 and 10 participate in the process of malignant transformation of actinic cheilitis, and absence or alteration of these biomarkers expression are indicative of dysplastic areas or invasive squamous cell carcinoma in the lower lip region.

Apesar da incidência relativamente baixa, as neoplasias de glândulas salivares (NGSs) constituem um grupo de doenças caracterizado pela notável heterogeneidade em sua apresentação clínica, histológica e comportamento biológico, abrigando um fator de extrema relevância a despeito de seu prognóstico: a ocorrência de metástases, considerada a principal intercorrência relacionada à mortalidade em pacientes oncológicos. Neste estudo, analisamos as características clínicas e histopatológicas de pacientes diagnosticados com metástases à distância de carcinomas de glândulas salivares tratados no A.C. Camargo Cancer Center, São Paulo, no período de 1970 a 2010. Investigamos alterações fenotípicas entre os tumores primários e tumores metastáticos de glândulas salivares, a partir do estudo da caracterização do padrão de expressão de citoqueratinas, procurando identificar a associação entre a expressão destes antígenos a fatores de risco, ou não, ao potencial metastático à distância e sua importância prognóstica. A casuística total compreendeu 43 pacientes diagnosticados com metástase à distância, com a média de idade de 42.5 anos sendo a glândula parótida o órgão primário mais acometido, e os pulmões o sítio metastático mais incidente. O tipo histológico mais comumente relacionado à metástase foi o carcinoma adenoide cístico. A análise imunoistoquímica da expressão de citoqueratinas fora realizada em 10 neoplasias primárias e 10 metastáticas, as quais revelaram um padrão fenotípico similar no tumor primário e no tumor metastático, apresentando a evidenciação majoritária de células luminais de estruturas ductais. A perda de expressão dos marcadores em áreas tumorais periféricas, adjacentes ao parênquima pulmonar, corrobora a predição de alteração do citoesqueleto das células neoplásicas, possivelmente promovendo uma maior capacidade de migração das células neoplásicas. / Salivary gland neoplasms consist of a notorious group of malignancies best known for its clinical features, biological behavior and histological heterogeneity. An important facto related to prognosis is distant metastasis (uncommon in salivary gland tumors). Cytokeratins (CK) are important differentiation markers frequently used for diagnosis processes. Its employment may be useful to identify metastatic disease. In this study, a retrospective evaluation was performed on patients undergoing treatment for metastatic salivary glands tumors between 1970 and 2010, from A.C. Camargo Cancer Center, São Paulo, Brazil. Demographic data and histopathological specimens were obtained from the medical records. Investigation of the expression of cytokeratins in primary and metastatic salivary gland neoplasms was performed and correlated with their phenotypical patterns. Forty three eligible patients were obtained from the database. Parotid gland represented the primary tumor site most frequently related to metastasis. Lung was the most common metastatic site. Adenoid cystic carcinoma was the histological type more frequently associated to metastatic disease. Immunohistochemistry analysis was performed in 10 primary salivary gland tumors and 10 metastatic salivary gland neoplasms. CK expression patterns were similar in both primary and metastatic neoplams, except for the tumor periphery in close contact with lung parenchyma. Cytokeratins were absent in the invasive front of metastatic lesions, and this may be related to an increased capacity of tumor cells migration and proliferation.

<p>Diss. (sammanfattning) Umeå : Umeå universitet, 2002</p> / digitalisering@umu

Immunohistologic investigations of wound healing in human oral mucosa require specific cell biological markers as well as consecutive small biopsies. Small specimens are ideally embedded in plastic (methylmethacrylate, MMA) resin due to their miniature size. This limits the use of antibodies for these markers. In this immunohistochemical study, the distribution of wound healing markers, e.g. cytokeratin (CK), laminin, collagen IV, vimentin, vinculin and fibronectin, were compared between semithin sections of plastic-embedded tissue and frozen sections of mucosal tissue in order to assess their use for future investigations. The antibodies against laminin, collagen IV and CK 1/2/10/11, 5/6, 13, 14, 17, 19 gave comparable staining patterns on cryostat sections of attached mucosa and on semithin sections of MMA-embedded attached mucosa. In the epithelial cell layers, the following distribution of CK immunostaining was observed: The basal cell layer was positive for CK 5/6, CK 14 and CK 19; the intermediate cell layer for CK 13, CK 17 and CK 1/2/10/11, and the superficial cell layer for CK 13 and CK 1/2/10/11. For most of these antibodies, enzyme digestion with 0.1% trypsin was adequate for demasking the antigens, except for anti-CK 14, anti-CK 17 and anti-laminin; predigestion with 0.4% pepsin in 0.01 N HCl gave similar staining results. The antibodies against vimentin, vinculin, fibronectin and CK 4 showed no affinity or a reciprocal reaction on the semithin sections. Therefore, the antibodies against CK 1/2/10/11; 5/6; 13; 14; 17, and 19, as well as the basement proteins laminin and collagen IV are deemed markers suitable on semithin sections of plastic-embedded attached oral mucosa. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

Immunohistologic investigations of wound healing in human oral mucosa require specific cell biological markers as well as consecutive small biopsies. Small specimens are ideally embedded in plastic (methylmethacrylate, MMA) resin due to their miniature size. This limits the use of antibodies for these markers. In this immunohistochemical study, the distribution of wound healing markers, e.g. cytokeratin (CK), laminin, collagen IV, vimentin, vinculin and fibronectin, were compared between semithin sections of plastic-embedded tissue and frozen sections of mucosal tissue in order to assess their use for future investigations. The antibodies against laminin, collagen IV and CK 1/2/10/11, 5/6, 13, 14, 17, 19 gave comparable staining patterns on cryostat sections of attached mucosa and on semithin sections of MMA-embedded attached mucosa. In the epithelial cell layers, the following distribution of CK immunostaining was observed: The basal cell layer was positive for CK 5/6, CK 14 and CK 19; the intermediate cell layer for CK 13, CK 17 and CK 1/2/10/11, and the superficial cell layer for CK 13 and CK 1/2/10/11. For most of these antibodies, enzyme digestion with 0.1% trypsin was adequate for demasking the antigens, except for anti-CK 14, anti-CK 17 and anti-laminin; predigestion with 0.4% pepsin in 0.01 N HCl gave similar staining results. The antibodies against vimentin, vinculin, fibronectin and CK 4 showed no affinity or a reciprocal reaction on the semithin sections. Therefore, the antibodies against CK 1/2/10/11; 5/6; 13; 14; 17, and 19, as well as the basement proteins laminin and collagen IV are deemed markers suitable on semithin sections of plastic-embedded attached oral mucosa. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.