Increased Expression of Type-I Collagen in Malignant Colorectal Lesion and Positive Correlations with Clinical Factors / Expressão aumentada do colágeno tipo I na lesão colorretal maligna e correlações positivas com fatores clínicos

Abstract Objective Type-I collagen (Col-I) is one of the main macromolecules of the extracellular matrix, and it is involved in the desmoplastic stromal reaction, an indicator of worse prognosis in cases of colorectal cancer (CRC). The purpose of the present study was to investigate Col-I expression in cases of CRC and adenoma and to correlate with the clinical data and the data regarding the lifestyle of the patients. Methods A retrospective study including 22 patients with adenoma and 15 with CRC treated at a coloproctology service. The clinical and lifestyle data were obtained through medical records, and Col-I expression was investigated by immunohistochemistry. Results Women represented most cases of adenoma (63.64%), whereas CRC was found mainly in men (73.33%) (p=0.0448). Immunoexpression of Col-I showed a basement membrane thickening in areas of lining of epithelium and around the glands in both lesions. The cases of CRC had a quite evident fibrosis process in the stroma. The quantitative analysis demonstrated a higher protein expression in CRCs compared to adenomas (p=0.0109), as well as in female patients (p=0.0214), patients aged ≥ 50 years (p=0.0400), and in those with a positive family history of colorectal disease (p=0.0292). These results suggested a remodeling of the microenvironment of the Worked developed at the Department of Morphology, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, ES, Brazil. Conclusion The immunohistochemical analysis encourages the performance of more comprehensive studies to ascertain if our results could be a tool for the diagnosis and monitoring of the patients.

Resumo Objetivo O colágeno tipo I (Col-I) é uma das principais macromoléculas da matriz extracelular, e está envolvido na reação desmoplástica estromal, um indicador de pior prognóstico em casos de câncer colorretal (CCR). O objetivo foi investigar a expressão do Col-I emcasos de CCR e adenoma, e correlacioná-la comdados clínicos e de estilo de vida dos pacientes. Metodologia Foi realizado umestudoretrospectivo com22pacientes comadenoma e 15 comCCR tratadosemumserviço de coloproctologia.Os dados dos pacientes foramobtidos dos prontuários médicos, e a expressão do Col-I foi investigada por imunohistoquímica. Resultados As mulheres representaram a maioria dos casos de adenomas (63,64%), enquanto o CCR (73,33%) (p=0,0448) foi mais comum entre os homens. A imunoexpressão de Col-I mostrou espessamento da membrana basal em áreas de revestimento do epitélio e em volta de glândulas em ambas as lesões. O CCR apresentou fibrose no estroma. As análises quantitativas demonstraram maior expressão proteica no CCR (p=0,0109), assim como em mulheres (p=0,0214), pacientes com idade ≥ 50 anos (p=0,0400), e em pacientes com histórico positivo de doença colorretal na família (p=0,0292). Estes resultados sugerem a remodelação do microambiente tumoral na carcinogênese do CCR. As correlações clínico-patológicas positivas mostram uma ligação plausível entre o perfil do paciente e os achados imunohistoquímcos, o que indica uma possível forma de estratificação dos pacientes. Conclusão As análises imunohistoquímicas estimulam a execução de estudos mais abrangentes para confirmar se nossos resultados poderão ser uma ferramenta para o diagnóstico e o monitoramento dos pacientes.

Immunohistochemical Study of Laminin-332 γ2 Chain and MMP-9 in High Risk of Malignant Transformation Oral Lesions and OSCC.

OBJECTIVES: Oral squamous cell carcinoma is associated with alterations in basement membrane. Laminin-332 is present in basal lamina and performs multiple biologic effects by γ2 chain. Matrix metalloproteinase acts disrupting extracellular components and was related to poor prognosis in cancer. Here, molecular profile of laminin-332 γ2 chain and matrix metalloproteinase-9 was assessed in oral lesions. MATERIAL AND METHODS: The expression of laminin-332 γ2 chain and matrix metalloproteinase-9 (MMP-9) was examined by immunohistochemistry in 10 patients with high risk of malignant transformation oral lesions and 26 cases of oral squamous cell carcinoma (OSCC). Associations between microscopic and clinicopathologic features were established. RESULTS: Immunostaining of laminin-332 γ2 chain in high risk oral lesions was most detected in basement membrane which is continuous, while the majority of OSCC cases showed a discontinuous membrane (P = 0.001). It was observed a positive reaction for γ2 chain in invasive fronts and a higher expression in epithelial compartment of smoking patients with OSCC (P < 0.0001). In epithelium, MMP-9 expression was presented in all layers with no difference between lesions. However, an elevated immunostaining in stromal cells was associated with male patients (P = 0.0054), older than 60 years (P = 0.0101) and with OSCC. CONCLUSIONS: Present study results support the hypothesis of changes in molecules expression in high risk oral lesions and oral squamous cell carcinoma. A relation between clinical and molecule profile was observed. Those molecules may represent a useful tool to predict oral cancer behaviour.

Could formaldehyde induce mutagenic and cytotoxic effects in buccal epithelial cells during anatomy classes?

BACKGROUND: Due to increased formaldehyde exposure, carcinogenic to humans, several researches have been studying the potential toxicity and the safe levels for human beings. The aim of this study was to investigate mutagenicity and cytotoxicity in buccal epithelial exfoliated cells (BEC) of students subjected to formaldehyde (FA) during anatomy classes. MATERIAL AND METHODS: BEC were collected periodically from 17 volunteers of undergraduate programs, who had participated in practical anatomy classes, before and after FA exposure. Cells were stained according to Feulgen method and then micronucleus test was applied. A total of 1,500 cells were assessed per individual in this study for the micronucleus frequency and other parameters of cytotoxicity. RESULTS: There was statistically significant increase in number of micronucleated BEC after FA exposure (after 1 month p=.034 and after 3.5 months p=.017). However, FA exposure caused no significant increase in other nuclear alterations closely related to cytotoxicity (p≥.05). CONCLUSIONS: FA induced mutagenicity during anatomy classes. Cell death increased, but it was not statistically significant. Efforts have to be made to improve air quality and reduce exposures during anatomy classes

Expressão da laminina 332 em vasos de carcinomas de células escamosas orais / Expression of laminin 332 in vessels of oral squamous cell carcinomas

Objetivo: Analisar expressão da cadeia γ2 da laminina 332 na membrana basal de vasos de carcinoma de células escamosas orais (CCE) e seu aspecto de continuidade para detecção de possíveis processos invasivos vasculares, importante evento na progressão da doença. Ainda, associar os aspectos clínicos do paciente e da lesão com os achados microscópicos. Material e Métodos: Foi realizada técnica imuno-histoquímica para detecção da cadeia γ2 da laminina 332 em 16 casos de CCE, sendo oito Bem Diferenciado e oito Moderadamente Diferenciado. Os aspectos microscópicos avaliados foram expressão da molécula na membrana basal de vasos e o seu aspecto de continuidade. Possíveis associações entre dados clínicos e achados microscópicos foram avaliadas. Resultados: Um total de 15 casos (93,75%) apresentaram vasos com expressão da cadeia γ2. Em CCE Bem Diferenciados, 85,71% das lesões apresentaram vasos com perda de continuidade da membrana basal, enquanto que em CCE Moderadamente Diferenciados 100% dos casos tiveram vasos com esse aspecto. Em relação às associações entre aspectos clínicos e integridade da membrana basal vascular, nos CCE Bem Diferenciados foram observadas correlações positivas da perda de continuidade com o gênero e idade acima de 60 anos (p < 0,0001). Para CCE Moderadamente Diferenciado, as associações foram com gênero masculino (p < 0,0001), hábito de fumar (p < 0,0001) e idade < 60 anos (p < 0,0001). Conclusão: Nossos dados sugerem que a expressão da cadeia γ2 em estruturas vasculares está presente em lesões de CCE e pode estar associada ao comportamento mais agressivo da doença.

Objective: 1) To analyze the γ2 chain expression of laminin 332 in the basement membrane of vessels of oral squamous cell carcinoma (OSCC), and the appearance of continuity, to identify possible vascular invasive procedures which are important events in the disease progression. 2) To examine the association between clinical aspects from patients and microscopic findings from lesions. Material and Methods: Immunohistochemistry was performed to detect γ2 chain of laminin 332 in 16 cases of OSCC; of which, eight were classified as being "Well Differentiated" and eight as being "Moderately Differentiated." Microscopic aspects were evaluated, including the expression of the molecule in the basement membrane of vessels and the appearance of continuity. Results: 15 cases (93.75%) showed vessels with γ2 chain expression. In OSCC defined as being "Well Differentiated," 85.71% of lesions presented with loss of vessel continuity of the basement membrane, whereas in OSCC defined as being "Moderately Differentiated," 100% of presented vessels with this aspect. Regarding associations between clinical aspects of patients and integrity of the vascular basement membrane, in OSCC defined as being "Well Differentiated," positive correlations were observed between loss of continuity and both gender and age >60 years (p < 0.0001). For OSCC defined as being "Moderately Differentiated," positive associations were observed for being male (p < 0.0001), currently smoking (p < 0.0001) and being <60 years old (p < 0.0001). Conclusion: Our data suggest that the γ2 chain expression in vascular structures is present in OSCC and is associated with more aggressive disease.

Fibrosis and Mast Cells in Colorectal Lesions: Significance in Adenoma-Colorectal Cancer Sequence and Association with Diet.

PURPOSE: Adenoma is the most common benign neoplasm with potential to progress into colorectal cancer (CRC), a disease responsible for high rates of mortality. However, it is not possible yet to establish which lesions will evolve into CRC. We aimed to investigate the intrinsic factors, diet profile, and microenvironment as factors involved in adenoma-CRC progression. METHODS: We evaluated nutritional profile and microscopic features. Adenomas and CRC lesions were stained with Mallory's trichrome to reveal fibrosis and Alcian blue to identify mast cells. RESULTS: A total of 143 patients was selected, 66 % diagnosed with normal mucosa, 28.4 % with benign lesion, and 5.6 % with CRC. A higher frequency of lesions was associated with patients older than 50 years (26.57 %) and women (18.18 %), but men were more likely to develop CRC. Regarding microscopic aspects, epithelium-connective tissue interface with disorganization feature was significantly higher in CRC when compared to adenoma (P = 0.0007). A greater fibrosis around vessels was also observed in CRC (P = 0.01). The statistical analysis indicated a significant correlation between mast cell population and patients older than 50 years (P < 0.01), male (P < 0.01), with right colon lesion (P < 0.05) and CRC. Low consumption of leafy greens (P < 0.001) and carotenoids (P < 0.01) and elevated consumption of red meat (P < 0.001) were associated with a high density of mast cells. CONCLUSIONS: An association between clinical, diet profile, and microscopic features in colorectal lesions was observed, with a progressive change in the microenvironment during adenoma-CRC sequence.

Oral lichen planus: clinical profile and determination of oral epithelial dysplasia / Liquen plano oral: perfil clínico y determinación de la displasia epitelial oral

Oral lichen planus (OLP) is the most common noninfectious oral disease and is considered a potentially malignant disorders (OPMDs). The etiology and pathogenesis of OLP are not completely understood and the malignant transformation remains under discussion. The aim was to asses the cytological and histological features of OLP, and establish relationship between clinical and microscopic profiles. A total of 10 patients diagnosed with OLP were investigated. Slides were prepared and stained with Hematoxylin­Eosin to identify 8 histological features and 8 cytological alterations. Finally, oral epithelial dysplasia degree was determined. Hyperplasia and loss of polarity of basal cells were detected in 90 % of cases. Anisonucleosis, nuclear pleomorphism and hyperchromasia affected 100 % of the sample. Six cases were classified as mild-grade OED, with no cases of severe-grade. Microscopic alterations were higher in women (P <0.0001) and moderate-grade OED was diagnosed only in this group (P <0.0014). The findings demonstrated that microscopic and clinical data association should be analyzed to a better understand of disease behavior. OED was absence only in one case, so the periodic follow-up of patients diagnosed with OLP is mandatory to avoid the malignant transformation.

El liquen plano oral (LPO) es la enfermedad no infecciosa más común y se considera un trastorno potencialmente maligno. La etiología y la patogénesis del LPO no se conoce y la transformación maligna sigue siendo objeto de debate. El objetivo de este estudio fue evaluar las características citológicas e histológicas del LPO y establecer la relación entre los perfiles clínicos y microscópicos. Un total de 10 pacientes diagnosticados con LPO fueron investigados. Secciones histológicas se prepararon y tiñeron con hematoxilina-eosina para identificar 8 características histológicas y 8 alteraciones citológicas. Finalmente, se determinó el grado de displasia epitelial. La hiperplasia y la pérdida de la polaridad de las células basales se detectaron en el 90 % de los casos. Anisonucleosis, pleomorfismo nuclear e hipercromasia afectaron al 100 % de las muestras. Seis casos fueron clasificados como displasia epitelial oral (DEO) de grado leve y no se reportaron casos de grado severo. Las alteraciones microscópicas fueron mayores en las mujeres (p <0,0001) y DEO de grado moderado fue diagnosticado sólo en este grupo (p <0,0014). Los resultados demostraron que la asociación de datos microscópicos y clínicos deben ser analizados para una mejor comprensión del comportamiento de la enfermedad. La DEO estuvo ausente sólo en un caso, por lo que el seguimiento periódico de los pacientes diagnosticados con LPO es necesario para evitar la transformación maligna.