Standardization of concentrated antibodies for use in automated immunohistochemistry

Background: Immunohistochemistry (IHC) is a method of identifying proteins in cells or tissues that is useful for diagnosis and research. In modern pathology, it has assumed an important supporting role in the molecular diagnosis of certain neoplasia, with an exponential contribution in personalized medicine. Automation in immunohistochemistry contributes to reduced test variability through standardization. The transition to automation is a process, and for a laboratory that already has a collection of stockconcentrated primary antibodies, ideally, these antibodies will be compatible with the chosen automated method, as antibodies are of high value to be discarded as a consequence of migration to automated immunohistochemistry. Methods: 78 concentrated antibodies were tested for use in Ventana Medical Systems’ Benchmark XT automation platform. Thirty-one human tissue samples containing the antigens of interest were used as positive control. Results: All antibodies tested showed good performance, indicating the feasibility of using these antibodies concentrated in the automation platform in question. The protocol most frequently used was the one with antigenic retrieval with Cell Conditioning 1 for 60 minutes and incubation in the primary antibody for 32 minutes at 42ºC. The dilutions of the primary antibodies in automation ranged from 1:20 to 1:4000. Conclusion: Under the aforementioned conditions, it was possible to take advantage of the portfolio of concentrated antibodies present in the laboratory at the time of transition from manual to automated immunohistochemistry

Avaliação do colágeno na cirrose hepática induzida por CCI4

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Prognostic value of the immunohistochemistry correlation of Ki-67 and p53 in squamous cell carcinomas of the larynx

Prognostic histological factors may contribute to determine the evolution of this neoplasia. AIM: To correlate p53 and Ki-67 immunohistochemical expression with age, histological degree, lymph node involvement and pathological staging in patients with laryngeal epidermoid carcinomas. METHODS: We assessed thirty consecutive cases of laryngeal epidermoid carcinomas submitted to immunohistochemistry to check the expression of p53 e Ki-67 antibodies. RESULTS: Mean age was of 56.2 years and the immunoexpression of the markers was observed in the group with more than 50 years of age, especially that o the ki-67 antibody (p=0.032). There was no relation between p53 and Ki-67 with lymph node involvement. Ki-67 was expressed in 70% of the high histology level cases and in 80% in the low histology ones; while p53 was of 70% only in the high level cases. Pathology staging showed that in the group of advanced carcinomas, p53 expression was of 61.5%, while Ki-67 proved positive for the early cases (100%) and advanced (73.1%). CONCLUSION: There were no significant differences between p53 and Ki-67 immunoexpression in laryngeal epidermoid carcinoma, except in the group of patients with more than 50 years of age, when Ki-67 expression was significantly higher.Os fatores prognósticos histológicos podem colaborar para determinar a evolução desta neoplasia. OBJETIVO: Correlacionar a expressão imunoistoquímica do p53 e Ki-67 com idade, grau histológico, comprometimento de linfonodos e estadiamento patológico de pacientes com carcinomas epidermóides de laringe. MÉTODO: Foram avaliados trinta casos consecutivos de carcinomas epidermóides de laringe, submetidos à técnica de imunoistoquímica para verificar a expressão dos anticorpos p53 e Ki-67. RESULTADO: A idade média foi de 56,2 anos e a imunoexpressão dos marcadores foi mais observada no grupo com mais de 50 anos de idade, especialmente a do anticorpo Ki-67 (p=0,032). Não houve relação do p53 e Ki-67 com o comprometimento de linfonodos. O Ki-67 se expressou em 70% nos casos de alto grau histológico e 80% nos de baixo, enquanto o p53 em 70% apenas nos de alto grau. O estadiamento patológico mostrou que no grupo dos carcinomas avançados a expressão do p53 foi de 61,5%, enquanto o Ki-67 mostrou positividade para os casos precoces (100%) e avançados (73,1%). CONCLUSÃO: Não houve diferenças significantes entre a imunoexpressão do p53 e do Ki-67 em carcinomas epidermóides de laringe, exceto no grupo de pacientes com mais de 50 anos, onde a expressão do Ki-67 foi significativamente maior.Universidade Luterana do Brasil Serviço de Anatomia PatológicaUniversidade Luterana do BrasilFFFCMPA Programa de Pós Graduação em PatologiaFFFCMPAUFRGSUniversidade Luterana do Brasil Serviço de PatologiaUNIFESPUniversidade de PittsburghUniversidade Luterana do Brasil Serviço de Otorrinolaringologia e Cirurgia de Cabeça e PescoçoUFRGS Departamento de Ciências MorfológicasUNIFESPSciEL

Postpartum incidental solid pseudopapillary tumor of the pancreas : case report

Solid pseudopapillary tumors of the pancreas are usually described as rare lesions with low malignant potential and unpredictable behavior. The prognosis may be favorable even with local or distant dissemination. Despite the steady increase of cases diagnosed, the pathogenesis of this neoplasm remains unclear. We present a case of an incidental tumor discovered because of an acute abdomen in a postpartum woman without previous evidence of a tumor during the pregnancy

O efeito da quercetina em mucosa gástrica e fígado de ratos com hipertensão porta experimental

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Mate (Ilex paraguariensis) as an etiological agent of neoplasia in the aerodigestive tract an experimental study

Existem estudos que consideram o “chimarrão” como fator de risco para desenvolvimento do câncer de orofaringe, esôfago e laringe (infusão de erva mate, Ilex paraguariensis). Comparar a histologia do trato aéro-digestivo de ratos submetidos ao consumo de erva mate Ilex paraguariensis, com um grupo controle. Setenta e cinco ratos Wistar adultos foram testados, 60 bebendo mate na temperatura ambiente e 15 bebendo água (grupo controle), durante 5 meses. A histologia do trato aéro-digestivo destes animais foi analisada. Os animais apresentaram alterações nos exames anátomo-patológicos havendo diferença significativa (p=0,02) entre os que tomaram mate e o grupo controle. Houve diferença estatística entre o peso corporal dos grupos em estudo e o grupo controle (p<0,001). Este estudo indica que o consumo de mate afeta o trato aerodigestivo superior e é responsável pela perda de peso dos animais estudados.There are studies that consider drinking “chimarrão” (an infusion of mate, Ilex paraguariensis) a risk factor for developing oropharynx, esophagus and larynx cancer. To make a histological comparison between the aerodigestive tracts of a group of rats submitted to the consumption of erva-mate Ilex paraguariensis, with a control group. Seventy-five adult Wistar rats were tested, 60 drinking mate at room temperature and 15 drinking water (control group), during a period of 5 months. The histology of the aerodigestive tracts of these animals was analyzed. The animals presented alterations in the anatomical-pathological exams. There was a significant difference (p=0.02) between those that were given mate and the control group. There is evidence to suggest that the consumption of mate affects the upper aerodigestive tract in the animals studied, but not inducing cancer

Standardization of concentrated antibodies for use in automated immunohistochemistry

Background: Immunohistochemistry (IHC) is a method of identifying proteins in cells or tissues that is useful for diagnosis and research. In modern pathology, it has assumed an important supporting role in the molecular diagnosis of certain neoplasia, with an exponential contribution in personalized medicine. Automation in immunohistochemistry contributes to reduced test variability through standardization. The transition to automation is a process, and for a laboratory that already has a collection of stockconcentrated primary antibodies, ideally, these antibodies will be compatible with the chosen automated method, as antibodies are of high value to be discarded as a consequence of migration to automated immunohistochemistry. Methods: 78 concentrated antibodies were tested for use in Ventana Medical Systems’ Benchmark XT automation platform. Thirty-one human tissue samples containing the antigens of interest were used as positive control. Results: All antibodies tested showed good performance, indicating the feasibility of using these antibodies concentrated in the automation platform in question. The protocol most frequently used was the one with antigenic retrieval with Cell Conditioning 1 for 60 minutes and incubation in the primary antibody for 32 minutes at 42ºC. The dilutions of the primary antibodies in automation ranged from 1:20 to 1:4000. Conclusion: Under the aforementioned conditions, it was possible to take advantage of the portfolio of concentrated antibodies present in the laboratory at the time of transition from manual to automated immunohistochemistry