Carcinosinumproduces phenotypic changes of 4T1 tumor cells in vitro

Considering that there are few published studies that specifically address the exclusive use of Carcinosinumin different potencies and, most of them focused on genotypic and clinical effects, the present study was proposed to identify possible phenotypic changes, including viability, expression of HER-2 and metastatic abilities, using 4T1 cells in vitroas a model. Carcinosinum was tested in different homeopathic potencies (12cH; 30cH; 200cH) mechanically prepared using sterile pure water. The time space between preparing the potencies and using them was 24 hours.The final dilutions were inserted into the culture medium in a volume equal to 10%, at the time of cell seeding. The same succussed vehicle used to prepare the drugs (70% ethanol) diluted 1:100 in sterile pure water was used as control. All treated cells were cultured in 25 mL flasks, with cell density of 5 x 105cells/mL. After 24 hours of treatment, cells were analyzed for apoptosis index using Annexin V kit and the Countess® system. The morphology of 4T1 cells was monitored by staining cell smears with hematoxylin-eosin and Giemsa methods. HER-2 expression was assessed by immunocytochemistry and metalloproteinase activity was assessed by zymography. The determination of the cytokine profile was performed using Cytometric Bead Array (CBA). The samples were evaluated in quadruplicate and the data were analyzed by one-way ANOVA. Carcinosinum30cH presented the highest apoptotic index and reduction of MMP-9-Pro expression; Carcinosinum200cH produced the highest positivity for HER-2 and no specific effect was seen after the treatment with Carcinosinum12cH. No change in cytokine expression was seen among treatments. We conclude that Carcinosinum30cH and 200cH can change phenotypic features important totumor development in vitro. The clinical meaning of these data deserves further investigation.

Lung cancer biopsy: Can diagnosis be changed after immunohistochemistry when the H&E-Based morphology corresponds to a specific tumor subtype?

OBJECTIVES: Advancements in non-small cell lung cancer treatment based on targeted therapies have made the differentiation between adenocarcinoma and squamous cell carcinoma increasingly important. Pathologists are challenged to make the correct diagnosis in small specimens. We studied the accuracy of an immunohistochemical panel in subclassifying non-small cell lung cancer in routine small biopsies and compared the results with the diagnosis from resected lung specimens, autopsy samples or biopsied/resected metastases. METHODS: In total, 340 lung cancer biopsies were investigated for the expression of CK5, TTF1, p63 and surfactant. RESULTS: We characterized 166 adenocarcinomas and 124 squamous cell carcinomas. Overall, 85% of cases displayed binary staining (TTF1 positive/p63 negative, and vice versa). The diagnoses of ten cases with a morphology that indicated a specific tumor subtype were changed after immunohistochemistry (IHC). A second specimen was available for 71 patients, and the first diagnosis at biopsy was confirmed in 95% of these cases. Most non-small cell lung cancer cases present a binary immunohistochemical profile in small biopsies, contributing to good diagnostic accuracy with routine markers. In a small proportion of cases, the diagnosis can be changed after IHC even when the morphological aspects indicate one specific tumor subtype. CONCLUSIONS: We recommend that routine small biopsies of lung cancer without classic morphology should be subjected to a minimum immunohistochemical panel to differentiate adenocarcinoma from squamous cell carcinoma.

Immunohistochemical Expression of the Tumor Suppressor Protein p16 INK4a in Cervical Adenocarcinoma / Expressão imunhistoquímica da proteína de supressão tumoral p16 INK4a em adenocarcinoma cervical

ABSTRACT Objective: To evaluate the diagnostic utility of the p16ink4a protein expression as a marker for adenocarcinoma of the cervix. Methods: In a cross-sectional study, p16ink4a expression was evaluated in 30 cervical biopsies from patients diagnosed with invasive adenocarcinoma from 2 reference clinics in Brazil, and compared with 18 biopsies of endocervical polyps (control cases). The performance of the tests for p16ink4a was evaluated using a conventional contingency table, and the Kappa (k) index was used to evaluate the agreement of the marker with the tissue diagnosis. Results: In total, 66% of the invasive adenocarcinoma cases were positive for p16ink4a. All of the adenomatous polyps cases used as negative controls were shown to be negative for p16ink4a. The marker showed a high sensitivity and a high negative predictive value. The Kappa index was good for p16ink4a (k 1/4 0.6). Conclusion: Considering the strong association between the p16ink4a marker and the cervical adenocarcinoma, its use represents an important tool for reducing incorrect diagnoses of adenocarcinoma and thereby avoiding overtreatment.

RESUMO Objetivo: Avaliar a utilidade diagnóstica da expressão da proteína p16ink4a como marcador de adenocarcinoma do colo. Métodos: Em estudo transversal, a expressão de p16ink4a foi avaliada em 30 biópsias cervicais de pacientes diagnosticadas com adenocarcinoma invasivo de colo uterino provenientes de dois serviços de referência no Brasil, comparando com achados em 18 biópsias de pólipos endocervicais (grupo de controle). Para avaliar a performance do teste, foi utilizada tabela de contingência convencional, e para avaliar a concordância com o diagnóstico, foi aplicado o índice de Kappa (k). Resultados: No total, 66% dos casos de adenocarcinoma invasivo foram positivos para p16ink4a. Todos os pólipos adenomatosos foram negativos para p16ink4a. O marcador mostrou uma alta sensibilidade e alto valor preditivo negativo. O índice de Kappa foi bom para p16ink4a (k 1/4 0.6). Conclusion: Considerando a forte associação entre o marcador p16ink4a e o adenocarcinoma cervical, seu uso representa uma ferramenta importante para reduzir o risco de diagnóstico incorreto de adenocarcinoma e, por conseguinte, evitar o excesso de tratamentos.

Characterization of reactive stroma in prostate cancer: involvement of growth factors, metalloproteinase matrix, sexual hormones receptors and prostatic stem cells

ABSTRACT Introduction and Objectives: Reactive Stroma (RStr) is observed in many human cancers and is related to carcinogenesis. The objectives of the present study were to stablish a relationship of the RStr microenvironment with prostate cancer (Pca) through a morphological and molecular characterization, and to identify a possible relationship between RStr with worse prognosis factors and occurrence of malignant prostatic stem cells. Materials and Methods: Forty prostatic samples were selected from men with Pca diagnosis submitted to radical prostatectomy; they were divided in two groups: Group-1 (n=20): samples without reactive stroma; Group-2 (n=20): samples of PCa with intense stroma reaction. Prostatic samples were evaluated for RStr intensity by Masson Trichromic stain and posteriorly submitted to histopathological and immunohistochemistry analysis for antigens: α-actin, vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA, AR, Erα and ERβ. Results: Reactive stroma with intense desmoplastic reactivity was significantly more frequent in intermediate (Gleason 7, 3+4) and high grade tumors (Gleason 7, 4+3). The group with intense stromal reactivity showed significant higher levels of Vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA and ERα. Conclusions: It can be concluded that RStr may be a predictive marker of Pca progression, since it was associated with increase of growth factors, imbalance of androgen and estrogen receptors and presence of malign prostatic stem cells.

Fiocruz as an actor in Brazilian foreign relations in the context of the Community of Portuguese-Speaking Countries: an untold story / A Fiocruz como ator da política externa brasileira no contexto da Comunidade dos Países de Língua Portuguesa: uma história revelada

Brazilian foreign policy paradigms and changes in the global scenario since the Cold War created conditions for stronger ties between Brazil and Portuguese-speaking African countries. Recently, Brazil took the lead in regional integration processes and in South-South cooperation initiatives. These strategies and Fiocruz's acknowledged technical expertise resulted in its direct involvement in Brazilian foreign public health policy in the Community of Portuguese-Speaking Countries. Fiocruz developed cooperation projects in various areas, sharing its know-how and best practices in the most critical fields in partner countries, consolidating "public health framework cooperation" and contributing to diversifying Brazil's partners and promoting Brazil as a global actor.

Os paradigmas da política externa brasileira e as mudanças no cenário global desde a Guerra Fria criaram as condições para aproximação do Brasil com os países africanos de língua portuguesa. Recentemente, o Brasil tomou a liderança nos processos de integração regional e nas iniciativas de cooperação Sul-Sul. Essas estratégias e a reconhecida expertise técnica da Fiocruz abriram espaço para o envolvimento direto da instituição na política externa do Brasil com a Comunidade de Países de Língua Portuguesa na área da saúde. A Fiocruz desenvolveu projetos de cooperação em áreas diversas, compartilhando seu know-how e melhores práticas em áreas prioritárias dos países parceiros, consolidando a "cooperação estruturante em saúde" e contribuindo para a diversificação de parceiros do país e promovendo o Brasil como ator global.

La "cultura de la sobrevivencia" y la salud pública internacional en América Latina: la Guerra Fría y la erradicación de enfermedades a mediados del siglo XX / The "culture of survival" and international public health in Latin America: the Cold War and the eradication of diseases in the mid-twentieth century

Este artículo analiza las principales campañas promovidas por agencias internacionales y organismos nacionales de salud dirigidas a erradicar enfermedad infecciosas en el ámbito rural latinoamericano de los años 1940 y 1950. Las dimensiones políticas del periodo han sido estudiadas pero todavía se ha prestado poca atención a sus dimensiones sanitarias. Este trabajo propone el concepto de "cultura de la sobrevivencia" para explicar los problemas de la salud pública oficial de Estados con políticas sociales limitadas que no permitieron el ejercicio de la ciudadanía. La salud pública, como parte de esta cultura de la sobrevivencia, buscaba ser una solución temporal sin enfrentarse a los problemas sociales que originaban las infecciones y dejó un legado en la salud pública de la región.

This article analyzes the main campaigns run by international agencies and national health bodies to eradicate infectious diseases in rural Latin America in the 1940s and 1950s. The political dimensions of the period have been studied but there has been little attention as yet to the health dimensions. This article proposes the concept of a "culture of survival" to explain the official public health problems of states with limited social policies that did not allow the exercise of citizenship. Public health, as part of this culture of survival, sought a temporary solution without confronting the social problems that led to infections and left a public health legacy in the region.

Pregnancy in women undergoing hemodialysis: case series in a Southeast Brazilian reference center / Gestação em mulheres em tratamento hemodialítico: série de casos em um centro de referência do Sudeste do Brasil

PURPOSE: To describe maternal and neonatal outcomes in pregnant women undergoing hemodialysis in a referral center in Brazilian Southeast side. METHODS: Retrospective and descriptive study, with chart review of all pregnancies undergoing hemodialysis that were followed-up at an outpatient clinic of high- risk prenatal care in Southeast Brazil. RESULTS: Among the 16 women identified, 2 were excluded due to follow-up loss. In 14 women described, hypertension was the most frequent cause of chronic renal failure (half of cases). The majority (71.4%) had performed hemodialysis treatment for more than one year and all of them underwent 5 to 6 hemodialysis sessions per week. Eleven participants had chronic hypertension, 1 of which was also diabetic, and 6 of them were smokers. Regarding pregnancy complications, 1 of the hypertensive women developed malignant hypertension (with fetal growth restriction and preterm delivery at 29 weeks), 2 had acute pulmonary edema and 2 had abruption placenta. The mode of delivery was cesarean section in 9 women (64.3%). All neonates had Apgar score at five minutes above 7. CONCLUSIONS: To improve perinatal and maternal outcomes of women undergoing hemodialysis, it is important to ensure multidisciplinary approach in referral center, strict control of serum urea, hemoglobin and maternal blood pressure, as well as close monitoring of fetal well-being and maternal morbidities. Another important strategy is suitable guidance for contraception in these women. .

OBJETIVOS: Descrever os resultados maternos e neonatais de mulheres grávidas que estavam em tratamento de hemodiálise em um centro de referência no Sudeste brasileiro. MÉTODOS: Estudo retrospectivo e descritivo, com revisão de prontuários de todas as gestações em hemodiálise, acompanhadas no pré-natal especializado da região Sudeste do Brasil. RESULTADOS: Entre as 16 mulheres identificadas, 2 foram excluídas devido à perda de seguimento. Das 14 descritas, a hipertensão foi a causa mais frequente de insuficiência renal crônica (50% dos casos). A maioria (71,4%) realizava tratamento de hemodiálise há mais de um ano e todas elas foram submetidas a 5 ou 6 sessões por semana. Onze mulheres tinham hipertensão crônica, 1 das quais também era diabética, e 6 eram fumantes. Em relação às complicações da gravidez, 1 das mulheres hipertensas desenvolveu hipertensão maligna (com restrição de crescimento fetal e parto prematuro com 29 semanas), 2 tiveram edema pulmonar agudo e 2 apresentaram descolamento prematuro de placenta. O tipo de parto foi cesariana em 9 mulheres (64,3%). Todos os recém-nascidos tiveram Apgar aos cinco minutos maior que 7. CONCLUSÕES: Para melhorar os resultados perinatais e maternos de mulheres em hemodiálise, é importante ter uma abordagem multidisciplinar em centro de referência, um controle rigoroso da uremia, hemoglobina e pressão arterial materna, bem como acompanhar de perto o bem-estar fetal e a morbidade materna. Outra estratégia importante é a orientação adequada para contracepção nessas mulheres. .

Transformation into large-cell neuroendocrine carcinoma associated with acquired resistance to erlotinib in nonsmall cell lung cancer

No abstract available.

A study on inorganic elements in psammomas from ovarian & thyroid cancer.

Background & objectives: Concentric lamellar calcifications known as psammoma bodies (PB) are found in benign and malignant tumours. Whether or not the inorganic element concentrations in psammomas are similar to serous adenocarcinoma of the ovary and thyroid papillary cancer tissues has not yet been ascertained. We undertook this retrospective study to establish if there is any difference in the concentrations of inorganic ions found in psammomas in serous adenocarcinoma of the ovary, and those found in thyroid papillary cancer tissue. Methods: PB samples from patients with adenocarcinoma of the ovary (n = 10) and with thyroid papillary cancer (n = 10) were analyzed through inductively-coupled plasma spectroscopy (ICP). Results: There were no significant differences in the concentrations of inorganic elements in PB from thyroid papillary cancer than in those PB from ovarian cancer. Interpretation & conclusions: Differences in the concentrations of inorganic elements may be due to the variation in environmental pollution. Our study had limitation of small sample size. Our results suggest that some inorganic elements can participate in the origin of psammoma bodies.

Expressão imunohistoquímica do marcador tumoral CD34 e P27 como fator prognóstico em adenocarcinoma de próstata clinicamente localizado após prostatectomia radical / Immunohistochemistry expression of tumor markers CD34 and P27 as a prognostic factor of clinically localized prostate adenocarcinoma after radical prostatectomy

OBJETIVO: Analisar a expressão imunoistoquímica do marcador CD34 e p27, como fator prognóstico em pacientes com neoplasia de próstata localizada. MÉTODOS: Análise de 100 casos de pacientes portadores de neoplasia prostática localizada submetida à cirurgia curativa. Realizou-se o preparo histológico habitual, seguido da reação imunoistoquímica para a detecção do acúmulo da proteína CD34 e p27 seguida de análise estatística. RESULTADOS: Na avaliação do marcador P27 e na correlação com as variáveis, observou-se diferença significativa no escore de Gleason com expressão positiva (P27 positivo) relacionada com PSA médio mais baixo (p=0,091), escore de Gleason mais baixo (p<0,0001) e menor área de tumor no CD34 (p=0,036). Correlacionando-se o marcador CD34 na área tumoral observou-se quanto menor o CD34 positivo menor é o valor do PSA (p<0,0001), e menor é o escore de Gleason (r=0,5726 ; p<0,0001) e quanto maior o CD34 positivo maior é o estadiamento (r=0,3305 ; p<0,0001) e a chance de recidiva (p=0,002). Os pacientes com estadiamento mais alto, também tinham maior área CD34 positivo (p<0,0001). CONCLUSÃO: Os marcadores P27 e CD34 estão associados com os eventos próprios ao câncer de próstata; contudo, apenas o CD34 foi capaz de determinar a possibilidade de recidiva bioquímica.

OBJECTIVE: to analyze the immunohistochemical expression of P27 and CD34 markers as prognostic factors in patients with localized prostate cancer. METHODS: analysis of 100 patients with localized prostate cancer submitted to curative surgery. We carried out the usual histological preparation, followed by immunohistochemistry to detect the accumulation of P27 and CD34 protein followed by statistical analysis. RESULTS: in the evaluation of P27 marker and on the correlation with the variables we found significant difference in Gleason score with positive expression (positive P27) related to lower mean PSA (p = 0.091), lower Gleason score (p < 0.0001) and smaller tumor area in CD34 (p = 0.036). Regarding the CD34 marker at the tumor area, it was observed that the smaller the positive CD34, the lower the PSA value (p < 0.0001) and lower the Gleason score (r = 0.5726, p < 0.0001), and the higher the positive CD34, the higher the staging (r = 0.3305, p <0.0001) and the chance of recurrence (p = 0.002). Patients with higher stage also displayed larger positive CD34 areas (p < 0.0001). CONCLUSION: the markers CD34 and P27 are associated with events specific to prostate cancer, however, only CD34 was able to determine the possibility of biochemical recurrence.

Immunohistochemical study of androgen, estrogen and progesterone receptors in salivary gland tumors

The aim of this work was to study the immunohistochemical expression of androgen receptor, estrogen receptor and progesterone receptor in pleomorphic adenomas, Warthin's tumors, mucoepidermoid carcinomas and adenoid cystic carcinomas of salivary glands. A total of 41 pleomorphic adenomas, 30 Warthin's tumors, 30 mucoepidermoid carcinomas and 30 adenoid cystic carcinomas were analyzed, and the immunohistochemical expression of these hormone receptors were assessed. It was observed that all cases were negative for estrogen and progesterone receptors. Androgen receptor was positive in 2 cases each of pleomorphic adenoma, mucoepidermoid carcinoma and adenoid cystic carcinoma. In conclusion, the results do not support a role of estrogen and progesterone in the tumorigenesis of pleomorphic adenomas, Warthin's tumors, mucoepidermoid carcinomas and adenoid cystic carcinomas. However, androgen receptors can play a role in a small set of salivary gland tumors, and this would deserve further studies.

Expressão da CASPASE-3 e CD-34 no adenocarcinoma de próstata / CASPASE-3 and CD-34 expression in prostate adenocarcinoma

OBJETIVO: 1. Avaliar em qual percentual as células tumorais se marcam com caspase-3 e CD-34; 2. quantificá-los nas células tumorais; 3. verificar correlação entre quantificação e grau de malignidade tumoral; 4. correlacioná-los entre si. MÉTODOS: Estudaram-se 38 blocos com adenocarcinoma, classificados por Gleason e marcação imunoistoquímica para caspase-3 e CD-34. As proteínas imunomarcadas foram quantificadas no software Immuno do Sistema Samba 4000 de citofotometria de imagem, pelo índice de marcagem e densidade óptica. RESULTADOS: Imunomarcou-se 25 lâminas para caspase-3 e 34 para CD-34. As quantificações da caspase-3 para o índice de marcagem foram acima de 50 em 76 por cento e, para densidade óptica, abaixo de 50 para 96 por cento. Em relação ao CD-34, índice de marcagem foi acima de 50 em 59 por cento e densidade óptica abaixo de 50 em 56 por cento. As correlações entre expressões dos marcadores e a gravidade do tumor, assim como entre os marcadores, não evidenciaram significância estatística. Não se mostrou relação de expressão com o score de Gleason. CONCLUSÃO: A presença caspase-3 e CD-34 foi de 73,5 por cento e 100 por cento, respectivamente; 2. caspase-3 e CD-34 apresentaram alta expressão do índice de marcagem, e baixa para densidade óptica; 3. não houve correlação entre as quantificações com a classificação de Gleason; 4. não houve correlação das expressões dos dois marcadores entre si.

OBJCTIVE: 1. To evaluate the percentage of caspase-3 and CD-34 expression on adenocarcinoma; 2. to quantify caspase-3 and CD-34 in tumor cells; 3. to verify the relationship between biomarkers and its malignancy; 4. to correlate biomarkers themselves. METHODS: Thirty-eight human malignant prostate specimens, Gleason's score, were immunohistochemically stained for caspase-3 and CD-34 protein. Quantification was done under Samba 4000 Immuno System reading, yielding two variables: label index and optical density. Statistical analyses were based on cross-methods involving univariate and bivariate as well as correlation factors among independent variables. RESULTS: Immunostaining was revealed in 25 plates for caspase-3 and 34 for CD-34. Caspase-3 expression for label index was over 50 in 76 percent, while for optical density was below 50 in 96 percent. CD-34 expression demonstrated label index over 50 in 59 percent and optical density below 50 in 56 percent. Correlation among expression and severity did not demonstrate to be statistically significant. There was no correlation between protein expression and Gleason's score. CONCLUSION: 1. Caspase-3 and CD-34 were present respectively in 73.5 percent and 100 percent of samples; 2. caspase-3 and CD-34 showed high expression regarding label index and low expression in optical density; 3. there was no statistical significance among expressions and tumor severity according to Gleason's score; 4. no significant correlation could be set between the biomarkers themselves.

Immunohistochemical expression of HER-2/NEU-CERBB-2 in patients with adenocarcinoma of the stomach / Expressão imunoistoquímica do HER-2/NEU-CERBB-2 em pacientes com adenocarcinoma do estômago

Objectives: To determine the prevalence of Her-2/Neu-cerbb-2 in the gastric mucosa of patients with gastric adenocarcinoma in a brazilian patient group. Methods: The immunohistochemical expression of Her-2/Neu was studied in 37 formalin-fixed paraffin–embedded tissue sections. Results: The immunohistochemical reaction produced by the anti-HER- 2/Neu antibody was positive in two cases (5.4%). Conclusion: The low prevalence of Her-2/Neu observed in these southern brazilian cases is probably due to the great number of poorly differentiated cancers in this serie.

Objetivo: Determinar a prevalência do Her-2/Neu-CerbB-2 na mucosa de pacientes com adenocarcinoma de estômago em um grupo de doentes brasileiros. Métodos: A expressão imunoistoquímica do Her-2/Neu foi estudada em 37 amostras de tecidosfixados em formalina e embebidos em parafina. Resultados: A reação imunoistoquímica produzida pelo anticorpo HER-2/Neu foi positiva em dois pacientes (5.4%). Conclusão: A baixa prevalência Her-2/Neu observada neste grupo de pacientes é provavelmente devida ao grande número de tumores pouco diferenciados encontrados nesta série.

Immunohistochemical evaluation for P53 and VEGF (Vascular Endothelial Growth Factor) is not prognostic for long term survival in end stage esophageal adenocarcinoma / Avaliação imunoistoquímica do P53 e VEGF (Fator de Crescimetno Endotelial Vascular) não representa fator prognóstico para sobrevida a longo prazo na fase terminal do adenocarcinoma do esôfago

OBJECTIVES: To correlate the expression of p53 protein and VEGF with the prognosis of patients submitted to curative resection to treat esophageal adenocarcinoma. METHODS: Forty-six patients with esophageal adenocarcinoma, submitted to curative resection, were studied. The expressions of p53 protein and VEGF were assessed by immunohistochemistry in 52.2 percent and 47.8 percent of tumors, respectively. RESULTS: P53 protein and VEGF expressions coincided in 26 percent of the cases, and no correlation between these expressions was observed. None of the clinicopathological factors showed a significant correlation with p53 protein or VEGF expressions. There was no significant association between p53 protein and VEGF expressions and long-term survival. CONCLUSION: The expression of p53 protein and VEGF did not correlate with prognosis in esophageal adenocarcinoma patients submitted to curative resection.

OBJETIVO: Correlacionar a expressão do p53 e VEGF com o prognóstico de pacientes submetidos à operação curativa para tratar adenocarcinoma do esôfago. MÉTODO: Foram estudados 46 pacientes com adenocarcinoma de esôfago, submetidos à ressecções curativas. As expressões do p53 e VEGF foram assessadas por imunoistoquímica em 52.2 por cento e 47.8 por cento dos tumors, respectivamente . RESULTADOS: As expressões de ambos coincidiram em 26 por cento dos casos sem correlação entre elas. Os fatores clinicopatológicos estudados não mostraram correlação significante. Não houve associação significante entre as expresses do p53 e VEGF na sobrevida a longo prazo. CONCLUSÃO: As expressões do p53 e VEGF não se correlacionaram com o prognóstico do adenocarcinoma do esôfago nos pacientes operados com ressecções curativas.

Expressão citofotométrica dos marcadores tumorais CD-34 e fator VIII no câncer de cólon / Cytophotometric expression of the tumor markers CD-34 and factor VIII in colorectal cancer

OBJETIVOS: Verificar a posssibilidade de quantificar a expressão dos marcadores tumorais CD-34 e Fator VIII no câncer de cólon; verificar se existe superioridade entre um marcador e outro para estudo da angiogênese; verificar se há correlação na análise do índice de marcagem e a densidade óptica média nos marcadores utilizados. MÉTODOS: Dezessete casos de adenocarcinoma colorretal recuperados de blocos de parafina e confirmados pela hematoxilina-eosina, foram submetidos à coloração imunoistoquímica pelo método da estreptoavidina-biotina-peroxidase e utilizados os marcadores tumorais CD-34 e Fator VIII. Após este processo as lâminas foram submetidas à leitura no sistema Samba 4000® e avaliadas pelo software Immuno®. Os parâmetros estudados foram: índice de marcagem e densidade óptica, expressos por médias, medianas, valores mínimos, valores máximos e desvios-padrão, analisados estatisticamente. RESULTADOS: Para o marcador CD-34 não houve normalidade dos dados em relação ao índice de marcagem e houve para a densidade óptica. Para o Fator VIII, houve normalidade de dados em relação ao índice de marcagem e para a densidade óptica. CONCLUSÃO: Foi possível quantificar a expressão dos marcadores tumorais CD-34 e Fator VIII através do índice de marcagem e da densidade óptica média; não houve diferença entre os marcadores em relação à média do índice de marcagem e da densidade óptica, não sendo possível definir superioridade entre um e outro; não foi observada tendência à correlação quando comparados densidade óptica e índice de marcagem do Fator VIII e do CD-34 isoladamente estudados; não houve correlação entre o índice de marcagem do Fator VIII quando comparado com o CD-34, bem como a densidade óptica do Fator VIII com o CD-34.

OBJECTIVES: In colorectal cancer, to describe the cytophotometric expression of the CD-34 and Factor VIII; to evaluate the degree of correlation between them; and to compare the CD-34 and Factor VIII expressions in relationship to label index and optical density. METHODS: Seventeen cases of colorectal adenocarcinoma recovered from paraffin-embedded archival tissue and confirmed by hematoxilin-eosin staining, were submitted to streptavidin-biotin-peroxidase immunohistochemical method. In this process was used the tumor markers CD-34 and Factor VIII. The obtained slides were analysed using the SAMBA 4000® system with Immuno® software. The results were evaluated into two parameters: label index and optical density, and expressed by averages, medians, minimum values, maximum values, and standard deviation values. The normality condition of the quantitative variables was investigated by using the Shapiro-Wilks test. In order to evaluate the degree of association between the expressions of the markers, Pearson's Correlation Coefficient or Spearman's Correlation Coefficient were applied. To evaluate the comparison degree of the markers expression, Student's t test or Wilcoxon's no parametric test were used. RESULTS: For the CD-34 there was no data normality for the label index and there was normality in the optical density. For the Factor VIII, there was data normality for the label index and for the optical density. CONCLUSION: When the expressions of CD-34 and Factor VIII markers were correlated, there was no difference between them in relationship to the average label index and average optical density. When the expressions of the CD-34 and Factor VIII were compared, there was no correlation between the two variables.

Bcl-2 expression in rectal cancer

BACKGROUD: Proteins involved in apoptosis process seem to play an important role in colorectal carcinogenesis AIM: To determine the prevalence of bcl-2 protein immunohistochemical expression and its relation with clinical and histopathological variables of rectal adenocarcinoma. PATIENTS AND METHODS: One hundred and thirty-two patients operated at "Hospital de Clínicas de Porto Alegre", Porto Alegre, RS, Brazil, between 1988 and 1999 were studied through immunohistochemical reaction using a monoclonal antibody anti-bcl-2 on formalin-fixed, paraffin-embedded tissue samples RESULTS: The prevalence of bcl-2 protein was 29.5 percent. There was a significant increased number of positive bcl-2 cases among women as compared to men. There was no significant association between bcl-2 and age, tumour site, histological grade, mucin production, depth of invasion, lymphatic involvement, distant metastasis or stage, despite a trend showing decreased immunoreactivity to bcl-2 among poorly and moderately differentiated tumours, as well as disseminated disease CONCLUSIONS: Analysis of bcl-2 protein expression in tumour tissues, as well as other oncoproteins, may have a role in predict therapeutic response and prognosis of colorectal cancer. However, the potential use of bcl-2 protein assessment in the clinical set for management of rectal cancer remains to be determined.

RACIONAL: As proteínas envolvidas no processo de apoptose parecem desempenhar papel importante na carcinogênese colorretal. OBJETIVOS: Determinar a prevalência da expressão imunoistoquímica da proteína bcl-2 e sua relação com variáveis clínicas e histopatológicas do câncer de reto. PACIENTES E MÉTODOS: Cento e trinta e dois pacientes operados no Hospital de Clínicas de Porto Alegre, RS, entre 1988 e 1999 foram estudados através de reação imunoistoquímica, utilizando um anticorpo monoclonal anti-bcl-2 em amostras teciduais fixadas em formalina e parafinizadas. RESULTADOS: A prevalência da proteína bcl-2 foi de 29,5 por cento. Houve aumento significativo no número de casos bcl-2 positivo entre mulheres quando comparado aos homens. Não houve associação significativa entre bcl-2 e idade, sítio do tumor, grau histológico, produção de muco, profundidade de invasão, envolvimento linfático, metástases distantes ou estágio, apesar de uma tendência demonstrando imunorreatividade ao bcl-2 diminuída entre os tumores pouco e moderadamente diferenciados, bem como para doença disseminada. CONCLUSÕES: A análise da expressão da proteína bcl-2 em tecidos tumorais, bem como outras oncoproteínas, pode ter um papel em predizer a resposta terapêutica e o prognóstico do câncer colorretal. Entretanto, o uso potencial da avaliação da proteína bcl-2 na prática clínica no manejo do câncer de reto permanece a ser determinado.

Assessment of staging, prognosis and mortality of colorectal cancer by tumor markers: receptor erbB-2 and cadherins

OBJETIVO: Avaliação da expressão tumoral das proteínas c-erbB-2 e E-caderina e sua relação com o prognóstico, estadiamento e grau de diferenciação celular, em doentes com câncer colo-retal . MÉTODOS: O estudo incluiu 117 doentes com média de idade de 63.1 anos e com acompanhamento médio de 28.1 meses. O intervalo livre de doença, sobrevida, índice de recidiva e mortalidade específica foram os parâmetros avaliados. Anticorpos anti-oncoproteína c-erbB-2 (Dako) foram utilizados pela técnica da estreptavidiva-biotina. Considerou-se como positiva a presença desta proteína quando mais de 10% das células tumorais estivessem coradas. A proteína E-caderina foi estudada pelo anticorpo anti-E-caderina (Dako), sendo computada como positiva a amostra que apresentasse 50% ou mais das células coradas. A análise estatística utilizou o teste do qui-quadrado de Pearson, o teste exato de Fischer, a curva de Kaplan-Meier, o teste de log-rank e o teste de Wilcoxon ( variante de Breslow),sendo estabelecido nível de significância de 5%( p<0,05). RESULTADOS: 52 de 108 doentes estudados para c-erbB-2 foram positivos (48,1%), 47 de 93 doentes estudados para E-caderina foram negativos (50,5%). Estes dados não mostraram relação com estadiamento TNM (tumor, nódulo e metástase), com o grau de diferenciação celular e índice de recidiva tumoral. O intervalo livre de doença para os doentes positivos para c-erbB-2 e negativos para E-caderina foi de 68.0 meses e não diferiu daqueles que foram negativos para c-erbB-2 e positivos para E-caderina ( 55.0 meses - p = 0.5510). A sobrevida média para os doentes positivos para c-erbB-2 e negativos para E-caderina foi 75 meses sem diferença estatisticamente significante com o outro grupo de comparação( 61 meses - p = 0.5256). A mortalidade específica foi de 20.0% dos casos e não se correlacionou com a expressão do c-erbB-2 (p=0,446) ou da E-caderina(p=0,883). CONCLUSÃO: A expressão das proteínas c-erbB-2 e E-caderina em doentes portadores de adenocarcinoma colo-retal não apresentou correlação com o estadiamento e grau de diferenciação celular. Não houve da mesma forma relação com o prognóstico, no que diz respeito ao índice de recidiva da doença, intervalo livre de doença, sobrevida e mortalidade específica.

Cyclin E, p27 and Mutant p53 do not Predict the Prognosis in AJCC Stage II Colorectal Carcinomas / 대한소화기학회지

BACKGROUND/AIMS: Carcinogenesis is characterized by the abnormal regulation of cell cycle. The abnormal expression of the regulators of cell cycle may be related to the prognosis. Since the clinical significance of the expression of the three proteins in colorectal carcinomas is still controversial, we evaluated the prognostic value of the expression of cyclin E, p27 and mutant p53 in stage II colorectal cancer. METHODS: The expression levels of cyclin E, p27 and mutant p53 proteins in 41 patients with stage II colorectal carcinomas were analyzed by immunohistochemistry. RESULTS: In the univariate analysis, the level of CEA at diagnosis was associated with disease relapse. In the multivariate analysis, the clinicopathological variables such as age, gender, site of primary tumor, tumor size, state of tumor differentiation and preoperative plasma CEA level were not associated with disease relapse. When Kaplan-Meier survival curves were constructed to determine the prognosis, cyclin E, p27 and mutant p53 expressions did not predict poor prognosis. CONCLUSIONS: Our results suggested that the expression of cyclin E, p27 and mutant p53 proteins did not predict the clinical outcome in the stage II colorectal carcinomas.

Apoptosis and Proliferation in Paired Primary Colorectal Adenocarcinomas and Their Liver Metastases

The proliferation potentials and the level of apoptosis were compared in paired primary colorectal adenocarcinomas and their liver metastases within each individual. From a total of 22 patients 44 specimens of paired primary and metastatic tumors were obtained for analysis. The levels of spontaneous apoptosis (a spontaneous apoptosis index, SAI: % apoptotic nuclei among a total of 1000 nuclei) and of proliferation (KI-67 index: % positively stained cells for KI-67 among a total of 1000 cells) were analyzed between primary and metastatic tumors. Survival rates and its relationship with the clinical parameters were also analyzed. The overall survival rate at 5 years was 16.9% with the median survival time of 45 months. T-stage (p=0.005) and time to liver metastasis (synchronous versus metachronous, p=0.03) showed statistical significance in relation to survival. The mean SAI of primary tumors was 1.35 +/- 0.25, which was not statistically different from the 1.58 +/- 0.18 of metastatic tumors (p=0.33). The mean KI-67 indices in primary and metastatic tumors were 23.9 +/- 3.4 and 16.4 +/- 2.5, respectively, and this difference was statistically significant (p=0.016). Subset analysis showed significant difference in the KI-67 index in the synchronous group but not in the metachronous group. No significant difference was shown in the relative ratios of apoptosis to proliferation between the primary tumor and the metastasis within each individual. The results in this study may partly explain the indolent behavior of liver metastasis from colorectal cancer and provides a rationale for the active treatment of metastatic tumors as well as of primary disease.

Utility of Thyroid Transcription Factor-1 and Cytokeratin 20 in Identifying the Origin of Metastatic Carcinomas of Cervical Lymph Nodes

The identification of primary location of a metastatic tumor is a difficult diagnostic problem and sometimes can be facilitated by the use of immunohistochemical markers. Thyroid transcription factor-1 (TTF-1) is a 38-kDa nuclear homeodomain transcription factor that is expressed specifically in lung or thyroid neoplasms. Cytokeratin 20 (CK20) is a 46-kDa low-molecular-weight cytokeratin that shows restricted expression in adenocarcinomas of the gastrointestinal tract (GIT) and transitional cell carcinomas of the urinary tract. We studied the immunohistochemical expression of TTF-1 and CK20 in 68 metastatic carcinomas in cervical lymph nodes. The primary sites were the lung in 29 cases, stomach in 13, colorectum in 3, and other sites in 23. TTF-1 expression was detected in 69.0% of metastatic lung carcinomas and none in metastatic GIT carcinomas, whereas CK20 expression was detected in 68.8% of metastatic GIT carcinomas and none of metastatic lung carcinomas. TTF-1 had a specificity of 0.95 and a sensitivity of 0.69 for metastatic lung carcinoma, whereas CK20 had a specificity of 1.00 and a sensitivity of 0.69 for metastatic GIT carcinoma. These results indicate that TTF-1 and CK20 should be the first choice as a component of antibody panel to prove or to exclude the lung and GIT origin, respectively, especially in patients presenting with metastatic carcinomas of unknown primary site.