Immunoexpression of Her-2 protein in fine-needle aspirates of breast carcinoma: correlation with surgical specimens

PURPOSE: to determine the expression of HER-2 protein in fine-needle aspirates obtained from infiltrating breast duct carcinoma and in the corresponding surgical specimens and to correlate HER-2 expression with histological grade. Methods: forty-eight smears fixed in alcohol and previously stained by the Papanicolaou technique and histological sections of corresponding paraffin-embedded tissue blocks were submitted to immunohistochemistry by the avidin-biotin-peroxidase method using a polyclonal antibody (A0485-Dako). The reactions were carried out simultaneously with antigen recovery. The results were analyzed by a semiquantitative method according to the HercepTest score. Scores of 3+ were considered to be positive. RESULTS: positivity was observed in 39.6% of the smears and in 35.4% of the respective histological sections, with almost perfect cytohistologic concordance (kappa 0.82). The method of evaluation of histological sections was found to be reproducible regarding the smears and standardized the interpretation of the results. The correlation between HER-2 positivity and histological grade was not statistically significant but there was a tendency towards HER-2 expression in less differentiated tumors. CONCLUSION: the positivity of the immunocytochemical reactions observed in smears was comparable to that obtained in the histological sections, indicating that the material obtained by fine-needle aspiration biopsies previously stained by the Papanicolaou technique can be used for the evaluation of HER-2 expression.OBJETIVO: avaliar a imunoexpressão da proteína HER-2 em punção biópsia aspirativa de carcinoma de ductos infiltrantes da mama e nos espécimes cirúrgicos correspondentes. Correlacionar a imunoexpressão da proteína HER-2 com o grau histológico. MÉTODO: foram examinados 48 esfregaços fixados em álcool, previamente corados pela técnica de Papanicolaou, e cortes histológicos de blocos de parafina do tecido correspondente. Todos foram submetidos à técnica de imuno-histoquímica, utilizando o método da avidina-biotina-peroxidase com um anticorpo policlonal (A0485-Dako). As reações foram realizadas simultaneamente com recuperação antigênica. A avaliação dos resultados foi feita por método semiquantitativo, segundo a graduação do HercepTest. Foram considerados positivos apenas os casos graduados em 3+. RESULTADOS: observou-se positividade de 39,6% nos esfregaços e 35,4% nos cortes histológicos respectivos, sendo a concordância cito-histológica nos casos positivos quase perfeita (kappa 0,82). A utilização do método de avaliação dos cortes histológicos mostrou-se reprodutível nos esfregaços e padronizou a interpretação dos resultados. A correlação entre a positividade para proteína HER-2 e o grau histológico não foi significante, mas houve tendência a imunoexpressão mais freqüente em tumores menos diferenciados. CONCLUSÃO: a positividade das reações imunocitoquímicas nos esfregaços foi comparável à obtida nos cortes histológicos, podendo portanto o material de punção aspirativa por agulha fina, previamente corado pela técnica de Papanicolaou, ser utilizado na avaliação da imunoexpressão da proteína HER-2.Universidade Federal de São Paulo (UNIFESP) Departamento de Patologia Escola Paulista de MedicinaInstituto Adolfo LutzUNIFESP, Depto. de Patologia Escola Paulista de MedicinaSciEL

Characterization of monocarboxylate transporter activity in hepatocellular carcinoma

AIM: To assess the immunoexpression of hypoxia-related markers in samples from cirrhosis and primary and metastatic hepatocellular carcinoma (HCC). METHODS: From a total of 5836 autopsies performed at the Pathology Department - University of Sao Paulo School of Medicine Hospital - from 2003 to 2009, 188 presented primary liver tumors. Immunohistochemical reactivity for monocarboxylate transporters (MCTs)-1, 2 and 4, CD147 and glucose transporter-1 (GLUT1) was assessed in necropsies from 80 cases of HCC. Data were stored and analyzed using the IBM SPSS statistical software (version 19, IBM Company, Armonk, NY). All comparisons were examined for statistical significance using Pearson's ? (2) test and Fisher's exact test (when n < 5). The threshold for significant P values was established as P < 0.05. RESULTS: Plasma membrane expression of MCT4 and overall expression of GLUT1 showed progressively higher expression from non-neoplastic to primary HCC and to metastases. In contrast, overall expression of MCT2 was progressively decreased from non-neoplastic to primary HCC and to metastases. MCT1 (overall and plasma membrane expression), MCT2 and CD147 plasma membrane expression were associated with absence of cirrhosis, while plasma membrane expression of CD147 was also associated with absence of HBV infection. MCT2 overall expression was associated with lower liver weight, absence of metastasis and absence of abdominal dissemination. Additionally, MCT4 plasma membrane positivity was strongly associated with Ki-67 expression. CONCLUSION: MCT4 and GLUT1 appear to play a role in HCC progression, while MCT2 is lost during progression and associated with better prognosis

A morphological protocol and guide-list on uterine cervix cytology associated to papillomavirus infection

The present study was designed to further assess the validity of the cytological description of morphological lesions said to be related to Papillomavirus (HPV) infections in senior women. The casuistic comprised 196 cervical smears from a group of women with no clinical or morphological evidence of neoplasia, collected simultaneously with samples submitted to detection of HPV DNA by PCR in a previous study. Three experienced cytologists studied each slide in two different conditions, with an interval of 20 months between them. The first approach was performed under routine laboratory standards, whereas the second was guided by a list of 16 well-defined parameters indicative of HPV-related cytological lesions. When suspicious cases of HPV-related alterations were grouped with positive cases, they showed on average: sensitivity of 25.5%, specificity of 84.4% and positive predictive value (PPV) of 26.8%. When suspicious cases were grouped with negative cases, sensitivity decreased, whereas specificity and PPV increased, as expected. In the second reading, which followed a “guide-list”, a decrease in sensitivity was observed, contrasting with a sharp increase of positive predictive value. Among the 16 cytomorphological criteria tested, “koilocytosis”, “mild koilocytosis” and “condylomatous parabasal cells” yielded the best predictive value for HPV DNA detection by PCR. In conclusion, despite the low sensitivity, cytopathologic assessment of cervico-vaginal smears leads to a highly specific diagnosis of HPV infection in menopausal women, with PPV of 91.0% when directed by a guide-list of well-defined morphologic criteria.O presente estudo analisou aspectos relacionados ao valor da citologia cervical na identificação de alterações cito-morfológicas relacionadas à infecção por Papilomavírus humano (HPV) em mulheres com idades avançadas. A casuística compreendeu 196 amostras cérvico-vaginais provenientes de uma população de mulheres sem evidências clínicas ou citológicas de neoplasia cervical. As amostras foram coletadas simultaneamente para pesquisas para DNA-HPV por Reação de Polimerização em Cadeia (PCR) e citologia. Três observadores experientes efetuaram análise das lâminas em duas etapas com intervalo de 20 meses: a primeira em condições de rotina laboratorial e a segunda dirigida por um roteiro de critérios pré-estabelecidos. Quando os casos suspeitos para alterações relacionadas ao HPV foram agrupados com os casos positivos, eles mostraram em média: 25,5% de sensibilidade, 84,4% de especificidade e valor preditivo positivo (VPP) de 26,8%. Quando os casos suspeitos foram agrupados com os negativos, a sensibilidade diminuiu, e a especificidade e o VPP aumentaram. Na segunda leitura, a sensibilidade diminuiu, contrastando com o aumento do VPP. Entre os 16 critérios cito-morfológicos avaliados, coilocitose, coilocitose leve e célula parabasal coilocitótica foram os que apresentaram melhor VPP para HPV em comparação ao PCR. Concluímos que, apesar da baixa sensibilidade, o painel de critérios cito-morfológicos poderá elevar a especificidade do teste de Papanicolaou para triagem das alterações relacionadas à infecção por HPV em pacientes de mais idade

Padronização da análise de critérios citomorfológicos de lesões associadas à infecção pelo HPV

The present study was designed to further assess the validity of the cytological description of morphological lesions said to be related to Papillomavirus (HPV) infections in senior women. The casuistic comprised 196 cervical smears from a group of women with no clinical or morphological evidence of neoplasia, collected simultaneously with samples submitted to detection of HPV DNA by PCR in a previous study. Three experienced cytologists studied each slide in two different conditions, with an interval of 20 months between them. The first approach was performed under routine laboratory standards, whereas the second was guided by a list of 16 well-defined parameters indicative of HPV-related cytological lesions. When suspicious cases of HPV-related alterations were grouped with positive cases, they showed on average: sensitivity of 25.5%, specificity of 84.4% and positive predictive value (PPV) of 26.8%. When suspicious cases were grouped with negative cases, sensitivity decreased, whereas specificity and PPV increased, as expected. In the second reading, which followed a "guide-list", a decrease in sensitivity was observed, contrasting with a sharp increase of positive predictive value. Among the 16 cytomorphological criteria tested, "koilocytosis", "mild koilocytosis" and "condylomatous parabasal cells" yielded the best predictive value for HPV DNA detection by PCR. In conclusion, despite the low sensitivity, cytopathologic assessment of cervico-vaginal smears leads to a highly specific diagnosis of HPV infection in menopausal women, with PPV of 91.0% when directed by a guide-list of well-defined morphologic criteria.O presente estudo analisou aspectos relacionados ao valor da citologia cervical na identificação de alterações cito-morfológicas relacionadas à infecção por Papilomavírus humano (HPV) em mulheres com idades avançadas. A casuística compreendeu 196 amostras cérvico-vaginais provenientes de uma população de mulheres sem evidências clínicas ou citológicas de neoplasia cervical. As amostras foram coletadas simultaneamente para pesquisas para DNA-HPV por Reação de Polimerização em Cadeia (PCR) e citologia. Três observadores experientes efetuaram análise das lâminas em duas etapas com intervalo de 20 meses: a primeira em condições de rotina laboratorial e a segunda dirigida por um roteiro de critérios pré-estabelecidos. Quando os casos suspeitos para alterações relacionadas ao HPV foram agrupados com os casos positivos, eles mostraram em média: 25,5% de sensibilidade, 84,4% de especificidade e valor preditivo positivo (VPP) de 26,8%. Quando os casos suspeitos foram agrupados com os negativos, a sensibilidade diminuiu, e a especificidade e o VPP aumentaram. Na segunda leitura, a sensibilidade diminuiu, contrastando com o aumento do VPP. Entre os 16 critérios cito-morfológicos avaliados, coilocitose, coilocitose leve e célula parabasal coilocitótica foram os que apresentaram melhor VPP para HPV em comparação ao PCR. Concluímos que, apesar da baixa sensibilidade, o painel de critérios cito-morfológicos poderá elevar a especificidade do teste de Papanicolaou para triagem das alterações relacionadas à infecção por HPV em pacientes de mais idade

Implementing human papillomavirus testing in a public health hospital : challenges and opportunities

Author's proof. The authors are indebted to Luciane Sarahyba and Nazareno dos Santos of the Molecular Biology Laboratory for their technical assistance.OBJECTIVES: Robust evidence now supports human papillomavirus (HPV) testing as a more effective option to screening and as more sensitive than cytology in detecting high-grade cervical intraepithelial neoplasia . Our goal was to analyze the performance of the Hybrid Capture II (HC2) assay for high-risk HPV (hrHPV) in women undergoing gynecological examination at a public health hospital as part of the evaluation of HPV screening as an alternative or complement to cytology. STUDY DESIGN: This analysis is a subset of a cross-sectional study carried out at a large public hospital serving a predominantly low-resource population. A total of 705 women were enrolled; the sensitivity and specificity of each test were estimated and compared. RESULTS: The analysis identified 272 hrHPV-positive women (mean age 36.3 years) and 433 hrHPV-negative women (mean age 41.2 years). HPV testing showed a significantly increased sensitivity of the HC2 assay versus cytology (84.5 vs. 69.7%; p < 0.0001) but a lower specificity (49.90 vs. 88.78%; p < 0.0001). CONCLUSION: The combination of both methods seems to be useful in improving detection of cervical lesions

Fatores preditivos para resposta da Lamivudine na hepatite crônica B

BACKGROUND: Lamivudine has been shown to be an efficient drug for chronic hepatitis B (CHB) treatment. AIM: To investigate predictive factors of response, using a quantitative method with high sensitivity. METHODS: We carried out a prospective trial of lamivudine in 35 patients with CHB and evidence for viral replication, regardless to their HBeAg status. Lamivudine was given for 12 months at 300 mg daily and 150 mg thereafter. Response was considered when DNA was undetectable by PCR after 6 months of treatment. Viral replication was monitored by end-point dilution PCR. Mutation associated with resistance to lamivudine was detected by DNA sequencing in non-responder patients. RESULTS: Response was observed in 23/35 patients (65.7%) but only in 5/15 (33.3%) HBeAg positive patients. Only three pre-treatment variables were associated to low response: HBeAg (p = 0.006), high viral load (DNA-VHB >; 3 x 10(6) copies/ml) (p = 0.004) and liver HBcAg (p = 0.0028). YMDD mutations were detected in 7/11 non-responder patients. CONCLUSIONS: HBeAg positive patients with high viral load show a high risk for developing drug resistance. On the other hand, HBeAg negative patients show a good response to lamivudine even with high viremia.INTRODUÇÃO: A Lamivudina tem-se mostrado útil no tratamento da hepatite crônica pelo vírus B (HC-VHB). OBJETIVO: Investigar os fatores preditivos da resposta à Lamivudina na HC-VHB. MATERIAL E MÉTODOS: Estudo prospectivo com Lamivudina em 35 pacientes com HC-VHB e evidência de multiplicação viral, independentemente do resultado do AgHBe. Administrou-se a Lamivudina na dose diária de 300 mg por 12 meses, seguida de 150 mg diários. Critério de resposta: DNA-VHB negativo (por técnica de PCR) aos 6 meses de tratamento. Nos pacientes não respondedores, pesquisaram-se mutações associadas com resistência à Lamivudina, através do sequenciamento do DNA viral. RESULTADOS: Observou-se resposta em 23/35 pacientes (65,7%). Dos 15 pacientes com AgHBe positivo antes do tratamento, apenas 5 (33,3%) responderam. As variáveis prévias ao tratamento que puderam prever uma má resposta foram: AgHBe positivo (p = 0,006), carga viral elevada (>; 3 x 10(6) genomas/ml) (p = 0,004) e AgHBc no tecido positivo (p = 0,0028). Mutações na região YMDD foram detectadas em 7/11 pacientes não respondedores. CONCLUSÕES: Pacientes com AgHBe positivo e com alta carga viral apresentam um alto risco de desenvolver resistência à Lamivudina. Por outro lado, pacientes com AgHBe negativo, mesmo com alta carga viral, mostraram uma boa resposta à Lamivudina

Aspectos moleculares da carcinogênese hepática

Agentes exógenos correlacionados com o carcinoma hepatocelular (HCC) têm sido identificados e bem caracterizados. Esses agentes, entre os quais se incluem os diferentes vírus que causam hepatite e cirrose hepática, podem provocar o aparecimento de nódulos regenerativos e nódulos displásicos/hiperplasia adenomatosa. Essas condições associadas com diversas alterações moleculares do hepatócito podem culminar com o aparecimento do HCC. Recentemente, grandes progressos têm ocorrido na identificação de mutações somáticas ou germinativas que estariam correlacionadas com o desenvolvimento do HCC, justificando ampla revisão do tema. Procuramos discutir nesta revisão os fatores envolvidos no processo de carcinogênese hepática, tal como a infecção pelos vírus das hepatites B e C, com ênfase nas alterações moleculares descritas nos últimos anos, assinalando áreas em que potenciais avanços na abordagem clínica poderão surgir em futuro próximo.Exogenous agents correlated with hepatocellular carcinoma (HCC) have been identified and well characterized. These agents, including the different viruses that cause chronic hepatitis and cirrhosis, can lead to regenerative nodules and dysplastic nodules/adenomatous hyperplasia. These conditions associated with several molecular alterations of hepatocyte ultimately culminate in hepatocellular carcinoma. Recently, there has been a great progress in the identification of somatic and germinative mutations that may be correlated with the development of HCC, justifying a review on the subject. Hence, the factors involved in the process of hepatic carcinogenesis, such as infection by the hepatitis B and C viruses, with a special focus in the molecular alterations described in recent years are discussed herein, pointing out areas potentially relevant for clinical development

Seroprevalence of human papillomavirus 6, 11, 16, and 18 in young primiparous women in Sao Paulo, Brazil

INTRODUCTION: Data on epidemiology of HPV infection are needed for the development of human papillomavirus (HPV) vaccine recommendations, especially in countries where HPV vaccination is not yet included in public vaccination programs. The aim of this study was to determine the prevalence of serum antibodies to HPV types 6, 11, 16, and 18 and associated factors among young women after birth of the first child. METHODS: This cross-sectional study was carried out in a large public maternity hospital in Sao Paulo, Brazil. Three hundred one women aged 15 to 24 years who gave birth to their first child were recruited between 43 and 60 days after delivery. Seroprevalence was performed using a type-specific enzyme-linked immunosorbent assay based on HPV Late protein 1 viruslike particles. The association of seroreactivity with these 4 HPV types with selected demographic and behavioral factors was assessed by Generalized Linear Model analysis. RESULTS: Fifty-eight (19.3%) women (95% confidence interval, 15.0%-24.2%) had antibodies to any of the 4 viruslike particles tested. The overall seroprevalence rates of the HPV types were: HPV16, 9.0%; HPV18, 7.0%; and HPV 6+11, 7.7%, which are targeted by the HPV prophylactic vaccines. In the multivariate analysis, only age (inversely, P = 0.044 for trend) and previous sexually transmitted disease (P = 0.008) were 2 factors independently associated with HPV seropositivity. CONCLUSIONS: These data offer additional information on the epidemiology of HPV in a group of young Brazilian women after first delivery and contribute to establish a baseline of HPV seroprevalence against which post-HPV vaccine era seroprevalence can be compared. PMID: 21051985 [PubMed - indexed for MEDLINE]This study was supported by a research grant from the Department of Immunization, Vaccines and Biologicals, World Health Organization (ref: V20-181-13)