Ana Paula Martins Sebastião

Estudo das atipias indeterminadas em relação à prevalência e ao percentual de discordância nos casos do Programa de Prevenção do Câncer Uterino do Paraná

BACKGROUND: The Bethesda Pap Smear System introduced two categories of undetermined atypias: atypical squamous cells of undetermined significance (ASCUS) and atypical glandular cells of undetermined significance (AGUS). The objective of this study is to report the prevalence and the disagreement percentile of ASCUS and AGUS as diagnosed by the Cervical Cancer Screening Program of Parana and revised by the Unit for External Monitoring of Cytology Quality (UEMCQ). METHODS: The UEMCQ revised 65,753 cervical smears during 25 months and all undetermined atypias were selected. These cases were classified according to their agreement or not and then prevalence was obtained. RESULTS: Before the UEMCQ review, the number of undetermined atypias was 4,067 (5.45%); this number decreased to 3,584 after revising the data. Considering the whole sample, the ASCUS rate was 4.91% (3,235) and AGUS corresponded to only 0.51% (338). Most of the reclassified cases presented negative diagnoses (57.32%). The primary grouped diagnoses, including CIN I and HPV, was responsible for 51.1% of undetermined atypias after reclassification. DISCUSSION AND CONCLUSION: Undetermined atypias were the second most frequent among disagreement diagnosis groups.

Does human papillomavirus play a role in endometrial carcinogenesis

To determine the relationship between human papillomavirus (HPV) and endometrial carcinogenesis by comparing data from women with endometrial carcinoma to those of women with normal endometrial tissue. The survey was conducted for 100 women (50 with endometrial carcinoma and 50 with normal endometrial tissue) through HPV-DNA testing of paraffin-embedded endometrial tissue sections by polymerase chain reaction. Age, cigarette consumption, squamous differentiation and tumor grade, endometrium trophism, and HPV types detected in endometrial tissues were studied. HPV estimated odds ratio was similar in endometrial carcinoma and in normal endometrial tissue. The presence of HPV was not associated with age, tobacco abuse, endometrial histology status, squamous differentiation, or tumoral grade. DNA sequences of HPV types 16 and 18 were the most frequently detected in both groups. An association between HPV and endometrial carcinoma was not observed.

Down regulation of ADAM33 as a Predictive Biomarker of Aggressive Breast Cancer

Breast cancer is a heterogeneous disease with differences in its clinical, molecular and biological features. Traditionally, immunohistochemical markers together with clinicopathologic parameters are used to classify breast cancer and to predict disease outcome. Triple-negative breast cancer (TNBC) is a particular type of breast cancer that is defined by a lack of expression of hormonal receptors and the HER2 gene. Most cases of TNBC also have a basal-like phenotype (BLBC) with expression of cytokeratin 5/6 and/or EGFR. A basal marker alone is insufficient for a better understanding of the tumor biology of TNBC. In that regard, the ADAM33 gene is silenced by DNA hypermethylation in breast cancer, which suggests that ADAM33 might be useful as a molecular marker. In the present study, we have produced monoclonal antibodies against the ADAM33 protein and have investigated the role of ADAM33 protein in breast cancer. We used 212 breast tumor samples and lower levels of ADAM33 were correlated with TNBC and basal-like markers. A lower level of ADAM33 was also correlated with shorter overall survival and metastasis-free survival and was considered an independent prognostic factor suggesting that ADAM33 is a novel molecular biomarker of TNBC and BLBC that might be useful as a prognostic factor.

Impact of preoperatory magnetic resonance imaging in oncoplastic surgery

Study carried out at Unidade de Mama do Hospital Nossa Senhora das Graças – Curitiba (PR), Brazil. 1Unidade de Mama, Hospital Nossa Senhora das Graças – Curitiba (PR), Brazil. 2Clínica de Diagnóstico Avançado por Imagem, DAPI – Curitiba (PR), Brazil. *Corresponding author: kfurlan@terram.com.br Conflict of interests: nothing to declare. Received in: 05/14/2017. Accepted in: 05/29/2017 Objetivo: Não há dados sobre o impacto da imagem de ressonância magnética (RM) na cirurgia oncoplástica. O objetivo deste trabalho foi avaliar o impacto da RM no planejamento cirúrgico e nas mudanças de conduta em pacientes com câncer de mama inicial e candidatas a realizar a cirurgia oncoplástica. Métodos: Trata-se de uma coorte prospectiva de 60 pacientes que foram candidatas à cirurgia oncoplástica entre janeiro de 2013 e julho de 2014. Todos elas foram submetidas a uma RM pré-operatória, além de mamografia (MG) e ultrassom (US). Qualquer tumor adicional na RM classificada como BIRADS 4-5 foi biopsiado ou marcado com carvão e ROLL para serem localizados durante a cirurgia. O impacto cirúrgico dos achados adicionais da RM foi avaliado quanto a mudanças para mastectomia ou ressecção mais ampla. Resultados: Das pacientes, 29/60 (48,3%) apresentaram achados adicionais na ressonância magnética, 16/29 (55%) foram tumores multifocais, 1/29 (3,4%) foi multicêntrico, 5/29 (17%) foram tumores contralaterais e 9/29 (31%) apresentaram tamanho de tumor maior que 10 mm na RM. Das 22 pacientes que apresentaram lesões adicionais na RM, 15 (68,2%) apresentaram carcinomas invasivos no exame anatomopatológico definitivo. A sensibilidade da RM foi maior na estimativa do tamanho do tumor. Das pacientes, 12/60 (20%) foram submetidas à mastectomia e 17/60 (28,3%) a ressecções mais amplas. Apenas 5% das pacientes apresentaram margens positivas em todo o grupo. No grupo de pacientes que apresentaram resultados adicionais na RM, apenas 3,4% tiveram margens positivas. Conclusões: A RM é melhor que a MG e o US na avaliação da extensão do tumor e na detecção de tumores multicêntricos, multifocais e bilaterais. Em consequência, contribuiu nesta série para um melhor planejamento cirúrgico na cirurgia oncoplástica com baixa taxa de margens comprometidas e reexcisão.

Comparison of cythopathology and frozen section methods in the intraoperative evaluation of breast sentinel lymph node

Introduction: Cytopathologic analysis (CP) and frozen section (FS) are available techniques for intraoperative evaluation of sentinel lymph node in surgeries for breast cancer treatment that will define the need for axillary lymphadenectomy. Objective: To compare CP and FS of axillary sentinel lymph nodes in metastasis detection of patients with breast cancer. Materials and methods: The electronic files from January 2010 and December 2014, from the Centro de Patologia de Curitiba, Parana, Brazil were reviewed, and were included all the cases in which the result of intraoperative exams by both methods, CP and FS, were recorded. The results of intraoperative exams were compared to the histopathology by hematoxylin and eosin (HE) staining. Results: A total of 183 sentinel lymph nodes from 94 patients were included. The mean age was 55 years and mean lymph node size was 11.70 mm. There was one false-positive case in both intraoperative methods and four false-negatives in FS, which were micrometastasis. FS’s sensibility, specificity, positive predictive value (PPV) and negative predictive value (NPV) and accuracy were respectively 80%, 99.38%, 94.11%, 96.42% and 97.26%. Regarding CP, there were six false-negative in which four were micrometastasis. CP’s sensibility, specificity, PPV and NPV and accuracy were respectively 70%, 99.38%, 93.33%, 96.42% and 97.26%. Conclusion: The results of the present study shows that both CP and FS are reliable techniques for metastasis detection in breast sentinel lymph nodes, and are equivalent in sensibility, specificity, accuracy, PPV and NPV.

Giant nodular pseudoangiomatous stromal hyperplasia of the breast with fbroadenomatoid myxoid changes: a potential pitfall in the differential diagnosis of phyllodes tumor

A 21-year old woman presented with a 17-cm left breast mass. Physical examination and ultrasound revealed the mass to be wellcircumscribed, homogeneous and freely mobile, suggestive of giant fibroadenoma or phyllodes tumor. The mass was surgically excised and initially interpreted as benign phyllodes tumor. Subsequent slide review established the diagnosis of nodular pseudoangiomatous stromal hyperplasia (PASH) associated with fibroadenomatoid areas and myxoid stromal changes. This case illustrates the difficulty encountered in recognizing nodular PASH. A thorough discussion of the histopathologic differential diagnosis of nodular PASH is provided.

Resultados do exame intraoperatório do linfonodo sentinela na Unidade de Mama do Hospital Nossa Senhora das Graças em Curitiba

A biopsia do linfonodo sentinela (LS) e o procedimento padrao para as pacientes com axila clinicamente negativa. O seu exame intraoperatorio ainda gera dificuldades na sua abordagem. Assim, o objetivo deste estudo foi avaliar a eficacia desse exame no câncer de mama. Foram avaliadas 342 pacientes que foram operadas na Unidade de Mama do Hospital Nossa Senhora das Gracas em Curitiba (PR), no periodo de 2000 a 2012. No exame intraoperatrorio eram rea lizados cortes longitudinais, ao longo do maior eixo, a cada 2 ou 3mm. Em seguida eram feitos imprints em cada face de cada fatia e, entao, realizavam se cortes histologicos em criostato em tres niveis. Tanto os imprints quanto os cortes eram corados com azul de toluidina. Em sua maio ria eram tumores T1c (n=151), e 60 (17,5%) delas apresentaram axila comprometida no exame definitivo. A acuracia foi de 92%, o valor preditivo negativo, de 91% e a taxa de falso negativo, de 8%. Nao foram encontrados fatores de risco significativos para falencia da tecnica dentro dos parâmetros estudados.(AU) Sentinel node (SN) biopsy is the standard of care for patients with clinically negative axilla. However, in traoperatory pathological exam remains as a controversial issue. So, the aim of this study was to evaluate its efficacy in 342 breast cancer patients operated at the Hospital Nossa Senhora das Gracas Breast Unit in Curitiba (PR), in the period between 2000 2012. In the intraoperatory evaluation, all SNs were cutted in the major axis, in three levels, combining frozen sections with imprints, using toluidin blue. The majority of patients were T1c (n=151), and 60 (17.5%) had positive axila in the definitive pathology evaluation. Accuracy was 92%, predictive negative value was 91%, and false negative rate was 8%. We did not find any significant risk factor for false negative SN in this series.(AU)

Abstract 3822: DNA copy number changes in the PI3K signaling pathway genes in triple negative breast cancer

Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL Background: Triple negative breast cancer (TNBC) is associated with poor prognosis and a high mortality rate when compared to the other breast cancer subtypes. These tumors lack the expression of ER, PR and HER2 receptors, and do not respond well to the currently available adjuvant therapies. The phosphatidylinositol 3-kinase (PIK3) signaling pathway mediates key cellular functions. Alterations of genes in this pathway have been shown to predict responsiveness to cancer targeted therapies. In this study, five genes involved in this signaling pathway, PIK3CA, AKT1, AKT2, PTEN and FGFR, were evaluated for DNA copy number changes in two sets of TNBC specimens. Methods: TNBC samples were obtained from the Hospital Nossa Senhora das Gracas, Curitiba, PR, Brazil. Two sets of TNBC samples were analyzed: the first set was composed of 30 formalin fixed-paraffin embedded tissue specimens and were analyzed for the PIK3CA, AKT1, AKT2, PTEN and FGFR genes by FISH. Locus-specific probes were prepared from BAC contigs for each gene and a centromeric probe for chromosome 3 was used as a control for copy number. The second set was composed of 17 frozen samples and were analyzed by array-CGH using a 44K-Agilent array platform. Real-Time PCR (TaqMan assay), was used to validate the array-CGH data for the above five genes. Results: FISH: PTEN loss and FGFR3 amplification were the most common findings occurring in 56% and 41% of the TNBC samples respectively. PIK3CA, AKT2 and AKT1 were amplified in 31%, 25% and 9.5% of the samples respectively. Array-CGH/Real time PCR: DNA copy number changes were observed by array-CGH in all, but one TNBC sample. The gene with highest frequency of amplification detected by array-CGH was PIK3CA (amplified in 77% of the cases). These findings were confirmed in 70% of the cases. FGFR2, AKT2 and PTEN were amplified in 35%, 24% and 24% of the cases respectively, These findings were confirmed for FGFR2 in 83% of the samples and for PTEN in all the samples. AKT2 amplification was only confirmed in 25% of the samples. Finally AKT1 was observed amplified in by array-CGH in 6% of the cases, which was confirmed by Real-time PCR.Conclusions: Based on our initial results, we conclude that TNBC presents DNA copy number alterations in genes involved in the PIK3 signaling pathway. Upon validation of these alterations in a larger number of cases, with correlation to prognostic parameters, their potential use as therapeutic targets can be considered for further investigation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3822. doi:10.1158/1538-7445.AM2011-3822

Study on the prevalence of human papillomavirus (HPV) in samples of endometrial tissue, both normal and with carcinoma, using the PCR technique

Objetivo: comparar a prevalencia da presenca do DNA do HPV pela tecnica de PCR em amostras de tecido endometrial normal e com carcinoma endometrial de mulheres submetidas a tratamento cirurgico (histerectomia) por carcinoma endometrial e doenca benigna e a sua correlacao com a idade, tabagismo, diferenciacao escamosa e grau de diferenciacao tumoral, tipo viral mais frequente e trofismo endometrial nas mulheres sem carcinoma. Metodos: trata-se de estudo observacional do tipo caso-controle em que foram avaliadas 100 mulheres (50 com endometrio normal e 50 com carcinoma endometrial) quanto a presenca do DNA do HPV em amostra tecidual conservada em blocos de parafina, pelo metodo de PCR. Resultados: o risco relativo estimado da presenca do HPV foi o mesmo nas mulheres com e sem carcinoma endometrial. A presenca do HPV nao esteve correlacionada com a idade das mulheres, tabagismo, trofismo endometrial, diferenciacao escamosa e grau de diferenciacao tumoral. O HPV 16 e 18 (5 dos casos com tipo 16 e 4 com o tipo 18) foram os virus mais frequentemente encontrados. Conclusao: o HPV esta presente no tecido endometrial de mulheres com carcinoma endometrial na mesma proporcao que nas com tecido endometrial normal, nao demonstrando a possivel associacao deste virus no desenvolvimento do carcinoma endometrial.

Pesquisa da prevalência do papilomavírus humano em amostras de tecido endometrial normal e com carcinoma pela técnica de PCR

OBJECTIVE: to compare the prevalence of DNA of human papillomavirus (HPV), in samples of normal endometrial tissue, and tissue with endometrial carcinoma of women submitted to surgical treatment (hysterectomy), or between endometrial carcinoma and benign disease, through the PCR technique. METHODS: this is an observational control-case study where 100 women (50 with endometrial carcinoma and 50 with normal endometrial tissue) were analyzed for the detection of HPV DNA in samples of endometrial tissue kept in paraffin blocks by the PCR technique. The cases of endometrial carcinoma with uncertain primary site of the lesion as well as the cases with previous or current history of pre-neoplasic lesions or carcinoma of the lower genital tract were excluded. Variables as age, smoking habit, endometrial trophism, squamous differentiation and degree of tumor differentiation were also evaluated. RESULTS: the estimated relative risk of the presence of HPV in the endometrial carcinoma and in the normal endometrial tissue was the same. HPV was detected in 8% of the cases of carcinoma and 10% in the normal endometrial tissue. In spite of HPV having been 3.5 times more detected in women with smoking habit in the group without carcinoma, there was no statistical difference. The presence of HPV was also not correlated with the womens age, endometrial trophism, squamous differentiation and degree of tumor differentiation. The HPV types 16 (5 cases) and 18 (4 cases) were the viruses most frequently found both in the normal endometrial tissue or in the tissue with carcinoma. No oncogenic low risk virus was detected in the samples. CONCLUSION: The same proportion of HPV is present in the endometrial tissue of women with endometrial cancer and with normal endometrium. It could not be demonstrated a possible correlation of DNA of HPV with the development of endometrial carcinoma.