Methylation profiling in promoter sequences of ATM and CDKN2A (p14ARF/p16INK4a ) genes in blood and cfDNA from women with impalpable breast lesions.

The objective of the present study was to evaluate the epigenetic changes occurring in early stages of breast cancer. The present study investigated the methylation profile of the ATM, p14ARF and p16INK4a promoters in total blood and plasma cell-free DNA (cfDNA) from women with impalpable breast lesions compared with in total blood of a control cohort of women without breast lesions. The samples were evaluated using the methylation-specific PCR method. The Fisher's exact test was used to evaluate statistical significance between the methylation and clinical variables. A total of 111 women were evaluated, including 56 women with impalpable breast cancer (39/56 also had paired plasma cfDNA) and 55 women in the control cohort (55 blood DNA). For blood DNA from women with malignant impalpable breast lesions, p16INK4a exhibited the greatest percentage of methylation (48%), followed by ATM (37.5%) and p14ARF (27%) promoters, regardless of age variation. For plasma cfDNA, the methylation rates for ATM, p14ARF and p16INK4a were 26, 26 and 10%, respectively. The methylation rates for the blood DNA of controls were the lowest for ATM (9%), p14ARF (7%) and p16INK4a (7%). The women with impalpable breast lesions (benign and malignant lesions) exhibited the highest methylation rate, regardless of age, compared with the paired plasma cfDNA and controls. This epigenetic change was statistically significant for the promoters of ATM (P=0.009) and p16INK4a (P=0.001) (impalpable breast lesions vs. control). The present study demonstrated that epigenetic changes occurring in the ATM and CDKN2A genes detectable in liquid biopsy were associated with the development of impalpable breast lesions.

Low Levels of Vitamin D in a Cohort of Women with Impalpable Breast Lesions from Rio de Janeiro/Brazil

Background: Low levels of vitamin D have been described as a risk factor for the development of breast cancer. The aim of this study was to evaluate the serum levels of vitamin D (25OHD) in patients with impalpable breast lesions comparing with a control group. Methods: Vitamin D quantification (25OHD) was assessed in the plasma of 65 patients with impalpable breast lesions and from 20 health controls using a chemiluminescent microparticle immunoassay. Pearson's chi-square test and nonparametric t-Student were used to evaluate statistical significance between the clinical variables and the means of quantification of vitamin D. The receiver operating characteristic (ROC) curve was used to evaluate the correlation between age and vitamin sufficiency for the cases and the controls. Results: The prevalence of vitamin D deficiency and/or insufficiency in women with malignant lesions was 84% and 60% for the control group. Using the chi-square or Fisher's exact test, the relationship between vitamin D levels and age presented significant association only for the control group (P=0.002). Using ROC curve, the plot area (0.778) for the control group defined a cut-off value of 45 years to age, with specificity and sensitivity of 60% and 50%, respectively. Thus, the odds ratio for vitamin D insufficiency in women over 45 years was 1.37 (P=0.011). For the case group, clinical characteristics, histological grade, and lymph node involvement did not show any significant association. Conclusion: The prevalence of vitamin D deficiency/insufficiency is high in women with impalpable breast lesions, as well as in the control group, even in a tropical city. According to the results the age advancement may be involved with the decrease in vitamin D levels in plasma, but there was no statistical association between low levels of Vitamin D and breast cancer.

Proteomic profile of saliva and plasma from women with impalpable breast lesions.

The present study evaluated the proteomic profile of saliva and plasma from women with impalpable breast lesions using nano-liquid chromatography-quadrupole-time-of-flight (nLC-Q-TOF) technology. Plasma and saliva from patients with fibroadenoma (n=10), infiltrating ductal carcinoma (n=10) and healthy control groups (n=8) were assessed by combinations of inter/intra-group analyses, revealing significant quantitative and qualitative differences. The major differentially-expressed proteins in the saliva of patients compared with the controls were α2-macroglobulin and ceruloplasmin, but the proteins that met the minimum fold-change and P-value cut-offs were leukocyte elastase inhibitor and α-enolase, and deleted in malignant brain tumors 1. Concerning plasma, α-2-macroglobulin and ceruplasmin were upregulated, while other proteins such as haptoglobin, hemopexin and vitamin D-binding protein were downregulated compared with the control. The changes in immune, molecular transport and signaling pathways were the most representative in the proteomic profile of the saliva and plasma. This is the first study to describe the proteome of saliva and plasma from the same women with impalpable breast lesions.

CDKN2A (p14(ARF)/p16(INK4a)) and ATM promoter methylation in patients with impalpable breast lesions.

Early detection of breast cancer increases the chances of cure, but the reliable identification of impalpable lesions is still a challenge. In spite of the advances in breast cancer detection, the molecular basis of impalpable lesions and the corresponding circulating biomarkers are not well understood. Impalpable lesions, classified by radiologists according to the Breast Imaging Reporting and Data System in the categories 3 and 4, can be either benign or malignant (slow growing or aggressive). In this article, we report the DNA methylation pattern in CDKN2A (p14(ARF)/p16(INK4a)) and in ATM gene promoters from 62 impalpable lesions, 39 peripheral blood samples, and 39 saliva samples, assessed by methylation-specific polymerase chain reaction method. ATM showed the greatest percentage of methylation in DNA from lesions (benign and malignant), blood (even with p16(INK4a)), and saliva, followed by p16(INK4a) and p14(ARF). Among the malignant cases, ATM promoter was the most hypermethylated in lesion DNA and in blood and saliva DNAs, and p14(ARF), the least. The highest percentage of p16(INK4a) methylation was found in the blood. Finally, our data are relevant because they were obtained using impalpable breast lesions from patients who were carefully recruited in 2 public hospitals of Rio de Janeiro.

Protein identification from dried nipple aspirate fluid on Guthrie cards using mass spectrometry.

Nipple aspirate fluid (NAF) requires investigation as a potential source of biomarkers for early diagnosis or risk assessment in breast cancer and other breast disorders. The present study demonstrated that proteins were easily extracted from dried NAF spots on Guthrie cards and were suitable for mass spectrometry analysis. NAF was obtained from 80 women, collected on Guthrie cards, between 2007 and 2010. The NAF-proteins were extracted from the card by incubating the card in water. These proteins were then quantified and separated using one-dimensional, 12% SDS-PAGE, gel electrophoresis and on high-resolution gradient gels at different concentrations (4-12, 8-16 and 4-20%). The bands with the most abundant proteins were excised from the gradient gels and the proteins were identified by liquid chromatography quadrupole time of flight. Immunoglobulins, Zn-α2-glicoprotein, apoliprotein D and prolactin inducible protein were among those identified. The NAF-Guthrie card collection method has not been applied previously, however, NAF proteins have been identified using other collecting techniques, confirming the feasibility of the NAF Guthrie card collection method for analyzing the proteins within NAF. The NAF-Guthrie card collecting method has multiple advantages, including being inexpensive, non-invasive, reliable and painless, and the cards can be stored at room temperature. Examining NAF may assist in identifying individuals at a higher risk of breast cancer and in improving patient prognosis.

Aspectos tomográficos da órbita aguda infecciosa: revisão de literatura / Tomographic findings of acute infections of the orbit: literature review

As doenças que acometem a órbita de forma aguda e não traumática por vezes não são conhecidas por médicos em centros de emergência. Essas condições são decorrentes de diversos fatores, como desordens imunológicas, congênitas, infecciosas, vasculares, entre outras. As causas infecciosas correspondem a mais de 50% de todos os casos e requer rápido diagnóstico e conduta para minimização de sequelas. A tomografia computadorizada (TC) é o exame de imagem de primeira linha nesses casos, sendo geralmente disponível nos centros de emergência e capaz de fornecer auxílio diagnóstico de forma rápida, precisa e eficaz. Esse artigo de revisão visa descrever os principais aspectos tomográficos da órbita aguda infecciosa, correlacionando-os com os dados da literatura.

The acute and nontraumatic diseases that involve the orbit are often little known by most physicians. These conditions are due to several factors, such as immune disorders, congenital, infections, vascular, among others disorders. The infectious causes correspond to more than 50% of all cases and require rapid diagnosis and management in order to minimize sequels. Computed tomography (CT) is the first line imaging method on these cases, generally being available in emergency centers and capable to provide an accurate, quick and effective diagnostic information. This review article aims to describe the main tomographic findings in acute orbit infections, correlating them with the literature data.

Segmenting mammographic microcalcifications using a semi-automatic procedure based on Otsu's method and morphological filters

INTRODUCTION: Breast cancer has the second highest world's incidence rate, according to the Brazilian National Cancer Institute (INCa). Clinical examination and mammography are the best methods for early diagnosis. Computer-aided detection (CADe) and computer-aided diagnosis (CADx) systems are developed to improve mammographic diagnosis. Basically, CADx systems have three components: (i) segmentation, (ii) parameters extraction and selection, (iii) lesion classification. The first step for a CADx system is segmentation. METHODS: A microcalcification segmentation method is proposed, based on morphological operators, Otsu's Method and radiologists' knowledge. Pre-processing with top-hat operators improves contrast and reduces background noise. The Otsu's method automatically selects the best grey-level threshold to segment microcalcifications, obtaining binary images. Following, inferior reconstruction and morphological dilatation operators are applied to reconstruct lost structure details and fill small flaws in the segmented microcalcifications. Finally, the Canny edge detection is applied to identify microcalcifications contour candidates for each region-of-interest (ROI). Two experienced radiologists intervene in this semi-automatic method, firstly, selecting the ROI and, then, analyzing the segmentation result. The method was assessed in 1000 ROIs from 158 digital images (300 dpi, 8 bits). RESULTS: Considering the radiologists opinion, the rates of ROIs adequately segmented to establish a diagnosis hypothesis were 97.8% for one radiologist and 97.3% for the other. Using the Area Overlap Measure (AOM) and the 2136 microcalcifications delineated by an experienced radiologist as gold standards, the method achieved an average AOM of 0.64±0.14, being 0.56±0.09 for small microcalcifications and 0.66±0.13 for the large ones. Moreover, AOM was 0.64±0.13 for the benign and 0.64±0.14 for the malignant lesions with no statistical differences between them. CONCLUSION: Based on these findings, the proposed method could be used to develop a CADx system that could help early breast cancer detection.

Detecção dos polimorfismos 213AG e 13494GA no gene TP53 em carcinoma ductal de mama: relato de caso / Detection of Polymorphisms 213AG and 13494GA in the TP53 Gene in Breast in Situ Ductal Carcinoma: Case Report

Introdução: A caracterização das alterações moleculares em lesões mamárias suspeitas para malignidade ainda não são bem definidas. Sabe-se que a detecção precoce do câncer de mama aumenta consideravelmente as chances de cura. Com isso, a busca por marcadores tumorais, a fim de auxiliar no diagnóstico precoce e predizer com confiança se essas lesões são benignas ou malignas, se faz necessária. No processo de carcinogênese, diversas são as alterações de expressão gênica, na qual envolve vários genes-chave que controlam o ciclo celular. Entre os genes, o TP53 tem sido amplamente pesquisado por apresentar mutações e variantes que podem estar envolvidas na carcinogênese mamária. Relato de caso: Paciente do sexo feminino, 45 anos, branca, casada, residente do Estado do Rio de Janeiro com carcinoma ductal in situ grau 2, multifocal. Lesão positiva para os receptores hormonais de estrogênio e progesterona, com ausência de mutação somática e com presença dos variantes 213A→G e 13494G→A no éxon 6 e intron 6 do gene TP53. Conclusão: Embora tenham sido considerados individualmente neutros, não existem estudos que tenham avaliado o efeito sinérgico dos variantes 213A→G e 13494G→A

Introduction:The characterization of molecular alterations in breast lesions suspicious for malignancy is not welldefined.it is known that early detection of breast cancer greatly increases the chances of cure. Thus, it is required the survey for tumor markers may help establish an early diagnosis and confidently predict whether these lesions arebenign or malignant.in the process of carcinogenesis, there are several changes in gene expression, which involvesseveral key genes that control the cell cycle.among the genes,TP53has been widely studied for its mutations andvariations, which may be involved in breast carcinogenesis...

Measurement of HER2 in saliva of women in risk of breast cancer.

HER2 amplification can be present in ductal carcinoma in situ (DCIS). The aim of the present study was to test the feasibility of measuring soluble HER2 in the saliva of patients at risk of breast cancer towards early diagnosis and prognosis. Women with lesions classified as 4 according to BIRADS and women with spontaneous nipple discharge (NAF) were recruited for this study. Quantification of soluble HER2 in saliva was performed using the enzyme immunoassay ELISA. Median values of HER2 were quantified in saliva of the control groups and in the patient groups. The statistical test nonparametric Mann-Whitney was applied for the evaluation of median differences. Although the medians increased with the severity of the clinical status, no significant difference was found in all possibilities (p > 0.05) when comparing the medians among the patients groups. Interestingly, inter-individual HER2 quantity variations in the saliva were detected in this study in some subjects from each group. Considering possible inter-individual variations, research on saliva-based circulating HER2 has to be reinforced to ensure its correct application in diagnosis, treatment and in follow-up of breast cancer patients. Older and current issues surrounding the controversy about the appropriate methods for HER2 evaluation are discussed.

Análise computacional da textura de tumores de mama em imagens por ultrassom de pacientes submetidas a cirurgia conservadora / Computer-assisted analysis of breast tumors texture on sonographic images of patients submitted to breast-conserving surgery

OBJETIVO: Avaliar as características de textura de lesões de mama em imagens por ultrassom de pacientes submetidas a cirurgia conservadora que apresentaram, ou não, recidiva. MATERIAIS E MÉTODOS: As imagens de ultrassom de 36 pacientes submetidas a cirurgia conservadora, com 12 tendo apresentado recidiva local e 24 que não apresentaram recidiva no local da cirurgia, foram divididas em: 3 malignas na mama oposta, 7 nódulos benignos, 5 hiperplasias atípicas e 9 alterações fibrocísticas. A textura das lesões foi quantificada utilizando-se dez parâmetros calculados da matriz de coocorrência e da curva de complexidade. Análise discriminante linear foi aplicada aos parâmetros para discriminação de lesões de mama em pacientes submetidas a cirurgia conservadora que apresentaram, ou não, recidiva. RESULTADOS: Avaliando-se a capacidade dos parâmetros em distinguir as recidivas do grupo composto por lesões não recidivas benignas e hiperplasias atípicas, obteve-se especificidade de 100 por cento, com valores de acurácia e sensibilidade superiores a 91 por cento. Num segundo teste, foi possível distinguir as cinco hiperplasias, das lesões não recidivas benignas. CONCLUSÃO: Apesar do número reduzido de casos, os resultados obtidos são encorajadores, sugerindo que o uso da quantificação da textura pode auxiliar na diferenciação entre lesões benignas, hiperplasias atípicas e lesões malignas de origem recidiva.

OBJECTIVE: The purpose of this study was to assess the features of breast lesion texture on sonographic images of patients submitted to breast-conserving surgery, with or without tumor recurrence. MATERIALS AND METHODS: Sonographic images of 36 patients submitted to conservative surgery for breast cancer, 12 of them with, and 24 without local recurrence, included 3 contralateral malignant lesions, 7 benign lumps (3 cysts and 4 fibroadenomas), 5 atypical hyperplasias and 9 fibrocystic changes. The quantification of features of breast lesion texture was based on ten parameters calculated from gray-level co-occurrence matrix and complexity curve. Linear discriminant analysis was applied to the texture parameters for differentiating between breast lesions in women submitted to conservative surgery with and without tumor recurrence. RESULTS: The assessment of performance of texture parameters in distinguishing lesion recurrences in a group including benign lumps and atypical hyperplasias demonstrated specificity of 100 percent, and accuracy and sensitivity > 91 percent. Another test demonstrated that texture parameters were useful in the differentiation between atypical hyperplasias and benign lumps. CONCLUSION: Despite the limited number of cases, the present results can be considered as promising, suggesting that texture parameters may help in the differentiation among benign lumps, atypical hyperplasias and recurrent malignant lesions.

Aplicação de operadores morfológicos na segmentação e determinação do contorno de tumores de mama em imagens por ultra-som / Application of morphological operators on the segmentation and contour detection of ultrasound breast images

Em pacientes com massa palpáveis e mamografias não-conclusivas, oexame da mama através de imagens por ultra-som são divididas em: forma, tamanho, contorno, ecotextura, ecogenicidade, reforço ou sombra posterior. Tumores malignos geralmente são infiltrantes e invadem os tecidos adjacentes, gerando, na imagem por ultra-som, um contorno irregular ou indefinido. Logo, analisando o contorno do tumor é possível iniciar uma hipótese diagnóstica indicativa de malignidade. Neste trabalho, o contorno de regiões hipoecóicas em imagens por ultra-som foi determinado usando-se a segmentação baseada em operadores morfológicos. A técnica foi aplicada na determinação do contorno de 22 tumores de mama em imagens por ultra-som. Os resultados obtidos foram comparados com os contornos delineados por radiologistas experientes usando-se dois parâmetros: a razão de superposição das áreas definicas pelos contornos e o valor médio quadrático residual normalizado. Para uma razão de superposição de 0,6 cerca de 90 por cento dos contornos dos tumores foram determinados corretamente, valor compatível com a literatura. Porém, testes testes-t(α = 0,05) demonstraram que o valor médio quadrático residual normalizado é mais sensível a variações no contorno,indicando que este parâmetro é mais adequado para diferenciar contornos regulares de irregulares...

Ressonância magnética das lesöes da mama: correlaçäo com a mamografia / Magnetic resonance imaging of breast lesions: mammographic correlation

Com relaçäo às lesöes da mama, malignas e benignas, as imagens por ressonância magnética (RM), utilizando-se meio de contraste paramagnético, têm alta especificidade na sua diferenciaçäo. Foi observado que a administraçäo do meio de contraste e novas técnicas de aquisiçäo de imagens permitem definir e delmitar melhor as lesöes malignas imprecisas na mamografia, para conduzir melhor a terapêutica. A análise das lesöes mamárias pela RM tem seu valor fundamental no estudo das próteses mamárias, de lesöes situadas posteriormente ao corpo glandular adjacente, ao plano muscular e na avaliaçäo de tumores em mulheres jovens

Radiologia da mama - qualidade em mamografia / Breast imaging - Mammography quality

Os autores fizeram um cadastramento dos Serviços de Mamografia no Brasil, que mostrou que, para um total de 585, quase todos são equipamentos dedicados(exclusivos). Demonstraram que a partir de 1990 os radiologistas começaram a adquirir novos equipamentos. A seguir, aproveitando o material do Programa de Controle e Manutenção da qualidade em Mamografia do Colégio Brasileiro de Radiologia, estudaram as doses de radiação, a qualidade das imagens mamográficas e a qualidade do diagnóstico, quando foi observado que os benefícios da mamografia superam amplamente os riscos de câncer radioinduzidos, que a má qualidade das mamografias prejudica a conclusão diagnóstica e isto, somado ao mau diagnóstico gera falsos-positivos e falsos-negativos, com enormes prejuizos à população feminina.

A análise das microcalcificações no diagnóstico das lesões infraclínicas da mama: uma contribuição para o program de detecção de câncer de mama

Este trabalho descreve a rotina do Serviço de Radiologia do Prof. Jean Louis Lamarque, no Hospital da Universidade de Montpellier, em Montepellier, na França, objetivando a discussão para a implantação de rotina em um Centro de Referência para o Ensino e pesquisa em Radiologia da Mama. Durante o período de sete meses de estágio, foram examinadas 1.449 pacientes, tendo sido selecionados 130 com presença de microcalcificações suspeitas. Essas pacientes foram submetidos a biópsia e citopunção,eletromamografia e análise morfométrica das microcalcificações, e seus dados foram cotejados com a literatura. A mamografia continua sendo o método com maior especificidade para o exame do seio, apesar de atualmente atuar em conjunto com uma série de outros. A análise morfométrica das microcalcificações, que tem lugar de destaque no diagnóstico de lesões infraclínicas da mama com presença de mircrocalcificações, mostrou ser, de fato, um auxiliar de grande valia na detecção de cânceres precoces.

Câncer da mama masculina / Breast cancer in men

Em estudo dos casos de câncer da mama masculina ocorridos no INCa-RJ, de 1983 a 1989, os autores apresentam os dados clínicos, radiológicos e histopatológicos mais encontrados, tendo sido observado o predomínio do carcinoma ductal infiltrante, 6 casos de implante secundário e 2 casos fatais. A importância deste trabalho está, principalmente, na oportunidade de reunir 11 ocorrências da lesäo, que representam apenas 0,2% dos tumores malignos no homem, e descrever suas manifestaçöes, chamando a atençäo do radiologista para esta entidade

Tumores duplos de mama / Double breast tumors

A reuniäo de 8 casos de tumores duplos de mama, ocorridos no período de 6 anos no INCa-RJ, serviu a aplicaçäo dos critérios de classificaçäo e diferenciaçäo diagnóstica apresentados nesse trabalho, que pretende oferecer um parâmetro de avaliaçäo que permita detectar e tratar precocemente este tipo de patologia

Aspectos radiológicos das lesöes mamárias com secreçäo papilar: importância da ductografia / Radiological aspects of the mammary injuries with papillary secretion: ductography importance

Por ser um exame fundamental para o estabelecimento do plano terapêutico em pacientes com descarga papilar espontânea, a ductografia, tal como é realizada no INCa, é descrita em seus procedimentos e análises. A partir da reuniäo de treze casos registrados naquela instituiçäo, os elementos definidores do correto diagnóstico, assim como os procedimentos complementares utilizados, säo demonstrados de modo simples e objetivo