Short tandem repeat analysis: a practical tool to identify specimen mix-ups in the pathology laboratory.

Despite all precautions in pathology laboratories, contaminations and specimen mix-ups still occur and can negatively impact both patients and institutions. We present two cases in which short tandem repeat (STR) analysis was used to assert the correct identity of specimens. The first patient had a biopsy diagnosis of triple negative invasive carcinoma of no special type of the breast. Sample mix-up with another biopsy was suspected, because in her post-chemotherapy mastectomy specimen, a hormone receptor-positive lobular carcinoma was diagnosed. STR analysis displayed a complete common loci profile of the patient's biopsy and mastectomy, supporting that no mix-up occurred. The second patient underwent hysterectomy due to cervix squamous cell carcinoma. A fragment of adenocarcinoma was identified and confirmed by STR profile to be a contaminant. STR analysis is a fast, easy-to-perform, and widely available technique which can clarify contaminations and specimen mix-ups in pathology laboratories.

Motor and cognitive outcomes of neonates with low birth weight in Brazil: a systematic review and meta-analysis.

BACKGROUND: Data on the outcomes of preterm newborns in South American countries are scarce. Given the great effect of low birth weight (LBW) and/or prematurity on children's neurodevelopment, it is extremely necessary to conduct studies on these phenomena in greater depth in more heterogeneous populations such as those ones from countries with limited resources. METHODS: We conducted a comprehensive literature search on databases including PubMed, the Cochrane Library, and Web of Science for articles published in Portuguese and English up to March 2021 involving children born and evaluated in Brazil. The analysis of the risk of bias was adapted from the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement and used to evaluate the methodology of the included studies. RESULTS: From the eligible trials, 25 articles were selected for qualitative synthesis, and 5 of those, for quantitative synthesis (meta-analysis). The meta-analyses showed that children born with LBW presented lower scores on motor development when compared with controls (standardized mean difference: -1.15; 95% confidence interval [95%CI]: -1.56--0.73]; I2: 80%) and also scored lower in terms of cognitive development (standardized mean difference: -0.71; 95% CI: -0.99--0.44; I2: 67%). CONCLUSION: The results of the present study reinforce that impaired motor and cognitive functions can be a significant long-term outcome of LBW. The lower the gestational age at delivery, the higher the risk of impairment in those domains. The study protocol was registered in the International Prospective Register of Systematic Reviews (PROSPERO) database under number CRD42019112403.

ANTECEDENTES: Dados sobre desfechos de recém-nascidos prematuros em países da América do Sul são escassos. Dado o grande efeito do baixo peso ao nascer (BPN) e/ou da prematuridade no neurodesenvolvimento das crianças, é extremamente necessária a realização de estudos que investiguem esses fenômenos com maior profundidade em populações mais heterogêneas. MéTODOS: Realizou-se uma busca da literatura em bases de dados, incluindo PubMed, Cochrane Library e Web of Science, por artigos publicados em português e inglês até março de 2021 envolvendo crianças nascidas e avaliadas no Brasil. A análise de risco de viés foi adaptada da declaração de Fortalecimento do Relato de Estudos Observacionais em Epidemiologia (Strengthening the Reporting of Observational Studies in Epidemiology, STROBE), que foi utilizada para avaliar a metodologia dos estudos. RESULTADOS: Dos estudos elegíveis, 25 artigos foram selecionados para síntese qualitativa, e 5 desses 25, para síntese quantitativa (metanálise). As metanálises mostraram que crianças nascidas com BPN apresentaram pontuação menor em desenvolvimento motor quando comparadas aos controles (diferença média padronizada, −1,15; intervalo de confiança de 95% [IC95%]: −1,56­−0,73]; I2: 80%) e pontuação também menor em termos de desenvolvimento cognitivo (diferença média padronizada, −0,71; IC95%: −0,992−0,44; I2: 67%). CONCLUSãO: Os resultados deste estudo reforçam que o comprometimento das funções motoras e cognitivas pode ser um desfecho significativo de longo prazo do BPN. Quanto menor a idade gestacional no momento do parto, maior o risco de prejuízo nesses domínios. O protocolo do estudo foi registrado no banco de dados International Prospective Register of Systematic Reviews (PROSPERO) sob o número CRD42019112403.

Motor and cognitive outcomes of neonates with low birth weight in Brazil: a systematic review and meta-analysis / Desfechos motores e cognitivos de recém-nascidos com baixo peso ao nascer no Brasil: uma revisão sistemática e metanálise

Abstract Background Data on the outcomes of preterm newborns in South American countries are scarce. Given the great effect of low birth weight (LBW) and/or prematurity on children's neurodevelopment, it is extremely necessary to conduct studies on these phenomena in greater depth in more heterogeneous populations such as those ones from countries with limited resources. Methods We conducted a comprehensive literature search on databases including PubMed, the Cochrane Library, and Web of Science for articles published in Portuguese and English up to March 2021 involving children born and evaluated in Brazil. The analysis of the risk of bias was adapted from the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement and used to evaluate the methodology of the included studies. Results From the eligible trials, 25 articles were selected for qualitative synthesis, and 5 of those, for quantitative synthesis (meta-analysis). The meta-analyses showed that children born with LBW presented lower scores on motor development when compared with controls (standardized mean difference: -1.15; 95% confidence interval [95%CI]: -1.56--0.73]; I2: 80%) and also scored lower in terms of cognitive development (standardized mean difference: -0.71; 95% CI: -0.99--0.44; I2: 67%). Conclusion The results of the present study reinforce that impaired motor and cognitive functions can be a significant long-term outcome of LBW. The lower the gestational age at delivery, the higher the risk of impairment in those domains. The study protocol was registered in the International Prospective Register of Systematic Reviews (PROSPERO) database under number CRD42019112403.

Resumo Antecedentes Dados sobre desfechos de recém-nascidos prematuros em países da América do Sul são escassos. Dado o grande efeito do baixo peso ao nascer (BPN) e/ou da prematuridade no neurodesenvolvimento das crianças, é extremamente necessária a realização de estudos que investiguem esses fenômenos com maior profundidade em populações mais heterogêneas. Métodos Realizou-se uma busca da literatura em bases de dados, incluindo PubMed, Cochrane Library e Web of Science, por artigos publicados em português e inglês até março de 2021 envolvendo crianças nascidas e avaliadas no Brasil. A análise de risco de viés foi adaptada da declaração de Fortalecimento do Relato de Estudos Observacionais em Epidemiologia (Strengthening the Reporting of Observational Studies in Epidemiology, STROBE), que foi utilizada para avaliar a metodologia dos estudos. Resultados Dos estudos elegíveis, 25 artigos foram selecionados para síntese qualitativa, e 5 desses 25, para síntese quantitativa (metanálise). As metanálises mostraram que crianças nascidas com BPN apresentaram pontuação menor em desenvolvimento motor quando comparadas aos controles (diferença média padronizada, -1,15; intervalo de confiança de 95% [IC95%]: -1,56--0,73]; I2: 80%) e pontuação também menor em termos de desenvolvimento cognitivo (diferença média padronizada, -0,71; IC95%: -0,992-0,44; I2: 67%). Conclusão Os resultados deste estudo reforçam que o comprometimento das funções motoras e cognitivas pode ser um desfecho significativo de longo prazo do BPN. Quanto menor a idade gestacional no momento do parto, maior o risco de prejuízo nesses domínios. O protocolo do estudo foi registrado no banco de dados International Prospective Register of Systematic Reviews (PROSPERO) sob o número CRD42019112403.

Increased Glucose Activity in Subgenual Anterior Cingulate and Hippocampus of High Performing Older Adults, Despite Amyloid Burden.

BACKGROUND: Individuals at 80 years of age or above with exceptional memory are considered SuperAgers (SA), an operationalized definition of successful cognitive aging. SA showed increased thickness and altered functional connectivity in the anterior cingulate cortex as a neurobiological signature. However, their metabolic alterations are yet to be uncovered. OBJECTIVE: Herein, a metabolic (FDG-PET), amyloid (PIB-PET), and functional (fMRI) analysis of SA were conducted. METHODS: Ten SA, ten age-matched older adults (C80), and ten cognitively normal middle-aged (C50) adults underwent cognitive testing and multimodal neuroimaging examinations. Anterior and posterior regions of the cingulate cortex and hippocampal areas were primarily examined, then subregions of anterior cingulate were segregated. RESULTS: The SA group showed increased metabolic activity in the left and right subgenual anterior cingulate cortex (sACC, p < 0.005 corrected, bilateral) and bilateral hippocampi (right: p < 0.0005 and left: p < 0.005, both corrected) as compared to that in the C80 group. Amyloid deposition was above threshold in 30% of SA and C80 (p > 0.05). The SA group also presented decreased connectivity between right sACC and posterior cingulate (p < 0.005, corrected) as compared to that of the C80 group. CONCLUSION: These results support the key role of sACC and hippocampus in SA, even in the presence of amyloid deposition. It also suggests that sACC may be used as a potential biomarker in older adults for exceptional memory ability. Further longitudinal studies measuring metabolic biomarkers may help elucidate the interaction between these areas in the cognitive aging process.

Evaluating subjective cognitive decline: a systematic review of tools available for evaluating cognitive complaints in Portuguese.

BACKGROUND: Subjective cognitive decline (SCD) is a perception that is not objectively measured in screening tests. Although many tools are available for evaluating SCD, no single gold standard is available for classifying individuals as presenting SCD, in the Portuguese-speaking population. The aim of this study was to systematically review the literature for tools used to evaluate SCD in the Portuguese-speaking population. METHODS: Four databases (Web of Science, SciELO, LILACS and MEDLINE) were primarily utilized in this study (Phase 1). Subsequently, we conducted a manual search of the literature (Phase 2). We then retrieved tools for critical evaluation (Phase 3). Studies that matched the inclusion criteria were analyzed. We summarized the features of each tool in terms of the number of questions, scoring system, benefits and deficiencies, translation and validity. RESULTS: A total of 30 studies utilizing four questionnaires and seven different single questions were found. The tools retrieved were the Memory Assessment Questionnaire (MAC-Q; 12/30 studies), single-question methods (7/30 studies), Subjective Memory Complaint Scale (SMC scale; 5/30 studies), Prospective and Retrospective Memory Questionnaire (PRMQ; 3/30 studies) and Memory Complaint Scale (MCS; 3/30 studies). Only two were formally translated and validated for the Portuguese speaking population (PRMQ and MCS). CONCLUSIONS: In summary, SCD is still underinvestigated in Portuguese-speaking countries. The MAC-Q was the most commonly used tool in Portuguese, despite its lack of formal translation and validation for the Portuguese-speaking population. Further studies are required in order to develop and validate a screening tool that includes questions for detecting SCD-plus features and affective symptoms, so as to improve its predictive value.

Left Hemisphere Lateralization of Epileptic Focus Can Be More Frequent in Temporal Lobe Epilepsy Surgical Patients with No Consensus Associated with Depression Lateralization.

Temporal lobe epilepsy (TLE) is considered to be the most common form of epilepsy, and it has been seen that most patients are refractory to antiepileptic drugs. A strong association of this ailment has been established with psychiatric comorbidities, primarily mood and anxiety disorders. The side of epileptogenic may contribute to depressive and anxiety symptoms; thus, in this study, we performed a systematic review to evaluate the prevalence of depression in TLE in surgical patients. The literature search was performed using PubMed/Medline, Web of Science, and PsycNet to gather data from inception until January 2019. The search strategy was related to TLE, depressive disorder, and anxiety. After reading full texts, 14 articles meeting the inclusion criteria were screened. The main method utilized for psychiatric diagnosis was Diagnostic and Statistical Manual of Mental Disorders/Structured Clinical Interview for DSM. However, most studies failed to perform the neuropsychological evaluation. For those with lateralization of epilepsy, focus mostly occurred in the left hemisphere. For individual depressive diagnosis, 9 studies were evaluated, and 5 for anxiety. Therefore, from the data analyzed in both situations, no diagnosis was representative in preoperative and postoperative cases. In order to estimate the efficacy of surgery in the psychiatry episodes and its relation to seizure control, the risk of depression and anxiety symptoms in epileptic patients need to be determined before surgical procedures. Rigorous preoperative and postoperative evaluation is essential for psychiatry conditions in patients with refractory epilepsy candidates for surgery.

Evaluating subjective cognitive decline: a systematic review of tools available for evaluating cognitive complaints in Portuguese / Avaliação de declínio cognitivo subjetivo: uma revisão sistemática de instrumentos disponíveis para avaliar queixa cognitiva em português

ABSTRACT Background: Subjective cognitive decline (SCD) is a perception that is not objectively measured in screening tests. Although many tools are available for evaluating SCD, no single gold standard is available for classifying individuals as presenting SCD, in the Portuguese-speaking population. The aim of this study was to systematically review the literature for tools used to evaluate SCD in the Portuguese-speaking population. Methods: Four databases (Web of Science, SciELO, LILACS and MEDLINE) were primarily utilized in this study (Phase 1). Subsequently, we conducted a manual search of the literature (Phase 2). We then retrieved tools for critical evaluation (Phase 3). Studies that matched the inclusion criteria were analyzed. We summarized the features of each tool in terms of the number of questions, scoring system, benefits and deficiencies, translation and validity. Results: A total of 30 studies utilizing four questionnaires and seven different single questions were found. The tools retrieved were the Memory Assessment Questionnaire (MAC-Q; 12/30 studies), single-question methods (7/30 studies), Subjective Memory Complaint Scale (SMC scale; 5/30 studies), Prospective and Retrospective Memory Questionnaire (PRMQ; 3/30 studies) and Memory Complaint Scale (MCS; 3/30 studies). Only two were formally translated and validated for the Portuguese speaking population (PRMQ and MCS). Conclusions: In summary, SCD is still underinvestigated in Portuguese-speaking countries. The MAC-Q was the most commonly used tool in Portuguese, despite its lack of formal translation and validation for the Portuguese-speaking population. Further studies are required in order to develop and validate a screening tool that includes questions for detecting SCD-plus features and affective symptoms, so as to improve its predictive value.

RESUMO Introdução: Declínio cognitivo subjetivo (DCS) é uma percepção não objetivamente mensurada em testes de rastreio. Apesar de muitos instrumentos estarem disponíveis para avaliação de DCS, nenhum padrão-ouro único é capaz de classificar um indivíduo com DCS em população falante de português. Este estudo objetivou revisar sistematicamente a literatura para instrumentos usados, para avaliar DCS em falantes de português. Métodos: Quatro bases de dados (Web of Science, SciELO, LILACS e MEDLINE) foram inicialmente usadas neste estudo (Fase 1). Em seguida, conduzimos uma busca manual (Fase 2) e os instrumentos coletados foram criticamente avaliados (Fase 3). Estudos que correspondiam aos critérios de inclusão foram analisados. Nós resumimos as características de cada instrumento em termos de números de questões, sistema de pontuação, vantagens e desvantagens, tradução e validação. Resultados: O total de 30 estudos utilizou 4, questionários e 7 diferentes questões para avaliar DCS. Os instrumentos avaliados foram Memory Assessment Questionnaire (MAC-Q, 12/30 estudos), método de questão única (7/30 estudos), Subjective Memory Complaint Scale (SMC-scale, 5/30 estudos), Prospective and Retrospective Memory Questionnaire (PRMQ, 3/30 estudos) e Memory Complaint Scale (MCS, 3/30 estudos). Apenas dois instrumentos foram formalmente traduzidos e validados para falantes de português (PRMQ e MCS). Conclusões: Em suma, DCS é ainda sub-representado em países lusofônicos. O MAC-Q foi o instrumento mais utilizado em português, apesar de sua falta de tradução e validação formal para a população falante de português. Mais estudos são necessários para desenvolver e validar um instrumento de rastreio que inclua questões sobre DCS-plus e sintomas afetivos, para aumentar seu poder preditivo.

The role of TP53 pathogenic variants in early-onset HER2-positive breast cancer.

Breast cancer is the most frequent event in Li-Fraumeni syndrome associated with germline TP53 variants. Some studies have shown that breast cancers in women with Li-Fraumeni syndrome are commonly HER2-positive, suggesting that HER2 amplification or over-expression in a young woman may be a useful criterion to test for germline variants in the TP53 gene. We assessed the prevalence of germline TP53 variants by Sanger sequencing or next-generation sequencing in 149 women with HER2-positive breast cancer diagnosed until age 40. The pattern of HER2 amplification was evaluated with dual-probe FISH in a subset of breast carcinomas from patients with germline TP53 variants as compared with those of noncarriers. Among 149 women tested, three presented a deleterious TP53 germline variant (2%), with one patient diagnosed at age 31 and the other two with bilateral breast cancer at ages 29/33 and 28/32, respectively. Three of the 36 patients (8.3%) with the first breast cancer diagnosed at age 31 or younger presented a pathogenic TP53 variant. Additionally, all TP53 deleterious variant carriers had a first degree relative diagnosed with different early-onset cancers (frequently not belonging to the Li-Fraumeni syndrome tumor spectrum) diagnosed at age 45 or younger. Higher levels of HER2 amplification were found in breast carcinomas of TP53 pathogenic variant carriers than in those of noncarriers. Deleterious germline TP53 variants account for a small proportion of early-onset HER2-positive breast cancers, but these seem to have higher HER2 amplification ratios. All TP53 pathogenic variant carriers found in this study had the first breast carcinoma diagnosed at age 31 or younger and a first-degree relative with early-onset cancer. Further studies are needed to clarify if HER2 status in early-onset breast cancer patients, in combination with other personal and/or familial cancer history, is useful to update the TP53 testing criteria.

Review of neuroimaging findings in congenital Zika virus syndrome and its relation to the time of infection.

BACKGROUND: Many original articles and case series have been published emphasizing the neuroimaging findings of congenital Zika virus (ZIKV) infection. The majority of these studies do not follow a neuroradiological methodology to describe malformations and brain abnormalities resulting from ZIKV infection. The cause-and-effect correlation between the gestational period of maternal infection and the severity of encephalic changes at birth has rarely been reported. A systematic literature review was conducted on the neuroimaging findings in children affected with microcephaly due to ZIKV. METHODS: PubMed, Cochrane Library and Web of Science were searched for full-text articles published up to July 2019. Duplicate entries were removed. Two independent reviewers performed a quality assessment of all the studies included. RESULTS: A total of 2214 publications were identified. Of these 2170 were excluded by analysis of titles and abstracts, resulting in the inclusion of only eight articles. Chi-square and Fisher's exact tests were performed with a 95% confidence interval to verify the statistically significant differences in the neuroradiological findings between the cases of ZIKV infection in the first or second trimester of gestation. The studies published so far have described image abnormalities at random, without utilizing any pre-established neuroradiological criteria, and imaging modalities with different sensitivity and accuracy have been used, which jeopardizes a reliable and adequate statistical analysis. CONCLUSIONS: Neuroimaging abnormalities are much more prevalent and severe when the infection by ZIKV is contracted in the first or second trimester of pregnancy.

Validação do instrumento "teste seu cérebro" para idosos: versão para iPad / Validation of the instrument "test your brain" for the elderly: iPad version

OBJETIVOS: Validar o aplicativo para iPad (Apple, Califórnia, EUA) "Teste seu Cérebro", bem como estabelecer ponto de corte entre idosos normais e com transtorno neurocognitivo leve. MÉTODOS: Em um estudo transversal prospectivo, idosos que frequentaram o ambulatório de neuropsicologia de um hospital terciário de saúde da região sul do Brasil e da comunidade em geral, foram submetidos a avaliação cognitiva por meio de dois instrumentos: Montreal Cognitive Assessment (teste padrão ouro) e "Teste seu Cérebro". Esses resultados serviram como parâmetro para validar o referido aplicativo a partir de um teste diagnóstico e estabelecer o ponto de corte entre idosos normais e com transtorno neuropsicológico leve; para tanto foram determinadas as seguintes medidas estatísticas: sensibilidade e especificidade; consistência interna e confiabilidade alcançadas pelo coeficiente Ômega de McDonald e coeficiente de correlação de Pearson, respectivamente. A classificação média do ponto de corte do "Teste seu Cérebro" para detectar os casos classificados como transtorno neurocognitivo leve pelo teste Montreal Cognitive Assessment, foi obtida através da curva ROC. As avaliações contemplam funções como: memória, atenção/orientação, fluência, linguagem e habilidades viso-espaciais... RESULTADOS: A amostra foi constituída por 104 participantes com média de idade de 70,3±6,6 anos, sendo a idade mínima de 60 e máxima de 87 anos. Foi alcançada uma confiabilidade aceitável para o aplicativo "Teste seu Cérebro" através da análise da consistência interna. Na comparação entre as pontuações gerais dos dois instrumentos (Teste seu Cérebro e Montreal Cognitive Assessment), o resultado apontou uma correlação estatisticamente significativa positiva e classificada como moderada. O ponto de corte das pontuações do "Teste seu Cérebro" que melhor discriminou os pacientes com transtorno neurocognitivo leve diagnosticados pelo Montreal Cognitive Assessment foi de 89,5%, ou seja, pontuações inferiores ou iguais a esse percentual alcançaram maiores sensibilidade e especificidade para o instrumento. Não foi identificada influência das variáveis sociodemográficas como sexo, idade e escolaridade sobre a relação de linearidade entre os instrumentos "Teste seu Cérebro" e Montreal Cognitive Assessment. CONCLUSÕES: Os resultados obtidos sugerem que o instrumento "Teste seu Cérebro" pode ser utilizado com segurança para identificar precocemente e com acurácia a presença de transtorno neurocognitivo leve na população idosa. Novos estudos serão direcionados à validação do instrumento "Teste seu Cérebro" na identificação de outros tipos de distúrbios cognitivos, além de Transtorno Neurocognitivo Leve.

AIMS: Validate the application for iPad (Apple, California, USA) "Teste seu Cérebro" as well as establish cutoff point between normal seniors and Mild Neurocognitive Impairment. METHODS: In a prospective cross-sectional study, elderly subjects who attended the neuropsychology clinic of a tertiary health hospital in the southern region of Brazil and the community in general underwent cognitive assessment using two instruments: the Montreal Cognitive Assessment (Gold Standard Test) and the "Teste seu Cérebro". These results served as a parameter to validate the said application from a diagnostic test and to establish the cutoff point between normal elderly and mild cognitive impairment; the following statistical measures were determined: sensitivity and specificity, internal consistency and reliability reached by the McDonald's Omega coefficient and Pearson's correlation coefficient, respectively. The average "Teste seu Cérebro" cutoff point to detect cases classified as mild neurocognitive impairment by the Montreal Cognitive Assessment was obtained through the ROC curve. Evaluations include functions such as memory, attention / orientation, fluency, language, and visuospatial skills. RESULTS: The sample consisted of 104 participants with mean age of 70.3 (standard deviation=6.6), with a minimum age of 60 and a maximum of 87 years. An acceptable reliability was achieved for the "Teste seu Cérebro" application by analyzing the internal consistency. In the comparison between the general scores of the two instruments (Teste seu Cérebro and Montreal Cognitive Assessment), where the result showed a statistically significant correlation, positive and classified as moderate. The cutoff point of the "Teste seu Cérebro" scores that best discriminated patients with mild neurocognitive impairment diagnosed by the Montreal Cognitive Assessment was 89.5%, that is, scores below or equal to that percentage reached higher sensitivity and specificity for the instrument. No influence of sociodemographic variables such as sex, age and schooling were identified on the linearity relationship between the "Teste seu Cérebro" and Montreal Cognitive Assessment instruments. CONCLUSIONS: The results suggest that the "Teste seu Cérebro" instrument can be safely used to identify early and accurately the presence of Mild Neurocognitive Impairment in the elderly population. New studies will be directed to the validation of the instrument "Teste seu Cérebro" in the identification of other types of cognitive disorders, in addition to Mild Neurocognitive Impairment.

Breast cancer in Angola, molecular subtypes: a first glance.

PURPOSE: The aim of this study was to investigate the breast cancer (BC) molecular subtypes according to its surrogate immunohistochemistry (IHC) markers. We conducted a preliminary study, to correlate the clinical pathological profiles and molecular subtypes of breast cancer in Luanda, Angola. METHODS: From January 2011 to 30 December 2014, 140 consecutive cases of microscopically confirmed invasive breast carcinoma were classified regarding histology and IHC (ER, PR, HER2, and Ki-67). Surrogate molecular subtypes were classified according to ESMO recommendations. RESULTS: All patients were female; the median age was 47 years (24-84 years). Invasive carcinoma NST was the most common type (91.4%) and grade 2 was prevalent (70.7%). Most tumours were locally advanced (stage III - 65% and stage IV - 3.6%). In 140 studied cases, 74 (52.8%) malignancies were hormone receptor positive; 25.7% were luminal A like, 19.3% luminal B and HER2 negative like, 7.9% luminal B and HER2-positive like, 15.7% HER2 positive, and 31.4% were triple negative. CONCLUSION: Women's BC in Luanda-Angola is diagnosed at a young age and at an advanced stage. The two predominant molecular subtypes are HR positive and triple negative. The percentage of HER2-positive BC cases was high. Determining the molecular subtype using surrogate IHC markers has important treatment and prognostic implications for Angolan women with BC. There is an urgent need to study a prospective BC series in order to confirm the present results.

Frequent copy number gains at 1q21 and 1q32 are associated with overexpression of the ETS transcription factors ETV3 and ELF3 in breast cancer irrespective of molecular subtypes.

Several ETS transcription factors are involved in the pathogenesis of human cancers by different mechanisms. As gene copy number gain/amplification is an alternative mechanism of oncogenic activation and 1q gain is the most common copy number change in breast carcinoma, we investigated how that genomic change impacts in the expression of the three 1q ETS family members ETV3, ELK4, and ELF3. We have first evaluated 141 breast carcinomas for genome-wide copy number changes by chromosomal CGH and showed that 1q21 and 1q32 were the two chromosome bands with most frequent genomic copy number gains. Second, we confirmed by FISH with locus-specific BAC clones that cases showing 1q gain/amplification by CGH showed copy number increase of the ETS genes ETV3 (located in 1q21~23), ELF3, and ELK4 (both in 1q32). Third, gene expression levels of the three 1q ETS genes, as well as their potential targets MYC and CRISP3, were evaluated by quantitative real-time PCR. We here show for the first time that the most common genomic copy number gains in breast cancer, 1q21 and 1q32, are associated with overexpression of the ETS transcription factors ETV3 and ELF3 (but not ELK4) at these loci irrespective of molecular subtypes. Among the three 1q ETS genes, ELF3 has a relevant role in breast carcinogenesis and is also the most likely target of the 1q copy number increase. The basal-like molecular subtype presented the worst prognosis regarding disease-specific survival, but no additional prognostic value was found for 1q copy number status or ELF3 expression. In addition, we show that there is a correlation between the expression of the oncogene MYC, irrespectively of copy number gain at its loci in 8q24, and the expression of both the transcriptional repressor ETV3 and the androgen respondent ELK4.

Validating the MSKCC nomogram and a clinical decision rule in the prediction of non-sentinel node metastases in a Portuguese population of breast cancer patients.

BACKGROUND: In order to assess the risk of non-sentinel node involvement in breast cancer patients, some prediction tools have been developed and tested. However, a wide range of results are observed. We tested a simplified clinical decision rule, against the complex nomogram created from the MSKCC sentinel node database. METHODS: Two single institutional consecutive series of patients with a positive SN, submitted to SN biopsy plus axillary dissection from June 1999 to October 2007, were evaluated. A receiver operating curve was drawn and the area under the curve was calculated as well as the negative predictive value for both tests, assuming discriminative values of 10 and 15%. RESULTS: Considering the derivation series, our results showed an area under the curve of 0.69 for both our clinical decision rule and the MSKCC nomogram. The analysis of the validation series showed an area under the curve of 0.65 for our clinical decision rule and of 0.67 for the MSKCC nomogram. The nomogram results are inferior to those found in the original population and are similar to our clinical decision rule results. CONCLUSIONS: Individual centres should develop and prospectively test their own clinical decision rules, based on their institutional Sentinel Node data.

High RASSF1A promoter methylation levels are predictive of poor prognosis in fine-needle aspirate washings of breast cancer lesions.

Previously, we reported that the accuracy of cytological diagnosis of breast lesions could be augmented through the quantitative assessment of DNA methylation of fine-needle aspirate (FNA) washings. Herein, we aimed at the evaluation of the prognostic value of quantitative promoter methylation at three gene loci (APC, CCND2, and RASSF1A) in a large series of FNA washings from breast lesions. Methylation levels of three gene promoters were assessed by quantitative methylation-specific PCR in bisulfite-modified DNA from 211 FNA washings, comprising 178 carcinomas and 33 benign lesions, both histopathologically confirmed. Receiver operator characteristic (ROC) curve analysis was used to determine the diagnostic performance of the gene panel in distinguishing cancer from non-cancerous lesions. Relevant clinicopathologic data and time to progression and/or death from breast cancer were correlated with methylation findings. Log-rank test and Cox-regression model identified independent predictors of prognosis. APC, CCND2, and RASSF1A methylation levels differed significantly between malignant and benign lesions. ROC curve analysis confirmed the diagnostic performance of the gene panel. In univariate analysis, stage was significantly associated with overall, disease-specific and disease-free survival, whereas tumor grade was associated with disease-specific and disease-free survival. Remarkably, RASSF1A methylation was significantly and independently associated with worse disease-free survival in the final multivariate analysis. We confirmed that quantitative gene promoter methylation augments the diagnostic performance of cytopathology. Importantly, and in addition to standard clinicopathologic parameters, RASSF1A high-methylation levels are independent predictors of worse outcome in breast cancer. Thus, epigenetic biomarkers provide valuable tools for breast cancer patient management.

Quantitative hypermethylation of a small panel of genes augments the diagnostic accuracy in fine-needle aspirate washings of breast lesions.

PURPOSE: We hypothesized that comprehensive breast cancer methylation profiling might provide biomarkers for diagnostic assessment of suspicious breast lesions using fine needle aspiration biopsy (FNA). EXPERIMENTAL DESIGN: Twenty-three gene promoters were surveyed by quantitative methylation-specific PCR in bisulfite-modified DNA from 66 breast carcinomas (BCa), 31 fibroadenomas (FB) and 12 normal breast (NT) samples to define a set of genes differentially methylated in malignant and non-malignant tissues. This set was tested in 78 FNA washings obtained pre-operatively (66 malignant, 12 benign), with histopathological diagnosis. Receiver operator characteristic (ROC) curve analysis identified a gene panel which might distinguish cancer from non-cancerous lesions. Finally, this panel was validated in an independent series of FNA washings (45 cases) in which cytomorphology did not reach definitive diagnosis. RESULTS: In tissue samples, 14-3-3-sigma, DAPK, CCND2, RASSF1A, CALCA, APC, HIN1, RARbeta2, TIG1, and GSTP1 methylation levels differed significantly among BCa, FB, and NT. ROC curve analysis identified a panel of four gene loci (CCND2, RASSF1A, APC, and HIN1) that discriminated BCa from benign lesions in a set of 78 FNA washings from histologically characterized breast lesions. When this panel was tested in the validation dataset of 45 FNA washings, breast cancer was identified with perfect specificity (100%) when 3 of 4 gene loci tested positive, providing estimated added information of 91% over cytomorphologic evaluation alone. CONCLUSIONS: Our data provide evidence that multigene methylation analysis augments diagnostic accuracy of cytological assessment of suspicious breast lesions, and might be a valuable ancillary tool for breast cancer diagnosis.