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BACKGROUND: The last five decades have seen a surge in viral outbreaks, particularly in tropical and subtropical regions like Brazil, where endemic arboviruses such as Dengue (DENV), Zika (ZIKV), and Chikungunya (CHIKV) pose significant threats. However, current diagnostic strategies exhibit limitations, leading to gaps in infection screening, arbovirus differential diagnoses, DENV serotyping, and life-long infection tracking. This deficiency impedes critical information availability regarding an individual's current infection and past infection history, disease risk assessment, vaccination needs, and policy formulation. Additionally, the availability of point-of-care diagnostics and knowledge regarding immune profiles at the time of infection are crucial considerations. OBJECTIVES: This review underscores the urgent need to strengthen diagnostic methods for arboviruses in Brazil and emphasizes the importance of data collection to inform public health policies for improved diagnostics, surveillance, and policy formulation. METHODS: We evaluated the diagnostic landscape for arboviral infections in Brazil, focusing on tailored, validated methods. We assessed diagnostic methods available for sensitivity and specificity metrics in the context of Brazil. RESULTS: Our review identifies high-sensitivity, high-specificity diagnostic methods for arboviruses and co-infections. Grifols transcription-mediated amplification assays are recommended for DENV, CHIKV, and ZIKV screening, while IgG/IgM ELISA assays outperform Rapid Diagnostic Tests (RDTs). The Triplex real-time RT-PCR assay is recommended for molecular screening due to its sensitivity and specificity. CONCLUSION: Enhanced diagnostic methods, on-going screening, and tracking are urgently needed in Brazil to capture the complex landscape of arboviral infections in the country. Recommendations include nationwide arbovirus differential diagnosis for DENV, ZIKV, and CHIKV, along with increased DENV serotyping, and lifelong infection tracking to combat enduring viral threats and reduce severe presentations.
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Infecções por Arbovirus , Arbovírus , Humanos , Brasil/epidemiologia , Infecções por Arbovirus/diagnóstico , Infecções por Arbovirus/epidemiologia , Arbovírus/imunologia , Arbovírus/classificação , Sensibilidade e Especificidade , Saúde Pública , Coleta de Dados , Dengue/diagnóstico , Dengue/epidemiologia , Infecção por Zika virus/diagnóstico , Infecção por Zika virus/epidemiologiaRESUMO
Abstract Background The last five decades have seen a surge in viral outbreaks, particularly in tropical and subtropical regions like Brazil, where endemic arboviruses such as Dengue (DENV), Zika (ZIKV), and Chikungunya (CHIKV) pose significant threats. However, current diagnostic strategies exhibit limitations, leading to gaps in infection screening, arbovirus differential diagnoses, DENV serotyping, and life-long infection tracking. This deficiency impedes critical information availability regarding an individual's current infection and past infection history, disease risk assessment, vaccination needs, and policy formulation. Additionally, the availability of point-of-care diagnostics and knowledge regarding immune profiles at the time of infection are crucial considerations. Objectives This review underscores the urgent need to strengthen diagnostic methods for arboviruses in Brazil and emphasizes the importance of data collection to inform public health policies for improved diagnostics, surveillance, and policy formulation. Methods We evaluated the diagnostic landscape for arboviral infections in Brazil, focusing on tailored, validated methods. We assessed diagnostic methods available for sensitivity and specificity metrics in the context of Brazil. Results Our review identifies high-sensitivity, high-specificity diagnostic methods for arboviruses and co-infections. Grifols transcription-mediated amplification assays are recommended for DENV, CHIKV, and ZIKV screening, while IgG/IgM ELISA assays outperform Rapid Diagnostic Tests (RDTs). The Triplex real-time RT-PCR assay is recommended for molecular screening due to its sensitivity and specificity. Conclusion Enhanced diagnostic methods, on-going screening, and tracking are urgently needed in Brazil to capture the complex landscape of arboviral infections in the country. Recommendations include nationwide arbovirus differential diagnosis for DENV, ZIKV, and CHIKV, along with increased DENV serotyping, and lifelong infection tracking to combat enduring viral threats and reduce severe presentations.
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Interleukin-33 (IL-33), a member of the interleukin-1(IL-1) family of cytokines, remains poorly understood in the context of human breast cancer and its impact on treatment outcomes. This study aimed to elucidate IL-33 expression patterns within tumor samples from a cohort of Brazilian female breast cancer patients undergoing neoadjuvant chemotherapy while exploring its correlation with clinicopathological markers. In total, 68 samples were meticulously evaluated, with IL-33 expression quantified through a quantitative polymerase chain reaction. The findings revealed a substantial upregulation of IL-33 expression in breast cancer patient samples, specifically within the Triple-negative and Luminal A and B subtypes, when compared to controls (healthy breast tissues). Notably, the Luminal B subtype displayed a marked elevation in IL-33 expression relative to the Luminal A subtype (p < 0.05). Moreover, a progressive surge in IL-33 expression was discerned among Luminal subtype patients with TNM 4 staging criteria, further underscoring its significance (p < 0.005). Furthermore, chemotherapy-naïve patients of Luminal A and B subtypes exhibited heightened IL-33 expression (p < 0.05). Collectively, our findings propose that chemotherapy could potentially mitigate tumor aggressiveness by suppressing IL-33 expression in breast cancer, thus warranting consideration as a prognostic marker for gauging chemotherapy response and predicting disease progression in Luminal subtype patients. This study not only sheds light on the intricate roles of IL-33 in breast cancer but also offers valuable insights for future IL-33-related research endeavors within this context.
Assuntos
Neoplasias da Mama , Humanos , Feminino , Neoplasias da Mama/metabolismo , Interleucina-33/genética , Interleucina-33/uso terapêutico , Terapia Neoadjuvante , Brasil , Resultado do Tratamento , Biomarcadores Tumorais , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Receptor ErbB-2/metabolismoRESUMO
BACKGROUND: RNA extraction is a step that precedes several molecular techniques. The fibrous tissue, more specifically the dura mater, has several limitations in routine protocols, and lacks optimization protocols to overcome these problems. OBJECTIVE: To test stock reagents and purification kits, optimizing commercial kit protocols for RNA extraction from the dura mater. METHODS: Dura mater samples were obtained from eight Wistar rats and maintained in two different stabilizers. The samples were purified using four different protocols, and the RNA was evaluated for the yield and purity in NanoDrop 2000 (Thermo Scientific, Wilmington, DE, United States). Beta-actin gene was used for analyzing gene expression, since is one of the most used reference genes. RESULTS: The RNA preservation was similar in both stabilizers. The addition of an incubation step prior the purification protocols allowed better tissue digestion and RNA recovery. The RNA purified using the protocols membrane-based showed higher quality than liquid-liquid purification. This impact was observed in the 3-week evaluation using RT-qPCR. CONCLUSION: Stabilizers are efficient for RNA preservation and membrane-based purification protocols are more suitable for RNA recovery from dura mater tissue, allowing the evaluation of gene expression in this type of tissue. Adaptations in the dura mater RNA extraction protocol differ from the pre-established protocols because it takes into account the peculiarity of fibrous tissue and low cellularity. In addition to providing a low-cost mechanism, based on techniques that are part of the laboratory routine, it is possible to improve the quality of the extracted material, ensuring greater efficiency in the use of subsequent techniques.
ANTECEDENTES: A extração de RNA é uma etapa que antecede várias técnicas moleculares. O tecido fibroso, mais especificamente a dura-máter, apresenta várias limitações nos protocolos de rotina e carece de protocolos de otimização para superar estes problemas. OBJETIVO: Testar reagentes de estoque e kits de purificação, otimizando protocolos de kits comerciais para extração de RNA da dura-máter. MéTODOS: Amostras de dura-máter foram obtidas de oito ratos Wistar e mantidas em dois estabilizadores diferentes. As amostras foram purificadas em quatro protocolos diferentes e o RNA foi avaliado quanto ao rendimento e pureza no NanoDrop 2000 (Thermo Scientific, Wilmington, DE, United States). O gene da beta-actina foi utilizado para analisar a expressão gênica, uma vez que é um dos genes de referência mais utilizados. RESULTADOS: A preservação do RNA foi semelhante em ambos os estabilizadores. A adição de uma etapa de incubação antes dos protocolos de purificação permitiu uma melhor digestão do tecido e recuperação de RNA. O RNA purificado pelos protocolos baseados em membrana apresentou qualidade superior ao da purificação líquido-líquido. Este impacto foi observado na avaliação de três semanas usando RT-qPCR. CONCLUSãO: Os estabilizadores são eficientes para preservação do RNA e os protocolos de purificação baseados em membrana são mais adequados para recuperação de RNA do tecido da dura-máter, permitindo a avaliação da expressão gênica neste tipo de tecido. As adaptações no protocolo de extração de RNA da dura-máter diferem dos protocolos preestabelecidos porque leva em consideração a peculiaridade do tecido fibroso e com baixa celularidade. Além de fornecer um mecanismo de baixo custo, baseado em técnicas que fazem parte da rotina laboratorial, é possível melhorar a qualidade do material extraído, garantindo maior eficácia no uso de técnicas subsequentes.
Assuntos
Dura-Máter , RNA , Animais , Ratos , Ratos Wistar , RNA/genética , RNA/análise , RNA/metabolismo , Dura-Máter/química , Dura-Máter/metabolismoRESUMO
Abstract Background RNA extraction is a step that precedes several molecular techniques. The fibrous tissue, more specifically the dura mater, has several limitations in routine protocols, and lacks optimization protocols to overcome these problems. Objective To test stock reagents and purification kits, optimizing commercial kit protocols for RNA extraction from the dura mater. Methods Dura mater samples were obtained from eight Wistar rats and maintained in two different stabilizers. The samples were purified using four different protocols, and the RNA was evaluated for the yield and purity in NanoDrop 2000 (Thermo Scientific, Wilmington, DE, United States). Beta-actin gene was used for analyzing gene expression, since is one of the most used reference genes. Results The RNA preservation was similar in both stabilizers. The addition of an incubation step prior the purification protocols allowed better tissue digestion and RNA recovery. The RNA purified using the protocols membrane-based showed higher quality than liquid-liquid purification. This impact was observed in the 3-week evaluation using RT-qPCR. Conclusion Stabilizers are efficient for RNA preservation and membrane-based purification protocols are more suitable for RNA recovery from dura mater tissue, allowing the evaluation of gene expression in this type of tissue. Adaptations in the dura mater RNA extraction protocol differ from the pre-established protocols because it takes into account the peculiarity of fibrous tissue and low cellularity. In addition to providing a low-cost mechanism, based on techniques that are part of the laboratory routine, it is possible to improve the quality of the extracted material, ensuring greater efficiency in the use of subsequent techniques.
Resumo Antecedentes A extração de RNA é uma etapa que antecede várias técnicas moleculares. O tecido fibroso, mais especificamente a dura-máter, apresenta várias limitações nos protocolos de rotina e carece de protocolos de otimização para superar estes problemas. Objetivo Testar reagentes de estoque e kits de purificação, otimizando protocolos de kits comerciais para extração de RNA da dura-máter. Métodos Amostras de dura-máter foram obtidas de oito ratos Wistar e mantidas em dois estabilizadores diferentes. As amostras foram purificadas em quatro protocolos diferentes e o RNA foi avaliado quanto ao rendimento e pureza no NanoDrop 2000 (Thermo Scientific, Wilmington, DE, United States). O gene da beta-actina foi utilizado para analisar a expressão gênica, uma vez que é um dos genes de referência mais utilizados. Resultados A preservação do RNA foi semelhante em ambos os estabilizadores. A adição de uma etapa de incubação antes dos protocolos de purificação permitiu uma melhor digestão do tecido e recuperação de RNA. O RNA purificado pelos protocolos baseados em membrana apresentou qualidade superior ao da purificação líquido-líquido. Este impacto foi observado na avaliação de três semanas usando RT-qPCR. Conclusão Os estabilizadores são eficientes para preservação do RNA e os protocolos de purificação baseados em membrana são mais adequados para recuperação de RNA do tecido da dura-máter, permitindo a avaliação da expressão gênica neste tipo de tecido. As adaptações no protocolo de extração de RNA da dura-máter diferem dos protocolos preestabelecidos porque leva em consideração a peculiaridade do tecido fibroso e com baixa celularidade. Além de fornecer um mecanismo de baixo custo, baseado em técnicas que fazem parte da rotina laboratorial, é possível melhorar a qualidade do material extraído, garantindo maior eficácia no uso de técnicas subsequentes.
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Abstract Background The neuropeptide Y (NPY) is one of the most abundant neurotransmitters in the nervous system. NPY acts as a potent stimulator of angiogenesis, inflammation, and adipogenesis, through the NPY 2 receptor (NPY2R). Changes in the NPY signaling pathway have been linked to Acute Coronary Syndrome (ACS). Objectives The purpose of this study is to determine the association between variants in the NPY and NPY2R genes, as well as the severity of acute coronary syndrome (ACS). Methods Approximately 221 ACS patients and 278 healthy controls were selected for this study. Four variants in NPY and two variants in NPY2R genes were genotyped using Taqman allelic discrimination and sequencing. The Chi-square and Fisher's exact tests were used to verify the genotype frequencies. The logistic regression analyses were used for the evaluation of the studied variables. Haplotype analysis was used to evaluate the linkage disequilibrium (LD) between the variants (p<0.05). Results An association of NPY c.20T>C variant was found with the ACS group when compared to the healthy group. In the analysis between variants and risk factors in the ACS group, NPY c.84G>A was associated with hypertension. The analysis between TIMI risk showed a significance for NPY c.20T>C between the low and intermediate/high TIMI risk groups. In the haplotype analysis, strong linkage disequilibrium (LD) was found between the variants NPY c.150G>A and NPY c.-485T>C. Conclusion The NPY c.20T>C variant appears to contribute to the development of ACS. The NPY2R c.-1116A>G variant may contribute to the early development of ACS and the NPY c.84G>A variant appears to contribute to the development of hypertension. In addition, the NPY c.20T>C is associated with a protective effect in ACS severity.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Adulto Jovem , Neuropeptídeo Y , Síndrome Coronariana Aguda/etiologia , Receptores de Neuropeptídeo Y , Polimorfismo de Nucleotídeo Único , Fatores de Risco de Doenças Cardíacas , HipertensãoRESUMO
OBJECTIVE: This study aimed to assess miRNA-195 expression in the tumor tissues from a cohort of Brazilian female breast cancer patients undergoing neoadjuvant chemotherapy (NAC) and evaluate its correlation with various clinicopathological markers. METHODS: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate the miRNA-195 expression in tumor tissues from a cohort of female breast cancer patients undergoing NAC. This expression was then correlated with the occurrence of several distinct breast cancer molecular subtypes and other clinicopathological variables. RESULTS: A total of 55 patients were included in this study, 28 (50.9%) of whom were treated using NAC. Tumor miRNA-195 expression was suppressed in breast cancer patients, regardless of their exposure to systemic treatments, histological grade, size, nodal status, and tumor-node-metastasis (TNM) staging. This was more pronounced in luminal and triple-negative patients, and patient's response to NAC was correlated with an increase in miRNA-195 expression. CONCLUSION: miRNA-195 is downregulated in the tumor tissues of Brazilian breast cancer patients regardless of NAC exposure; this reinforces its role as a tumor suppressor and a potential biomarker for chemotherapy response.
Assuntos
Neoplasias da Mama , MicroRNAs , Protocolos de Quimioterapia Combinada Antineoplásica , Biomarcadores Tumorais/genética , Brasil , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Feminino , Humanos , MicroRNAs/genética , Terapia Neoadjuvante , Estadiamento de Neoplasias , PrognósticoRESUMO
OBJECTIVE: This study aimed to assess miRNA-195 expression in the tumor tissues from a cohort of Brazilian female breast cancer patients undergoing neoadjuvant chemotherapy (NAC) and evaluate its correlation with various clinicopathological markers. METHODS: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate the miRNA-195 expression in tumor tissues from a cohort of female breast cancer patients undergoing NAC. This expression was then correlated with the occurrence of several distinct breast cancer molecular subtypes and other clinicopathological variables. RESULTS: A total of 55 patients were included in this study, 28 (50.9%) of whom were treated using NAC. Tumor miRNA-195 expression was suppressed in breast cancer patients, regardless of their exposure to systemic treatments, histological grade, size, nodal status, and tumor-node-metastasis (TNM) staging. This was more pronounced in luminal and triple-negative patients, and patient's response to NAC was correlated with an increase in miRNA-195 expression. CONCLUSION: miRNA-195 is downregulated in the tumor tissues of Brazilian breast cancer patients regardless of NAC exposure; this reinforces its role as a tumor suppressor and a potential biomarker for chemotherapy response.
Assuntos
Humanos , Feminino , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Neoplasias da Mama/tratamento farmacológico , MicroRNAs/genética , Prognóstico , Brasil , Protocolos de Quimioterapia Combinada Antineoplásica , Biomarcadores Tumorais/genética , Terapia Neoadjuvante , Estadiamento de NeoplasiasRESUMO
Objective: This study sought to identify the presence of HPV infection and the risk factors related to susceptibility to cervical cancer in asymptomatic women. Methods: It is a cross-sectional study with 428 users attended Basic Health Units, in Arapiraca, Alagoas, Brazil. Sociodemographic, behavioral variables, and cytopathological reports were collected. Molecular detection of the HPV virus was performed by Nested-PCR. Statistical analysis was conducted with SPSS version 22.0. Results: A total of 428 women were studied, HPV DNA detected in 39.2% (n = 168), with a mean age of 41 years old. There was an association of HPV with use of oral contraceptives (p <0.016) and alcoholism (p <0.038). It was showed a higher frequency of positive HPV in women older than 25 years old (88.7%), up to 5 sexual partners (93.4%), up to 3 pregnancies (71.4%), and with the cytopathologic results within the limits of normality (61.9%). HPV was identified in 40.3% (104/258) of the women with results within the limits of normality. Conclusion: Our results suggest that the use of oral contraceptives and alcoholism may be considered as possible risk factors related to cervical oncogenesis. With this, it is necessary to propose interventions aimed at the health education of this population, actions of prevention, and early detection.
Objetivo: Este estudo buscou identificar a presença de infecção pelo HPV e os fatores de risco relacionados à suscetibilidade ao câncer do colo do útero em mulheres assintomáticas. Métodos: Trata-se de um estudo transversal com 428 usuários atendidos em Unidades Básicas de Saúde, em Arapiraca, Alagoas, Brasil. Foram coletados relatórios sociodemográficos, variáveis comportamentais e citopatológicos. A detecção molecular do vírus HPV foi realizada por Nested-PCR. A análise estatística foi realizada com SPSS versão 22.0. Resultados: Foram estudadas 428 mulheres, com DNA de HPV detectado em 39,2% (n =168), com média de idade de 41 anos. Houve associação do HPV com o uso de anticoncepcional oral (p<0,016) e alcoolismo (p <0,038). Foi evidenciada maior frequência de HPV positivo em mulheres maiores de 25 anos (88,7%), até cinco parceiros sexuais (93,4%), até três gestações (71,4%) e com resultados citopatológicos dentro dos limites da normalidade (61,9%). O HPV foi identificado em 40,3% (104/258) das mulheres com resultados dentro dos limites da normalidade. Conclusão: Nossos resultados sugerem que o uso de anticoncepcionais orais e o alcoolismo podem ser considerados como possíveis fatores de risco relacionados à oncogênese cervical. Com isso, é necessário propor intervenções voltadas para a educação em saúde dessa população, ações de prevenção e detecção precoce.
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Papillomaviridae , Infecções por Papillomavirus , Vírus , Mulheres , Neoplasias do Colo do Útero , Saúde , Educação em Saúde , Fatores de Risco , Saúde da Mulher , Prevenção de Doenças , Teste de PapanicolaouRESUMO
The NOS3 gene polymorphisms T-786C, G894T and VNTR 4b/a are associated with a predisposition to the development of Metabolic Syndrome (MetS). The NOS3 gene contributes to a normal pregnancy and fetal development. According to their birthweight, newborns can be classified as: small (SGA), adequate (AGA) or large (LGA) for gestational age. The SGA and LGA present a higher risk of developing disorders related to MetS, both during childhood and adulthood. Therefore, the aim of this work is to relate the incidence of G894T, T-786C and VNTR 4b/a on SGA and LGA newborns and their mothers. 204 blood samples were collected from mothers (102) and the umbilical cords of 102 newborns (SGA = 12; AGA = 47; LGA = 43). The genotyping was performed through PCR-RFLP to evaluate presence of the G894T, T-786C and VNTR 4b/a polymorphisms. A significant difference was found between the groups of newborns in the genotypic frequency of T-786C, but without Hardy-Weinberg equilibrium. The VNTR 4b/a and the G894T polymorphisms showed no significance between the groups. The haplotype analysis showed that the SGA newborns presented the higher frequency of 4aGT (9.8%) and of the 4aTT combination (25.4%), while LGA newborns presented the higher frequency of the 4bTT haplotype (23%). Only the SGA newborns and their mothers presented the 4aTC haplotype. In conclusion, the NOS3 polymorphisms do not appear to be a factor to inadequate birth weight. However, the G894T and VNTR 4b/a polymorphisms, and the haplotype 4aTC, seem to influence the occurrence of SGA.
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Peso ao Nascer/genética , Repetições Minissatélites/genética , Óxido Nítrico Sintase Tipo III/genética , Polimorfismo de Nucleotídeo Único , Adulto , Alelos , Feminino , Frequência do Gene , Genótipo , Idade Gestacional , Haplótipos , Humanos , Recém-Nascido , Masculino , Síndrome Metabólica/genética , Fatores de RiscoRESUMO
INTRODUCTION: MicroRNA-21 (miRNA-21) has been described as one of the most significantly upregulated miRNAs in human breast cancer. However, limited knowledge exists on miRNA-21 expression in breast cancer tissue after neoadjuvant chemotherapy (NAC). PURPOSE: The aim of this study was to assess miRNA-21 expression in the tumor tissues of Brazilian patients with breast cancer who underwent NAC and its correlation with clinicopathological variables. PATIENTS AND METHODS: Utilizing qRT-PCR, miRNA-21 expression in tumor tissue was measured in a cohort of female patients with breast cancer who underwent NAC. The correlation of miRNA-21 expression with breast cancer molecular subtypes and other clinicopathological variables was also assessed. RESULTS: A total of 55 patients were included in the study, and 28 (50.9%) underwent NAC. miRNA-21 was upregulated in patients with breast cancer, regardless of previous exposure to chemotherapy, molecular subtypes, tumor-node-metastasis (TNM) staging and lymph node status of the axilla. miRNA-21 expression did not differ between patients with breast cancer who achieved a pathologic complete response after NAC and healthy controls. CONCLUSION: miRNA-21 was upregulated in the tumor tissue of Brazilian patients with breast cancer regardless of NAC treatment, which reinforces its role as an "oncomiR" and a potential biomarker.
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Simulação de Dinâmica Molecular , Papillomaviridae , Sítios de Ligação , DNA , Ligação ProteicaRESUMO
Human epidermal growth factor receptor-2 (HER2) is an important biomarker in the diagnosis and prognosis of breast cancer. This work aimed to develop an aptasensor to detect HER2 in human serum. HER2 aptamer was immobilized by electrostatic adsorption on the surface of a homemade screen-printed electrode modified with poly-L-lysine. Measurements were made by differential pulse voltammetry using methylene blue as a redox indicator. A calibration curve was constructed (R2 = 0.997) using different concentrations of HER2 protein (10-60 ng/mL) in PBS buffer (pH 7.4), with a detection limit of 3.0 ng/mL. The aptasensor showed good reproducibility with relative standard deviations (RSDs) of 3% and remained stable for 3 days with an RSD around 2%. When the tests were performed with serum from a healthy woman, a peak of 6.72 µA was found without the addition of the protein. When we tested the serum of a woman with HER2+ breast cancer, we obtained a signal of 2.65 µA; the same pattern was found when adding to protein in serum control, i.e., the higher the concentration of protein, the lower the signal. The aptasensor was characterized by scanning electron microscopy and isothermal titration calorimetry (ITC), showing excellent interaction between aptamer and target protein. The results revealed a promising and sensitive tool capable of detecting HER2 protein in human serum with albumin depletion, aiding in the molecular diagnosis of breast cancer. Graphical abstract.
Assuntos
Aptâmeros de Nucleotídeos/química , Neoplasias da Mama/sangue , Receptor ErbB-2/sangue , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Neoplasias da Mama/diagnóstico , Detecção Precoce de Câncer , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Eletrodos , Desenho de Equipamento , Feminino , Humanos , Limite de Detecção , Receptor ErbB-2/análise , Reprodutibilidade dos TestesRESUMO
Breast and cervical cancer are the first and fourth cancer types with the highest prevalence in women, respectively. The developmental profiles of cancer in women can vary by genetic markers and cellular events. In turn, age and lifestyle influence in the cellular response and also on the cancer progression and relapse. The human DEK protein, a histone chaperone, belongs to a specific subclass of chromatin topology modulators, being involved in the regulation of DNA-dependent processes. These epigenetic mechanisms have dynamic and reversible nature, have been proposed as targets for different treatment approaches, especially in tumor therapy. The expression patterns of DEK vary between healthy and cancer cells. High expression of DEK is associated with poor prognosis in many cancer types, suggesting that DEK takes part in oncogenic activities via different molecular pathways, including inhibition of senescence and apoptosis. The focus of this review was to highlight the role of the DEK protein in these two female cancers.
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Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/metabolismo , Proteínas Cromossômicas não Histona/metabolismo , Proteínas Oncogênicas/metabolismo , Proteínas de Ligação a Poli-ADP-Ribose/metabolismo , Neoplasias do Colo do Útero/metabolismo , Saúde da Mulher , Animais , Biomarcadores Tumorais/química , Biomarcadores Tumorais/genética , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Neoplasias da Mama/terapia , Proteínas Cromossômicas não Histona/química , Proteínas Cromossômicas não Histona/genética , Epigênese Genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Proteínas Oncogênicas/química , Proteínas Oncogênicas/genética , Proteínas de Ligação a Poli-ADP-Ribose/química , Proteínas de Ligação a Poli-ADP-Ribose/genética , Prognóstico , Conformação Proteica , Relação Estrutura-Atividade , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/terapiaRESUMO
Objectives: The objective of the study was to compare the postoperative staging and clinical development outcomes in a period of three years with the histopathological and immunohistochemical characteristics considered prognostic and/or predictive factors in patients being treated for triple negative type of breast cancer in the Barão Lucena Hospital, Recife, Pernambuco. Method: The study was conducted with 125 female patients suffering from triple negative breast cancer who underwent surgical treatment in the mastology service of Barão Lucena Hospital from 2009 to 2012. The clinical and pathological features of the tumors were studied and correlated with basal and non-basal subtypes. A descriptive data analysis was carried out using tables and/or graphs for qualitative variables. Association analysis was performed using χ2 test for independence. In tables that showed expected frequency lower than 5, in more than 20% of cells, we used the Fisher's exact test. In addition, the odds ratio (OR) and the confidence interval (CI) for OR were calculated. In the entire analysis, a 5% significance level was considered. Results: M ean a ge w as 4 9 y ears; r egarding r ace, b lack w as p resent i n 8 3 ( 66.4%). The most common histological type was ductal, in 111 (88.8%). The pathological stage I/II was the most common, in 87 (69.6%) patients. A total of 71 patients (56.8%) showed no axillary metastasis. Regarding the type of surgery, the conservative one was performed in 57 (45.6%), including sectorectomy and oncoplastic surgery. The recurrence was present in 30 patients, basal in 16 (53.3%) patients and 14 (46.7%) in the non-basal, and bone metastasis was the most frequent. Conclusion: In this triple-negative tumor sample, the most important facts related to survival were: being aged less than 40 years, histological type, cytokeratin CK5/6 and higher significance level of the factors EGFR and KI-67.
Objetivos: O objetivo do estudo foi correlacionar o estadiamento pós-cirúrgico e a evolução clínica em um período de três anos com as características histopatológicas e imunoistoquímicas consideradas fatores prognósticos e/ou preditivos nas pacientes em tratamento de câncer de mama do tipo triplo negativo do Hospital Barão de Lucena, Recife, Pernambuco. Método: O estudo foi feito com 125 pacientes do sexo feminino portadoras de câncer de mama triplo negativo e que foram submetidas a tratamento cirúrgico no serviço de mastologia do Hospital Barão de Lucena no período de 2009 a 2012. Nessas pacientes foram estudadas as características clínicas e patológicas dos tumores, as quais foram correlacionadas com os subtipos basal e não basal. A análise descritiva dos dados foi feita através de tabelas e/ou gráficos para variáveis qualitativas. Para análise de associação, foi utilizado o teste do χ2 para independência. Nas tabelas que apresentaram frequência esperada menor que 5, em mais de 20% das caselas, foi utilizado o teste exato de Fisher. Além disso, foi calculada a razão de chance (OR) e o intervalo de confiança (IC) para OR. Em toda a análise foi considerado nível de significância de 5%. Resultados: A média de idade foi de 49 anos; com relação à raça, tivemos a cor negra em 83 (66,4%) delas. O tipo histológico mais comum foi o ductal, em 111 (88,8%) pacientes. O estágio patológico I/II foi o mais comum, em 87 (69,6%) delas. Um total de 71 (56,8%) pacientes não demonstrou comprometimento axilar. Com relação ao tipo de cirurgia, a conservadora foi utilizada em 57 (45,6%) pacientes, incluindo setorectomia e técnicas de oncoplastia. A recorrência esteve presente em 30 pacientes, sendo basal em 16 (53,3%) e não basal 14 (46,7%) delas, nas quais a metástase óssea foi a mais frequente. Conclusão: Nessa amostra de tumores triplo negativo, os fatos mais importantes associados à sobrevida foram a idade abaixo de 40 anos, o tipo histológico, a citoqueratina CK5/6 e o grau de significância maior dos fatores EGFR e KI-67.
RESUMO
BACKGROUND: Cervical cancer is one of the most common female cancers and is caused by human papillomavirus (HPV). Viral infection leads to cell cycle deregulation by inactivating p53 and retinoblastoma protein by viral oncoproteins E6 and E7, respectively. Then, nuclear proteins such as DNA topoisomerase type IIa (TOP2A) and Ki-67 show increased expression because of increased cell division. These molecules are used as biomarkers for immunohistochemistry analysis of cervical tissue. METHODS: In this cross-sectional study, we recruited 110 women receiving regular gynecological surveillance at public health centers in Olinda - PE, Brazil. Cervicovaginal cells were collected to determine the presence of cytological abnormalities and HPV infection. Pap smear slides were used to evaluate the expression of TOP2A and Ki-67 using immunocytochemistry techniques. RESULTS: Of the 110 women, 75.4 % showed HPV-DNA(+) infection (83/110) and 29.1 % showed cellular abnormalities (32/110). Two atypical cells of undetermined significance, one low-grade squamous intraepithelial lesion, and one high-grade squamous intraepithelial lesion samples showed no HPV-DNA. TOP2A was positive in 71.9 % of samples, while Ki-67 was positive in 81.2 %. Immunocytochemistry results were positive in 4 of 5 atypical cells of undetermined significance samples. In HPV-DNA(+) samples with cytological abnormalities, immunocytochemistry results were positive 96.4 % of samples (p < 0.0001; odds ratio = 28.0). Among the samples infected with HR-HPV, TOP2A(+) was effective in 71 % samples, while and Ki-67(+) was 77.4 %. Ki-67 and TOP2A were positive for all samples infected with HPV6, HPV11, and HPV18. Ki-67 was also positive for all HPV16 samples, except for one negative sample in cytopathology analysis. CONCLUSIONS: TOP2A and Ki-67 antibodies may be used in combination for cervical cancer screening in immunocytochemistry assays.
Assuntos
Alphapapillomavirus , Antígenos de Neoplasias/metabolismo , DNA Topoisomerases Tipo II/metabolismo , Proteínas de Ligação a DNA/metabolismo , Antígeno Ki-67/metabolismo , Proteínas de Neoplasias/metabolismo , Infecções por Papillomavirus/metabolismo , Neoplasias do Colo do Útero/metabolismo , Esfregaço Vaginal , Adolescente , Adulto , Idoso , Anticorpos Antineoplásicos/química , Feminino , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Infecções por Papillomavirus/patologia , Proteínas de Ligação a Poli-ADP-Ribose , Neoplasias do Colo do Útero/patologiaRESUMO
UNLABELLED: CYP2D6 is a high polymorphic enzyme from P450, responsible for metabolizing almost 25% of drugs. The distribution of different mutations among CYP2D6 alleles has been associated with poor, intermediate, extensive and ultra-metabolizers. AIM: To evaluate how missenses mutations in CYP2D6*7 and CYP2D6*14A poor metabolizer alleles affect CYP2D6 stability and function. MATERIALS & METHODS: CYPalleles database was used to collect polymorphisms data present in 105 alleles. We selected only poor metabolizers alleles that presented exclusively missenses mutations. They were analyzed through seven algorithms to predict the impact on CYP2D6 structure and function. RESULTS: H324P, the unique mutation in CYP2D6*7, has high impact in enzyme function due to its occurrence between two alpha-helixes involved in active site dynamics. G169R, a mutation that occurs only in CYP2D6*14A, leads to the gain of solvent accessibility and severe protein destabilization. CONCLUSION: Our in silico analysis showed that missenses mutations in CYP2D6*7 and CYP2D6*14A cause CYP2D6 dysfunction.
Assuntos
Citocromo P-450 CYP2D6/genética , Mutação de Sentido Incorreto/genética , Tamoxifeno/metabolismo , Alelos , Antineoplásicos Hormonais/metabolismo , Antineoplásicos Hormonais/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Domínio Catalítico/genética , Feminino , Genótipo , Humanos , Polimorfismo de Nucleotídeo Único/genética , Tamoxifeno/uso terapêuticoRESUMO
Dengue fever is the most prevalent vector-borne disease in the world, with nearly 100 million people infected every year. Early diagnosis and identification of the pathogen are crucial steps for the treatment and for prevention of the disease, mainly in areas where the co-circulation of different serotypes is common, increasing the outcome of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Due to the lack of fast and inexpensive methods available for the identification of dengue serotypes, herein we report the development of an electrochemical DNA biosensor for the detection of sequences of dengue virus serotype 3 (DENV-3). DENV-3 probe was designed using bioinformatics software and differential pulse voltammetry (DPV) was used for electrochemical analysis. The results showed that a 22-m sequence was the best DNA probe for the identification of DENV-3. The optimum concentration of the DNA probe immobilized onto the electrode surface is 500 nM and a low detection limit of the system (3.09 nM). Moreover, this system allows selective detection of DENV-3 sequences in buffer and human serum solutions. Therefore, the application of DNA biosensors for diagnostics at the molecular level may contribute to future advances in the implementation of specific, effective and rapid detection methods for the diagnosis dengue viruses.
Assuntos
Técnicas Biossensoriais/métodos , DNA Viral/genética , Vírus da Dengue/genética , Sondas de DNA , DNA Viral/análise , DNA Viral/sangue , Dengue , Técnicas Eletroquímicas , Guanina , Humanos , Limite de Detecção , OxirreduçãoRESUMO
BACKGROUND: H19 is a strong candidate gene for influencing birth weight variation and is exclusively imprinted maternally. In an attempt to understand the relationship of this gene polymorphism with low birth weight children, we investigated association of H19/RsaI polymorphism with low birth weight and normal birth weight in children and their mothers. OBJECTIVES: The aim of our study was to establish the association between H19 gene polymorphism and LW in children born in Pernambuco, state of Brazil. PATIENTS AND METHODS: It were selected 89 children, 40 low birth weight (LW) and 49 normal birth weight (NW) and 71 mothers (40 mothers of newborns NW and 31 mothers of newborns LW) attended at Dom Malan Hospital, Petrolina, Pernambuco - Brazil. Peripheral blood samples were collected from patients and genomic DNA was extracted and detected by electrophoresis agarose gel, stained by Blue Green Loading Dye. DNA PCR amplification was done using the primers H1 (sense) and H3 (antisense). PCR products were digested with RsaI and electrophoresed on agarose gel stained by ethidium bromide. Statistical analyses were performed using the program BioEstat version 5.0. RESULTS: The RsaI polymorphism in the H19 gene showed that genotype frequencies did not differ statistically between low birth weight (AA = 12.5%, AB = 45%, BB = 42.5%) and control (AA = 8.6% AB = 36.73%, BB= 55.10% groups) and the allele frequencies were not significantly different (P = 0.2897). We also did not observe any association between maternal H19 allele polymorphism and low birth weight newborns (P =0.7799) or normal birth weight children (P = 0.8976). CONCLUSIONS: The small size of sample may be the explanation for these results; future studies with more patients are needed to confirm the effect of H19/RsaI polymorphism on birth weight of LW newborns.