The gene expression of GPER1 is low in fresh samples of papillary thyroid carcinoma (PTC), and in silico analysis.

Papillary thyroid cancer (PTC), whose incidence has been increasing in the last years, occurs more frequently in women. Experimental studies suggested that estrogen could be an important risk factor for the higher female incidence. In fact, it has been demonstrated that 17ß-estradiol (E2) could increase proliferation and dedifferentiation in thyroid follicular cells. Genomic estrogen responses are typically mediated through classical estrogen receptors, the α and ß isoforms, which have been described in normal and abnormal human thyroid tissue. Nevertheless, effects mediated through G protein estrogen receptor 1 (GPR30/GPER/GPER1), described in some thyroid cancer cell lines, could be partially responsible for the regulation of growth in normal cells. In this study, GPER1 gene and protein expression are described in non-malignant and in papillary thyroid cancer (PTC), as well as its association with clinical features of patients with PTC. The GPER1 expression was lower in PTC as compared to paired non-malignant thyroid tissues in fresh samples of PTC and in silico analysis of GEO and TCGA databases. In PTC cases of TCGA database, low GPER1 mRNA expression was independently associated with metastatic lymph nodes, female gender, and BRAF mutation. Besides, GPER1 mRNA levels were positively correlated with mRNA levels of thyroid differentiation genes. These results support the hypothesis that GPER1 have a role in PTC tumorigenesis and might be a potential target for its therapy. Further studies are needed to determine the functionality of these receptors in normal and diseased thyroid.

Activity of ecto-5'-nucleotidase (NT5E/CD73) is increased in papillary thyroid carcinoma and its expression is associated with metastatic lymph nodes.

The incidence of papillary thyroid carcinoma (PTC) has been increasing, which raised the interest in its molecular pathways. Although the high expression of ecto-5'-nucleotidase (NT5E) gene expression and NT5E enzymatic activity in several types of cancer is associated with tumor progression, its role in PTC remains unknown. Here, we investigated the AMP hydrolysis in human normal thyroid cells and PTC cells, in primary culture, and the association of NT5E expression with clinical aspects of PTC patients. AMPase activity was higher in thyroid cells isolated from PTC, as compared to normal thyroid (P = 0.0063). Significant correlation was observed between AMPase activity and NT5E levels in primary thyroid cell cultures (r = 0.655, P = 0.029). NT5E expression was higher in PTC than in the adjacent non-malignant thyroid tissue (P = 0.0065) and were positively associated with metastatic lymph nodes (P = 0.0007), risk of recurrence (P = 0.0033), tumor size (P = 0.049), and nodular hyperplasia in the adjacent thyroid parenchyma, when compared to normal thyroid or lymphocytic thyroiditis (P = 0.0146). After adjusting for potential confounders, the malignant/non-malignant paired expression ratio of NT5E mRNA was independently associated with metastatic lymph nodes (P = 0.0005), and tumor size (P=0.0005). In addition, the analysis of PTC described in the TCGA database also showed an association between higher expression of NT5E and metastatic lymph nodes, and tumor microinvasion. These results support the hypothesis that NT5E have a role in PTC microenvironment and might be a potential target for PTC therapy.

Validation of reference genes for normalization gene expression in reverse transcription quantitative PCR in human normal thyroid and goiter tissue.

Reverse transcription quantitative polymerase chain reaction (RT-qPCR) has been recognized as the most accurate method for quantifying mRNA transcripts, but normalization of samples is a prerequisite for correct data interpretation. So, this study aimed to evaluate the most stable reference gene for RT-qPCR in human normal thyroid and goiter tissues. Beta-actin (ACTB); glyceraldehyde-3-phosphate dehydrogenase (GAPDH); succinate dehydrogenase, subunit A, flavoprotein (Fp) (SDHA); hypoxanthine phosphoribosyltransferase I (HPRTI); tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide (YWHAZ); and beta-2-microglobulin (B2M) were evaluated in 14 thyroid tissue samples (7 normal and 7 goiter tissues) by RT-qPCR. The mean Cq and the maximum fold change (MFC) and NormFinder software were used to assess the stability of the genes. As a result, ACTB gene was more stable than GAPDH, SDHA, HPRTI, YWHAZ, and B2M. In conclusion, ACTB could be used to normalize RT-qPCR data in normal thyroid and goiter tissues.

Role of VEGF-A and its receptors in sporadic and MEN2-associated pheochromocytoma.

Pheochromocytoma (PHEO), a rare catecholamine producing tumor arising from the chromaffin cells, may occurs sporadically (76%-80%) or as part of inherited syndromes (20%-24%). Angiogenesis is a fundamental step in tumor proliferation and vascular endothelial growth factor (VEGF-A) is the most well-characterized angiogenic factor. The role of angiogenic markers in PHEO is not fully understood; investigations were therefore made to evaluate the expression of VEGF-A and its receptors in PHEO and correlate to clinical parameters. Twenty-nine samples of PHEO were evaluated for VEGF-A, VEGF receptor-1 (VEGFR-1) VEGFR-2 expression and microvessel density (MVD) by immunohistochemistry. Clinical data were reviewed in medical records. The mean age of patients was 38±14 years, and 69% were woman. VEGF-A, VEGFR-1 and VEGFR-2 staining were detected in nearly all PHEO samples. No significant correlation was observed between VEGF-A, VEGFR-1, VEGFR-2 expression or MVD and age at diagnosis, tumor size or sporadic and hereditary PHEO. However, the levels of expression of these molecules were significantly higher in malignant PHEO samples (p=0.027, p=0.003 and p=0.026, respectively).VEGF-A and its receptors were shown to be up-regulated in malignant PHEO, suggesting that these molecules might be considered as therapeutic targets for unresectable or metastatic tumors.

Reduced tissue inhibitor of metalloproteinase-2 expression is associated with advanced medullary thyroid carcinoma.

Matrix metalloproteinases (MMPs) are enzymes for extracellular matrix remodeling that are involved in tumor growth, progression and metastasis. Among them, MMP-9 has been implicated in tumor angiogenesis. Tissue inhibitor of matrix metalloproteinase (TIMP)-2, a member of the family of MMP inhibitors, induces apoptosis and inhibits various stages of angiogenesis. Previous studies analyzing the expression of MMP-9 and TIMP-2 in medullary thyroid carcinoma (MTC) are scarce. The aims of the current study were to evaluate MMP-9 and TIMP-2 expression in MTC samples and correlate the results with clinical parameters. Paraffin-embedded samples from 77 MTC patients were evaluated for expression by immunohistochemistry. The clinical data in medical records were retrospectively reviewed. In total, 77 patients aged 35.6±17.1 years were enrolled. Of these patients, 36 had hereditary disease (46.8%). Immunohistochemical staining for MMP-9 and TIMP-2 was detected in 89.6 and 93.5% of the samples, respectively. The expression of MMP-9 was not found to correlate with clinical parameters, although, a trend toward a correlation between MMP-9 and distant metastasis was observed (P=0.053). By contrast, TIMP-2 staining was found to correlate with age at diagnosis (P=0.026) and negatively correlate with tumor size and tumoral stage (P=0.002 and P=0.001, respectively). Notably, the highest levels of TIMP-2 expression were observed in patients with intrathyroidal disease. The MMP-9 enzyme involved in extracellular matrix remodeling is overexpressed in MTC lesions and may contribute to tumor vascularization and growth. Reduced levels of TIMP-2 expression may be implicated in tumor progression and spread of disease.

Weighted calibration with reduced number of signals by weighing factor modelling: application to the identification of explosives by ion chromatography.

The linear weighted regression model (LW) can be used to calibrate analytical instrumentation in a range of quantities (e.g. concentration or mass) wider than possible by the linear unweighted regression model, LuW (i.e. the least squares regression model), since this model can be applied when signals are not equally precise through the calibration range. If precision of signals varies within the calibration range, the regression line should be defined taking into account that more precise signals are more reliable and should count more to define regression parameters. Nevertheless, the LW requires the determination of the variation of signals precision through the calibration range. Typically, this information is collected experimentally for each calibration, requiring a large number of replicate collection of signals of calibrators. This work proposes reducing the number of signals needed to perform LW calibrations by developing models of weighing factors robust to daily variations of instrument sensibility. These models were applied to the determination of the ionic composition of the water soluble fraction of explosives. The adequacy of the developed models was tested through the analysis of control standards, certified reference materials and the ion balance of anions and cations in aqueous extracts of explosives, considering the measurement uncertainty estimated by detailed metrological models. The high success rate of the comparisons between estimated and known quantity values of reference solutions, considering results uncertainty, proves the validity of developed metrological models. The relative expanded measurement uncertainty of single determinations ranged from 1.93% to 35.7% for calibrations performed along 4 months.

Increased expression of vascular endothelial growth factor and its receptors, VEGFR-1 and VEGFR-2, in medullary thyroid carcinoma.

BACKGROUND: Vascular endothelial growth factor (VEGF-A) expression is upregulated in the majority of human tumors, where it stimulates proliferation, migration, and survival of endothelial cells. Studies have suggested that VEGF inhibitors can be used as an alternative therapy in medullary thyroid carcinoma (MTC), but data about expression of VEGF-A and its receptor in this tumor are scarce. The aims of this study were to evaluate VEGF-A, VEGF receptor (VEGFR)-1, VEGFR-2, and microvessel density (MVD) expression in MTC samples and correlate it with clinical parameters. METHODS: Paraffin-embedded samples from 38 MTC patients were evaluated for VEGF-A, VEGFR-1, VEGFR-2, and MVD expression by immunohistochemistry. Clinical data were retrospectively reviewed in medical records. RESULTS: Thirty-eight patients aged 31.8 +/- 17.1 years were enrolled. Twenty-seven patients had hereditary disease (71.1%). Twenty-five of them were found to have multiple endocrine neoplasia (MEN) 2A and two were found to have MEN 2B. VEGF-A immunohistochemical staining was detected in 95% (36/38), VEGFR-1 in 96% (36/37), and VEGFR-2 in 91% (31/34) of MTC samples. Age at surgery was positively correlated with VEGFR-2 (p = 0.003). There was no correlation between VEGF-A, VEGFR-2, and tumor stage (tumor node metastasis). Nevertheless, VEGFR-1 was found to be inversely correlated with tumor node metastasis (p = 0.034). We also observed a trend toward an association between VEGFR-1 signal intensity and cure of disease, although this did not reach statistical significance (p = 0.054). Neither VEGF-A nor VEGFR-2 was associated with disease outcome after a median follow-up period of 5 years (p = 0.882 and p = 0.236, respectively). As expected, MVD was correlated with age at surgery (p = 0.005) and tumor size (p = 0.03). Patients with the hereditary form of the disease had a stronger intensity for VEGFR-1 (p = 0.039), whereas patients with sporadic disease displayed higher MVD counts (44 [27-63] vs. 21 [9-49], p = 0.018). CONCLUSION: The VEGF-A, VEGFR-1, and VEGFR-2 immunoreactive proteins are overexpressed in MTC lesions and might be implicated in tumor progression. It is not clear, however, if expression of these molecules provides prognostic information regarding the spread or outcome of MTC.

Indicadores da assistência ao paciente oncológico no Hospital Nossa Senhora da Conceição: desenvolvimento de ferramentas de tecnologia da informação para a gestão

O presente projeto descreve as etapas da elaboração de indicadores da assistência prestada ao paciente com câncer no Hospital Nossa Senhora da Conceição com auxílio de ferramentas da teconologia da informação. A necessidade de avaliação do conjunto de serviços oferecidos através de indicadores impõe-se face à gravidade da doença, seu alto custo de diagnóstico e tratamento e a relação entre determinados padrões de qualidade e possibilidade de cura de muitos tipos de tumores. A opção pela elaboração de sistemas eletrônicos que subsidiem a elaboração de indicadores deu-se pelo grande número de casos atendidos( cerca de 3000 diagnósticos por ano) e por seu caráter difuso dentro do hospital, onde diferentes especialidades realizam tais atendimentos em ambientes clínicos e cirúrgicos. Também contribui para a escolha desta modalidade o grande número de pacientes encaminhados já com diagnóstico realizado em outras instituições, não havendo para o conjunto de pacientes uma etapa obrigatória onde sua identificação para posterior acompanhamento fosse possível. Os sistemas desenvolvidos permitirão, a partir de informações obrigatórias solicitadas em etapas críticas dos processos assistenciais e administrativos, cruzar informações de forma a recuperar todos os pacientes com câncer atendidos na instutição, com suas características e datas de eventos críticos, permitindo a elaboração de indicadores que avaliem características da doença, seu estágio e tempo de evolução aliado ao desempenho institucional no diagnóstico e tratamento da patologia, correlacionando tais dados com o tempo de sobrevida e sistemas do prontuário eletrônico do paciente, tendo além da gestão, também a finalidade de otimizar os processos assistenciais, aumentar a segurança e a recupração de informações, padronizar os tratamentos oferecidos e avaliar a efetividade do conjunto de serviços oferecidos ao público alvo.

[Cancer in thyroid nodules: evaluation of ultrasound and fine-needle aspiration biopsy]. / Nódulos de tireóide: valor da ultra-sonografia e da biópsia por punção aspirativa no diagnóstico de câncer.

BACKGROUND: Thyroid nodules are a very common clinical problem and fine-needle aspiration biopsy (FNAB) is its main diagnostic test. Recently, it has been shown that some ultrasonographic features are associated to an increased risk of cancer, but a comparison between FNAB and ultrasonography (US) or its combination is not available. The aim of this study was to evaluate FNAB and US performance, isolated or combined, for cancer diagnosis in thyroid nodules. METHODS: Eighty patients, with palpable solitary thyroid nodules or multinodular goiters, who were to undergo surgery, were selected among 289 patients seen in our clinic between March, 1995 and July, 1997. US and FNAB were performed by one of us. US was considered positive for cancer when microcalcifications, absent halo or hypoechogenicity were seen. FNAB was considered positive for cancer when it was either suspicious or compatible with malignancy. Both methods (FNAB and US) were evaluated in parallel combination (positive when FNAB or US was positive) and in serial combination (positive when both, FNAB and US, were positive). RESULTS: Sensitivity and specificity of FNAB and US for cancer diagnosis in thyroid nodules were, respectively, 87% and 62%, and 81% and 70%. Serial combination of FNA and US had sensitivity of 69% and specificity of 91%, and parallel combination resulted in sensitivity of 94% and specificity of 52%. CONCLUSION: The US had good accuracy in the evaluation of thyroid nodules, and can help us take some decisions in the clinical practice, supplementary to FNA. Nevertheless, the quality of the ultrasound examination depends on the experience of the examiner and on the characteristics of the equipment, so new studies are necessary to evaluate these factors to the current usage of ultrasonographic characteristics in the evaluation diagnostic of thyroid nodules.

Nódulos de tireóide: valor da ultra-sonografia e da biópsia por punçäo aspirativa no diagnóstico de câncer / Cancer in thyroid nodules: evaluation of ultrasound and fine-needle aspiration biopsy

Os nódulos de tireóide säo um problema clínico comum e a punçäo aspirativa por agulha fina (PAAF) é o principal exame utilizado para seu diagnóstico diferencial. Recentemente, foi demonstrado que algumas características ultra-sonográficas dos nódulos estäo associadas a um maior risco de câncer. Entretanto, näo encontramos estudos que avaliassem o desempenho dos dois exames de forma comparativa e/ou complementar. OBJETIVO: Avaliar o desempenho da ultra-sonografia (US), da PAAF e de sua combinaçäo no diagnóstico de câncer em nódulos de tireóide. MÉTODOS: Oitenta pacientes com nódulos únicos ou dominantes, com indicaçäo cirúrgica, foram selecionados dentre 289 pacientes com nódulos tireoidianos atendidos entre março de 1995 e julho de 1997. Os pacientes foram submetidos à PAAF e à US de tireóide por um dos autores. A PAAF foi considerada positiva quando os achados citológicos eram suspeitos ou diagnósticos de câncer. A US foi considerada positiva quando os nódulos apresentavam pelo menos um dos seguintes achados: microcalcificações, hipoecogenicidade ou halo ausente. PAAF e US foram combinados em paralelo (o resultado positivo em qualquer um dos dois exames era considerado indicativo de malignidade) e em série (somente o resultado positivo de ambos os testes indicava presença de malignidade). RESULTADOS: A sensibilidade e a especificidade da PAAF na detecçäo do câncer em nódulos de tireóide foram, respectivamente, 87 por cento e 62 por cento, e a sensibilidade e a especificidade da US foram, respectivamente, 81 por cento e 70 por cento. A combinaçäo em série dos dois exames (PAAF e US) resultou em sensibilidade de 69 por cento e especificidade de 91 por cento, e a combinaçäo em paralelo (PAAF ou US) apresentou sensibilidade de 94 por cento e especificidade de 52 por cento. CONCLUSÄO: A US pode ser útil na avaliaçäo dos nódulos de tireóide, alternativamente ou em combinações com a PAAF. Como o resultado da US depende da experiência do operador e das características do equipamento, säo necessários estudos em diferentes centros para o seu uso na prática clínica

Punçäo aspirativa de tireóide com agulha fina em um hospital geral: estudo de 754 punçöes / Aspiration puncture of thyroid with fine needle in a geral hospital: study of 754 punctures

A punçäo aspirativa com agulha fina (PAAF) é considerada o método diagnóstico inicial e o de melhor relaçäo custo-benefício na avaliaçäo da doença nodular da tireóide. Apesar das vantagens do procedimento, o sucesso do método depende da experiência do patologista e da habilidade técnica na obtençäo de amostras adequadas. Neste trabalho avaliamos a eficácia do procedimento em um hospital geral de ensino. Foram realizadas 754 punçöes em um período de 3 anos. Cada nódulo palpável foi aspirado por médicos residentes de endocrinologia com agulha fina por 4 a 6 vezes. O material foi corado pelos métodos de Giemsa e de Papanicolaou. O resultado anatomopatológico foi obtido de 93 pacientes. A PAAF foi realizada em 663 pacientes (613F/50M), com idade média de 50ñ14 anos (12 a 93). Nos exames cito patológicos, os diagnósticos foram; bócio colóide (58,5 por cento), neoplasia folicular ou suspeito (8,2 por cento), tireoidite de Hashimoto (5,3 por cento), carcinoma papilar (2,3 por cento), carcinoma anaplásico (0,4 por cento), carcinoma medular (0,1 por cento) e insatisfatório (25,2 por cento). Dos 93 pacientes operados, 41 (44 por cento) apresentavam neoplasia e, entre as malignas, o carcinoma papilar foi o mais freqüente (65,6 por cento). Na neoplasia folicular, a percentagem de carcinoma foi 24 por cento. A análise dos dados da PAAF demonstrou: sensibilidade 92 por cento (IC= 78,6 a 98,3 por cento), especificidade 80 por cento (IC= 65,4 a 90,4 por cento), valor preditivo positivo 79,5 por cento (IC= 64,7 a 90,2 por cento), valor preditivo negativo 92,3 por cento (IC= 79,1 a 98,4 por cento) e acurácia diagnóstica 86 por cento (IC= 76,1 a 92,3 por cento). A PAAF estabeleceu um diagnóstico em 66,6 por cento dos nossos pacientes, diminuindo os custos e o número de cirurgias. A taxa de detecçäo à cirurgia de carcinomas de tireóide aumentou de 19,4 por cento para 31,2 por cento em 11 anos em nosso hospital.

Carcinoma sarcomatóide de bexiga: relato de caso / Sarcomatoid carcinoma of the bladder: a case report

Os carcinomas sarcomatóides de bexiga são neoplasias raras, representando apenas 0,3 por cento das patologias malignas da bexiga. Por apresentarem marcado grau de malignidade e agressividade, devem ser sempre consideradas no diagnóstico diferencial dos tumores da bexiga. Os autores revisam a literatura e relatam o caso de um paciente de 62 anos de idade, branco e tabagista, com queixa de hematúria macroscópica, investigada por cistocopia e biópsia. Um exame histológico do tumor sugeriu carcinoma sarcomatóide de bexiga. O diagnóstico foi confirmado por um exame imunohistoquímico. Como o paciente apresentava severa disfunção ventilatória, foi tratado com ressecção transuretral do tumor, recusou a indicação de terapia adjuvante evoluindo para óbito 5 meses após o diagnóstico