ABSTRACT Objective The transcriptional repressor DREAM is involved in thyroid-specific gene expression, thyroid enlargement and nodular development, but its clinical utility is still uncertain. In this study we aimed to investigate whether DREAM mRNA levels differ in different thyroid tumors and how this possible difference would allow the use of DREAM gene expression as molecular marker for diagnostic and/or prognosis purpose. Materials and methods We quantified DREAM gene mRNA levels and investigated its mutational status, relating its expression and genetic changes to diagnostic and prognostic features of 200 thyroid tumors, being 101 malignant [99 papillary thyroid carcinomas (PTC) and 2 anaplastic thyroid carcinomas] and 99 benign thyroid lesions [49 goiter and 50 follicular adenomas (FA)]. Results Levels of mRNA of DREAM gene were higher in benign (0.7909 ± 0.6274 AU) than in malignant (0.3373 ± 0.6274 AU) thyroid lesions (p < 0.0001). DREAM gene expression was able to identify malignancy with 66.7% sensitivity, 85.4% specificity, 84.2% positive predictive value (PPV), 68.7% negative predictive value (NPV), and 75.3% accuracy. DREAM mRNA levels were also useful distinguishing the follicular lesions FA and FVPTC with 70.2% sensitivity, 73.5% specificity, 78.5% PPV, 64.1% NPV, and 71.6% accuracy. However, DREAM gene expression was neither associated with clinical features of tumor aggressiveness, nor with recurrence or survival. Six different genetic changes in non-coding regions of DREAM gene were also found, not related to DREAM gene expression or tumor features. Conclusion We suggest that DREAM gene expression may help diagnose thyroid nodules, identifying malignancy and characterizing follicular-patterned thyroid lesions; however, it is not useful as a prognostic marker.

OBJETIVO: O isolamento adequado de ácidos nucleicos a partir de sangue periférico, lâmina corada de punção aspirativa por agulha fina, tecido fixado em formalina e emblocado em parafina e tecido fresco é fundamental para assegurar o sucesso de técnicas aplicadas em endocrinologia molecular, principalmente quando lidamos com amostras estocadas por longos períodos ou quando há impossibilidade de nova coleta de amostra de pacientes que perderam o seguimento. Neste trabalho, objetivamos otimizar as metodologias clássicas para a extração de DNA (salting-out) e RNA. MATERIAIS E MÉTODOS: Utilizamos proteinase K, choque térmico, dentre outras modificações, com o objetivo de aumentar a quantidade e a qualidade do material recuperado a partir das amostras descritas acima. RESULTADOS: Isolamos com sucesso DNA e RNA de tais amostras e o material obtido foi adequado para a realização de PCR, perfil de metilação, PCR em tempo real e sequenciamento de DNA. CONCLUSÃO: As técnicas aplicadas neste estudo para isolar ácidos nucleicos permitiram a realização posterior de análises moleculares consistentes e confiáveis. Arq Bras Endocrinol Metab. 2012;56(9):618-26

OBJECTIVE: Adequate isolation of nucleic acids from peripheral blood, fine-needle aspiration cells in stained slides, and fresh and formalin-fixed/paraffin-embedded tissues is crucial to ensure the success of molecular endocrinology techniques, especially when samples are stored for long periods, or when no other samples can be collected from patients who are lost to follow-up. Here, we evaluate several procedures to improve current methodologies for DNA (salting-out) and RNA isolation. MATERIALS AND METHODS: We used proteinase K treatment, heat shock, and other adaptations to increase the amount and quality of the material retrieved from the samples. RESULTS: We successfully isolated DNA and RNA from the samples described above, and this material was suitable for PCR, methylation profiling, real-time PCR and DNA sequencing. CONCLUSION: The techniques herein applied to isolate nucleic acids allowed further reliable molecular analyses. Arq Bras Endocrinol Metab. 2012;56(9):618-26

OBJECTIVE: To analyze cytokine gene expression in keratinocytes from patients with systemic lupus erythematosus (SLE). INTRODUCTION: Keratinocytes represent 95% of epidermal cells and can secrete several cytokines. METHODS: Keratinocytes were obtained by laser microdissection from 21 patients with SLE (10 discoid and 11 acute lesions) at involved and uninvolved sites. All patients were receiving a low/moderate prednisone dose and 18 were receiving chloroquine diphosphate. IL-2, IL-5, TNF-α and IFN-γ gene expression was evaluated by real-time PCR and expressed as the ratio (R) to a pool of skin samples from 12 healthy volunteers. RESULTS: Heterogeneity in cytokine gene expression was found among patients with SLE. Eighteen of 38 valid SLE samples (47%) presented overexpression (R>1) of at least one cytokine. Lesional skin samples tended to show higher cytokine expression than samples from uninvolved skin (p = 0.06). IL-5 and IFN-γ were the most commonly overexpressed cytokines. Samples with cytokine overexpression corresponded to more extensive and severe lesions. Prednisone dose did not differ between samples without cytokine overexpression (15.71±3.45 mg/day) and those with overexpressed cytokines (12.68±5.41 mg/day) (p = 0.216). Samples from all patients not receiving diphosphate chloroquine had at least one overexpressed cytokine. CONCLUSIONS: The heterogeneous keratinocyte cytokine gene expression reflects the complex immunological and inflammatory background in SLE. Patients with severe/extensive skin lesions showed a higher frequency of cytokine gene overexpression. Increased IFN-γ and IL-5 expression suggests that Th1 and Th2 cells are involved in SLE skin inflammation. The possibility that prednisone and antimalarial drugs may have contributed to low cytokine gene expression in some samples cannot be ruled out.

OBJECTIVE: To analyze glucose transporter 1 expression patterns in malignant tumors of various cell types and evaluate their diagnostic value by immunohistochemistry. INTRODUCTION: Glucose is the major source of energy for cells, and glucose transporter 1 is the most common glucose transporter in humans. Glucose transporter 1 is aberrantly expressed in several tumor types. Studies have implicated glucose transporter 1 expression as a prognostic and diagnostic marker in tumors, primarily in conjunction with positron emission tomography scan data. METHODS: Immunohistochemistry for glucose transporter 1 was performed in tissue microarray slides, comprising 1955 samples of malignant neoplasm from different cell types. RESULTS: Sarcomas, lymphomas, melanomas and hepatoblastomas did not express glucose transporter 1. Fortyseven per cent of prostate adenocarcinomas were positive, as were 29% of thyroid, 10% of gastric and 5% of breast adenocarcinomas. Thirty-six per cent of squamous cell carcinomas of the head and neck were positive, as were 42% of uterine cervix squamous cell carcinomas. Glioblastomas and retinoblastomas showed membranous glucose transporter 1 staining in 18.6% and 9.4% of all cases, respectively. Squamous cell carcinomas displayed membranous expression, whereas adenocarcinomas showed cytoplasmic glucose transporter 1 expression. CONCLUSION: Glucose transporter 1 showed variable expression in various tumor types. Its absence in sarcomas, melanomas, hepatoblastomas and lymphomas suggests that other glucose transporters mediate the glycolytic pathway in these tumors. The data suggest that glucose transporter 1 is a valuable immunohistochemical marker that can be used to identify patients for evaluation by positron emission tomography scan. The function of cytoplasmic glucose transporter 1 in adenocarcinomas must be further examined.

Canais iônicos auxiliam diferentes funções celulares, principalmente na transdução de sinal. Nas células endócrinas, esses canais têm funções importantes na secreção hormonal, sinalização do Ca2+, transporte transepitelial, regulação da motilidade, volume e conteúdo iônico celular e da acidificação do compartimento lisossomal (pH). Como esperado, as alterações nos canais iônicos podem causar distúrbios endocrinológicos, como pseudo-hipoaldosteronismo tipo 1, síndrome de Liddle, síndrome de Bartter, hipoglicemia hiperinsulinêmica da infância, diabetes melito neonatal, fibrose cística, doença de Dent, hipomagnesemia com hipocalcemia secundária, diabetes insípido nefrogênico e paralisia periódica tirotóxica hipocalêmica. Este artigo propõe uma breve revisão das canalopatias endócrinas conhecidas, com foco particular nos recentes progressos no conhecimento dos mecanismos fisiopatológicos adquirido a partir das alterações funcionais encontradas.

Ion channels serve diverse cellular functions, mainly in cell signal transduction. In endocrine cells, these channels play a major role in hormonal secretion, Ca2+-mediated cell signaling, transepithelial transport, cell motility and growth, volume regulation and cellular ionic content and acidification of lysosomal compartments. Ion channel dysfunction can cause endocrine disorders or endocrine-related manifestations, such as pseudohypoaldosteronism type 1, Liddle syndrome, Bartter syndrome, persistent hyperinsulinemic hypoglycemia of infancy, neonatal diabetes mellitus, cystic fibrosis, Dent's disease, hypomagnesemia with secondary hipocalcemia, nephrogenic diabetes insipidus and, the most recently genetically identified channelopathy, thyrotoxic hypokalemic periodic paralysis. This review briefly recapitulates the membrane action potential in endocrine cells and offers a short overview of known endocrine channelopathies with focus on recent progress regarding the pathophysiological mechanisms and functional genetic defects.

OBJETIVOS: Os sarcomas sinoviais são tumores raros e agressivos que acometem adultos jovens, com sobrevida doençaespecífica em cinco anos de 57 a 63%. O presente estudo analisa a experiência institucional com este tumor, dando ênfase à associação entre variáveis clínicas, padrões de recorrência e sobrevida. MÉTODO: Entre 1970 e 2001 foram identificados 57 pacientes com sarcomas sinoviais. Fatores demográficos, clínicos e anatomopatológicos foram pesquisados. Associações entre variáveis clínicas e a sobrevida livre de recidiva local, livre de metástases e doença-específica em cinco anos foram calculadas. RESULTADOS: A idade mediana dos pacientes foi 26 anos, 56% eram masculinos, 79% eram brancos. Localizavam-se em membro inferior em 74%, proximalmente em 53%. O sintoma mais comum foi a presença de tumor em 42%. Na admissão 18% eram intactos, 42% manipulados e 40% recidivados. A maioria era maior que 5 cm., e três pacientes apresentavam metástase linfonodal. A cirurgia mais freqüente foi ressecção ampla, 30% necessitou amputação. Margens amplas foram obtidas em 65%, 51% eram tumores bifásicos. Neoadjuvância foi utilizada em 46% e adjuvância em 58% dos casos. As sobrevidas livre de recidiva local , metástases e doença específica em cinco anos foram 60±8%, 47±7% e 58±7%. A localização proximal do tumor associou-se com preservação de membro (p=-0,001), margens inadequadas (p=0,006) e subtipo bifásico (p=0,047). CONCLUSÕES: Os dados confirmam a hipótese de tratar-se de tumor agressivo, com altos índices de recidiva local e à distância. Os resultados do tratamento são comparáveis a outros centros especializados. Tratamento fora destes centros deve ser desencorajado.

BACKGROUND: Synovial sarcoma is a rare and aggressive tumor more frequently affecting young adults and has a 5 year disease specific survival of 57-63%. The present study describes the institutional experience with the treatment of this tumor, emphasizing associations between clinical variables, patterns of recurrence and survival. METHODS: Between 1970 and 2001, 57 patients were identified with synovial sarcomas. Demographic, clinical and histological factors were analyzed. Associations between clinical variables and 5 year local recurrence free, metastasis free and disease specific survival were calculated. RESULTS: Median patient age was 26 years, 56% were male and 79% caucasians. Tumors were located in the lower limb in 74%, proximally in 53%.The most common symptom was the presence of tumor in 42%. At admission, 18% were intact, 42% manipulated and 40% recurrent. Most of them were larger than 5 cm, 3 patients had lymph node metastasis. The most common procedure was extensive resection, 30% were amputated. Wide margins were obtained in 65%, 51% were biphasic tumors. Neoadjuvant therapy was performed in 46%, 56% had adjuvant therapy. The 5 year local recurrence free, metastasis free and disease specific survival were 60±8%, 47±7% e 58±7%. Proximal location was associated with limb sparing procedures (p=0.001), close margins (p=0.006) and biphasic tumors (p=0.047). CONCLUSION: Data support the hypothesis that this is an aggressive tumor, with high rates of local and remote recurrences. The survival obtained was comparable to other reference centers. Treatment outside these centers should be discouraged.

Um caso de teratoma imaturo gigante do sistema nervoso central em um natimorto é relatado. Durante a gravidez, a mãe desenvolveu poli-hidrâmnio abrupto na 25ª semana de gestação e dor pélvica intensa, que levou a indicação de cesárea. O natimorto pesava 2750g, apresentava macrocefalia evidente e o conteúdo craniano pesou 1350g. Este era representado por tecido tumoral em sua maioria e apenas uma pequena rima de tecido encefálico normal. A histologia mostrou tratar-se de um teratoma imaturo. Teratomas intracranianos gigantes congênitos são neoplasias raras, de histogênese incerta e não há explicação para o crescimento contínuo e rápido durante a vida intra-uterína. O diagnóstico pré-natal pode ser realizado por ultra-sonografia, tomografía computadorizada ou ressonância magnética.

A case of massive intracranial immature teratoma in a female stillborn is reported. She was the product of the second pregnancy of a 25-year-old healthy woman. The pregnancy was unremarkable until the 25th week of gestation when the mother noticed a rapid enlargement of her abdomen and intense pelvic pain. Because of the pain, a cesarean section was indicated, and a stillborn weighing 2750g with macrocephaly was delivered. The cranial contents weighed 1350g and showed a huge tumoral mass with only a rim of normal brain. A histologic diagnosis of immature teratoma was made. Massive intracranial teratomas are rare tumors and their occurrence in intrauterine life is even rarer. Their histogenesis is unknown, and there is no explanation for their continuous growth during embryogenesis. A prenatal diagnosis of this rare condition can be made by ultrasound, computed tomography, or magnetic resonance imaging.