AGK-BRAF gene fusion is a recurrent event in sporadic pediatric thyroid carcinoma

Thyroid cancer is the fastest increasing cancer worldwide in all age groups. Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer in both adults and children. PTC genomic landscape has been extensively studied in adults, but information regarding sporadic pediatric patients is lacking. Although BRAF V600E mutation is highly prevalent in adults, this mutation is uncommon in pediatric cases. As adult and pediatric PTC is a mitogen-activated protein kinase-driven cancer, this altered pathway might be activated by different genetic events. The aim of this study was to investigate the occurrence of AGK-BRAF fusion gene, recently described in radiation-exposed pediatric PTC, in a cohort of exclusively sporadic pediatric PTC. The series consisted of 30 pediatric PTC younger than 18 years of age at the time of diagnosis and 15 matched lymph node metastases (LNM). Primary tumors and matched LNM were screened for the presence of the AGK-BRAF fusion transcript by RT-PCR. To confirm the identity of the amplified products, randomly selected samples positive for the presence of the fusion transcripts were sequenced. Moreover, BRAF dual-color, break-apart probes confirmed BRAF rearrangement. Overall, the AGK-BRAF fusion gene was detected in 10% (3/30) of primary tumors. For one of these cases, paired LNM was also available, which also shows the presence of AGK-BRAF fusion gene. This study described, for the first time, the presence of AGK-BRAF in sporadic pediatric PTC. Understanding the molecular events underlying pediatric PTC may improve preoperative diagnosis, allow molecular prognostication and define a therapeutic approach toward sporadic PTC patients.Sao Paulo State Research Foundation (FAPESP)CNPqFAPESP scholarUniv Fed Sao Paulo, Dept Morphol & Genet, Div Genet, Genet Bases Thyroid Tumors Lab, Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Dept Pathol, Sao Paulo, SP, BrazilIrmandade Santa Casa Misericordia Sao Paulo, Dept Pediat, Sao Paulo, SP, BrazilIrmandade Santa Casa Misericordia Sao Paulo, Dept Med, Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Dept Morphol & Genet, Div Genet, Genet Bases Thyroid Tumors Lab, Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Dept Pathol, Sao Paulo, SP, BrazilFAPESP: 2012/02902-9FAPESP: 2013/03867-5FAPESP: 2014/06570-6CNPq: 470441/2013-5Web of Scienc

Sistema Glutation-S-Transferase como fator prognóstico no carcinoma papilífero da tireoide

OBJETIVO: Analisar se existe relação entre os fatores moleculares dos genes GTS e a mortalidade dos pacientes com câncer de tireoide dado pelo índice AMES de prognóstico clínico. MÉTODOS: Foram coletadas amostras da tireoide de 66 pacientes com carcinoma papilífero (53 mulheres e 13 homens), de modo a permitir extração do material genético das enzimas. Foram constituídos dois grupos, segundo os fatores prognósticos clínicos de alto e baixo risco, de acordo a classificação AMES. Cada grupo foi avaliado pela presença ou não do genótipo nulo para as enzimas estudadas, correlacionando-os com os fatores prognósticos clínicos (AMES). RESULTADOS: Foram analisados os resultados de 17 doentes com alto risco (grupo A) e 49 com baixo (grupo B). Todas combinações de genótipos do GSTT1 e GSTM1 foram encontrados. O genótipo nulo dos dois genes do grupo de alto risco foi encontrado em 5,8% e no de baixo risco em 6,1%. CONCLUSÃO: A presença ou deleção dos genes GST (GSTT1 e GSTM1) não são bom fatores prognósticos no câncer papilífero da tireoide.PURPOSES: Analyze the relationship between the AMES classification and molecular factors from Glutation-S-Transferase System, specifically the GSTT1 and GSTM1 in patients with well differentiated thyroid cancer. METHODS: Samples of thyroid tissue of 66 patients with papillary thyroid carcinoma were obtained (53 women and 13 men). Patients were divided in two groups (high and low risk) according to the AMES classification. In each group, presence of the null genotype of both GST enzymes system was studied. These results were compared with the AMES classification. Samples were obtained in the operating room immediately after thyroidectomy, placed in cryotubes, immersed in liquid nitrogen and stored in a freezer at -80ºC. DNA of this enzymes was extracted by the fenol-cloroformium method. RESULTS: There were 17 high risk patients and 49 low risk patients. The null genotype of the high risk group was 5.8% and in the other group was 6.1%. CONCLUSION: There was no relationship between absence of genes GSTT1 and GSTM1 and prognosis of the papillary thyroid carcinoma when compared to the AMES classifications

Insuficiência adrenal leve causada por mutação do gene NR0B1 (DAX1) em menino com uma associação de hipogonadismo hipogonadotrófico, baixa estatura final e transtorno de déficit de atenção

Mutation on NROB1 (DAX1) gene can cause different phenotypes of adrenal insufficiency in infancy. Long-term evolution of these patients shows that it is possible to have an association with hypogonadotropic hypogonadism. In this article we describe the evolution of a patient with NROB1 gene mutation, diagnosed with a mild form of adrenal insufficiency, and we highlight the presence of hypogonadotropic hypogonadism and short stature, besides the presence of attention deficit disorder. Such associations should make physicians aware during the follow-up of patients with this disease.FCMSCSP, Unidade Endocrinol Pediat, Dept Pediat, Sao Paulo, BrazilFCMSCSP, Dept Ciencias Fisiol, Sao Paulo, BrazilWeb of Scienc

Atualização no diagnóstico e acompanhamento do diabetes melito relacionado à fibrose cística (DRFC)

Cystic fibrosis (CF) is the most common recessive autosomal disease among Caucasian. Children with CF have benefitted from advances in medical and nutritional treatments, and this can be gleaned from the improvement in the survival of these patients. The increase in the survival rate brought with it the appearance of co-morbidities related to CF. Nowadays cystic fibrosis-related diabetes (CFRD) is considered the most common complication associated with CF. It can appear as early as infancy or adolescence, and its prevalence can be as high as 50% in adult patients. Because of its high prevalence, difficulties in early detection and the risks involved, in recent years several studies and consensuses have focused on this condition, adding information about the epidemiology, pathophysiology, prognosis and treatment of CFRD. The main aspects of these new concepts, as well as the current recommendations for its diagnosis and follow-up, will be presented in this study. Arq Bras Endocrinol Metab. 2011;55(8):613-21Santa Casa de Misericordia Sao Paulo, Dept Pediat, Pediat Endocrinol Unit, BR-0122102 Sao Paulo, BrazilFCM SCSP, Sao Paulo, BrazilHosp Santa Casa Sao Paulo, Dept Pediat, Pediat Endocrinol Unit, Sao Paulo, BrazilHosp Santa Casa Sao Paulo, Dept Pediat, Pediat Pneumol Unit, Sao Paulo, BrazilDept Clin Med, Discipline Endocrinol, Sao Paulo, BrazilWeb of Scienc

Freqüência de mutação no códon 12 do gene K-ras no carcinoma ductal invasivo de mama

Objetivo: pesquisar a freqüência de mutação pontual no códon 12 do gene K-ras, em espécimes cirúrgicos de pacientes portadoras de carcinoma ductal invasivo de mama. Material e Métodos: foram utilizados cortes de 50 espécimes cirúrgicos incluídos em blocos de parafina, de pacientes portadoras de carcinoma ductal invasivo de mama, com graus histológicos II e III. Os cortes destinados ao estudo foram desparafinizados e submetidos a extração do DNA, por meio do emprego da proteinase K. Para a amplificação do fragmento a ser analisado, utilizou-se a reação em cadeia da polimerase, seguida por clivagem com o emprego de enzima de restrição de comprimento variável (RFLP). A verificação da presença de mutação nas amostras foi feita com o emprego de eletroforese em gel de agarose, com marcador de peso molecular "Ladder 123" (GIBCO-BRL), e a documentação dos resultados, mediante fotografia, utilizando-se luz ultravioleta transmitida. Resultados: em cinco dos 50 carcinomas ductais invasivos de mama estudados (10%) constatou-se a presença de mutação no códon 12 do gene K-ras, sendo todas elas polimórficas para esse caráter. As afetadas pelos tumores, que apresentavam a referida mutação, encontravam-se na pós-menopausa. Em quatro dos cinco casos em que se constatou a mutação, o grau histológico dos tumores era II e no caso restante III

GST genes expression as prognostic factor in papillary thyroid cancer

PURPOSES: Analyze the relationship between the AMES classification and molecular factors from Glutation-S-Transferase System, specifically the GSTT1 and GSTM1 in patients with well differentiated thyroid cancer. METHODS: Samples of thyroid tissue of 66 patients with papillary thyroid carcinoma were obtained (53 women and 13 men). Patients were divided in two groups (high and low risk) according to the AMES classification. In each group, presence of the null genotype of both GST enzymes system was studied. These results were compared with the AMES classification. Samples were obtained in the operating room immediately after thyroidectomy, placed in cryotubes, immersed in liquid nitrogen and stored in a freezer at -80ºC. DNA of this enzymes was extracted by the fenol-cloroformium method. RESULTS: There were 17 high risk patients and 49 low risk patients. The null genotype of the high risk group was 5.8% and in the other group was 6.1%. CONCLUSION: There was no relationship between absence of genes GSTT1 and GSTM1 and prognosis of the papillary thyroid carcinoma when compared to the AMES classifications.OBJETIVO: Analisar se existe relação entre os fatores moleculares dos genes GTS e a mortalidade dos pacientes com câncer de tireoide dado pelo índice AMES de prognóstico clínico. MÉTODOS: Foram coletadas amostras da tireoide de 66 pacientes com carcinoma papilífero (53 mulheres e 13 homens), de modo a permitir extração do material genético das enzimas. Foram constituídos dois grupos, segundo os fatores prognósticos clínicos de alto e baixo risco, de acordo a classificação AMES. Cada grupo foi avaliado pela presença ou não do genótipo nulo para as enzimas estudadas, correlacionando-os com os fatores prognósticos clínicos (AMES). RESULTADOS: Foram analisados os resultados de 17 doentes com alto risco (grupo A) e 49 com baixo (grupo B). Todas combinações de genótipos do GSTT1 e GSTM1 foram encontrados. O genótipo nulo dos dois genes do grupo de alto risco foi encontrado em 5,8% e no de baixo risco em 6,1%. CONCLUSÃO: A presença ou deleção dos genes GST (GSTT1 e GSTM1) não são bom fatores prognósticos no câncer papilífero da tireoide.27928

Efeitos da suplementação do 17beta-estradiol no dano oxidativo cardíaco de ratas submetidas à privação dos hormônios ovarianos Effects of 17beta-estradiol replacement on cardiac oxidative damage in rats submitted to ovarian hormone deprivation

OBJETIVO: avaliar o estresse oxidativo em tecido cardíaco de ratas ooforectomizadas, com ou sem terapia hormonal. MÉTODOS: ratas Wistar foram divididas em três grupos: grupo controle (GC), grupo ooforectomizada (GO) e grupo ooforectomizada + suplementação hormonal (GOS). A privação estrogênica foi obtida pela ooforectomia bilateral. Uma semana após a ooforectomia, um pellet de 1,5 mg de 17beta-estradiol foi implantado nos animais do grupo GOS. Nove semanas após a ooforectomia, o tecido cardíaco foi obtido para a análise do estresse oxidativo por meio da medida da quimiluminescência e da atividade das enzimas antioxidantes catalase (CAT), superóxido dismutase (SOD) e glutationa peroxidase (GPx). RESULTADOS: a quimiluminescência estava aumentada no GO (7348±312 cps/mg proteína) quando comparado ao GC (6250±41 cps/mg proteína, p<0,01), mas não houve diferença significante entre GC e GOS (6170±237 cps/mg proteína). A ooforectomia reduziu a atividade da SOD (22%, p<0,001) e da CAT (35%, p<0,05) no GO comparado ao GC. A terapia hormonal normalizou a atividade das enzimas antioxidantes no GOS. Não houve significância estatística na atividade da GPx quando os grupos estudados foram comparados. CONCLUSÕES: a privação dos hormônios ovarianos induziu aumento do estresse oxidativo e redução das defesas antioxidantes no tecido cardíaco. No entanto, a terapia hormonal preveniu o estresse oxidativo após a ooforectomia, provavelmente devido a um aumento das enzimas CAT e SOD no músculo cardíaco. Esses achados sugerem uma importante participação do estresse oxidativo nas disfunções cardiovasculares observadas em mulheres após a menopausa, reforçando a importância da terapia hormonal para o manejo dos riscos de doenças cardiovasculares neste grupo de mulheres.<br>PURPOSE: to evaluate oxidative stress in cardiac tissue of ovariectomized rats, with and without hormonal therapy. METHODS: female Wistar rats were divided in three groups: control group (CG); ovariectomized group (OG); ovariectomized group with estrogen supplementation (ESG). The estrogen deprivation was done through bilateral ovariectomy. After one week from the ovariectomy, a pellet of 1.5 mg of 17beta-estradiol was implanted in the ESG animals. Nine weeks after the ovariectomy, cardiac tissue was obtained for the analysis of the oxidative stress through CL (chemiluminescence), and measurement of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and gluthatione peroxidase (GPx). RESULTS: CL was increased in the OG (7348±312 cps/mg protein) when compared with the CG (6250±41 cps/mg protein, p<0.01), but there was no significant difference between the CG and the ESG (6170±237 cps/mg protein). Ovariectomy reduced SOD (35%, p<0.05) and CAT (22%, p<0.001) activities in the OG as compared with the CG. Hormonal therapy normalized antioxidant enzymes activities in the ESG. There was no statistically significant difference in GPx activity among the groups studied. CONCLUSIONS: ovarian hormone deprivation induced an increase of oxidative stress with reduction of antioxidant defenses in the cardiac tissue. However, hormonal therapy prevented oxidative stress after ovariectomy, probably due to an increase of the CAT and SOD activities in the cardiac muscle. These findings suggest an important oxidative stress contribution in cardiovascular dysfunctions observed in women after menopause, reinforcing the importance of hormonal therapy in the management of cardiovascular diseases risk in this group of women

Predictive Potential of Twenty-Two Biochemical Biomarkers for Coronary Artery Disease in Type 2 Diabetes Mellitus

We investigated the potential of a panel of 22 biomarkers to predict the presence of coronary artery disease (CAD) in type 2 diabetes mellitus (DM2) patients. The study enrolled 96 DM2 patients with (n = 75) and without (n = 21) evidence of CAD. We assessed a biochemical profile that included 22 biomarkers: total cholesterol, LDL, HDL, LDL/HDL, triglycerides, glucose, glycated hemoglobin, fructosamine, homocysteine, cysteine, methionine, reduced glutathione, oxidized glutathione, reduced glutathione/oxidized glutathione, L-arginine, asymmetric dimethyl-L-arginine, symmetric dimethyl-L-arginine, asymmetric dimethyl-L-arginine/L-arginine, nitrate plus nitrite, S-nitrosothiols, nitrotyrosine, and n-acetyl-β-glucosaminidase. Prediction models were built using logistic regression models. We found that eight biomarkers (methionine, nitratate plus nitrite, n-acetyl-β-glucosaminidase, BMI, LDL, HDL, reduced glutathione, and L-arginine/asymmetric dimethyl-L-arginine) along with gender and BMI were significantly associated with the odds of CAD in DM2. These preliminary findings support the notion that emerging biochemical markers might be used for CAD prediction in patients with DM2. Our findings warrant further investigation with large, well-designed studies