Alterações nos genes da E-caderina e β-catenina em adenoma pleomórfico e carcinoma adenóide cístico: estudo molecular e imuno-histoquímico

Pleomorphic adenoma and adenoid cystic carcinoma represent a benign and malignant salivary gland neoplasm, respectively, that shares the same histological origin, however with distinct biological behavior. The aim of the present study was identify the -160 C/A polymorphism in the gene CDH1, mutational analysis of CTNNB1 gene and evaluation the expression of the E-cadherin and β-catenin in pleomorphic adenomas and adenoid cystic carcinomas. Furthermore, it was proposed correlate the immunochemistry staining patterns with the polymorphism and mutations. Twenty-four pleomorphic adenomas and 24 adenoid cystic carcinomas were retrieved. The polymorphism analysis was performed by restriction fragment length polymorphism (RFLP), using the restriction enzymes HphI or AflIII and the mutational screening was performed by PCR-single strand conformational polymorphism (PCR-SSCP). The immunohistochemical analysis was taken by the counting of cells, recorded as the Hscore index, and considering the presence or absence, intensity, distribution and localization of proteins expression. Comparing the two neoplasms, the results demonstrated statistically significant difference for the E-cadherin and β-catenin expression, with pleomorphic adenoma presenting weaker immunostaining. Was observed statistical correlation between E-cadherin and β-catenin expression. CDH1 heterozigotic polymorphism was seen in two cases and 13 cases displayed abnormal mobility electrophoretic shifts, suggesting CTNNB1 gene mutation. The immunohistochemical expression was not statistically correlated with the polymorphism or suggested mutations. In conclusion this study supports that the E-cadherin/β-catenin complex immunohistochemical expression might be related with the myoepithelial component amount and differentiation neither the tumor biological behavior. The cases that showed E-cadherin gene polymorphism presented reduced protein expression and, moreover, CTNNB1 suggested mutations seem not influence in the β-catenin protein expression

Estudo da expressão imuno-histoquímica das MMPs -2, -7, -9 e -26 e TIMPs -1 e -2 em adenomas pleomórficos e carcinomas adenóides císticos de glândulas salivares menores

The balance between the expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) has been related to various physiological and pathological processes, including salivary gland morphogenesis and tumor invasion and metastasis processes. Pleomorphic adenoma (PA) and adenoid cystic carcinoma (ACC) respectively represent benign and malignant neoplasias of salivary glands. Although they share the same cell origin, they present distinct biological behavior. The aim of this study was to compare the immunohistochemical expression of MMPs -2, -7, -9 and -26, and of TIMPs -1 and -2, in cases of PA and ACC of minor salivary glands. Twenty cases of PA and twenty cases of ACC were assessed according to the presence, intensity and location of MMPs and TIMPs in the tumor parenchyma. Most of the PAs and ACCs presented a high expression of MMP -2, -7, -9 and -26 and of TIMP -1 and -2, predominantly located in tumor cells. There was no significant difference in the expression of MMPs -2 (p=0.359), -7 (p=0.081) and -26 (p=0.553), as well as of TIMPs -1 (p=0.657) and -2 (p=0.248), between the parenchyma of PAs and ACCs. However, MMP-9 showed a significant difference of expression between the two tumors, with the ACC showing more intense marking for this gelatinase (p=0.041). The strong expression of MMP-9 observed in the parenchyma suggests that this gelatinase may play an important role in the biological behavior of these tumors. On the other hand, although there was no significant difference between the marking of MMP -2, 7 and 26 in the studied tumors, the data, when analyzed as a whole, suggest that these proteases may take part in the process of tissue remodeling in both tumors, but do not present a direct relation with the pattern of aggressiveness of ACC. Nonetheless, matrilisins may indirectly influence the behavior of this tumor due to their capacity of activating MMP-9, strongly expressed in the parenchyma of ACC

Expressão imuno-histoquímica das integrinas a2ß1, a3ß1 e a5ß1 em adenoma pleomórfico de glândula salivar menor e maior e carcinoma adenóide cístico

Pleomorphic adenoma and adenoid cystic carcinoma (ACC) consist benign and malignant neoplasm from salivary gland, respectively. These neoplasms share some characteristics, such as cellular origin and considerable production of extracellular matrix, however, with distinct biological behavior. The aim of the present study was to compare the expression of D2E1, D3E1 e D5E1 integrins in pleomorphic adenoma from minor and major salivary glands and ACCs. Furthermore, it was investigated possible differences in the expression of these integrins according to histological subtypes of ACC. Fourteen cases of pleomorphic adenoma from major salivary gland, fourteen cases from minor salivary gland and ten cases of ACC were selected. It was taken into consideration the presence or absence, localization and intensity of integrin immunoexpression. The cases of pleomorphic adenoma were grouped in order to compare the expression between the distinct neoplasms. It was observed a highly significant difference (p<0,0001) in relation to D2E1 integrin between the neoplasms since pleomorphic adenoma showed a pronounced immunostaining. It was not possible to perform statistical tests considering the D2E1 integrin expression; nevertheless, it could be observed a tendency of higher staining in pleomorphic adenoma. For comparative reasons the cases ACCs were divided in two groups: solid and tubular/cribriform. It was not detected significant differences in regard to D2E1 integrin; and statistical analysis could not be realized in relation to D3E1 and D5E integrin expression. However, it was also verified a tendency of absence or reduced expression in the solid subtype. It can be concluded that the reduced D2E1 integrin expression observed in CACs may be related to a lesser degree of cell differentiation in this neoplasm and the reduced D5E1 integrin expression can be associated with aggressive biological behavior. Moreover, the absence and/or reduced expression of the studied integrins in solid ACC suggests a role in pathogenesis and more aggressive biological behavior of this histological subtype

Glia-Pinealocyte Network: The Paracrine Modulation of Melatonin Synthesis by Tumor Necrosis Factor (TNF)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Lymphatic drainage on healthy and neoplasic mammary glands in female dogs: Can it really be altered?

The purpose of this research was to study the mammary lymphatic drainage under a macroscopic and mesoscopic view, comparing the vascular pattern of healthy and neoplasic mammary glands injected with drawing ink alcoholic and fluorescein solutions, in 46 mongrel female dogs. The results pointed out that the thoracic gland is drained by the axillary lymph centre, but in mammary neoplasia either superficial cervical or ventral thoracic lymph centres can be involved. Cranial and caudal abdominal glands may be drained by the axillary, inguinofemoral and popliteal lymph centres. However, the popliteal drainage is specific for the healthy caudal abdominal mammary gland. The inguinal gland can be drained by both inguinofemoral and popliteal lymph centres in both neoplasic and healthy conditions. Regarding the mammary lymphatic communications, this research demonstrated that neoplasic glands present more types of anastomosis (40.9%), than healthy glands (33.33%), and an increase in contralateral anastomosis (50%) compared with healthy ones (33%). Given the data, the mammary neoplasia can change the lymphatic drainage pattern in terms of lymph centres and vascular arborization, thus forming new drainage channels and recruiting a larger number of lymph nodes. Lastly, some comments were made about the severity of a specific neoplasic mammary gland and conditions to be considered before making a decision in terms of the most adequate operative procedure, and suggestions for further investigations.

Thyroid cell proliferation in Graves' disease - Use of MIB-1 monoclonal antibody

OBJECTIVE: To measure thyroid cell proliferation in patients with Graves' disease (GD) before and during treatment with antithyroid drugs.STUDY DESIGN: Patients were assessed by fine needle aspiration biopsy before (n=20) and after 4 (n=19) and 12 months of treatment (n=15) with propylthiouracil or methimazole. Cell proliferation index (CPI) was estimated by immunocytochemistry using MIB-1. CPI was studied in relation to the cytologic parameters of the smears; clinical parameters, such as Wayne's Clinical Index (WCI) and time without treatment; laboratory parameters, such as (131)Iuptake and dosage of serum free thyroxin and thyroid-stimulating hormone; and thyroid ultrasound.RESULTS: CPI varied from 0.00% to 25.00% before treatment, 0.00% to 23.00% at 4 months and 0.00% to 14.84% at 12 months. CPI median values were 6.50%, 4.30% and 3.30%, respectively (before and after 4 months and 12 months of treatment). CPI had a positive correlation with WCI and FT4 at 12 months of treatment.CONCLUSION: Thyroid CPI in GD varies from case to case. However, due to its decreasing pattern during follow-up and its positive correlation with thyrotoxicosis severity, CPI may indicate the functional status of the gland and contribute to a better understanding of GD.

A novel mutation (Q40P) in PAX8 associated with congenital hypothyroidism and thyroid hypoplasia: Evidence for phenotypic variability in mother and child

Congenital hypothyroidism associated with thyroid hypoplasia can be caused by several genetic defects, including mutations in the TSH beta -subunit, the TSH receptor, the G(A)alpha -subunit, and the transcription factor PAX8. Four girls with sporadic congenital hypothyroidism and hypoplastic thyroid glands were analyzed for mutations in PAX8 and TTF2 (FKHL15). Mutations in the coding region of the TSH beta -subunit gene, the TSH receptor gene, and exons 8 and 9 of G(mu)alpha had been excluded previously. Serum TSH concentrations were 150 mU/liter or more, TG levels were within normal limits, and thyroid autoantibodies were absent. Technetium scintigraphies did not reveal the presence of thyroid tissue, but ultrasonography documented hypoplastic, normally located glands.One patient was found to harbor a heterozygous transversion 119A -->C in exon 3 of PAX8 replacing a conserved glutamine by proline in the paired box domain (Q40P). Analysis of her family members revealed that her mother, who has a thyroid gland of normal size and mild, adult-onset autoimmune hypothyroidism, is also heterozygous for this mutation. Functional analyses of the PAX8 Q40P mutation showed impaired binding to a PAX8 response element and absent transactivation of a thyroid peroxidase promoter luciferase reporter gene.These findings confirm the important role of PAX8 in the development of the thyroid, but they indicate that PAX8 gene mutations may have a variable penetrance or expressivity. The absence of mutations in the coding sequences of the analyzed genes in the three other patients supports the concept that the pathogenesis of congenital hypothyroidism associated with thyroid hypoplasia is diverse.

Tireóide ectópica cervical lateral: Relato de caso e revisão da literatura

Tireóide ectópica é qualquer tecido tireoideano localizado fora de sua topografia habitual, podendo apresentar-se na linha mediana do pescoço ou, mais raramente, na região cervical lateral. Algumas teorias tentam explicar a origem do tecido tireoideano ectópico: 1. falha na descida da glândula; 2. seqüestro de nódulos tireoideanos; 3. presença de tecido tireoideano na cápsula de linfonodos cervicais; 4. formação teratomatosa; 5. Secundário a anomalias branquiais. Na abordagem diagnóstica, diversos exames têm sido utilizados, sendo o diagnóstico definitivo algumas vezes fornecido apenas pelo estudo histopatológico. Apesar das controvérsias em relação às abordagens terapêuticas apresentadas na literatura, é necessário um planejamento rigoroso para evitar iatrogenias. Conclui-se que a presença de tecido tireoideano ectópico deve ser lembrada no diagnóstico diferencial de massas cervicais laterais, e sua origem histológica considerada, sendo na maioria das vezes metástase de um carcinoma tireoideano oculto. Os autores relatam um caso de tecido tireoideano ectópico lateral no pescoço, em paciente do sexo feminino com bócio colóide mergulhante. Foram realizadas considerações importantes sobre dismorfogênese tireoideana, métodos diagnósticos e opções de tratamento, com revisão da literatura das últimas cinco décadas.

Perfil eletroforético e concentração de imunoglobulinas G (IgG) do soro sanguíneo de cabras Saanen com mastite experimental induzida por Staphylococcus aureus suplementadas com vitamina E

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hipertireoidismo na gravidez: repercussões materno-fetais

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Could zinc prevent reproductive alterations caused by cigarette smoke in male rats?

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Pituitary Apoplexy After a Single Dose of Long-Acting Octreotide

Pituitary apoplexy (PA) is a rare and potentially life-threatening syndrome resulting from an acute infarction or hemorrhage of the pituitary gland. Although the pathogenesis is not fully understood, some predisposing factors such as pituitary stimulation tests, diabetes mellitus, anticoagulant or antiplatelet aggregation therapy, head trauma, and high blood pressure may play a role in its pathophysiology. Octreotide is the mainstay of medical treatment for acromegaly. The majority of reported complications of octreotide therapy are gastrointestinal. We report the case of a 51-year-old acromegalic woman who developed pituitary apoplexy within the context of high blood pressure and a single dose of long-acting octreotide. Our data suggest that the combination of hypertension and octreotide therapy enhances the risk of pituitary apoplexy.

Controle de qualidade e dosimetria em equipamentos de tomografia computadorizada

OBJETIVO: Avaliação de condições dos equipamentos e dosimetria em setores de tomografia computadorizada utilizando protocolos de cabeça, abdome e coluna lombar em pacientes adultos (em três equipamentos distintos) e pediátricos com até um ano e meio de vida (em um dos equipamentos avaliados). MATERIAIS E MÉTODOS: Foram estimados o índice de dose em tomografia computadorizada e a dose média em cortes múltiplos, em exames com pacientes adultos, em três distintos equipamentos. Ainda foram estimadas as doses na superfície de entrada e as doses absorvidas em exame de cabeça para pacientes adultos e pediátricos em um dos equipamentos avaliados. RESULTADOS: Foram realizados testes de controle de qualidade, mecânicos, demonstrando que os equipamentos satisfazem as especificações de uso estabelecidas pelas normas vigentes. Os resultados da dosimetria mostraram que valores de dose média em cortes múltiplos excederam em até 109,0% os valores de níveis de referência, apresentando consideráveis variações entre os equipamentos avaliados neste estudo. As doses absorvidas obtidas com protocolos pediátricos são inferiores aos de pacientes adultos, apresentando redução de até 51,0% na tireoide. CONCLUSÃO: Neste estudo foram avaliadas as condições de operação de três equipamentos tomográficos, estabelecendo quais parâmetros devem ser trabalhados para a implantação de um programa de controle de qualidade nas instituições onde esta pesquisa foi desenvolvida.

Serum and urinary measurements of prostatic acid phosphatase (PAP) and prostatic specific antigen (PSA) in dogs

Realizaram-se mensurações sérica e urinária de fosfatase ácida prostática (PAP) e antígeno prostático específico (PSA) de 20 cães. Os testes de PAP e PSA foram feitos em um equipamento automatizado, com o uso de kits comerciais para humanos. A média de PAP sérico foi de 0,7U/l e urinário 0,U/l. As médias do PSA sérico e urinário foram 0,005ng/dL e 0,004ng/dl, respectivamente. A determinação do dois biomarcadores in vivo é uma nova opção de diagnóstico na medicina veterinária e os valores obtidos devem ser correlacionados com a lesão morfológica da próstata.

Óxido nítrico, GSTP-1 e p53: qual o papel desses biomarcadores nas lesões prostáticas do cão?

Confeccionou-se um microarranjo de tecido (TMA) com 146 amostras de lesões prostáticas caninas. Este continha 17,2% de hiperplasia prostática benigna (HPB), 32,4% de atrofia inflamatória proliferativa (PIA), 2,6% de prostatite, 8,6% de focos de neoplasia intraepitelial prostática (PIN), 29,1% de carcinomas e 9,3% de próstatas normais. Cortes histológicos sequenciais foram feitos e utilizados para reação de imunoistoquímica com os anticorpos primários anti-p-53, anti-NOS-2 e anti-GSTP. Avaliou-se de cada core o escore de células marcadas para cada anticorpo utilizado. Os resultados foram tabulados por grupo diagnóstico e submetidos ao teste Tuckey. Os carcinomas prostáticos do cão e a PIA apresentaram maior número de amostras (41) com mais de 75% das células positivas para NOS-2, demonstrando a influência do estresse oxidativo no desenvolvimento dessas lesões. As próstatas normais e as afecções desta glândula, HPB, PIA, PIN, prostatite e carcinoma, expressaram a proteína GSTP-1, o que conferiu proteção ao tecido prostático canino a danos oxidativos. A proteína p53 estava presente em todas as amostras estudadas, incluindo o tecido prostático normal, porém as lesões prostáticas apresentaram maior número de amostras com escores mais elevados de marcação (escores três e quatro), presente em 95% dos focos de PIA e carcinoma. Concluiu-se que o aumento de expressão de óxido nítrico nas lesões prostáticas no cão e a expressão de GSTP-1 podem ter protegido o tecido prostático canino e que a expressão de p53 foi positiva e uniforme nas próstatas normais e com lesões hiperplásicas e displásicas.