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

Imunoexpressão de vegf-c, vegfr-3 e hif-1α e mensuração da densidade linfática em carcinomas epidermóides de lábio inferior metastáticos e não-metastáticos: uma relação com parâmetros clinicopatológicos e prognósticos

Squamous cell carcinoma of the lower lip is among the most common malignant tumors of the oral and maxillofacial region, with good prognosis in more than 90% of patients with 5-year survival. In these carcinomas, the development of lymph node metastasis decreases the prognosis and it has been associated with the formation of new lymphatic vessels. It has been suggested the important role of vascular endothelial growth factor-C (VEGF-C), the receptor type 3 VEGF (VEGFR-3) and hypoxia-induced factor 1 (HIF-1) in this process. The aim of this study was to evaluate the immunoexpression of VEGF-C, VEGFR-3 and HIF-1α and correlate with intra and peritumoral lymphatic density in squamous cell carcinomas of the lower lip metastatic and non-metastatic. The sample consisted of 50 cases of squamous cell carcinoma of lower lip, of which 25 had regional lymph node metastasis and 25, absence of metastasis. The percentages of cells immunostained for VEGF-C, VEGFR-3 and HIF-1α in front of tumor invasion and in the center of tumor were evaluated. Microvessel density lymphatic (MDL) was determined by the counting of lymph microvessels immunostained by the anti-D2-40 in five fields (200×), in an area of evaluation with 0.7386 mm2. The invasion of the lymph vessels by malignant cells was also evaluated. Immunostaining was correlated with the presence and absence of metastasis, TNM clinical stage, local recurrence, disease outcome (remission of injury or patient death) and histological grading. The analysis of intra and peritumoral lymphatic density showed no significant association with clinicopathological parameters and immunoexpressions of VEGF-C, VEGFR-3 and HIF-1α (p > 0,05). There was a weak positive correlation, significant, between intra and peritumoral lymphatic density (r = 0,405; p = 0,004). VEGF-C showed no significant association with clinicopathological and prognosis parameters (p > 0,05). For VEGFR-3, there was scarce membrane staining and intense and homogenous cytoplasmic staining in neoplastic cells. Percentage of positive cytoplasmic VEGFR-3 in center of tumor, exhibited a statistically significant association with metastasis (p = 0,009), patient death (p = 0,008) and histological grades of malignancy proposed by Bryne et al. (1992) (p = 0,002) and World Health Organization (p = 0,003). A low positive correlation was statistically significant between the immunoreactivity of VEGFC and VEGFR-3 cytoplasmic (r = 0,358; p = 0,011) and between the percentage of positive cytoplasmic VEGFR-3 in front of tumor invasion and in the center of the tumor (r = 0,387; p = 0,005) was also demonstrated. There was no association between HIF-1α, clinicopathological and prognosis parameters, and VEGF-C and VEGFR-3. The percentage of nuclear positivity for HIF-1α was significantly higher in cases without invasion of peritumoral lymphatic (p = 0,040). Based on the results we can conclude that most cytoplasmic expression of VEGFR-3 in center of tumor in metastatic cases, high degree of malignancy and poorly differentiated, contributes to poor outcome of squamous cell carcinoma of the lower lip, including patient death. Intra and peritumoral lymphatic density seems to be not associated with lymph node metastasis in these carcinomas

Estudo in vitro dos efeitos da BMP-2 e do seu antagonista Noggin sobre a proliferação e migração celulares em carcinoma epidermóide de língua

Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the oral cavity and reach a large number of individuals, has become an important public health problem. Studies have demonstrated changes in pathway components BMP in various types of cancers as prostate, colon, breast, gastric and OSCCs. Is the current knowledge that these proteins may exert pro-tumor effect in more advanced stages of neoplastic development coming to favor progression and invasion tumor. The inhibition of the signaling pathway BMP-2 through its antagonists, have shown positive results of antitumor activity and use of Noggin may be a novel therapeutic target for cancer. Given this evidence and the few studies with BMP-2, Noggin and OSCC, the objective of this research was to evaluate the effect of BMP-2 and its antagonist Noggin on proliferation and migration cell in line of cell cultures of human tongue squamous cell carcinoma (SCC25). The study was divided in three groups, a control group, where SCC25 cells suffered no treatment, a BMP-2 group, in which cells were treated with 100ng/ml of BMP-2 and a group of cells that were treated with 100ng/ml of Noggin. For the proliferation assay and cell cycle were established three time intervals (24, 48 and 72 hours). Proliferative activity was investigated by trypan blue and cell cycle analysis by staining with propidium iodide flow cytometry. The potential for migration / invasion of SCC25 cells was performing by a cell invasion assay using Matrigel in a 48-hour interval. The proliferation curve showed a higher proliferation in cells treated with BMP-2 in 72 hours (p < 0.05), and lower overgrowth and cell viability in Noggin group. Recombinant proteins favored a greater percentage of cells in cell cycle phase Go/G1 with a statistically significant difference in the interval of 24 hours (p < 0.05). BMP- 2 produced a greater invasion of cells studied as well as its antagonist Noggin inhibits invasion of cells (p < 0.05). Thus, these results indicate that BMP-2 promotes malignant phenotype, dues stimulates proliferation and invasion of SCC25 cells and, its antagonist Noggin may be an alternative treatment, due to inhibit the tumor progression

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

Parâmetros parasitológicos e desempenho produtivo de ovinos naturalmente infectados com nematoides gastrintestinais mantidos em diferentes pastagens tropicais

The control of gastrointestinal nematodes of small ruminants is usually done with anthelmintics. However, due to the emergence of ever-increasing parasite resistance to these drugs, looking up an alternative control parasites. One of this is sought in pasture management, as these are the sources of animals` infection by L3 infective larvae of helminths. The aim of this study was to evaluate the performance of sheep to natural infections by gastrointestinal nematodes grazing Panicum maximum cv. Massai and cv. Aruana, and Brachiaria brizantha cv. Piatã and cv. Marandu. The work was conducted from May to August-2011 with 48 male sheeps SRD versus Santa Inês breed. The animals were naturally infected with gastrointestinal nematodes, and maintained in four different cultivars of tropical forage grasses, naturally contaminated with eggs and larvae of gastrointestinal nematodes. Each week the animals were phenotypically characterized by parasitological examinations (eggs per gram of feces - EPG, and feces culture), hematological (packed cell volume PCV, and blood eosinophil count) method to evaluate the Famacha© colorof ocular mucosa, and the measures of body condition score and weight. In pastures was made the recovery of infective larvae in order to determine the quantity of L3 present in the pasture. The experimental design was a randomized completed block with two replications and before the entry of animals in the paddocks, they have been wormed. The experiment was ended when the animals reached 32.0 kg liveweight, and then were slaughtered and autopsies performed for the recovery and identify parasites of the abomasum, small intestine and large intestine. Results were subjected to analysis of variance, t test and Tukey`s test. The animals kept on pastures of Marandu grass had lower EPG counts, higher percentage of packed cell volume and higher average weight; those who remained in the Piatã pasture had lower eosinophil counts per microliter of blood. About the Famacha©, the highest prevalence was Famacha 2, and the body condition score ranged between 2 and 3. The results of feces cultures and recovery of larvae on pastures showed the presence of larvae of Trichostrongylus sp., and at the necropsy too. This way, it was concluded that the grass cultivars influences the sheep parasite load; the Famacha, together with EPG and packed cell volume are important indicators for use in controlling gastrointestinal nematode infections. The Trichostrongylus sp. was the most prevalent parasite in sheep during the rainy season

Contribuição ao estudo da vascularização arterial do testículo de búfalos da raça Murrah

Estudamos o comportamento da artéria testicular, o número e a distribuição dos vasos penetrantes em 30 pares de testículos de búfalos da raça Murrah (Bubalus bubalis), com idade entre 8 meses e 5 anos, procedentes da região de Ilha Solteira, no Estado de São Paulo, mediante a análise de moldes obtidos pela injeção de vinil, na artéria testicular e em seguida corrosão do órgão em ácido sulfúrico a 30%. Encontramos mais freqüentemente (68,4%) a artéria testicular cedendo dois ramos, o cranial e o caudal, com participação equivalente de ambos os ramos na vascularização do órgão (35,0%), ou com predominância do ramo cranial (21,7%) ou do ramo caudal (11,7%). em outros arranjos, a artéria testicular cede três ramos: o cranial, o médio e o caudal (20,0%) ou ainda número variável de ramos craniais (4 a 7) e caudais (3 a 5) (11,6%) para a vascularização arterial do testículo.

Adubação nitrogenada e potássica no desenvolvimento de mudas de maracujazeiro

Este trabalho objetivou avaliar os efeitos da adubação com N e K no desenvolvimento de mudas de maracujazeiro-amarelo (Passiflora edulis) cultivadas em Latossolo Vermelho distrófico. O delineamento experimental foi o inteiramente casualizado, com três repetições, num esquema fatorial 4 x 4, com 4 doses de N (zero, 150, 300 e 600mg de N dm-3) e quatro de K (zero, 75, 150 e 300mg de K dm-3), parceladas em quatro vezes. A unidade experimental foi constituída por vasos de 3,3L. Após 84 dias da germinação, foram avaliados: altura da planta, o diâmetro do caule, o número de folhas e a área foliar. O melhor desenvolvimento das mudas de maracujazeiro ocorreu com as doses de 150mg de N dm-3 e de 300mg de K dm-3.

Efeitos alelopáticos de extratos vegetais na germinação, colonização micorrízica e crescimento inicial de milho, soja e feijão

Restos vegetais e liteira podem interferir no desenvolvimento de plantas. Este trabalho objetivou avaliar os efeitos alelopáticos de extratos aquosos de Pinus sp., milheto (Pennisetum americanum (L.) Leeke) e mucuna (Stizolobium aterrimum Piper & Tracy) sobre a germinação, colonização micorrízica e crescimento inicial de milho (Zea mays L.), soja (Glycine max L.) e feijão (Phaseolus vulgaris L.). Os extratos foram elaborados utilizando folhas trituradas. O experimento in vitro empregou papel Germitest umedecido com extratos ou água destilada por sete dias. O experimento em casa de vegetação teve esquema fatorial 3 x 3 x 4, com quatro repetições: três espécies vegetais (soja, milho e feijão), três extratos aquosos (Pinus, milheto e mucuna) e quatro doses de extrato (0,0; 0,5; 1,0; e 2,0 kg L-1). O substrato foi Latossolo Vermelho coletado no município de Selvíria-MS, no bioma Cerrado. Após a semeadura, os vasos receberam, a cada cinco dias, por 45 dias, 50 mL dos extratos. Para a soja, extratos de mucuna e milheto diminuíram o comprimento do hipocótilo e da radícula e os de Pinus aumentaram esses comprimentos. em feijão, o extrato de Pinus diminuiu o comprimento do hipocótilo e da radícula, mas os extratos de mucuna e milheto aumentaram-no. O extrato do milheto reduziu a percentagem e a velocidade de germinação em feijão. Todos os extratos reduziram a colonização micorrízica e o número de esporos de fungos micorrízicos arbusculares em soja, milho e feijão.

Efeitos de baixas pressões no balonete da máscara laríngea na mucosa faringolaríngea do cão

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

Uso de dexmedetomidina em neurocirurgia

JUSTIFICATIVA E OBJETIVOS: Os fármacos alfa2-agonistas são a cada dia mais utilizados em Anestesiologia, seja como adjuvantes ou como agentes anestésicos únicos. Atualmente, o emprego da dexmedetomidina vem se popularizando devido à sua maior seletividade aos receptores alfa2 e, também, ao seu perfil farmacocinético. O objetivo desta revisão foi fazer uma análise do emprego da dexmedetomidina em neurocirurgia. CONTEÚDO: Além das considerações e revisão da literatura quanto ao emprego da dexmedetomidina especificamente em procedimentos neurocirúrgicos, foi realizada descrição dos efeitos do fármaco nos diversos sistemas do organismo. CONCLUSÕES: A dexmedetomidina tem perfil farmacocinético e farmacodinâmico que favorece seu emprego em diversos procedimentos neurocirúrgicos. A utilização clínica em procedimentos cirúrgicos com craniotomia para pinçamento de aneurisma e remoção de tumores é crescente. Além disso, seu uso em intervenções cirúrgicas funcionais é promissor.

Peritoneal pseudomyxoma associated with synchronic malignant mucinous neoplasias of the cecum, appendix and rectum: case report and review of the literature

O pseudomixoma peritoneal é uma condição patológica que acomete o peritoneo, caracterizada pela produção de grandes quantidades de líquido mucinoso, que progressivamente preenche a cavidade peritoneal, tendo em geral como origem tumores mucinosos apendiculares ou de ovário. Relatamos a ocorrência de um pseudomixoma peritoneal associado a adenocarcinoma mucinoso do apêndice sincrônico e adenocarcinoma do reto em paciente de 44 anos, cujo diagnóstico inicial foi de adenocarcinoma do reto. A neoplasia do apêndice e o pseudomixoma peritonial foram achados incidentais, intra-operatórios. Enfocamos as principais características anatomo-patológicas das lesões, o diagnóstico e tratamento, através de ampla revisão da literatura.

Melanoma anorretal: relato de 2 casos e revisão da literatura

Os tumores malignos do canal anal e do anus são muito raros, não ultrapassam 2% de todos os tumores do colo, reto e anus; segundo os principais levantamentos os melanomas não ultrapassam a incidência de 0,1 a 1,2% dos tumores malignos. Os autores descrevem 2 casos de melanoma, discutindo os principais dados da literatura, enfocando os aspectos diagnósticos, tratamento, evolução e prognostico. Os indicies de cura s são baixos e com elevados índices de mortalidade a curto prazo.

Quilopericárdio idiopático primário: relato de caso

O acúmulo de quilo no espaço pericárdico ou quilopericárdio é uma condição que, com maior frequência, ocorre após trauma, cirurgia cardíaca e torácica ou associado a tumores, tuberculose ou linfoangiomatose. Quando não é possível a identificação precisa da etiologia, o quilopericárdio é denominado primário ou idiopático. Essa é uma situação clínica rara. Descrevemos um caso em paciente do sexo feminino, com 20 anos de idade, tratada cirurgicamente. A propósito do caso, apresentamos breve revisão da literatura e comentários sobre quadro clínico, etiopatogenia, exames diagnósticos complementares e opções de tratamento.

Origem anômala de artéria coronária esquerda do tronco pulmonar: síndrome de Bland-White-Garland

A origem anômala da artéria coronária esquerda a partir do tronco pulmonar, conhecida como síndrome de Bland-White-Garland (BWG), é uma doença rara, que, habitualmente, leva à morte antes do primeiro ano de vida. Os autores relatam o caso de uma criança branca, com 2 anos e 6 meses de idade, portadora da síndrome de BWG, e revisam a apresentação clínica, a fisiopatologia, o diagnóstico e o tratamento desses pacientes.