BUBR1 expression in benign oral lesions and squamous cell carcinomas: Correlation with human papillomavirus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Assessment of DNA damage induced by extracts, fractions and isolated compounds of Davilla nitida and Davilla elliptica (Dilleniaceae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Development of praziquantel-loaded PLGA nanoparticles and evaluation of intestinal permeation by the everted gut Sac model

Praziquantel has been shown to be highly effective against all known species of Schistosoma infecting humans. Spherical nanoparticles made of poly(D,L-lactide-co-glycolide) acid with controlled size were designed as drug carriers. Praziquantel, a hydrophobic drug, was entrapped into the polymeric nanoparticles with 30% (w/w) of theoretical loading. The nanoparticles size was approximately of 350 nm with 66% of encapsulation efficiency. The everted gut sac model shows to be efficient to evaluate the drug permeation through the intestinal membrane. The results show that free praziquantel presents 4-fold times more permeation than praziquantel-loaded PLGA nanoparticles and physical mixture. For this drug, in special, this result can be interesting, since the nanoparticulate system can behave as a drug reservoir and/or to have a more localized effect in intestinal membrane for a prolonged period of time, since great amounts of parasites can be usually found in the mesenteric veins.

Dermatophyte-host relationship of a murine model of experimental invasive dermatophytosis

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

Estudo da imunoexpressão dos transportadores de glicose 1 e 3 em carcinomas epidermóides de lábio inferior e sua relação com parâmetros clínico-patológicos

The Epidermoid Carcinoma (EC) is the most common lesions located in the region of the head and neck and, despite advances in treatment modalities, the prognosis is still poor. The malignant cells show an increase in glucose uptake, process mediated by glucose transporters (GLUTs). Increased expression of GLUT 1 and GLUT 3 is related to the aggressive behavior of this lesion. The aim of this study was to evaluate, through immunohistochemistry, the expression of GLUTs 1 and 3 in EC of the lower lip. The sample consisted of 40 cases of EC of the lower lip, of which 20 had regional lymph node metastasis and the remaining 20 with absence of metastasis. The percentages of immunostained cells in front of tumor invasion and in the center of tumor were evaluated. These results were related to the presence and absence of lymph node metastasis, TNM classification and histological grading. The percentage of cytoplasmic/membranous expression of GLUT 1 ranged from 77.35% to 100%, while for GLUT 3 this value ranged from 0.79% to 100%. As for nuclear staining for GLUT 1, this percentage ranged from 0 to 0.42%, however. GLUT 3 showed only one case with nuclear staining. Despite the significant expression of tumor cells related to the proteins studied, we observed no statistically significant relationship between the variables and the antibodies analyzed, regardless of the region evaluated. However, there was a moderate positive correlation between cytoplasmic/membranous immunoexpressions of GLUT 1 in invasion front and in the tumor center (r = 0.679, p <0.001). Similarly, moderate positive correlation was found between the nuclear immunoexpressions of GLUT 1 in the invasion front and in the tumor center (r = 0.547, p <0.001). For GLUT 3, was also observed a moderate statistically significant positive correlation between cytoplasmic/membranous expression in tumor invasion front and in tumor center (r = 0.589, p <0.001). We also observed that the immunoreactivity for GLUT 1 was higher than GLUT 3 expression in invasion front (p <0.001) and tumor center (p <0.001). From these results, this study suggests that tumor hypoxia is a remarkable characteristic of the EC of the lower lip and GLUT 1 may be primarily responsible for glucose uptake into the interior of the malignant cells

Estudo da influência de linfócitos tcd-8 e das células nk em casos de carcinoma epidermóide de lábio inferior sem metástase e com metástase e sua relação com a progressão da lesão

The presence of inflammatory cells within the tumor microenvironment plays a dual role that may contribute both to the progression and for inhibition of tumor growth. Recent studies suggest that the quality, not the quantity, of the inflammatory infiltrate is the most important determinant for prognosis. Therefore, TCD8 cells and natural killer cells are the main effector cells in combating cancer. The aim of this study was to assess, through the immunohistochemical study, the expression of TCD8 lymphocytes and NK cells in epidermoid carcinoma (EC) of the lower lip. The sample consisted of 32 specimens of EC of the lower lip, of which 16 had regional lymph node metastasis, and the 16 remaining, free of metastases. The total number of positive cells at the front of invasion were evaluated quantitatively and the results were related to clinical TNM staging, histological grade of malignancy and prognostic factors. It was observed for the group with metastasis, prevalence of stages III and IV (p<0.0001). Most patients with metastasis, had a high grade of malignancy (p=0.006). Most cases classified as high grade of malignancy had stages III and IV (p=0.032). Of the total sample, there were three cases of recurrence and five with death, however these variables were not statistically significant when associated with clinicopathological parameters. The immunostaining of CD8 and CD57, respectively, showed no statistically significant association with any of the clinicopathological parameters studied, metastasis (p=0.346, p=0.622), TNM classification (p=0.146, p=0.576), histological grade of malignancy (p=0.936, p=936), recurrence (p=0.075, p=0.075) and death (p=0.897, p=0.856). Believing in the function of the immunological system against malignant cells, it is concluded that the TD8 lymphocytes and NK cells, would be acting in the control of the progression of malignant neoplasms, but not in isolated manner

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

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.

Electromyographical, ultrasonographical and morphological modifications in semitendinous muscle after transposition as ventral perineal muscle flap

The aim of this study was to evaluate modifications occurring in semitendinous muscle after transposition as a ventral perineal muscle flap using electromyography, ultrasonography, and morphological studies. Ten male crossbreed dogs of 3-4 year old were used. The left semitendinous muscle was cut close to the popliteus lymph node, rotated and sutured at the perineal region. The contralateral muscle was considered as control. Motor nerve conduction studies of both sciatic-tibial nerves, and electromyographic and ultrasonographic examinations of both semitendinous muscles were performed before surgery and 15, 30, 60, and 90 days postoperatively. Semitendinous muscle samples were collected for morphological analysis 90 days after surgery. No alterations were observed in clinical gait examinations, or in goniometrical and electroneuromyographical studies in pelvic limbs after surgery. Electromyography demonstrated that the transposed muscle was able to contract, but atrophy was detected by ultrasonography and morphological analysis.

Estudo da ação do extrato de Ginkgo biloba e amido hidroxietílico hipertônico na atenuação de alterações decorrentes de isquemia e reperfusão de órgãos esplâncnicos em ratos

A oclusão e reperfusão das artérias esplâncnicas ocasiona choque circulatório, causado principalmente pelo aumento de permeabilidade vascular e pela agressão celular provocada por radicais livres derivados do oxigênio. Este estudo tem por finalidade verificar a ação do extrato de Ginkgo biloba (Egb-761) e do amido hidroxietílico (AHH) na prevenção do choque circulatório produzido pela isquemia e reperfusão de órgãos esplâncnicos. O Egb-761 tem propriedades antioxidantes relatadas na literatura. O AHH, tem sido utilizado como recurso terapêutico do choque hipovolêmico. Ratos anestesiados receberam infusão contínua de Egb-761 ou AHH, sendo submetidos à isquemia (oclusão do tronco celíaco, artéria mesentérica superior e artéria mesentérica inferior por 30 minutos) e reperfusão (por 90 minutos) dos órgãos esplâncnicos. Foram feitas: análise histopatológica ileal, dosagem de malondialdeído ileal e determinação contínua da pressão arterial média (PAM). A PAM ao final do período de reperfusão foi significativamente mais elevada nos animais tratados com Egb-761 e AHH, que no grupo controle (F=18,29; p<0,001). Não houve diferença entre os grupos tratados e controle quanto à dosagem de MDA (H=4,61; p>0,10) e quanto às alterações histológicas (H=6,003; p>0,10). em conclusão, houve melhora nas condições hemodinâmicas, com atenuação do choque nos ratos que receberam Egb-761 ou AHH. Novos estudos serão necessários para se avaliar melhor as alterações histológicas e para esclarecer a formação de produtos finais da peroxidação lipídica.

Effects of pentoxifylline and n-acetylcysteine on injuries caused by ischemia and reperfusion splanchnic organs in rats

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

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.

Efeito do sítio de venopunção nos parâmetros hematológicos em tigre-d'água-americano, Trachemys scripta elegans

O objetivo do presente estudo foi realizar a comparação entre dois sítios de coleta sanguínea em 24 exemplares de tigre-d'água-americano (Trachemys scripta elegans) oriundos de um criadouro comercial, localizado no município de Antonina, litoral do Paraná, Brasil. Os animais foram submetidos a contenção física e as venopunções foram realizadas no seio supraocciptal e na veia coccígea dorsal. As amostras heparinizadas foram identificadas e refrigeradas para posterior análise laboratorial. A contagem total de eritrócitos e leucócitos foi realizada pela técnica de hemocitometria. O hematócrito (Ht) e a hemoglobina (Hb) foram determinados pelo método de microhematócrito e cianometahemoglobina, respectivamente. A proteína plasmática total (PPT) foi determinada por refratometria e a contagem diferencial de leucócitos foi realizada através da técnica de Shilling. Houve diferença significativa no número de leucócitos e no valor da proteína plasmática total, e em ambos os casos os valores encontrados nas amostras provenientes da veia coccígea dorsal foram inferiores. A diferença encontrada no número de leucócitos provavelmente foi devido à contaminação por linfa, que também justifica o menor valor na concentração da proteína plasmática total. Conclui-se que é mais indicada a venopunção no seio supraocciptal quando comparado a veia coccígea dorsal.

Paracoccidioidomycosis in a Dog: Case Report of Generalized Lymphadenomegaly

Paracoccidioidomycosis (PCM) is a severe systemic mycosis, endemic in Latin America and highly prevalent in Brazil, where it ranks eighth as a mortality cause among infectious and parasitic diseases in humans. The disease in animals has been little explored. It is observed that armadillos can harbor the fungus at high frequencies, although the active disease has not been well documented in this wild mammal. Dogs are susceptible to experimental infection, and the naturally acquired PCM-disease was reported only recently in a dog from Brazil. The present work reports the second case of naturally acquired PCM in a 6-year-old female dog that presented emaciation, lymphadenomegaly, and hepatosplenomegaly. Biochemical and pulmonary radiographic evaluation did not reveal any abnormalities. PCM was diagnosed by clinical findings, culturing, immunohistochemistry, and histopathology of popliteal lymph node. The fungus was recovered from popliteal lymph node, and the molecular analysis showed respective sequencing similarities of 99 and 100% for 803 nucleotides of the Gp43 gene and 592 nucleotides from the ITS-5.8S region of Paracoccidioides brasiliensis. Immunohistochemistry revealed severe lymphadenitis and presented numerous yeasts, which reacted against the gp43 antibody. Histopathology revealed a severe granulomatous lymphadenitis associated with numerous single or multiple budding yeasts. After diagnosis, the dog was successfully treated with itraconazol for 2 years. Veterinarians should be aware of the importance of considering PCM for differential diagnosis, especially in dogs from PCM-endemic areas, whose monophagocytic system involvement is evident.

Association between CD31 expression and histopathologic features in stage IB squamous cell carcinoma of the cervix

This study was undertaken to evaluate the association between the expression of CD31 in the tumor and the histopathologic findings in patients with carcinoma of the cervix. This study included prospectively 30 women, aged 46.6 +/- 10.7 years, with stage IB squamous cell carcinoma of the cervix submitted to radical hysterectomy from November 2001 to September 2002. Samples from the tumor were taken and immunohistochemically evaluated by a monoclonal antibody for CD31. Clinicopathologic characteristics such as stage, tumor size, grade of differentiation, lymphatic vascular space invasion (LVSI), parametrial involvement, and status of pelvic lymph nodes were also recorded. The clinical stage (FIGO) was IB1 in 22 patients (73.3%) and IB2 in 8 patients (26.7%). The expression of CD31 was significantly associated with tumor size and the presence of LVSI, but not with grade of differentiation and vaginal or parametrial involvement (P= 0.03, P= 0.032, P= 0.352, P= 0.208, and P= 0.242, respectively). on univariate analysis, the presence of pelvic lymph node metastasis was influenced by LVSI (P= 0.003) and CD31 expression (P= 0.032). However, on multivariate analysis, the presence of LVSI (P= 0.007) was the only independent predictor of pelvic lymph node metastasis. The CD31 expression in tumor is significantly associated with LVSI and tumor size in patients with early-stage squamous cell carcinoma of the cervix.