Estudo clínico da aplicação de matriz inorgânica de osso associado a peptídeo sintético de adesão celular (MIO/P-15), PepGen P-15®, em lesões periodontais avançadas de cães

Os mecanismos biológicos desenvolvidos para aumentar a qualidade da regeneração óssea e da reparação tecidual de sítios periodontais específicos continuam a ser um desafio e têm sido complementado pela capacidade de adesão celular do colágeno do tipo I, promovida por um peptídeo sintético de adesão celular (P-15), associado a uma matriz inorgânica de osso (MIO) para formar MIO/P-15. O objetivo deste estudo foi avaliar a perda do nível clínico de inserção e a resposta da bolsa periodontal em dentes após 3 e 6 meses da aplicação de enxerto com MIO/P-15. Vinte e um cães do Hospital Veterinário da Universidade de São Paulo foram anestesiados para realização de tratamento periodontal e 132 faces dentais com perda de nível clínico de inserção foram tratadas, sendo que 36,4% (48 faces) receberam o peptídeo de adesão celular e 63,6% (84 faces) compuseram o grupo controle que recebeu tratamento convencional (retalho muco-gengival e aplainamento radicular). O procedimento foi documentado através de radiografia intra-oral e todas as sondagens de bolsas periodontais foram fotografadas. Depois de 3 e de 6 meses, os animais foram re-anestesiados a fim de se obter novas avaliações, radiografias, fotografias e sondagens periodontais. As 48 faces com perda de nível clínico de inserção que receberam material de enxertia apresentaram taxa de 40% de recuperação do nível clínico de inserção após 6 meses. O grupo controle de faces dentais não apresentou alteração do nível clínico de inserção. A face palatina foi a que apresentou melhor taxa de regeneração (40%) e os dentes caninos e molares mostraram as melhores respostas (57,14% e 65%, respectivamente). Não houve sinais de infecção pós-cirúrgica relacionadas à falta de higienização oral dos animais. Pode-se concluir que o MIO/P-15 auxilia na regeneração e re-aderência das estruturas periodontais, incluindo osso alveolar. Sua aplicação mostrou-se fácil e prática e a incidência de complicações pós-cirúrgicas foi baixa. Ainda assim, mais estudos e pesquisas são necessários para que se avalie a quantidade e a qualidade do osso e do ligamento periodontal formados.

Padronização do eletrorretinograma de campo total em cães

Eletrorretinograma (ERG) é o meio diagnóstico objetivo e não-invasivo para avaliar a função retiniana e detectar precocemente, em várias espécies, lesões nas suas camadas mais externas. As indicações mais comuns para ERG em cães são: avaliação pré-cirúrgica de pacientes com catarata, caracterização de distúrbios que causam cegueira, além de servir como importante modelo para o estudo da distrofia retiniana que acomete o homem. Vários são os fatores que podem alterar o ERG tais como: eletrorretinógrafo, fonte de estimulação luminosa, tipo do eletrodo, tempo de adaptação ao escuro, tamanho pupilar, opacidade de meios e protocolo de sedação ou anestesia; além da espécie, raça e idade. Objetivou-se com este estudo padronizar o ERG para cães submetidos à sedação, seguindo o protocolo da International Society for Clinical Electrophysiology of Vision (ISCEV), utilizando Ganzfeld e eletrodos Burian Allen. Foram realizados 233 eletrorretinogramas em cães, 147 fêmeas e 86 machos, com idades entre um e 14 anos. Dos 233 cães examinados, 100 apresentavam catarata em diferentes estágios de maturação, 72 eram diabéticos e apresentavam catarata madura ou hipermadura, 26 apresentaram eletrorretinograma compatível com degeneração retiniana progressiva, três apresentaram eletrorretinograma compatível com síndrome da degeneração retiniana adquirida subitamente e 32 não apresentaram lesão retiniana capaz de atenuar as respostas do ERG, sendo considerados normais quanto à função retiniana. A sedação foi capaz de produzir boa imobilização do paciente sem rotacionar o bulbo ocular, permitindo adequada estimulação retiniana bilateralmente, com auxílio do Ganzfeld. O sistema eletrodiagnóstico Veris registrou com sucesso e simultaneamente de ambos os olhos, as cinco respostas preconizadas pela ISCEV. Como o ERG de campo total tornou-se exame fundamental na rotina oftalmológica, sua padronização é indispensável quando se objetiva comparar resultados de laboratórios distintos. A confiabilidade e reprodutibilidade deste protocolo foi demonstrada com a obtenção de registros de ótima qualidade utilizando protocolo padrão da ISCEV, eletrorretinógrafo Veris, Ganzfeld e eletrodos Burian Allen nos cães submetidos à sedação.

AT1-receptor mediated vascular damage in myocardium, kidneys and liver in rats

The systemic aspect of vascular damage induced by angiotensin II (ANG II) has been poorly explored in the literature. Considering the presence of ANG II and its specific receptor AT1, in several organs, all tissues might be potentially affected by its effects. The aims of this study were: To evaluate the early histological changes in the heart, liver and kidneys, produced by ANG II infusion, to evaluate the protective effect of losartan. Wistar rats were distributed into three groups: control (no treatment), treated with ANG II, and treated with ANG II + losartan. ANG II was continuously infused over 72 hours by subcutaneous osmotic pumps. Histological sections of the myocardium, kidneys and liver were stained and observed for the presence of necrosis. There were ANG II-induced perivascular inflammation and necrosis of the arteriolar wall in the myocardium, kidney, and liver by, which were partially prevented by losartan. There was no significant correlation between heart and kidney damage. Tissue lesion severity was lower than that of vascular lesions, without statistical difference between groups. ANG II causes vascular injury in the heart, kidneys and liver, indicating a systemic vasculotoxic effect; the mechanisms of damage/protection vary depending on the target organ; perivascular lesions may occur even when anti-hypertensive doses of losartan are used.

Achados clínico patológicos durante um surto de disenteria de inverno em bovinos no Rio Grande do Sul

Descreve-se um surto de disenteria de inverno que afetou 10 vacas leiteiras de uma propriedade localizada em Viamão, Rio Grande do Sul. O quadro clínico caracterizou-se por uma diarréia inicialmente líquida esverdeada com estrias de sangue e muco, evoluindo, em alguns animais, para uma diarréia de coloração marrom escura à sanguinolenta, que persistiu, em média, cinco dias. Drástica diminuição na produção de leite e no consumo de alimentos, além de graus variados de depressão também foram observados. Apenas um dos 10 animais afetados morreu. Durante a necropsia, observaram-se mucosas pálidas, conteúdo sanguinolento com presença de grande quantidade de coágulos, principalmente no cólon espiral e petéquias na mucosa do cólon. Os principais achados histológicos foram encontradas no cólon espiral, onde havia criptas dilatadas, sem epitélio de revestimento ou revestidas por epitélio pavimentoso e/ou cuboidal, por vezes com núcleos grandes e nucléolos proeminentes. Algumas criptas eram preenchidas por debris necróticos e polimorfonucleares. Na imuno-histoquímica com anticorpo monoclonal para coronavírus bovino (8F2) em cortes do cólon espiral, havia marcações positivas no citoplasma de enterócitos das criptas, nos debris necróticos destas e em macrófagos na lâmina própria.

Experimental infection of dogs with a Brazilian strain of Rickettsia rickettsii: clinical and laboratory findings

The bacterium Rickettsia rickettsii is the etiological agent of an acute, severe disease called Rocky Mountain spotted fever in the United States or Brazilian spotted fever (BSF) in Brazil. In addition to these two countries, the disease has also been reported to affect humans in Mexico, Costa Rica, Panama, Colombia and Argentina. Like humans, dogs are also susceptible to R. rickettsii infection. However, despite the wide distribution of R. rickettsii in the Western Hemisphere, reports of R. rickettsii-induced illness in dogs has been restricted to the United States. The present study evaluated the pathogenicity for dogs of a South American strain of R. rickettsii. Three groups of dogs were evaluated: group 1 (G1) was inoculated ip with R. rickettsii; group 2 (G2) was infested by R. rickettsii-infected ticks; and the control group (G3) was infested by uninfected ticks. During the study, no clinical abnormalities, Rickettsia DNA or R. rickettsii-reactive antibodies were detected in G3. In contrast, all G1 and G2 dogs developed signs of rickettsial infection, i.e., fever, lethargy, anorexia, ocular lesions, thrombocytopenia, anemia and detectable levels of Rickettsia DNA and R. rickettsii-reactive antibodies in their blood. Rickettsemia started 3-8 days after inoculation or tick infestation and lasted for 3-13 days. Our results indicate that a Brazilian strain of R. rickettsii is pathogenic for dogs, suggesting that canine clinical illness due to R. rickettsii has been unreported in Brazil and possibly in the other South American countries where BSF has been reported among humans.

Comparison of three decontamination methods for Mycobacterium bovis isolation

Sixty samples of tissue fragments with lesions suggestive of tuberculosis from bovine abattoirs, kept in saturated solution of sodium borate, were subjected to four treatments: 4% NaOH (Petroff Method), 12 % H2SO4 and 1.5% HPC (1-Hexadecylpyridinium Chloride) decontamination, and physiological saline solution (control). The HPC method showed the lowest contamination rate (3%) when compared to control (88%, p<0.001), NaOH (33%, p<0.001) and H2SO4 (21.7%, p<0.002). Regarding the isolation success, the HPC method was better (40%) than the control (3%, p<0.001), NaOH (13%, p=0.001) and H2SO4 (1.7%, p<0.001) methods. These results indicate that HPC is an alternative to the Petroff method.

Detection of Mycobacterium avium in pet birds

The present study is a report on the presence of Mycobacterium avium in four birds of the psittaciform order kept as pets. Anatomopathological diagnosis showed lesions suggestive of the agent and presence of alcohol-acid resistant bacilli (AARB) shown by the Ziehl-Neelsen staining. The identification of Mycobacterium avium was performed by means of PRA (PCR Restriction Analysis). DNA was directly extracted from tissue of the lesions and blocked in paraffin. The role of this agent in pet bird infection is discussed, as well as its zoonotic potential.

Remyelination in experimentally demyelinated connexin 32 KnockOut mice

The aim of this study was to evaluate the role of connexin 32 (Cx 32) during remyelination of the peripheral nervous system, through a local injection of either 0,1% ethidium bromide solution or saline in the sciatic nerve of Cx 32 knockout mice. Euthanasia was performed ranging from 1, 2, 3, 7, 15, 21 to 30 days after injection. Histochemical, immunohistochemical, immunofluorescence and transmission electron microscopical techniques were used to analyze the development of the lesions. Within the sciatic nerves, Schwann cells initially showed signs of intoxication and rejected their sheaths; after seven days, some thin newly formed myelin sheaths with uneven compactness and redundant loops (tomacula) were conspicuous. We concluded that the regeneration of lost myelin sheaths within the PNS followed the pattern already reported for this model in other laboratory species. Therefore, these results suggest that absence of Cx 32 did not interfere with the normal pattern of remyelination in this model in young mice.

Hepatotrophic factors reduce hepatic fibrosis in rats

CONTEXT: Hepatic fibrosis occurs in response to several aggressive agents and is a predisposing factor in cirrhosis. Hepatotrophic factors were shown to stimulate liver growth and to restore the histological architecture of the liver. They also cause an improvement in liver function and accelerate the reversion of fibrosis before it progresses to cirrhosis. OBJECTIVE: To test the effects of hepatic fibrosis solution composed by amino acids, vitamins, glucose, insulin, glucagon and triiodothyronine on hepatic fibrosis in rats. METHODS: Fibrosis was induced in rats by gastric administration of dimethylnitrosamine (10 mg/kg) for 5 weeks. After liver biopsy, the rats received either hepatotrophic factors solution (40 mg/kg/day) or saline solution for 10 days by intraperitoneal injection. Blood samples and liver fragments were collected for hepatic function analysis, standard histopathology evaluation, and morphometric collagen quantification. RESULTS: Rats in the hepatotrophic factors group showed a decrease of the histopathological components of fibrosis and an increase of their hepatic mass (12.2%). There was no development of neoplasic lesions in both groups. Compared with the saline group, the hepatotrophic factors group also had a decrease of blood levels of hepatic-lesion markers (AST, ALT) and a decrease of collagen content in the portal spaces (31.6%) and perisinusoidal spaces (42.3%), as well as around the hepatic terminal vein (57.7%). Thus, hepatotrophic factors administration in the portal blood promoted a regenerative hepatic response, with an overall reduction of the volumetric density of collagen, improved hepatic function, and a general improvement in the histopathological aspects of fibrosis. CONCLUSION: Taken together, these results suggest the potential therapeutic use of this hepatotrophic factors solution to treat chronic liver diseases.

Evaluation of the efficacy of hydrated sodium aluminosilicate in the prevention of aflatoxin-induced hepatic cancer in rainbow trout

The use of aluminum silicates for decontaminating animal feed containing aflatoxins has yielded encouraging results in chicken and turkey poults. In contrast, very few studies have tested these substances in aquaculture. In this work, we investigated the efficacy of a trout diet containing 0.5% hydrated sodium aluminosilicate (HSAS) in protecting against contamination with aflatoxin B1. Trout were reared on these diets for one year and the experimental groups were examined monthly for hepatic presumptive preneoplastic and neoplastic lesions. Regardless of the presence of HSAS, all of the fish that received aflatoxin in their diet have shown hepatic lesions indicative of a carcinogenic process, presenting also the development of cancer in some fish. The concentration of HSAS used in this study was ineffective in preventing the onset of hepatic lesions induced by aflatoxin B1 in rainbow trout.

Expression and distribution of connexin 32 in rat liver with experimentally induced fibrosis

The connexin 32 (Cx32) is a protein that forms the channels that promote the gap junction intercellular communication (GJIC) in the liver, allowing the diffusion of small molecules through cytosol from cell-to-cell. Hepatic fibrosis is characterized by a disruption of normal tissue architeture by cellular lesions, and may alter the GJIC. This work aimed to study the expression and distribution of Cx32 in liver fibrosis induced by the oral administration of dimethylnitrosamine in female Wistar rats. The necropsy of the rats was carried out after five weeks of drug administration. They presented a hepatic fibrosis state. Sections from livers with fibrosis and from control livers were submitted to immunohistochemical, Real Time-PCR and Western-Blot analysis to Cx32. In fibrotic livers the Cxs were diffusely scattered in the cytoplasm, contrasting with the control livers, where the Cx32 formed junction plaques at the cell membrane. Also it was found a decrease in the gene expression of Cx32 without reduction in the protein quantity when compared with controls. These results suggest that there the mechanism of intercellular communication between hepatocytes was reduced by the fibrotic process, which may predispose to the occurrence of a neoplastic process, taken in account that connexins are considered tumor suppressing genes.

The effects of the experimental onfection from a focus of transmissopn of Schistosoma mansoni in a population of Biomphalaria tenagophila (D'Orbigny, 1835) in the region of "Vale do Ribeira de Iguape", Brazil

Com objetivo de observar os efeitos da infecção parasitária na biologia de B. tenagophila, foram realizadas infecções experimentais em populações de campo e laboratório, ambas procedentes de Itariri, Vale do Ribeira, Brasil. Cada molusco recebeu 10 miracídios de Schistosoma mansoni (linhagem SJ), sendo observado durante o desenvolvimento dos parasitos. As variáveis biológicas foram comparadas segundo os critérios "grupo" e "fase de infecção". Os principais danos decorrentes do parasitismo se manifestaram na reprodução, na longevidade e em lesões na concha dos moluscos na fase patente. Foi encontrada uma taxa de infecção de 58,8 por cento. O estudo microanatômico da glândula digestiva e do ovoteste do molusco revelou a presença de formas larvárias em evolução e cercárias. Concluiu-se que os efeitos da infecção parasitária, sobre as duas populações, foram moderados, uma vez que os danos não impediram a reprodução e a eliminação de cercárias que se manteve por um longo período, apesar da baixa sobrevivência dos moluscos parasitados

Prevalência do HPV em mulheres rastreadas para o câncer cervical

OBJETIVO: Analisar a prevalência da infecção genital por papilomavírus humano (HPV) de alto risco por faixa etária e fatores associados. MÉTODOS: Estudo transversal com amostra de 2.300 mulheres (15-65 anos) que buscaram rastreamento para o câncer cervical entre fevereiro de 2002 e março de 2003 em São Paulo e Campinas, estado de São Paulo. Aplicou-se questionário epidemiológico e realizou-se coleta cervical para citologia oncológica e teste de captura híbrida II. As análises estatísticas empregadas foram teste de qui-quadrado de Pearson e análise multivariada pelo método forward likelihood ratio. RESULTADOS: A prevalência total da infecção genital por HPV de alto risco foi de 17,8%, distribuída nas faixas etárias: 27,1% (<25 anos), 21,3% (25-34 anos), 12,1% (35-44 anos), 12,0% (45-54 anos) e de 13,9% (55-65 anos). Participantes com maior número de parceiros sexuais durante a vida apresentaram maior freqüência da infecção. Relacionamento estável, idade de 35 a 44 anos e ex-fumantes foram associados à proteção da infecção. A infecção genital por HPV de alto risco ocorreu em 14,3% das citologias normais, em 77,8% das lesões escamosas de alto grau e nos dois (100%) casos de carcinoma. CONCLUSÕES: A prevalência da infecção genital por HPV de alto risco na amostra estudada foi alta. Houve predomínio de casos abaixo dos 25 anos e tendência a um novo aumento após os 55 anos, com maior freqüência naqueles com maior número de parceiros sexuais durante a vida

Echocardiographic study of paediatric patients with mucopolysaccharidosis

Principle Mucopolysaccharidosis is an inborn error of metabolism causing glucosaminoglycans tissue storage. Cardiovascular involvement is variable but contributes significantly towards the morbidity and mortality of the patients. Objective To characterise the echocardiographic abnormalities in children and adolescents with different types of mucopolysaccharidosis. Method Echocardiograms and medical records of 28 patients aged 2–14 years, seen from 2003 to 2005, were revised. At that time, the enzymatic replacement therapy was still not available in our institution.Results Echocardiographic alterations were detected in 26 patients (93 per cent), whereas 16 (57 per cent) had abnormal auscultation, and only 6 (21 per cent) presented with cardiovascular complaint. Mitral valve thickening with dysfunction (regurgitation, stenosis, or double lesion) was diagnosed in 60.8 per cent, left ventricular hypertrophy in 43 per cent and aortic valve thickening with regurgitation in 35.8 per cent of the patients. There was no systolic dysfunction and mild left diastolic dysfunction was shown in 21.5 per cent of the patients. Pulmonary hypertension was present in 36 per cent of the patients, causing the only two deaths recorded. There was a strong association between the accumulation of dermatan sulphate and the presence of mitral valve dysfunction (p = 0.0003), aortic valve dysfunction (p = 0.006), and pulmonary hypertension (p = 0.006). Among individuals with two or more examinations, 82 per cent had a worsening evolution. Conclusions Echocardiographic alterations in children with Mucopolysaccharidosis are frequent and have a progressive character Left valve lesions, ventricular hypertrophy, and pulmonary hypertension were the most common findings and there was an association between the accumulation of dermatan sulphate and cardiovascular involvement. Unlike in adults, pulmonary hypertension was the main cause of death, not left ventricle systolic dysfunction

Classification of breast masses using a committee machine of artificial neural networks

This work proposes a new approach using a committee machine of artificial neural networks to classify masses found in mammograms as benign or malignant. Three shape factors, three edge-sharpness measures, and 14 texture measures are used for the classification of 20 regions of interest (ROIs) related to malignant tumors and 37 ROIs related to benign masses. A group of multilayer perceptrons (MLPs) is employed as a committee machine of neural network classifiers. The classification results are reached by combining the responses of the individual classifiers. Experiments involving changes in the learning algorithm of the committee machine are conducted. The classification accuracy is evaluated using the area A. under the receiver operating characteristics (ROC) curve. The A, result for the committee machine is compared with the A, results obtained using MLPs and single-layer perceptrons (SLPs), as well as a linear discriminant analysis (LDA) classifier Tests are carried out using the student's t-distribution. The committee machine classifier outperforms the MLP SLP, and LDA classifiers in the following cases: with the shape measure of spiculation index, the A, values of the four methods are, in order 0.93, 0.84, 0.75, and 0.76; and with the edge-sharpness measure of acutance, the values are 0.79, 0.70, 0.69, and 0.74. Although the features with which improvement is obtained with the committee machines are not the same as those that provided the maximal value of A(z) (A(z) = 0.99 with some shape features, with or without the committee machine), they correspond to features that are not critically dependent on the accuracy of the boundaries of the masses, which is an important result. (c) 2008 SPIE and IS&T.