980 resultados para IMMUNOFLUORESCENCE


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Endo-oligopeptidase A, EC 3.4.22.19, converts small enkephalin-containing peptides into the corresponding enkephalins in vitro. We investigated the presence of endooligopeptidase A in the retina and its possible colocalization with enkephalins in retinal neurons. The specific activity of endo-oligopeptidase A found in pigeon retinae (30.3 +/- 7.3 mU/mg, mean +/- standard deviation) was four times higher than in rabbit retinae (7.0 +/- 1.1 mU/mg). The enzyme activity was not modified by EDTA, but it was enhanced by dithiothreitol and inhibited by zinc and 5,5'-dithiobis(2-nitrobenzoic acid). Immunohistochemical experiments with a purified antiserum against rabbit endo-oligopeptidase A revealed labeled neurons in both the inner nuclear layer and the ganglion cell layer of pigeon and rabbit retinae. Double-labeling immunofluorescence experiments demonstrated that about 90% of neurons containing endo-oligopeptidase A-like immunoreactivity also contained [Leu5]-enkephalin-like immunoreactivity. These colocalization results may represent an important step toward the demonstration of the possible involvement of endo-oligopeptidase A in enkephalin generation in vivo.

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Leishmaniasis is a disease emerging or re-emerging worldwide distribution (Sousa, 2008), a major impact on public health. The canine visceral leishmaniasis is an infectious parasitic zoonosis of worldwide distribution (Troncarelli, 2009), caused by a flagellate protozoan called Leishmania chagasi (Costard, 2009). Osteomyelitis can develop when the parasite reaches the bone tissue of the host via blood or continuity of adjacent soft tissue infection (Baltenperger, 2004). The histopathology of the lesions has 100% specificity when viewing the amastigote form of the parasite (Feitosa et al., 2000). A dog breed poodle, female, five years, with chronic lameness four months ago was attended by a veterinary, proceeded to the general clinical examination, radiographic evaluation of the hindquarters and the laboratory tests of enzyme immunoassay (ELISA) and indirect immunofluorescence (RIFI) for Leishmania sp. On examination, besides the enlargement of lymph nodes, the animal showed pain on flexion and extension of hind limbs. In radiographic lytic bone lesions were observed in bilateral ischial board and greater trochanter of the femur, suggestive of osteomyelitis. In specific laboratory tests for diagnosis of leishmaniasis ELISA reagent and RIFI reagent 1:40. As recommended by the Ministry of Health, the animal was euthanized. No macroscopic findings were reported during the necropsy, usually associated with leishmaniasis. The ischium bone fragments were sent for histopathological examination. There was intense proliferation of mononuclear inflammatory cells, mainly macrophages and lymphocytes. Amastigotes of Leishmania sp, were identified in the cytoplasm of some macrophages and bone tissue. In endemic areas for canine leishmaniasis, dogs with a history of intermittent lameness, and radiographic lytic bone lesions suggestive of osteomyelitis should be directed to realization of the histopathology and serologic tests for the differential diagnosis of Leishmania sp. Even without evidence of cutaneous or visceral lesions, usually associated with this disease.

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The feline leukemia virus (FeLV) was described in 1964 by William Jarrett and collaborators wen find viral particles attached to the membrane of lymphoblasts in cat with lymphoma. The virus belongs to the family Retroviridae, subfamily oncornavirus. With worldwide distribution, the occurrence of FeLV has 1.6% in healthy cats and 10.8% in sick cats in Brazil. The mortality of persistently viremic animals in catteries is about 50% in two years and 80% in three years. In catteries that have endemic feline Coronavirus (FCoV), FeLV and / or Feline Immunodeficiency Virus (FIV), the FeLV infection has greater contribution to mortality. The test for infection and FeLV positive cats segregation is the main way to prevent the spread of infection. The diagnostic methods are based on clinical signs and changes compatible with FeLV infection observed by physical examination, complete blood count, X-ray, bone marrow aspirate and biochemical. The viral p27 protein is produced in infected cells in high amounts and is found in abundance in the cytoplasm and in body fluids enabling diagnosed methods such as enzyme-linked immunosorbent assay - ELISA and direct immunofluorescence, detection of viral genome (Chain Reaction Polymerase - PCR) and detection of the virus by virus isolation. Although diagnostic tests are highly sensitive, it should be made more than a confirmatory test, especially serological due to variable characteristic of the progress of infection

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Porcine parvovirus (PPV) is associated with reproductive failure and it has been found worldwide, including Brazil. Many diagnostic procedures are used for its detection, for example, immunofluorescence, HA, HI and PCR and this is an important technique because it is very specific and sensitive. In this work, the presence of PPV in fetuses from swine farms with reproductive problems was detected by PCR. All of 170 samples from aborted fetuses, mummies or stillborns were sampled by PCR with primers designed to VP2 region of PPV and c-myc (endogenous control). Only 142 samples (83,53%) were positive for c-myc and among them six samples (4,22%) were positive for PPV which were tested in HA. In this test, erythrocytes suspension 1% was used and three samples (50%) agglutinated but they presented low titer (4). For this work, porcine parvovirus was detected in the samples analyzed by PCR and HA. It is important to emphasize the use of endogenous control when material with elevated degree of autolysis is examined

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Pós-graduação em Medicina Veterinária - FCAV

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Pós-graduação em Medicina Veterinária - FCAV

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Respiratory syncytial virus is the major cause of acute lower respiratory tract illness in infants and young children. Because there is currently no licensed vaccine for RSV, there is a substantial interest in the identification and development of RSV specific inhibitory agents. There are clinical evidences that glycosaminoglycans (GAGs) are potential inhibitors of viral infection. In this study, the performance of two GAGs (heparin and dextran sulfate) were compared for their antiviral and virucidal activities on RSV. Analysis was performed using an in vitro infection model where, previously to infection, Hep-2 cells or RSV were incubated with heparin or dextran sulfate. The presence of viral particles was analyzed by Reverse Transcriptase-Polimerase Chain Reaction (RT-PCR) and indirect immunofluorescence assays (IFA). The results showed that viral infection was more efficiently inhibited when Hep-2 cells were pre-incubated with heparin or, when viral particles were pre-incubated with dextran sulfate. Our study suggest that, in the absence of cellular death, heparin and dextran sulfate reduce RSV infection by different mechanisms, antiviral and virucidal ones, respectively. These data contribute for recent medical, microbiology and biochemical studies which suggest that the use of antiviral and virucidal compounds as more effective treatment to control virus infections.

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