Made available in DSpace on 2016-12-23T13:53:38Z (GMT). No. of bitstreams: 1 Marcel Arcanjo Silva Azevedo.pdf: 869015 bytes, checksum: cfeb9681dc674d9ccb6415fdfbdee8ee (MD5) Previous issue date: 2013-07-31 / The bovine enzootic haematuria (BEH) is a chronic disease caused by Pteridium aquilinum and is characterized by the formation of non-neoplastic and neoplastic lesions in the bladder of the affected animals and has no specific treatment. In the southern region of the Espírito Santo, Brazil, this disease has a high prevalence and is responsible for major economic losses in dairy cattle, resulting from late diagnosis of the disease. Cytology associated with the use of biomarkers could aid in the early diagnosis of this disease. The aim of the study was to evaluate lesions bladders of cattle associated with enzootic hematuria through cytological techniques, immunocytochemistry and comet assay, for the diagnosis of early cellular damage caused by the disease. The study was divided into two experiments. In experiment 1, to standardize the harvesting technique, obtain samples and perform cytology of the bovine washing urinary bladder with BEH, were used 10 cattle, adult females divided into two groups. In group A was recovered all the liquid infused in the bladder, in group B, only the last liquid was recovered. In experiment 2, the cells obtained by bovine washing urinary bladder and were evaluated by immunocytochemistry using anti-p53 and comet assay and were used 10 cattle, adult females, divided in two groups of five animals, five healthy and five with BEH. The material was fixed and subjected to tests cytological, immunocytochemical and comet assay and evaluated microscopically. Data were analyzed using descriptive statistics, analysis of variance and nonparametric Kolmogorov-Smirnov test. Regarding the number of inflammatory cells and epithelial cells obtained per sample revealed that the two groups of experiment 1, all animals had more inflammatory cells than epithelial cells, however, there was no difference between the type of washing taken. The epithelial cells were found in 60% of cases and the alterations observed were discretes and cannot classify any sample as hyperplastic or neoplastic. In experiment 2, we observed positive immunostaining of p53 in only 20% of the samples, one of healthy animal and another animal with BEH. The comet assay revealed that both the samples of healthy animals as well as in animals positive for BEH not observed migrating nuclear fragments. Data from this study showed that the cytological examination of bovine washing urinary bladder may aid in the diagnosis of BEH and the two harvesting methods employed were adequate for obtaining viable samples. Cytology allowed the identification of non-neoplastic lesions predominantly inflammatory and immunocytochemistry technique with the expression of p53 as well as comet assay revealed no important cell damage since the animals used in the experiment had no neoplastic lesions

Pacientes submetidos ao transplante de medula óssea, apresentando hematúria/cistite hemorrágica, tiveram amostras de urina analisadas pela microscopia eletrônica. Esta foi a técnica escolhida de pesquisa viral pela sua confiabilidade. Noventa em 402 pacientes submetidos ao transplante de medula óssea neste serviço apresentaram hematúria/cistite hemorrágica (incidência de 22%). O estudo por microscopia eletrônica foi realizado em 72 destes pacientes com hematúria/cistite hemorrágica (grupo de estudo), identificando 55,6% (40/72) de positividade viral. Foram também estudadas amostras de urina de 12 pacientes submetidos ao transplante de medula óssea sem hematúria/cistite hemorrágica (grupo controle); houve associação significante entre a presença de vírus e hematúria/cistite hemorrágica (p<0,01 - Teste exato de Fisher). No grupo com hematúria/cistite hemorrágica, 65% (26/40) dos vírus detectados pertenciam à família Poliomaviridae, 30% (12/40) à Adenoviridae, e 5% (2/40) foram positivos para ambas as famílias. Houve associação entre a positividade para vírus e a presença da doença enxerto-contra-hospedeiro (p=0,05-x2) e o de início tardio (>dia+21) da H/CH (P=0,04-x2), bem como entre a doença enxerto-contra-hospedeiro e a severidade da H/CH (p=0,04-x2). O presente estudo mostra que a microscopia eletrônica é uma ferramenta útil para detecção de vírus na urina de pacientes submetidos ao transplante de medula óssea apresentando hematúria/cistite hemorrágica e que o vírus provavelmente tem um papel no desenvolvimento do sangramento urinário, o qual é agravado pelo doença enxerto-contra-hospedeiro. A microscopia eletrônica deveria ser realizada rotineiramente para guiar as outras técnicas de detecção viral, como a imunocitoquímica e biologia molecular. / Patients submitted to bone marrow transplantation presenting hematuria/hemorrhagic cystitis had the urine analyzed by electron microscopy. It was the elected technique for viral search because its reliability. Ninety from 402 patients submitted to bone marrow transplantation at this service showed hematuria/hemorrhagic cystitis (incidence of 22%). Electron microscopy study was performed in 72 of these patients with hematuria/hemorrhagic cystitis (study group), identifying 55.5% (40/72) of viral positivity. It was also study the urine of 12 patients submitted to bone marrow transplantation without hematuria/hemorrhagic cystitis (control group); there was a significant association between the presence of virus and hematuria/hemorrhagic cystitis (p<0.01). In the hematuria/hemorrhagic cystitis group, 65% (26/40) of the virus belonged to Poliomaviridae family, 30% (12/40) to Adenoviridae, and 5% (2/40) to both families. There was association between the status of graft-versus-host disease and positivity for virus (p=0.05), as well as between graft-versus-host disease and the severity of hematuria/hemorrhagic cystitis (p=0.04). The present study shows that electron microscopy is a useful tool for detection of virus in urine of patients submitted to bone marrow transplantation presenting hematuria/hemorrhagic cystitis and that the virus probably play a role in the development of the urinary bleeding, which is aggravated by graft-versus-host disease. Electron microscopy should be performed routinely to guide the other techniques for viral detection, like immunocitochemistry and molecular biology.

Pacientes submetidos ao transplante de medula óssea, apresentando hematúria/cistite hemorrágica, tiveram amostras de urina analisadas pela microscopia eletrônica. Esta foi a técnica escolhida de pesquisa viral pela sua confiabilidade. Noventa em 402 pacientes submetidos ao transplante de medula óssea neste serviço apresentaram hematúria/cistite hemorrágica (incidência de 22%). O estudo por microscopia eletrônica foi realizado em 72 destes pacientes com hematúria/cistite hemorrágica (grupo de estudo), identificando 55,6% (40/72) de positividade viral. Foram também estudadas amostras de urina de 12 pacientes submetidos ao transplante de medula óssea sem hematúria/cistite hemorrágica (grupo controle); houve associação significante entre a presença de vírus e hematúria/cistite hemorrágica (p<0,01 - Teste exato de Fisher). No grupo com hematúria/cistite hemorrágica, 65% (26/40) dos vírus detectados pertenciam à família Poliomaviridae, 30% (12/40) à Adenoviridae, e 5% (2/40) foram positivos para ambas as famílias. Houve associação entre a positividade para vírus e a presença da doença enxerto-contra-hospedeiro (p=0,05-x2) e o de início tardio (>dia+21) da H/CH (P=0,04-x2), bem como entre a doença enxerto-contra-hospedeiro e a severidade da H/CH (p=0,04-x2). O presente estudo mostra que a microscopia eletrônica é uma ferramenta útil para detecção de vírus na urina de pacientes submetidos ao transplante de medula óssea apresentando hematúria/cistite hemorrágica e que o vírus provavelmente tem um papel no desenvolvimento do sangramento urinário, o qual é agravado pelo doença enxerto-contra-hospedeiro. A microscopia eletrônica deveria ser realizada rotineiramente para guiar as outras técnicas de detecção viral, como a imunocitoquímica e biologia molecular. / Patients submitted to bone marrow transplantation presenting hematuria/hemorrhagic cystitis had the urine analyzed by electron microscopy. It was the elected technique for viral search because its reliability. Ninety from 402 patients submitted to bone marrow transplantation at this service showed hematuria/hemorrhagic cystitis (incidence of 22%). Electron microscopy study was performed in 72 of these patients with hematuria/hemorrhagic cystitis (study group), identifying 55.5% (40/72) of viral positivity. It was also study the urine of 12 patients submitted to bone marrow transplantation without hematuria/hemorrhagic cystitis (control group); there was a significant association between the presence of virus and hematuria/hemorrhagic cystitis (p<0.01). In the hematuria/hemorrhagic cystitis group, 65% (26/40) of the virus belonged to Poliomaviridae family, 30% (12/40) to Adenoviridae, and 5% (2/40) to both families. There was association between the status of graft-versus-host disease and positivity for virus (p=0.05), as well as between graft-versus-host disease and the severity of hematuria/hemorrhagic cystitis (p=0.04). The present study shows that electron microscopy is a useful tool for detection of virus in urine of patients submitted to bone marrow transplantation presenting hematuria/hemorrhagic cystitis and that the virus probably play a role in the development of the urinary bleeding, which is aggravated by graft-versus-host disease. Electron microscopy should be performed routinely to guide the other techniques for viral detection, like immunocitochemistry and molecular biology.