Application of the mammalian glyceraldehyde-3-phosphate dehydrogenase gene for sample quality control in multiplex PCR for diagnosis of leishmaniasis

Leishmaniasis is a neglected disease endemic in five continents. It is a severe disease that may lead to death, and its early detection is important to avoid severe damage to affected individuals. Molecular methods to detect Leishmania are considered alternatives to overcome the limitations presented by conventional methods. The aim of this study was to develop multiplex PCR systems able to detect small amounts of target DNA of Leishmania infantum and Leishmania braziliensis, and the gene coding for glyceraldehyde-3-phosphate dehydrogenase (G3PD) in mammals, enabling quality evaluation of the sample simultaneously with detection of the specific target. The systems created for G3PD recognition were combined with detection systems for L. infantum and L. braziliensis to compose multiplex PCR systems for visceral (mVL) and cutaneous (mACL) leishmaniasis diagnosis. The multiplex PCR systems developed were assessed in blood samples from five different species of mammal reservoirs involved in the disease cycle in Brazil, and 96 and 52 human samples from patients with suspected visceral leishmaniasis (VL) and cutaneous leishmaniasis (ACL), respectively. Three G3PD detection systems were created (G3PD1, G3PD2 and G3PD3) with different product sizes, G3PD2 was chosen for the formation of multiplex PCR systems. The two multiplex PCR systems (mVL and mACL) were reproducible in all species evaluated. Results of test samples (sensitivity, specificity and efficiency) suggest its use in routine diagnosis, research activities in medicine and veterinary medicine. Additionally, the systems designed to detect the G3PD gene are capable of combining with other targets used for molecular diagnosis of infectious diseases. Concerning leishmaniasis, the multiplex PCR systems can be used in epidemiological studies for the detection of new and classic reservoirs, which may contribute to the reliability of results and development of actions to control the disease.(AU)

Efeitos do amitraz e da xilazina sobre algumas características fisiológicas de cavalos / Effect of amitraz and xylazine on some physiological variables of horses

Avaliaram-se os efeitos das injeçöes intravenosa (iv) de amitraz (0,1mg/kg) e xilazina (1mg/kg), em cavalos, sobre a atividade cardíaca, freqüência respiratória, atividade motora intestinal, temperatura retal, sudorese e freqüência de apreensäo de alimentos. O amitraz causou diminuiçäo significativa da atividade cardíaca, da freqüência respiratória e da movimentaçäo intestinal, mas esses efeitos näo foram täo pronunciados quanto os causados pela xilazina. O amitraz causou, também, relaxamento significativo da musculatura lisa retal, e um aparente aumento da sudorese e da freqüência de cavalos flagrados mastigando feno. A temperatura retal näo foi influenciada pelo amitraz. Os resultados indicam que o amitraz, na dose utilizada, näo causou efeitos colaterais severos em cavalos

Hepatic regeneration after partial hepatectomy in mice infected with Schistosoma mansoni, at the acute and chronic phases of the disease

Outbred male albino mice normal or infected with 30 cercariae of Schistosoma mansoni (LE strain) were submitted to 65 percent hepatectomy during the acute (70 days) and chronic phase (160 days) phases of the disease. A group of the infected animals was treated with 400 mg/kg of oxamniquine during the acute phase before hepatectomy. Non-infected, infected and treated but not hepatectomized animals were kept as controls. Hepatic regeneration was evaluated by incorporation of tritiated thymidine, intraperitoneally injected into non-hepatectomized and hepatectomized animals, 24 hours after surgery. The results showed that removal of 65 percent of the hepatic parenchyma, during the acute phase, led to a statistically significant increase of thymidine incorporation, when compared with the uninfected hepatectomized controls. This phenomenon was not observed at the chronic phase. Treatment with oxamniquine administered during the acute phase led to a decrease in thymidine incorporation rate 160 days after infection (90 days after treatment) and 24 hours after hepatectomy. The data suggest that infection with S. mansoni represents a considerable stimulus for the regenerative capacity of the liver during the acute, but not the chronic phase of disease

Percepçäo e registro de abuso de álcool e de doenças relacionadas num hospital geral universitário / Perception and registration of alcohol abuse and alcohol related diseases by nurses and medical staff of an university general hospital

Objetivo. Avaliar a associaçao entre o resultado do teste CAGE aplicado em pacientes internados num hospital geral universitário (Hospital de Clínicas de Porto ALegre - HCPA) e a percepçao e registro de abuso de álcool e de doenças relacionadas pelas equipes de saúde. Pacientes e Métodos. O delineamento envolveu três fases: a) aplicaçao do teste CAGE aos 385 pacientes adultos e adolescentes internados considerados elegíveis para o estudo; b) revisao dos prontuários dos pacientes CAGE-positivo e de um número igual de pacientes CAGE-negativo, estratificados por área de internaçao e sorteados, para identificar hábitos alcoólicos e a presença de doenças relacionadas ao uso de álcool; c) entevista com os membros das equipes médicas responsáveis para avaliar sua percepçao sobre o hábito alcoólico em seu paciente. As diferenças encontradas foram analisadas através do teste Qui-quadrado e do teste t de Student, com significância estatística em nível de 5 por cento bicaudal. Resultados. Foram encontrados 51 pacientes CAGE-positivo (13 por cento). A equipe médica registrou abuso de álcool no prontuário de 51 por cento dos pacientes CAGE-positivo e 10 por cento dos CAGE-negativo, e a equipe de enfermagem fez este registro no prontuário de 31 por cento dos pacientes CAGE-positivo e 8 por cento dos CAGE-negativo (p=.724). Nao houve diferença entre o registro e a percepçao da equipe médica (p=.184). A prevalência de doenças relacionadas ao abuso de álcool foi 27 por cento quando se examinou os prontuários médicos e 26 por cento quando se entrevistou a equipe médica (p=.861), sendo maior entre os pacientes CAGE-poistivo (43 por cento) do que entre os CAGE-negativo (10 por cento) (p=.0003). Conclusoes. O abuso de bebidas alcoólicas e a presença de doenças associadas ao mesmo sao condiçoes prevalentes nos pacientes admitidos ao HCPA. Entretanto, mais de metade dos casos nao foram detectados pela equipe de saúde, havendo correspondência entre os registros de prontuário e a entrevista.

Atherosclerosis and acute arterial thrombosis in rabbits: a model using ballon desendothelization without dietary intervention

Acute thrombosis can be induced in rabbits by a triggering protocol using Russell's viper venom and histamine given after 8 months of a 1 per cent cholesterol diet and balloon desendothelization. In the present study, we tested the hypothesis that aortic desendothelization performed 4 months before the triggering protocol without a high cholesterol diet is a highly effective and less expensive way of producing arterial atherosclerosis and thrombosis. Nineteen male New Zealand white rabbits on a normal diet were studied. The control group (N = 9) received no intervention during the 4-month observation period, while the other group (N = 10) was submitted to aortic balloon desendothelization using a 4F Fogarty catheter. At the end of this period, all animals were killed 48 h after receiving the first dose of the triggering treatment. Eight of 10 rabbits (80 per cent) in the balloon-trauma group presented platelet-rich arterial thrombosis while none of the animals in the control group had thrombus formation (P<0.01). Thus, this model, using balloon desendothelization without dietary manipulation, induces arterial atherosclerosis and thrombosis and may provide possibilities to test new therapeutic approaches.