Modelo experimental comportamental e histológico da regeneração do nervo facial em ratos

O estabelecimento de modelos experimentais é o passo inicial para estudos de regeneração neural. OBJETIVO: Estabelecer modelo experimental de regeneração do nervo facial. MATERIAIS E MÉTODOS: Ratos Wistar com secção completa e sutura do tronco do nervo facial extratemporal, com análise comportamental e histológica até 9 semanas. FORMA DE ESTUDO: Estudo prospectivo experimental. RESULTADOS: Progressiva recuperação clínica e histológica dos animais. CONCLUSÃO: Estabelecemos um método aceitável para o estudo de regeneração do nervo facial em ratos.

A JOINT EXPERIMENTAL ANALYSIS OF INVESTOR BEHAVIOR IN IPO PRICING METHODS

This article jointly examines the differences of laboratory versions of the Dutch clock open auction, a sealed-bid auction to represent book building, and a two-stage sealed bid auction to proxy for the “competitive IPO”, a recent innovation used in a few European equity initial public offerings. We investigate pricing, seller allocation, and buyer welfare allocation efficiency and conclude that the book building emulation seems to be as price efficient as the Dutch auction, even after investor learning, whereas the competitive IPO is not price efficient, regardless of learning. The competitive IPO is the most seller allocative efficient method because it maximizes offer proceeds. The Dutch auction emerges as the most buyer welfare allocative efficient method. Underwriters are probably seeking pricing efficiency rather than seller or buyer welfare allocative efficiency and their discretionary pricing and allocation must be important since book building is prominent worldwide.

Infecção experimental de Calomys callosus (Rodentia Cricetidae) com Trypanosoma cruzi

São apresentados resultados sobre a infecção experimental de Calomys callosus (Rodentia) e duas cepas (Y e Berenice) de Trypanosoma cruzi, isoladas de casos humanos. O estudo da evolução foi feito comparado com Mus musculus albino cepa "Swiss", quanto a prepatência, parasitemia, patência e letalidade. Análise histopatológica foi também conduzida em C. callosus, com o objetivo de verificar o tropismo tissular e agressividade das cepas neste roedor. Os experimentos mostraram que a evolução da infecção em C. Callosus foi diferente para as duas cepas de T. cruzi. A cepa Y apresentou maior parasitemia do que a cepa Berenice. O período prepatente variou com as doses utilizadas tendo sido mais curto nos animais inoculados com a cepa Y (2, 2-5, 2 dias) do que naquelas com a cepa Berenice (3, 2-7 dias). Embora as duas cepas inoculadas nos C. callosus tenham-se mostrado miotrópicas, as alterações tissulares foram mais acentuadas com a Y. Os resultados obtidos abrem perspectivas quanto à possibilidade do uso de C. callosus como animal experimental para T. cruzi.

Investigación experimental de la equinococosis canina a partir de quiste hidatídico de origen porcino en México

OBJETIVO: Para evaluar la infección y obtener el estado adulto del cestodos, se buscó reproducir la equinococosis en perros a partir de quiste hidatídico de origen porcino. MÉTODOS: Se formaron 2 grupos, uno de 5 y otro de 3 perros, a cada animal del grupo experimental se le dió 2 g de membrana germinativa de quíste hidatídico fértil por vía oral, el segundo grupo fue testigo. Ambos grupos fueron evaluados clínica, serológica y parasitológicamente, en el grupo experimental se sacrificó un animal el día 35 de la infección y los siguientes cada 5 dias hasta el 55, en el segundo grupo todos se sacrificaron el día 55. Se observaron huevos del cestodos en heces a partir del dia 51 postinfección. La evaluación morfológica se realizó mediante observación microscópica del raspado de mucosa intestinal. RESULTADOS: De 50 cestodos analizados, 10 de cada uno de los perros infectados, 49 (98%) presentaron 3 proglótidos y 1 (2%) tenía 4; 18 (36%) de los cestodos presentaban un proglótido grávido. La longitud de los estróbilos varió de 1,6 a 2,6 mm. El número promedio de los ganchos largos y cortos fue de 31 y 34 respectivamente. La longitud de los ganchos largos varió entre 0,081 y 0,09 mm, los ganchos cortos fluctuaron entre 0,034 y 0,041 mm. En los perros evaluados clínicamente, el número de leucocitos y la cantidad de proteínas plasmáticas fue significativamente mayor en el grupo testigo (P < 0,05); la cantidad de alfa globulinas fue mayor en el grupo infectado (P < 0,05). CONCLUSIONES: Los resultados permiten confirmar el ciclo perro-cerdo y una infección subclínica en los huéspedes definitivos, lo que dificulta su diagnóstico y control en una especie intimamente relacionada con el hombre.

Evaluation of the respiratory motion effect in small animal PET images with GATE Monte Carlo simulations

The rapid growth in genetics and molecular biology combined with the development of techniques for genetically engineering small animals has led to increased interest in in vivo small animal imaging. Small animal imaging has been applied frequently to the imaging of small animals (mice and rats), which are ubiquitous in modeling human diseases and testing treatments. The use of PET in small animals allows the use of subjects as their own control, reducing the interanimal variability. This allows performing longitudinal studies on the same animal and improves the accuracy of biological models. However, small animal PET still suffers from several limitations. The amounts of radiotracers needed, limited scanner sensitivity, image resolution and image quantification issues, all could clearly benefit from additional research. Because nuclear medicine imaging deals with radioactive decay, the emission of radiation energy through photons and particles alongside with the detection of these quanta and particles in different materials make Monte Carlo method an important simulation tool in both nuclear medicine research and clinical practice. In order to optimize the quantitative use of PET in clinical practice, data- and image-processing methods are also a field of intense interest and development. The evaluation of such methods often relies on the use of simulated data and images since these offer control of the ground truth. Monte Carlo simulations are widely used for PET simulation since they take into account all the random processes involved in PET imaging, from the emission of the positron to the detection of the photons by the detectors. Simulation techniques have become an importance and indispensable complement to a wide range of problems that could not be addressed by experimental or analytical approaches.

Atividade antiparasitária do artemether na esquistossomose mansônica experimental

OBJETIVO: Avaliar o efeito da administração intramuscular de artemether a camundongos infectados experimentalmente por Schistosoma mansoni no momento da infecção, durante a maturação dos esquistossômulos e após iniciada a oviposição. MÉTODOS: Oitenta camundongos Balb/c, fêmeas adultas, foram divididos em oito grupos com 10 animais cada. Sete grupos foram infectados por S. mansoni empregando-se 60 cercárias para cada animal, inoculadas por via subcutânea; o grupo restante foi mantido sem infecção. Entre os sete grupos infectados, seis foram tratados com artemether, segundo o seguinte esquema: três grupos receberam dose correspondente a 100 mg/kg no dia 0, 20 ou 60 após inoculação das cercárias; os demais receberam 50 mg/kg de artemether, no mesmo período que os lotes anteriores. Da 9ª, 10ª e 11ª semanas após infecção os camundongos infectados por S. mansoni foram submetidos a exames de fezes pela técnica de Kato-Katz. No 80º dia do experimento, os animais sobreviventes foram sacrificados e submetidos à perfusão do sistema porta para recuperação de vermes. Determinaram-se, nessa ocasião, os pesos corporal, hepático e esplênico de cada animal. RESULTADOS: Observou-se queda na oviposição e no número de vermes recuperados entre os camundongos tratados com artemether (50 ou 100 mg/kg) no 20º dia após infecção. A diminuição do número de vermes foi mais expressiva no caso de fêmeas de S. mansoni. Verificou-se, ainda, diminuição significativa nos pesos hepático e esplênico entre os animais tratados com 50 e 100 mg/kg de artemether no 20º dia e também entre os que receberam a droga na dose de 50 mg/kg 60 dias após infecção. CONCLUSÕES: Ficou evidenciada a atividade anti-Schistosoma do artemether, mesmo ao se empregar dose correspondente a 50 mg/kg, quando a droga foi administrada durante o período de maturação dos esquistossômulos no sistema porta do hospedeiro vertebrado.

O Mastomys natalensis como modelo alternativo nos estudos da esquistossomose mansoni experimental

Foram estudados aspectos da infecção e da imunidade experimentais em um roedor — o Mastomys natalensis — inoculado com a cepa LE (Belo Horizonte) do Schistosoma mansoni, comparando os resultados com os do camundongo albino, animal tido como melhor modelo experimental. Os parâmetros estudados ofereceram resultados em tudo semelhantes aos encontrados em camundongos, com o M. natalensis se mostrando bastante sensível à infecção pelo S. mansoni, sendo de grande utilidade para os estudos da biologia do parasito e/ou observações da imu-nologia advinda da infecção. Somando-se a tal fato a grande reprodutividade do animal em cativeiro, a criação fácil, o manejo e a manutenção simples em laboratório, sugere-se que seja o M. natalensis usado como modelo experimental alternativo nos diversos estudos da esquistossomose mansoni.

Comportamiento experimental del Sporothrix schenckii y la Leishmania mexicana en el hamster

La descripción macroscópica del proceso de patogénesis en hamsters inoculados subcutáneamente en nariz con Sporothrix schenckii ó Leishmania mexicana spp. proporcionó bases para diferenciar estos dos microorganismos en un modelo animal utilizado comunmente para estudiarlos. Observaciones secuenciales durante 150 días permitieron afirmar que en las infecciones causadas por estos patógenos se presentaron edema y eritema como signos primarios, seguidos de alopecia, necrosis y ulceración. La producción de pus fué una característica distintiva para el S. schenckii. Estos signos clínicos se observaron más temprano en la esporotricosis que en la infección por L. mexicana, mostrando diferencias estadísticas significantes en días promedio de aparición. El presente trabajo muestra que las lesiones producidas tanto por el S. schenckii como la L. mexicana en este modelo experimental comparten signos clínicos, pero el tiempo de aparición de los mismos y su frecuencia relativa permiten diferenciarlas. Las condiciones de inoculación como: cepa de los microorganismos, dosis del inóculo, sitio y vía de inoculación, deben tenerse presentes en la evaluación de su comportamiento experimental.

A critical evaluation of the expression of parasitemia in experimental Chagas' disease

This paper aims to study the best way to express the parasitemia of Trypanosoma cruzi's experimentally infected animals. Individual scores may have a great variability, not emphasized by the majority of the authors. A group of 50 rats infected with 1x10(6) trypomastigotes of T. cruzi Y strain was used and the parasitemia was estimated by BRENER' s method. The results showed that the median can avoid false results due to very high or low parasitemias but it does not have the mathematic properties necessary for analysis of variance. The comparison of the means of the original and transformed data, with their respective coefficients of variability (CV), showed that the logarithmic mean (Mlog) have the minor value of CV. Therefore, the Mlog is the best way to express the parasitemia when the data show great variability. The number of the animal for group did not affect the variability of data when the Mlog and CV were used.

Repercussões crónicas nas vias aéreas da hiperóxia neonatal : modelo experimental

RESUMO: O objectivo deste trabalho foi avaliar se a exposição crónica neonatal à hiperóxia mo-derada induz alterações funcionais e estruturais persistentes nas vias aéreas. Desenvolveu-se um modelo animal, no rato, a partir do qual se retiraram implicações para a compreensão das repercussões crónicas da hiperóxia neonatal sobre as vias aéreas de displasia broncopulmonar (DBP), em duas fases distintas: imediatamente após a exposi-ção neonatal a 50%O2 (grupo 50%O2) e após três semanas de recuperação em ar ambiente (grupo 50%O2+Ar).Compararam-se os resultados da resposta do músculo liso de traqueia (MLT) à esti-mulação in vitro com metacolina e salbutamol e avaliaram-se as alterações quantitativas da área de MLT, bem como as alterações qualitativas da estrutura da traqueia. Demonstrou-se que a exposição a 50% de oxigénio não tinha repercussões imediatas sobre a resposta in vitro do MLT à estimulação colinérgica, mas que induzia um aumento do relaxamento em resposta ao salbutamol. A contractilidade do MLT em resposta à estimula-ção com metacolina no grupo 50%O2+Ar foi significativamente superior à do grupo de con-trolo da mesma idade e também superior à observada no grupo 50%O2, enquanto que a resposta ao salbutamol se voltou a aproximar dos valores de controlo após a recuperação em normóxia. Não se observaram diferenças estatisticamente significativas na área de MLT entre os grupos experimental e de controlo, o que se deve provavelmente ao número reduzido de amostras avaliadas e à variabilidade deste parâmetro no grupo de controlo; contudo, verifi-cou-se um aumento médio de 15% imediatamente após a exposição à hiperóxia que persis-tiu após o período de recuperação.As alterações qualitativas sobre a arquitectura da traqueia, avaliadas por microscopia óptica, revelaram no grupo 50%O2 aumentos da espessura da matriz extracelular e da den-sidade de mastócitos desgranulados na submucosa e adventícia vizinhas do MLT, sem outras alterações relativamente ao grupo de controlo com 15 dias. As alterações da matriz extrace-lular foram reversíveis após a recuperação em ar ambiente. A densidade de mastócitos per-maneceu superior à do grupo de controlo de 36 dias de idade, apresentando-se em maior contiguidade com o MLT relativamente ao grupo 50%O2. Em síntese, demonstrou-se que a hiperóxia neonatal crónica em níveis moderados in-duz alterações da resposta contráctil do MLT e da estrutura da traqueia que podem ter ex-pressão funcional após a exposição ter cessado. Assim, o contributo original do presente trabalho foi o desenvolvimento de um modelo animal que permite avaliar os mecanismos pelos quais a hiperóxia é capaz de induzir, isoladamente, alterações crónicas da contracti-lidade, do relaxamento do ML e da estrutura das vias aéreas que podem ser responsáveis pela HRB persistente em doentes sujeitos a oxigenioterapia neonatal.-------------ABSTRACT: The aim of this work was to evaluate whether chronic neonatal exposure to hyperoxia in-duces persistent structural and functional airway changes. An animal model was developed, using neonatal rats, in order to understand the chronic effects of neonatal hyperoxia on the airways, in bronchopulmonary dysplasia, in two distinct phases: immediately after neonatal exposure to 50%O2 (50%O2 group) and after three weeks of recovery at ambient air (50%O2+Ar group).The results from the tracheal smooth muscle (TSM) response to in vitro stimulation with metacholine and salbutamol were compared and quantitative changes in TSM area, as well as qualitative changes in tracheal structure were evaluated. It was demonstrated that while exposure to 50% oxygen had no immediate effects on in vitro TSM response to cholinergic stimulation, it induced an increase in relaxation as a result of salbutamol administration. TSM contractility as a result of methacholine administration in the 50%O2 + Ar group was significantly higher than that of the same-age control group, and also higher than the one observed in the 50%O2 group, whereas the response to salbutamol admini-stration was once again closer to the control values after recovery in normoxia. There were no statistically significant differences in the TSM area between the experi-mental and control groups, which is most likely due to the reduced number of samples evalu-ated and to the variability of this parameter in the control group. However, there was an aver-age increase of 15% immediately after exposure to hyperoxia, which persisted after the recov-ery period. Qualitative changes in tracheal architecture, evaluated by optic microscopy, revealed that the 50%O2 group suffered an increase in the thickness of the extracellular matrix and degranu-lated mast cell density in the submucosa and adventitia adjacent to the TSM, without further changes when compared with the control group at 15 days of age. The changes in extracellular matrix were reversible after recovery in ambient air. Mast cell density remained higher than that of the control group at 36 days of age, and more contiguous to TSM than the 50%O2 group. In conclusion, it has been demonstrated that moderate levels of chronic neonatal hyperoxia in-duce changes in TSM contractile response and tracheal structure, which may be functionally ex-pressed after discontinuation of exposure. Therefore, the original contribution of the present work was the development of an animal model which allows the evaluation of the mechanisms through which hyperoxia alone can induce chronic changes in contractility and relaxation of SM and also in airway structure that can be responsible for the persistent airway hyperrespon-siveness found in patients who were submited to neonatal oxygen therapy.

Experimental infection and horizontal transmission of Bartonella henselae in domestic cats

In order to study B. henselae transmission among cats, five young cats were kept in confinement for two years, one of them being inoculated by SC route with B. henselae (10(5) UFC). Only occasional contact among cats occurred but the presence of fleas was observed in all animals throughout the period. Blood culture for isolation of bacteria, PCR-HSP and FTSZ (gender specific), and BH-PCR (species-specific), as well as indirect immunofluorescence method for anti-B. henselae antibodies were performed to confirm the infection of the inoculated cat as well as the other naive cats. Considering the inoculated animal, B. henselae was first isolated by blood culture two months after inoculation, bacteremia last for four months, the specific antibodies being detected by IFI during the entire period. All contacting animals presented with bacteremia 6 months after experimental inoculation but IFI did not detect seroconversion in these animals. All the isolates from these cats were characterized as Bartonella (HSP and FTSZ-PCR), henselae (BH-PCR). However, DNA of B. henselae could not be amplified directly from peripheral blood by the PCR protocols used. Isolation of bacteria by blood culture was the most efficient method to diagnose infection compared to PCR or IFI. The role of fleas in the epidemiology of B. henselae infection in cats is discussed.

Treatment with mebendazole is not associated with distal migration of adult Angiostrongylus costaricensis in the murine experimental infection

Abdominal angiostrongyliasis is a zoonotic infection produced by a metastrongylid intra-arterial nematode, Angiostrongylus costaricensis. Human accidental infection may result in abdominal lesions and treatment with anti-helminthics is contra-indicated because of potential higher morbidity with excitement or death of worms inside vessels. To evaluate the effect of mebendazole on localization of the worms, male Swiss mice, 5 week-old, were infected with 10 third stage larvae per animal. Twelve infected mice were treated with oral mebendazol, at 5 mg/kg/day, for 5 consecutive days, begining 22 days after inoculation. As control groups, 12 infected but non-treated mice and other 12 non-infected and non-treated mice were studied. The findings at necropsy were, respectively for the treated (T) and control (C) groups: 92% and 80% of the worms were inside the cecal mesenteric arterial branch; 8% and 10% were located inside the aorta. Only in the group C some worms (10%) were found inside the portal vein or splenic artery. These data indicate that treatment with mebendazole does not lead to distal or ectopic migration of A. costaricensis worms.

Susceptibility of Cebus apella monkey (Primates: Cebidae) to experimental Leishmania (L.) infantum chagasi-infection

In Amazonian Brazil, the Cebus apella monkey (Primates: Cebidae) has been associated with the enzootic cycle of Leishmania (V.) shawi, a dermotropic parasite causing American cutaneous leishmaniasis (ACL). It has also been successfully used as animal model for studying cutaneous leishmaniasis. In this work, there has been investigated its susceptibility to experimental Leishmania (L.) infantum chagasi-infection, the etiologic agent of American visceral leishmaniasis (AVL). There were used ten C. apella specimens, eight adult and two young, four males and six females, all born and raised in captivity. Two experimental infection protocols were performed: i) six monkeys were inoculated, intra-dermal via (ID), into the base of the tail with 2 x 10(6) promastigotes forms from the stationary phase culture medium; ii) other four monkeys were inoculated with 3 x 10(7) amastigotes forms from the visceral infection of infected hamsters by two different via: a) two by intravenous via (IV) and, b) other two by intra-peritoneal via (IP). The parameters of infection evaluation included: a) clinical: physical exam of abdomen, weigh and body temperature; b) parasitological: needle aspiration of the bone-marrow for searching of amastigotes (Giemsa-stained smears) and promastigotes forms (culture medium); c) immunological: Indirect fluorescence antibody test (IFAT) and, Delayed-type hypersensitivity (DTH). In the six monkeys ID inoculated (promastigotes forms) all parameters of infection evaluation were negative during the 12 months period of follow-up. Among the four monkeys inoculated with amastigotes forms, two IV inoculated showed the parasite in the bone-marrow from the first toward to the sixth month p.i. and following that they cleared the infection, whereas the other two IP inoculated were totally negative. These four monkeys showed specific IgG-antibody response since the third month p.i. (IP: 1/80 and IV: 1/320 IgG) toward to the 12th month (IP: 1/160 and IV: 1/5120). The DTH-conversion occurred in only one IV inoculated monkey with a strong (30 mm) skin reaction. Considering these results, we do not encourage the use of C. apella monkey as animal model for studying the AVL.

PARASITOLOGICAL AND MOLECULAR DIAGNOSIS IN EXPERIMENTAL Strongyloides venezuelensis INFECTION

Strongyloides venezuelensis is a parasitic nematode of rats which is frequently used as a model to study human and animal strongyloidiasis. The aim of this study was to evaluate the correlation between parasitological and molecular diagnosis in Strongyloides venezuelensis infection. PCR assays were used to detect S. venezuelensis DNA in fecal samples obtained from experimentally infected Rattus norvegicus. The results showed a higher sensitivity of the PCR assay in detecting the infection compared to parasitological methods.

ANIMAL MODELS FOR THE STUDY OF LEISHMANIASIS IMMUNOLOGY

Leishmaniasis remains a major public health problem worldwide and is classified as Category I by the TDR/WHO, mainly due to the absence of control. Many experimental models like rodents, dogs and monkeys have been developed, each with specific features, in order to characterize the immune response to Leishmania species, but none reproduces the pathology observed in human disease. Conflicting data may arise in part because different parasite strains or species are being examined, different tissue targets (mice footpad, ear, or base of tail) are being infected, and different numbers (“low” 1×102 and “high” 1×106) of metacyclic promastigotes have been inoculated. Recently, new approaches have been proposed to provide more meaningful data regarding the host response and pathogenesis that parallels human disease. The use of sand fly saliva and low numbers of parasites in experimental infections has led to mimic natural transmission and find new molecules and immune mechanisms which should be considered when designing vaccines and control strategies. Moreover, the use of wild rodents as experimental models has been proposed as a good alternative for studying the host-pathogen relationships and for testing candidate vaccines. To date, using natural reservoirs to study Leishmania infection has been challenging because immunologic reagents for use in wild rodents are lacking. This review discusses the principal immunological findings against Leishmania infection in different animal models highlighting the importance of using experimental conditions similar to natural transmission and reservoir species as experimental models to study the immunopathology of the disease.