Tamoxifen as a potential antileishmanial agent: efficacy in the treatment of Leishmania braziliensis and Leishmania chagasi infections

The aim of this study was to evaluate the efficacy of tamoxifen in vivo in experimental models of cutaneous (CL) and visceral leishmaniasis (VL) caused by Leishmania braziliensis and Leishmania chagasi, respectively. Drug activity was assessed against intracellular amastigotes by treating infected macrophage cultures and evaluating the number of infected cells. In vivo efficacy of tamoxifen was tested in L. braziliensis-infected BALB/c mice and in L. chagasi-infected hamsters. Treatment with 20 mg/kg/day tamoxifen was administered for 15 days by the intraperitoneal route. Efficacy was evaluated through measurements of lesion size, parasite burden at the lesion site or liver and spleen and survival rate. Tamoxifen killed L. braziliensis and L. chagasi intracellular amastigotes with 50% inhibitory concentrations (IC(50)) of 1.9 +/- 0.2 and 2.4 +/- 0.3 mu M, respectively. Treatment of L. braziliensis-infected mice with tamoxifen resulted in significant reductions in lesion size and 99% decrease in parasite burden, compared with mock-treated controls. L. chagasi-infected hamsters treated with tamoxifen showed significant reductions in liver parasite load expressed as Leishman-Donovan units and 95% to 98% reduction in spleen parasite burden. All animals treated with tamoxifen survived while 100% of the mock-treated animals had died by 11 weeks after the interruption of treatment. Tamoxifen is effective in the treatment of CL and VL in rodent models.

Comparative genomics of the neglected human malaria parasite Plasmodium vivax

The human malaria parasite Plasmodium vivax is responsible for 25 - 40% of the similar to 515 million annual cases of malaria worldwide. Although seldom fatal, the parasite elicits severe and incapacitating clinical symptoms and often causes relapses months after a primary infection has cleared. Despite its importance as a major human pathogen, P. vivax is little studied because it cannot be propagated continuously in the laboratory except in non- human primates. We sequenced the genome of P. vivax to shed light on its distinctive biological features, and as a means to drive development of new drugs and vaccines. Here we describe the synteny and isochore structure of P. vivax chromosomes, and show that the parasite resembles other malaria parasites in gene content and metabolic potential, but possesses novel gene families and potential alternative invasion pathways not recognized previously. Completion of the P. vivax genome provides the scientific community with a valuable resource that can be used to advance investigation into this neglected species.

Enhancement of Th1 Lung Immunity Induced by Recombinant Mycobacterium bovis Bacillus Calmette-Guerin Attenuates Airway Allergic Disease

Mycobacterium bovis Bacillus Calmette-Guerin (BCG) has been shown to down-regulate experimental allergic asthma, a finding that reinforced the hygiene hypothesis. We have previously found that recombinant BCG (rBCG) strain that express the genetically detoxified Si subunit of pertussis toxin (rBCG-S1PT) exerts an adjuvant effect that enhances Th1 responses against BCG proteins. Here we investigated the effect of this rBCG-S1PT on the classical ovalbumin-induced mouse model of allergic lung disease. We found that rBCG-S1PT was more effective than wild-type BCG in preventing Th2-mediated allergic immune responses. The inhibition of allergic lung disease was not associated with increased concentration of suppressive cytokines or with an increased number of pulmonary regulatory T cells but was positively correlated with the increase in IFN-gamma-producing T cells and T-bet expression in the lung. In addition, an IL-12-dependent mechanism appeared to be important to the inhibition of lung allergic disease. The inhibition of allergic inflammation was found to be restricted to the lung because when allergen challenge was given by the intraperitoneal route, rBCG-S1PT administration failed to inhibit peritoneal allergic inflammation and type 2 cytokine production. Our work offers a nonclassical interpretation for the hygiene hypothesis indicating that attenuation of lung allergy by rBCG could be due to the enhancement of local lung Th1 immunity induced by rBCG-S1PT. Moreover, it highlights the possible use of rBCG strains as multipurpose immunomodulators by inducing specific immunity against microbial products while protecting against allergic asthma.

Phylogenetic, morphological and behavioural analyses support host switching of Trypanosoma (Herpetosoma) lewisi from domestic rats to primates

We characterized four Brazilian trypanosomes isolated from domestic rats and three from captive nonhuman primates that were morphologically similar to T. lewisi, a considered non-pathogenic species restricted to rodents and transmitted by fleas, despite its potential pathogenicity for infants. These isolates were identified as T. lewisi by barcoding using V7V8 SSU rDNA sequences. In inferred phylogenetic trees, all isolates clustered tightly with reference T. lewisi and T. lewisi-like trypanosomes from Europe, Asia and Africa and despite their high sequence conservation formed a homogeneous clade separate from other species of the subgenus T. (Herpetosoma). With the aim of clearly resolving the relationships between the Brazilian isolates from domestic rats and primates, we compared sequences from more polymorphic ITS rDNA. Results corroborated that isolates from Brazilian rats and monkeys were indeed of the same species and quite close to T. lewisi isolates of humans and rats from different geographical regions. Morphology of the monkey isolates and their behaviour in culture and in experimentally infected rats were also compatible with T. lewisi. However, infection with T. lewisi is rare among monkeys. We have examined more than 200 free-ranging and 160 captive monkeys and found only three infected individuals among the monkeys held in captivity. The findings of this work suggest that proximity of monkeys and infected rats and their exposure to infected fleas may be responsible for the host switching of T. Iewisi from their natural rodent species to primates. This and previous studies reporting T. lewisi in humans suggest that this trypanosome can cause sporadic and opportunistic fleaborne infection in primates. (C) 2010 Elsevier B.V. All rights reserved.

Beteendestudier och miljöberikning av gnagare

It is often challenging to create a new enrichment program for any animal. This is especially true with the additional challenge that I wanted to use only naturalistic enrichment, and I’m working with animals some of which are nocturnal. Even with a many hurdles, a little bit of enrichment can do a big difference and make a big difference in activity levels of the animals. Eskilstuna parkens zoo have a house called the MUSeum housing four kinds of rodents. Rodents are generally much more active at night and spend much of the day sleeping out of sight of the public. One of the goals of starting an enrichment program was to increase the animal’s activity throughout the day to provide an improved guest experience. Many of the guests would stop briefly at the exhibit only seeing the empty exhibit, unaware of that there were animals there. Before starting an enrichment program, I wanted to determine how the rodent was spending their time. By observing the animals’ behaviour for 15 minutes a day at various times throughout the day, for a total of 15 observations, I was able to construct an activity budget for their behaviour. The results obtained were, as expected, that the rodents spent a lot of time resting. By enrichment both with food and the temperature we were able to take the activity to another level, and several new behaviours were observed.

Avaliação da genotoxicidade induzida por emissões de veículos automotores - Ctenomys minutus como organismo bioindicador

Existe um número crescente de componentes químicos lançados ao meio ambiente, muitos dos quais são capazes de induzir efeitos danosos adversos à saúde de animais e humanos, representando uma causa importante de preocupação por seus possíveis efeitos a longo prazo. O impacto ecológico e os riscos a saúde dos organismos associados com a exposição a poluentes ambientais são extremamente difíceis de se avaliar devido a muitos desses componentes serem parte de misturas complexas. Os gases produzidos pelos motores dos veículos à combustão contém diversos poluentes sabidamente genotóxicos, como óxidos de nitrogênio (NOX), monóxido de carbono (CO), óxidos de enxofre (SOx), hidrocarbonetos (HC) e seus derivados, bem como particulados, e metais (cádmio, cromo, cobre, níquel, vanádio, zinco e chumbo). Todos esses compostos isolados ou associados a outros elementos são tóxicos ou de efeito danoso aos organismos, de forma não totalmente esclarecida. Este estudo teve como objetivo verificar o possível efeito genotóxico das emissões dos automóveis em roedor nativo Ctenomys minutus cronicamente exposto, através do Ensaio Cometa (EC), comparando os resultados com o Teste de Micronúcleos (MN), ambos em sangue periférico. Levando em consideração alguns fatores que pudessem influenciar os resultados dos testes de genotoxicidade, este trabalho ainda teve como objetivos: identificar a presença de alguns agentes envolvidos na poluição gerada pelos veículos; verificar possíveis diferenças sazonais, como temperatura e ventos; e se existe influência da idade e sexo dos roedores. Os C. minutus (Octodontidae-Rodentia), foram capturados em dois campos diferentes, ambos ao lado da estrada RS/030, na cidade de Osório, Estado do Rio Grande do Sul (RS): (a) Amaral, e (b) Weber. Animais para controle externo foram capturados no Campo Maribo à cerca de 3 km de distância de outra estrada (RS/389-Osório/RS), conseqüentemente afastada das emissões dos veículos. No final do período desse estudo, foram capturados 123 animais (73 fêmeas e 50 machos).

Slow Oscillations in the Mouse Hippocampus Entrained by Nasal Respiration

Different types of network oscillations occur in different behavioral, cognitive, or vigilance states. The rodent hippocampus expresses prominentoscillations atfrequencies between 4 and 12Hz,which are superimposed by phase-coupledoscillations (30 –100Hz).These patterns entrain multineuronal activity over large distances and have been implicated in sensory information processing and memory formation. Here we report a new type of oscillation at near- frequencies (2– 4 Hz) in the hippocampus of urethane-anesthetized mice. The rhythm is highly coherent with nasal respiration and with rhythmic field potentials in the olfactory bulb: hence, we called it hippocampal respiration-induced oscillations. Despite the similarity in frequency range, several features distinguish this pattern from locally generatedoscillations: hippocampal respiration-induced oscillations have a unique laminar amplitude profile, are resistant to atropine, couple differentlytooscillations, and are abolished when nasal airflow is bypassed bytracheotomy. Hippocampal neurons are entrained by both the respiration-induced rhythm and concurrent oscillations, suggesting a direct interaction between endogenous activity in the hippocampus and nasal respiratory inputs. Our results demonstrate that nasal respiration strongly modulates hippocampal network activity in mice, providing a long-range synchronizing signal between olfactory and hippocampal networks.

A microestrutura e o desenvolvimento dos órgãos genitais masculinos de preás (Galea spixii) criados em cativeiro

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fases do desenvolvimento e diferenciação testicular em cutias (Dasyprocta aguti) criadas em cativeiros

A cutia (Dasyprocta aguti) é um roedor silvestre encontrado amplamente na região Nordeste do Brasil. É uma espécie muito utilizada pela população humana de baixa renda como fonte alternativa de proteína na alimentação. Foram utilizadas 31 cutias, machos, provenientes da Universidade Federal do Piauí (FUFPI), Estado do Piauí e da Escola Superior de Agricultura de Mossoró Estado do Rio Grande do Norte. Os animais foram divididos em grupos etários desde o nascimento até os 14 meses de idade. O diâmetro nuclear médio foi obtido pela medida de 10 núcleos do tipo celular estudado em cada testículo, no estágio 1 do ciclo do epitélio seminífero. Nos animais que não apresentaram o epitélio organizado em estágios bem definidos em virtude da idade, foram feitas medidas em secções transversais escolhidas somente pelo contorno circular. O início da assincronia do processo espermatogênico foi observado a partir dos seis meses de idade. A puberdade, na cutia Dasyprocta aguti, foi definitivamente estabelecida a partir dos nove meses de idade, pois estavam presentes todos os tipos celulares e espermatozóides liberados no lume tubular em grande parte do parênquima testicular.

Caracterização morfológica e frequência dos estádios do ciclo do epitélio seminífero em preás (Galea spixii Wagler, 1831) criados em cativeiro

Estudos baseados nas características testiculares estão altamente relacionados com a eficiência reprodutiva de varias espécies. Assim, o projeto desenvolvido teve como objetivo identificar as células do epitélio seminífero, caracterizar histologicamente suas associações, que formam os estádios, e determinar a frequência destes. Os fragmentos de testículos, com 30, 45, 60, 75, 90, 105, 120, 150 dias foram coletados no Centro de Multiplicação da Universidade Federal Rural do Semi-Árido (UFERSA), Mossoró/ RN. Passando pelos processos de fixação, lavagens em soluções de concentrações crescentes de álcoois (70-100%), desidratação em xilol, inclusão em Histosec®, preparação das lâminas histológicas, colorações em Hematoxilina e Eosina (HE) e suas fotomicrografias para a caracterização dos núcleos celulares do epitélio germinativo e a definição dos oitos estágios do ciclo do epitélio seminífero (CES) baseados no Método da Morfologia Tubular. Das faixas etárias analisadas todos os animais de 90-150 dias de idade apresentaram todos os estádios do CES. Os estádios I e III foram os que apresentaram maior e menor freqüência, respectivamente. Os animais caracterizados como pré-púberes (30 dias), púberes (45-90 dias de idade) e pós-púberes (105150 dias de idade) apresentaram os estádios I, VIII e IV com uma maior freqüência, respectivamente.

Ovine leptospirosis in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morfologia do útero de cutias nulíparas e não nulíparas

Descreveu-se a morfologia do útero de cutias nulíparas e não nulíparas por meio de ovariossalpingoisterectomia. A análise macroscópica do útero foi realizada in loco e na peça retirada. Segmentos das porções proximal, média e distal do órgão foram colhidas, fixadas e preparadas histologicamente, sendo as amostras analisadas à microscopia de luz, e realizada a histomorfometria das camadas uterinas. Topograficamente, o útero localiza-se na região sublombar, caudalmente aos rins, em continuação aos ovários e às tubas uterinas, estendendo-se até à entrada da pelve, onde se posiciona dorsalmente à bexiga. Caracteriza-se como do tipo duplo, embora culmine em apenas um óstio cervical externo. Microscopicamente, a mucosa uterina é formada por variações do epitélio, de cilíndrico a pseudoestratificado, que se apoia em tecido conjuntivo frouxo, onde se observam glândulas endometriais revestidas por epitélio cilíndrico, além da presença de vasos sanguíneos. A camada muscular subdivide-se em interna ou submucosa, média ou vascular e externa ou subserosa. A camada serosa é composta por tecido conjuntivo e mesotélio. Na histomorfometria, verificou-se que a espessura uterina total e a espessura da camada mucosa, em média, foram maiores nas fêmeas não nulíparas.

The locus coeruleus and central chemosensitivity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Análise morfológica da placenta da paca (Agouti paca, Linnaeus, 1766: Estudo ao microscópio de luz e à microscopia eletrônica de transmissão

Foram estudados os aspectos morfológicos de nove placentas de paca (Agouti paca, L., 1766) mediante análises em microscopia de luz e eletrônica de transmissão dos fragmentos teciduais correspondentes à porção de maior conexão placentária em diferentes fêmeas gestantes, nos estágios intermediário e final da prenhez. Realizamos este estudo, pois, aliada à necessidade da procura de novas espécies que atuem como modelos experimentais adequados, havia a disponibilidade deste roedor em nosso meio; por outro lado, o melhor conhecimento dos aspectos reprodutivos destes animais oferece subsídios ao estabelecimento de criatórios racionais desta espécie, uma vez que a preservação deste vertebrado é necessária, além do grande interesse comercial em torno de sua carne. Os resultados mostraram que este roedor possui uma placenta do tipo vitelina e outra do tipo corioalantoidiana, sendo este órgão do tipo hemocorial, labiríntico, que se apresenta histologicamente composto por lóbulos divididos em três regiões distintas: o centro do lóbulo, o labirinto e o interlóbulo. Na região do centro do lóbulo, verificou-se a presença de artérias e veias; e em sua região periférica estavam presentes dois sistemas tubulares arranjados de forma paralela, onde as lacunas sangüíneas e os capilares estavam em íntimo contato, formando a região do labirinto. O interlóbulo era composto de artérias e veias. O trofoblasto era o principal componente da placenta, que, independentemente da região onde se encontrava, mostrava-se de natureza sincicial. Ultra-estruturalmente a barreira placentária da paca foi classificada como hemomonocorial.

Descrição anátomo-topográfica do coração da paca (Agouti paca)

Background: The domestic animals heart is a conical hollow viscera, surrounded by pericardium, laterally compressed, accompanying the thorax shape. Atriums constituted the heart basis and their auricles partially bound the initial portion of the aorta and pulmonary trunk. In mammals, heart is kept suspended in the thoracic cavity and the pericardic sac is fixed dorsally by great veins and arteries roots, and ventrally fixed to the sternum, although its fixation to the diaphragm varies among species. This paper aimed to describe morphological aspects of the heart of the paca, the second biggest Brazilian rodent.Materials, Methods & Results: There were used 12 hearts of adult pacas for this study, obtained from the UNESP, Jaboticabal, SP, which died due fights or anesthesia during bandages or radiograph exams. The thoracic aorta was filled with colored latex and the animal was set in a 10% formaldehyde solution for at least 72 hours. The thoracic cavity was dissected and hearts individualized and measured with a paquimeter, lateromedially, craniocaudally and dorsoventrally. The paca heart is placed between the first and fifth intercostal space (ICS), in a craniocaudal oblique position; its basis is craniodorsally positioned, on the middle third between the first and second ICS and its apex is located near the sternodiaphragmatic joint, on the fifth ICS, tilted to the left antimere. The heart is surrounded by pericardium, which from ventrocaudally is originated the sternopericardic ligament, that continues as phrenopericardic ligament. At the heart basis, the rising of the pulmonary trunk was observed and the conus arteriosus formed a typical projection. The aorta also rised from the heart basis and its arch, which was caudally curved, crossed dorsally the pulmonary trunk; the right cranial and caudal cava veins drained to the right atrium. There is a left cranial cava vein, which surrounded the left atrium and joined the right caudal cava vein on the right atrium. The azygos vein joins the right cranial cava vein and four pulmonary veins drained to the left atrium. At palpation, a hard structure on the rising of the aorta was observed, similarly to a cartilaginous tissue, which would be part of the cardiac skeleton. The left and right coronary arteries were observed in all hearts.Discussion: The paca heart is anatomica and topographically similar to those of domestic mammals, differing from them for being placed one intercostal space more cranial and due to the presence of two cranial cava veins, the left and the right ones, besides the presence of the caudal cava vein. This vascular description is similar to that of small rodents, as rats and mice. In paca heart, the sinus venous, the terminal crest, the oval fossa, the atrioventricular valvae, the papillary muscles and tendinous cords, besides smooth atriums and auricles covered by pectinate muscles, were observed. The sternopericardic ligament, which is dorsally elongated as phrenopericardic ligament, is similar to the one present in humans, pigs, castors, and different from the one observed in carnivorous, that presents the phrenopericardic ligament and from the one of horses and ruminants, which present the sternopericardic ligament.