Avaliação do efeito anti-inflamatório e antiasmático da 15-deoxy-delta-12,14-prostaglandina J2 em modelos murinos de asma

A asma é um distúrbio crônico pulmonar caracterizado por inflamação, obstrução e remodelamento brônquico, levando a sintomas como sibilo, tosse e falta de ar. A terapia antiasmática consiste em corticosteroides inalados e agonistas β2 de curta ou longa duração. O tratamento é limitado por efeitos colaterais e refratariedade de alguns pacientes, justificando a necessidade de novas terapias. Estudos demonstram que a 15-deoxy-delta- 12,14-prostaglandina J2 (15d-PGJ2), um ligante endógeno de receptores ativados por proliferadores de peroxissomos do tipo gama (PPAR-γ), é capaz de reduzir a expressão de citocinas pró-inflamatórias, o que pode resultar em benefícios no tratamento de doenças com esse perfil. O objetivo deste estudo foi avaliar o potencial anti-inflamatório e antiasmático da 15d-PGJ2 em modelos experimentais de asma. Camundongos A/J machos foram sensibilizados nos dias 0 e 7 através de injeção subcutânea (s.c.), contendo ovoalbumina (OVA) e Al(OH)3, e desafiados com 4 instilações intranasais (i.n.) de OVA em intervalos semanais. O tratamento com 15d-PGJ2 (30 e 100 g/Kg, s.c.) foi realizado 30 min antes dos desafios a partir da terceira provocação antigênica. Em outro modelo, camundongos A/J foram desafiados intranasalmente com extrato de ácaro 3 vezes por semana durante 3 semanas. As administrações de 15d-PGJ2 (30, 70 e 100 g/Kg, s.c. e 0,65; 1,5 e 2,3 g/animal, i.n.) foram realizadas a partir da 3 semana, 30 min antes dos desafios. As análises ocorreram 24 h após o último desafio. Nossos resultados mostraram que, em camundongos previamente sensibilizados e desafiados com OVA, a administração de 15d-PGJ2 limitou significativamente o influxo peribrônquico de eosinófilos e neutrófilos, bem como a produção de muco por células caliciformes e fibrose sub-epitelial, além da hiperreatividade das vias aéreas e produção de IL-5. A redução do epitélio brônquico e das citocinas IL-13 e TNF-α foram observadas somente na maior dose administrada. No modelo HDM a inflamação e o remodelamento foram atenuados em todas as doses administradas do composto, enquanto que a hiperresponssividade brônquica foi inibida apenas nas doses de 70 e 100 μg/Kg (via sistêmica) e na dose intermediária dada topicamente (1,5 μg/animal, i.n.). Os níveis de citocinas foram atenuados pelo tratamento subcutâneo, porém somente os níveis de IL-17, eotaxina-1 e TNF-α foram inibidos com a dose intranasal de 0,65 g/animal. O aumento da expressão de NF-κB, induzido por provocação com HDM também foi reduzido significativamente pela administração de 15d-PGJ2. Em conjunto, nossos dados indicam que o tratamento com 15d-PGJ2 inibe alterações cruciais associadas à patogênese da asma, em modelos experimentais distintos da doença, demonstrando possuir grande potencial para controlar e reverter inflamação, hiperreatividade e remodelamento pulmonar desencadeados por provocação alérgica.

Anti-grazing properties of the toxic dinoflagellate Karlodinium veneficum during predator-prey interactions with the copepod Acartia tonsa

Karlodinium veneficum (syn. Karlodinium micrum, Bergholtz et al. 2006; J Phycol 42:170–193) is a small athecate dinoflagellate commonly present in low levels in temperate, coastal waters. Occasionally, K. veneficum forms ichthyotoxic blooms due to the presence of cytotoxic, hemolytic compounds, putatively named karlotoxins. To evaluate the anti-grazing properties of these karlotoxins, we conducted food removal experiments using the cosmopolitan copepod grazer Acartia tonsa. Wild-caught, adult female A. tonsa were exposed to 6 monoalgal or mixed algal diets made using bloom concentrations of toxic (CCMP 2064) and non-toxic (CSIC1) strains of K. veneficum. Ingestion and clearance rates were calculated using the equations of Frost (1972). Exposure to the toxic strain of K. veneficum did not contribute to an increased mortality of the copepods and no significant differences in copepod mortality were found among the experimental diets. However, A. tonsa had significantly greater clearance and ingestion rates when exposed to a monoalgal diet of the non-toxic strain CSIC1 than when exposed to the monoalgal diet of toxic strain CCMP 2064 and mixed diets dominated by this toxic strain. These results support the hypothesis that karlotoxins in certain strains of K. veneficum deter grazing by potential predators and contribute to the formation and continuation of blooms.

Investigation of anti-lock braking strategies for heavy goods vehicles

An articulated lorry was instrumented in order to measure its performance in straight-line braking. The trailer was fitted with two interchangeable tandem axle sub-chassis, one with an air suspension and the other with a steel monoleaf four-spring suspension. The brakes were only applied to the trailer axles, which were fitted with anti-lock braking systems (ABS), with the brake torque controlled in response to anticipated locking of the leading axle of the tandem. The vehicle with the air suspension was observed to have significantly better braking performance than the steel suspension, and to generate smaller inter-axle load transfer and smaller vertical dynamic tyre forces. Computer models of the two suspensions were developed, including their brakes and anti-lock systems. The models were found to reproduce most of the important features of the experimental results. It was concluded that the poor braking performance of the steel four-spring suspension was mainly due to interaction between the ABS and inter-axle load transfer effects. The effect of road roughness was investigated and it was found that vehicle stopping distances can increase significantly with increasing road roughness. Two alternative anti-lock braking control strategies were simulated. It was found that independent sensing and actuation of the ABS system on each wheel greatly reduced the difference in stopping distances between the air and steel suspensions. A control strategy based on limiting wheel slip was least susceptible to the effects of road roughness.

An experimental semi-active anti-roll system

The use of anti-roll bars to provide additional roll stiffness and therefore to reduce the trade-off between ride and rollover performance has previously been studied. However, little work has been carried out to investigate the benefits of a switchable roll stiffness. Such a semi-active anti-roll system has the ability to have a low roll stiffness during straight-ahead driving for improved ride performance and high roll stiffness during cornering for improved roll performance. Modelling of such a system is conducted and the model is validated against a semi-active anti-roll system fitted to an experimental vehicle. Experimental and theoretical investigations are used to investigate the performance of such a system with several different strategies employed to switch to the high-stiffness state. The use of an air suspension on the vehicle to roll into corners is also investigated, as is the possibility of exploiting the road layout by allowing the vehicle to be in a low-roll-stiffness configuration during a corner, and then to switch to the high-roll-stiffness configuration midcorner, hence 'locking in' a roll angle. The best rollover performance improvement that was achieved was 12.5 per cent. © IMechE 2008.

Molecular characterization of Trimeresurus stejnegeri venom L-amino acid oxidase with potential anti-HIV activity

A novel L-amino acid oxidase, named TSV-LAO, has been purified and cloned from the snake Trimeresurus stejnegeri. Fifty percentage cytotoxic concentrations (CC50) of TSV-LAO on C8166 cells were 24 and 390 nM in the absence or presence of catalase (400nM), respectively. However, at concentrations that showed little effect on cell viability, TSV-LAO displayed dose dependent inhibition on HIV-1 infection and replication. The antiviral selectivity indexes (CC50/EC50) were 16 and 6, respectively, corresponding to the measurements of syncytium formation and HIV-1 p24 antigen expression. Interestingly, the presence of catalase resulted in an increase of its antiviral selectivity to 52 and 38. Under the same conditions, no anti-HIV-1 activity was observed by exogenous addition of H2O2. The complete amino acid sequence of TSV-LAO, as deduced from its cDNA, exhibits a high degree of sequence identity with other snake venom LAOs. (C) 2003 Elsevier Inc. All rights reserved.