Antigen-presenting cells transfected with Hsp65 messenger RNA fail to treat experimental tuberculosis

In the last several years, the use of dendritic cells has been studied as a therapeutic strategy against tumors. Dendritic cells can be pulsed with peptides or full-length protein, or they can be transfected with DNA or RNA. However, comparative studies suggest that transfecting dendritic cells with messenger RNA (mRNA) is superior to other antigen-loading techniques in generating immunocompetent dendritic cells. In the present study, we evaluated a new therapeutic strategy to fight tuberculosis using dendritic cells and macrophages transfected with Hsp65 mRNA. First, we demonstrated that antigen-presenting cells transfected with Hsp65 mRNA exhibit a higher level of expression of co-stimulatory molecules, suggesting that Hsp65 mRNA has immunostimulatory properties. We also demonstrated that spleen cells obtained from animals immunized with mock and Hsp65 mRNA-transfected dendritic cells were able to generate a mixed Th1/Th2 response with production not only of IFN-γ but also of IL-5 and IL-10. In contrast, cells recovered from mice immunized with Hsp65 mRNA-transfected macrophages were able to produce only IL-5. When mice were infected with Mycobacterium tuberculosis and treated with antigen-presenting cells transfected with Hsp65 mRNA (therapeutic immunization), we did not detect any decrease in the lung bacterial load or any preservation of the lung parenchyma, indicating the inability of transfected cells to confer curative effects against tuberculosis. In spite of the lack of therapeutic efficacy, this study reports for the first time the use of antigen-presenting cells transfected with mRNA in experimental tuberculosis

Equações de volume para Eucalyptus dunnii Maiden, determinadas para a depressão central do Estado do Rio Grande do Sul Equations for Eucalyptus dunnii maiden volume determination on the central depression of State Rio Grande do Sul

O estudo foi realizado para definir equações de volume total com casca e comercial sem casca para Eucalyptus dunnii Maiden, na idade de 6,5 anos, implantado em espaçamento 3 x 2m. Foram cubadas, pelo método de Smaliam, 80 árvores distribuídas em classe diamétrica e de forma proporcional em quatro hortos florestais. As equações de volume total com casca e comercial sem casca foram determinadas através do procedimento "stepwise" de regressão, de forma independente, para cada local e, posteriormente, foi estudada a possibilidade de agrupamento destas equações em um modelo único. A análise de covariância aplicada aos dados indicou a possibilidade do emprego de uma única equação para estimar o volume da espécie em toda a região. Selecionou-se pelo alto coeficiente de determinação e baixo erro padrão da estimativa a equação log v = - 9,06566 + 0,93016 . log(d² h) para o volume total com casca e log v = -14,14078 + 1,39608 . log(d² h) para o volume comercial sem casca. Além disso, não observou-se diferenças na forma do tronco e por conseqüência no volume das árvores nos quatro locais estudados.<br>This study has been carried out to define total bulk volume equations for Eucalyptus dunni Maiden, 6.5 years old, lined up along a 3 by 2 meter-spaced planted setting. These equations were supposed to both trees with and without bark. Eighty trees were cubed by Smalian's method distributed according to diameter classes and following a proportional way, over four forest sites. Equations were determinated through stepwise regression procedure, following an independent process for each site. A second step was evaluate the possiblility of these equations to be reduced into a single one. The covariance analysis applied to data showed the possibility of using an unique one equation to estimate the tree bulk volume to the region. Two equations were selected: one to calculate the total volume of trees with bark and another to determine the commercial volume without bark. The equation tom determine total volume was equal to: log v = -9.06566 + 0.93016 . log(d²h) and the equation to determine comercial volume was equal to: log v = -14.14078 + 1.39608 . log(d²h). Besides, no differences in the trunk shape and, consequently, in the bulk volume of trees in the four sites studied were observed

Concentrações plasmáticas de testosterona, triiodotironina (T3) e tiroxina (T4) em bodes submetidos ao estresse calórico Plasma concentrations of testosterone, triiodothyronine (T3), and thyroxine (T4) in bucks submitted to heat stress

Para verificar o efeito do estresse calórico (EC) nas concentrações plasmáticas de testosterona, triiodotironina (T3) e tiroxina (T4), oito bodes, das raças Saanen (n=4) e Alpina (n=4), foram mantidos em câmara bioclimática, sob condições de termoneutralidade (13,0ºC a 26,7ºC) durante 30 dias e, após um período (60 dias) de descanso, submetidos ao EC (23,7ºC a 34,0ºC) por 30 dias. Para minimizar as variações sazonais nos perfis hormonais devido ao fotoperíodo, durante toda fase experimental, incluindo a de adaptação em condições de termoneutralidade (30 dias), o fotoperíodo foi controlado utilizando-se alternância de dias longos (16h de luz e 8h de escuro) e de dias curtos (8h de luz e 16h de escuro) a cada 30 dias. As amostras de sangue foram coletadas duas vezes por semana durante cinco semanas. No conjunto das raças, o EC não influenciou (P>0,05) as concentrações de testosterona (1,8±0,2 vs 1,3±0,2ng/ml) e nem a de T4 (52,7±2,8 vs 50,0±2,8ng/ml). Houve declínio (P<0,01) das concentrações de T3 nos animais submetidos ao experimento (1,3±0,1 vs 1,0±0,1ng/ml), mas a redução foi observada somente nos bodes Saanen. Em ambas as raças, as concentrações de T3 e T4 variaram (P<0,01) conforme o dia da coleta das amostras de sangue. O EC foi suficiente para produzir uma resposta fisiológica com redução das concentrações plasmáticas de T3 em bodes das raças Saanen, mas não da raça Alpina, assim como não foi capaz de alterar os níveis plasmáticos de testosterona e nem de T4.<br>To verify the effect of heat stress (HS) on plasma testosterone, triiodothyronine (T3), and thyroxine (T4) concentrations, eight Saanen (n=4) and Alpine Brown (n=4) bucks were kept in climate chamber under thermal neutral conditions (13.0ºC to 26.7ºC) for 30 days. After a resting period (60 days), the same bucks were submitted to heat stress (23.7ºC to 34.0ºC) for another 30 days. To neutralize the seasonal variations of hormonal profiles throughout the period, the photoperiod was controlled every 30 days altering long (16 hours of light and 8 hours of darkness) and short days (8 hours of light and 16 hours of darkness). The blood samples were collected twice a week during five weeks. In both breeds, there was no effect of HS (P>0.05) on plasma concentrations of testosterone (1.8±0.2 vs 1.3±0.2ng/ml) and T4 (52.7±2.8 vs 50.0±2.8ng/ml). There was a decline (P<0.01) of plasma T3 concentrations (1.3±0.1 vs 1.0±0.1ng/ml) after HS treatment, but this reduction was only evident in Saanen bucks. In both breeds, the plasma concentrations of T3 and T4 varied (P<0.01) according to the day of blood sample collection. The HS was sufficient to provoke a physiological response with reduction of plasma concentrations of thyroid hormones mainly of T3 in Saanen bucks, but not in Alpine ones. The HS did not affect the plasma testosterone and T4 levels

Correlação entre heterose e divergência genética estimadas por cruzamentos dialélicos e marcadores moleculares rapd em populações de milho-pipoca Correlation between heterosis and genetic divergence estimated of diallel crosses and rapd molecular markers in populations of popcorn

Com o objetivo de correlacionar a heterose, estimada através de cruzamentos dialélicos, com a divergência genética obtida pelo uso de marcadores moleculares RAPD, oito populações de milho-pipoca (1-PASHA, 2-PAPA, 3-PAAPC, 4-PO, 5-ZL, 6-CMS 042, 7-RS 20 e 8-CMS 43) foram intercruzadas em esquema dialélico completo, sem recíprocos, no ano agrícola de 2002/2003, gerando 28 híbridos. A avaliação dos híbridos foi realizada no ano agrícola de 2003/2004, em Londrina e Ponta Grossa, PR, em um ensaio com trinta e oito tratamentos, constituídos de vinte e oito combinações híbridas, oito parentais e duas testemunhas (IAC 112 e IAC TC01). O delineamento experimental utilizado foi o de blocos casualizados com três repetições. Foram avaliados seis caracteres: massa de grãos, capacidade de expansão, altura de planta, altura de espiga, prolificidade e florescimento feminino. Foi utilizada a técnica de RAPD para a obtenção das estimativas de distâncias genéticas entre as populações. Os resultados inferem em correlações positivas e significativas entre a divergência genética detectada pelos marcadores RAPD e massa de grãos, altura de plantas, altura de espiga e prolificidade, dos vinte e oito híbridos avaliados no dialelo em estudo. Para capacidade de expansão, florescimento e heterose percentual não foi detectada correlação significativa com a divergência genética.<br>The objective of this study was to correlate the heterosis evaluated by diallel complete design with the genetic divergence estimated through the use of RAPD markers. Eight popcorn populations (1-PASHA, 2-PAPA, 3-PAAPC, 4-PO, 5-ZL, 6-CMS 42, 7-RS 20 and 8-CMS 43) were intercrossed on a complete diallel scheme, without reciprocal crosses, during 2002/2003 summer season, resulting in 28 hybrids. Hybrid evaluations were accomplished in the 2003/2004 summer season, at Londrina and Ponta Grossa, PR, in a trial with thirty-eight treatments, including all hybrid combinations, eight parentals and two controls (IAC 112 and IAC TC01). Complete randomized blocks design with tree replications was used. Traits evaluated were grain yield, popping expansion, plant height, ear height, ear prolificity and female flowering. RAPD was also used to estimate genetic distances among populations. The results infer in positive and significant correlations among the genetic divergence detected by RAPD molecular markers and the traits grain yield, plants height, ear height and ear prolificity of all hybrids evaluated on present diallel. For popping expansion, flowering and percentage heterosis no significant correlations with genetic divergence were detected