Interactions of maize bushy stunt phytoplasma with the leafhopper vector, Dalbulus maidis (Delong and Wolcott) (Hemiptera: Cicadellidae) and associated microbiota

Phytoplasmas are bacteria with a persistent propagative transmission by insect vectors that generates direct and indirect interactions among them. In order to understand these interactions for maize bushy stunt phytoplasma (MBSP) and the leafhopper vector Dalbulus maidis (Hemiptera: Cicadellidae), two research lines were addressed. The first one aimed to determine the indirect effects of maize infection by MBSP on some biological and behavioral parameters of the vector, whereas a second line investigated direct interactions of the phytoplasma with D. maidis during its movement through the vector body following acquisition from plants, and associated microbiota. Indirect effects were investigated in choice experiments in which alighting and oviposition preferences by D. maidis were compared on healthy vs. MBSP-infected plants with variable incubation time (diseased plants with early and advanced symptoms, or still asymptomatic). Likewise, indirect effect of MBSP on the D. maidis biology was determined in two life table experiments in which the vector was reared on healthy vs. MBSP-infected plants expressing advanced disease symptoms or still asymptomatic. Choice experiments showed that alighting and oviposition preferences of D. maidis on MBSP-infected plants compared to healthy plants depend on the pathogen incubation period in the plant. The leafhopper preferred MBSP-infected plants over healthy ones during the asymptomatic phase of the disease, but rejected infected plants with advanced symptoms. The vector was able to acquire MBSP from asymptomatic infected plants shortly (3 days) after inoculation, but transmission efficiency increased when acquisition occurred at later stages of the pathogen incubation period (≥14 days) in the source plants and the test plants showed disease symptoms faster. These results suggest that MBSP modulates D. maidis preference for asymptomatic infected plants in the early stages of the crop, allowing rapid spread of this pathogen. Maize infection by the phytoplasma had a neutral effect on most life table parameters of D. maidis; a lower net reproductivity rate (Ro) was observed in the cohort reared on MBSP-infected plants with advanced symptoms, which was compensated to some extent by a higher sexual ratio. MBSP acquisition by all vector nymphal stadia was confirmed by PCR, and the pathogen as detected in both male and female reproductive organs. Concerning direct MBSP-vector interactions, transmission electron microscopy analyses showed phytoplasma-like cells in the midgut lumen, microvilli and epithelial cells, suggesting that MBSP enters the epithelium midgut through the microvilli wall. Within the epithelial cells, mitochondria and bacteria-like cells (possibly endosymbionts) were observed together with masses of phythoplasma-like cells. In the hemocoel, phytoplasma-like cells grouped into a matrix were also observed in association with bacteria-like cells similar to those observed in the midgut epithelium. Similar associations were found in the salivary gland. Interestingly, in-situ hybridization (FISH) technique revealed a variation in diversity and abundance of the microbiota in intestine and salivary glands of D. maidis adults over time after MBSP acquisition from plants. Sulcia sp., Cardinium sp. and eubacteria increased their abundance over time, whereas Rickettsia sp. decreased. The frequent association of the vector microbiota with the phytoplasma in some tissues of D. maidis suggests that endosymbiotic bacteria may play some role in MBSP-vector interactions.

Influence of different genotypes in the pattern of selenoprotein expression in response to Brazil nut supplementation

The micronutrient selenium is essential to human physiology. As the amino acid selenocysteine, it is inserted into selenoproteins with a wide range of functions including antioxidant capacity, thyroid hormone metabolism, improvement of immune system, brain function, fertility and reproduction. Low selenium status has been associated with increased risk for chronic diseases, such as cancer, type-2 diabetes and cardiovascular disease. In this context, several studies have been conducted in order to investigate if selenium supplementation could reduce the risk of such diseases. However, genetic variations may interfere in the response of individuals to a dietary intervention and must be considered as a important source of inter-individual variation. Therefore, this study was conducted was conducted to investigate the influence of genetic variations in selenoproteins genes on the response to an intervention with Brazil nuts, the richest source of selenium known in nature. The study included 130 healthy volunteers with both genders, aged 20 to 60 years old selected in University of São Paulo. They received nuts for 8 weeks, eating one nut a day, and did a washout period for more 8 weeks. All volunteers had a blood sampling collection every 4 weeks during 4 months, in a total of 5. The following analysis were done: anthropometric measurements, lipid profile, plasma malondialdehyde, plasma and erythrocyte Se, selenoprotein P, plasma and erythrocyte GPx activity, gene expression of GPX1, SEPP1, SELS and SEP15. The volunteers were also genotyped for SNPs rs1050450, rs3811699, rs1800699, rs713041, rs3877899, rs7579, rs34713741 and rs5845. Each unit of Brazil nut provided an average of 300 µg of selenium. All 130 volunteers completed the protocol. The concentrations of total cholesterol and glucose decreased after 8 weeks of supplementation. Moreover, HDL concentrations were higher for carriers of the variant T allele for GPX4_rs713041. The frequencies of the variant genotypes were 5,4% for rs1050450, rs3811699 e rs1088668, 10% for rs3877899, 19,2% for rs713041 e rs7579, 11,5% for rs5845 and 8,5% for rs34713741. The levels of the five biomarkers increased significantly after supplementation. In addition, erythrocyte GPx activity was influenced by rs1050450, rs713041 and rs5845; erythrocyte selenium was influenced by rs5845 and plasma selenium by rs3877899. Gene expression of GPX1, SEPP1 and SEP15 were higher after supplementation. The SNP rs1050450 influenced GPX1 mRNA expression and rs7579 influenced SEPP1 mRNA expression. Therefore, it can be concluded that the supplementation with one of Brazil nut for 8 weeks was efficient to reduce total cholesterol and glucose levels and to increase the concentrations of the main biomarkers of selenium status in healthy adults. Furthermore, our results suggest that GPX4_rs713041 might interfere on HDL concentrations and GPx1 activity, GPX1_rs1050450 might interfere on GPx1 activity, SEP15_rs5845 might interfere on GPx1 activity and erythrocyte selenium and SEPP1_3877899 might interfere on plasma Se levels. Therefore, the effect of genetic variations should be considered in future nutritional interventions evaluating the response to Brazil nut supplementation.

Proteção compartilhada e restauração de tráfego em redes ópticas de segmentação espectral flexível

A busca de sistemas de comunicação eficientes e econômicos é cada vez maior, principalmente com o aumento da demanda por tráfego e banda de transmissão. Conhecidas pela utilização eficiente de largura de banda, as redes ópticas de segmentação espectral flexível têm sido intensamente estudadas e são consideradas boas candidatas para a próxima geração de redes. Tais redes flexíveis utilizam formatos de modulação multinível e multiplexação multiportadora. Sistemas com alta capacidade de transmissão devem ser extremamente confiáveis, pois a falha em um dispositivo ou enlace da rede acarreta enorme perda de dados. Por isso, as técnicas de proteção e restauração do tráfego devem ser eficientes, de modo a garantir a sobrevivência da rede. Neste trabalho, foi desenvolvido um algoritmo capaz de lidar com cada requisição de conexão, encontrar um caminho óptico para transmissão e reservar outro caminho para recuperação do tráfego em caso de falha na rede. A seleção dos caminhos primário e de proteção utiliza a técnica smart-fit, que escolhe a solução com menor custo final, definido pela distância da rota percorrida somada ao custo do índice inicial da faixa espectral alocada, buscando equilibrar a escolha entre a posição no espectro e rota escolhida. Além disso, são definidos custos diferentes para slots de frequência livres e compartilhados, a fim de incentivar o compartilhamento espectral em caminhos de proteção. Dentre as técnicas de alocação espectral, a varredura com janela espectral varre o espectro em todos os enlaces da rota, slot por slot, em busca de uma faixa livre com tamanho suficiente para atender uma demanda. Neste trabalho, foi desenvolvida uma técnica chamada busca lógica, que lida simultaneamente com todos os enlaces da rota ao realizar operações lógicas com seus espectros. Em seguida, é realizada uma convolução entre a janela espectral, com tamanho da demanda, e o espectro resultante. A partir desse resultado, é possível saber as posições no espectro onde a demanda poderá ser alocada. Tal técnica, como será demonstrado, é mais veloz que a varredura com janela espectral. Para garantir a eficácia e confiabilidade do algoritmo, utilizando o software MATLAB, avaliou-se a probabilidade de bloqueio e probabilidade de bloqueio de banda, a fragmentação espectral média na rede, o grau de compartilhamento, a taxa de utilização espectral e a taxa de capacidade reservada. O algoritmo desenvolvido foi capaz de organizar o tráfego na rede de modo ordenado e pouco fragmentado, além de utilizar de maneira eficiente os recursos destinados à proteção.