Microbial Symbionts in Insects Influence Down-Regulation of Defense Genes in Maize

Diabrotica virgifera virgifera larvae are root-feeding insects and significant pests to maize in North America and Europe. Little is known regarding how plants respond to insect attack of roots, thus complicating the selection for plant defense targets. Diabrotica virgifera virgifera is the most successful species in its genus and is the only Diabrotica beetle harboring an almost species-wide Wolbachia infection. Diabrotica virgifera virgifera are infected with Wolbachia and the typical gut flora found in soil-living, phytophagous insects. Diabrotica virgifera virgifera larvae cannot be reared aseptically and thus, it is not possible to observe the response of maize to effects of insect gut flora or other transient microbes. Because Wolbachia are heritable, it is possible to investigate whether Wolbachia infection affects the regulation of maize defenses. To answer if the success of Diabrotica virgifera virgifera is the result of microbial infection, Diabrotica virgifera virgifera were treated with antibiotics to eliminate Wolbachia and a microarray experiment was performed. Direct comparisons made between the response of maize root tissue to the feeding of antibiotic treated and untreated Diabrotica virgifera virgifera show down-regulation of plant defenses in the untreated insects compared to the antibiotic treated and control treatments. Results were confirmed via QRT-PCR. Biological and behavioral assays indicate that microbes have integrated into Diabrotica virgifera virgifera physiology without inducing negative effects and that antibiotic treatment did not affect the behavior or biology of the insect. The expression data and suggest that the pressure of microbes, which are most likely Wolbachia, mediate the down-regulation of many maize defenses via their insect hosts. This is the first report of a potential link between a microbial symbiont of an insect and a silencing effect in the insect host plant. This is also the first expression profile for a plant attacked by a root-feeding insect

Cell-Surface Marker Signatures for the Isolation of Neural Stem Cells, Glia and Neurons Derived from Human Pluripotent Stem Cells

Neural induction of human pluripotent stem cells often yields heterogeneous cell populations that can hamper quantitative and comparative analyses. There is a need for improved differentiation and enrichment procedures that generate highly pure populations of neural stem cells (NSC), glia and neurons. One way to address this problem is to identify cell-surface signatures that enable the isolation of these cell types from heterogeneous cell populations by fluorescence activated cell sorting (FACS).We performed an unbiased FACS- and image-based immunophenotyping analysis using 190 antibodies to cell surface markers on naïve human embryonic stem cells (hESC) and cell derivatives from neural differentiation cultures. From this analysis we identified prospective cell surface signatures for the isolation of NSC, glia and neurons. We isolated a population of NSC that was CD184(+)/CD271(-)/CD44(-)/CD24(+) from neural induction cultures of hESC and human induced pluripotent stem cells (hiPSC). Sorted NSC could be propagated for many passages and could differentiate to mixed cultures of neurons and glia in vitro and in vivo. A population of neurons that was CD184(-)/CD44(-)/CD15(LOW)/CD24(+) and a population of glia that was CD184(+)/CD44(+) were subsequently purified from cultures of differentiating NSC. Purified neurons were viable, expressed mature and subtype-specific neuronal markers, and could fire action potentials. Purified glia were mitotic and could mature to GFAP-expressing astrocytes in vitro and in vivo.These findings illustrate the utility of immunophenotyping screens for the identification of cell surface signatures of neural cells derived from human pluripotent stem cells. These signatures can be used for isolating highly pure populations of viable NSC, glia and neurons by FACS. The methods described here will enable downstream studies that require consistent and defined neural cell populations

Comparação do Índice de Torg obtido por meio de radiografia e ressonância magnética nos pacientes com mielopatia cervical espondilótica Comparación del Índice de Torg, obtenido por medio de radiografía y resonancia, en los pacientes con mielopatía espondilótica cervical Comparison of the Torg Index obtained by radiography and magnetic resonance in patients with spondylotic cervical myelopathy

OBJETIVO: A Mielopatia Cervical Espondilótica (MCE) é uma disfunção da medula espinhal relacionada à degeneração típica do envelhecimento. No estudo de imagem podemos obter a medida do Índice de Torg, para estimar a estenose cervical. Objetivamos, assim, medir o Índice de Torg através de radiografia e ressonância magnética (RM), possibilitando verificar possíveis discrepâncias entre os métodos de medida. MÉTODOS: Realizada mensuração do Índice de Torg na radiografia e na RM da coluna cervical, sendo obtido através da relação entre a superfície posterior do corpo vertebral e o ponto mais próximo à linha laminar correspondente, dividido pelo diâmetro sagital do corpo vertebral. RESULTADOS: Participaram 29 pacientes, sendo 10 mulheres e 19 homens, com médias de idade 48,1 ± 11 anos, de peso 68,7 ± 5 Kg e de altura 1,68 ± 0,6 m. Houve diferença significativa entre o Índice de Torg calculado através de radiografia e RM, sendo menores os índices observados na RM (radiografia: 0,73 ± 0,17 vs. RM: 0,48 ± 0,14, p< 0,05). No entanto, ambas as aferições traduzem o mesmo resultado: estenose cervical absoluta (Índice de Torg < 0,8). CONCLUSÕES: Nossos resultados corroboram relatos de outros autores que acreditam que o Índice de Torg medido através da radiografia, como preconizado, minimiza a real estenose do canal cervical. Sugerimos em nosso estudo que a RM permite melhor estimativa do grau de estenose do canal, muito embora nossos resultados em relação ao grau de estenose cervical tenham sido semelhantes estatisticamente.<br>OBJETIVO: La Mielopatía Cervical Espondilótica (MCE) es un trastorno relacionado con la degeneración de la médula espinal, típica del envejecimiento. En el estudio de imagen se puede obtener la medida del Índice de Torg para estimar la estenosis cervical. Nuestro objetivo es, por tanto, medir el índice de Torg mediante rayos-X y resonancia magnética (RM), posibilitando verificar posibles discrepancias entre los métodos de medición. MÉTODOS: Se realiza la medición del Índice de Torg en la radiografía y en la resonancia magnética de la columna cervical, siendo obtenido mediante la determinación de la relación entre la superficie posterior del cuerpo vertebral y el punto más cercano a la línea de la lámina correspondiente, dividido por el diámetro sagital del cuerpo vertebral. RESULTADOS: Participó un total de pacientes 29, siendo 10 mujeres y 19 hombres, edad promedio de 48,1 ± 11 años, peso 68,7 ± 5 Kg y altura de 1,68 ± 0,6 m. No hubo diferencias significativas entre el Índice de Torg calculado por la radiografía y el de la resonancia magnética, siendo menores los índices observados en la RM (radiografía: 0,73 ± 0,17 vs RM: 0,48 ± 0,14, p <0,05). Sin embargo, ambas mediciones reflejan el mismo resultado: estenosis cervical absoluta (Índice de Torg <0,8). CONCLUSIONES: Nuestros resultados confirman los informes de otros autores quienes creen que el Índice de Torg medido por radiografía, como se recomienda, minimiza la estenosis real del canal cervical. En nuestro estudio se sugiere que la RM permite hacer una mejor estimativa del grado de estenosis del canal, aunque nuestros resultados, sobre el grado de estenosis cervical, fueron estadísticamente similares.<br>OBJECTIVE: Cervical spondylotic myelopathy (SCM) is a dysfunction related to spinal cord degeneration typical of aging. In the imaging studies can achieve a measure of the Torg index to estimate cervical stenosis. We aim therefore to measure the Torg index in X-rays and Magnetic Resonance (MR), to investigate possible discrepancies between the methods of measurement. METHODS: The Torg Index was measured on radiographs and MR of the cervical spine, obtained by determining the relationship between the posterior surface of the vertebral body and the nearest point to the corresponding laminar line, divided by the sagittal diameter of vertebral body. RESULTS: A total 29 patients has participated, 10 women and 19 men, with mean age of 48.1 ± 11 years, weight 68.7 ± 5kg and height 1.68 ± 0.6m. There were significant differences between the Torg Index calculated by radiography and MR, and lower rates observed on MR (radiography: 0.73 ± 0.17 vs. RM: 0.48 ± 0.14, p <0.05). However, both measurements reflect the same result: absolute cervical stenosis (Torg index < 0.8). CONCLUSIONS: Our results confirm reports by other authors who believe that the Torg Index measured by radiography, as recommended, minimizes the actual cervical canal stenosis. Our study suggests that MRI allows better estimate of the degree of stenosis, although our results regarding the degree of cervical stenosis were statistically similar