Differential expression of microRNAs in mouse pain models

<p>Abstract</p> <p>Background</p> <p>MicroRNAs (miRNAs) are short non-coding RNAs that inhibit translation of target genes by binding to their mRNAs. The expression of numerous brain-specific miRNAs with a high degree of temporal and spatial specificity suggests that miRNAs play an important role in gene regulation in health and disease. Here we investigate the time course gene expression profile of miR-1, -16, and -206 in mouse dorsal root ganglion (DRG), and spinal cord dorsal horn under inflammatory and neuropathic pain conditions as well as following acute noxious stimulation.</p> <p>Results</p> <p>Quantitative real-time polymerase chain reaction analyses showed that the mature form of miR-1, -16 and -206, is expressed in DRG and the dorsal horn of the spinal cord. Moreover, CFA-induced inflammation significantly reduced miRs-1 and -16 expression in DRG whereas miR-206 was downregulated in a time dependent manner. Conversely, in the spinal dorsal horn all three miRNAs monitored were upregulated. After sciatic nerve partial ligation, miR-1 and -206 were downregulated in DRG with no change in the spinal dorsal horn. On the other hand, axotomy increases the relative expression of miR-1, -16, and 206 in a time-dependent fashion while in the dorsal horn there was a significant downregulation of miR-1. Acute noxious stimulation with capsaicin also increased the expression of miR-1 and -16 in DRG cells but, on the other hand, in the spinal dorsal horn only a high dose of capsaicin was able to downregulate miR-206 expression.</p> <p>Conclusions</p> <p>Our results indicate that miRNAs may participate in the regulatory mechanisms of genes associated with the pathophysiology of chronic pain as well as the nociceptive processing following acute noxious stimulation. We found substantial evidence that miRNAs are differentially regulated in DRG and the dorsal horn of the spinal cord under different pain states. Therefore, miRNA expression in the nociceptive system shows not only temporal and spatial specificity but is also stimulus-dependent.</p

Narrar histórias para se constituir educador ambiental pela pesquisa

Research, along with narrative, is presented as an educational principle which can transform the reality in environmental teacher education. The research exercise described in this paper was developed by a discussion group - GDP - about research in environmental teacher education at the IV EPEA, a seminar on research in Environmental Education. We believe that narrative may transform environmental educators in their learning process.Apresenta-se a pesquisa como princípio educativo agregada à narrativa como elemento transformador da realidade da formação de educadores ambientais. O exercício de pesquisa apresentado foi desenvolvido no grupo de discussão - GDP - sobre pesquisa na formação de educadores ambientais no IV EPEA - Encontro de Pesquisa em Educação Ambiental. Aposta-se na possibilidade da narrativa na constituição transformadora de educadores ambientais

Osteoartropatia hipertrófica idiopática: relato de caso e revisão da literatura Primary hypertrophic osteoarthropathy: case report and literature review

A osteoartropatia hipertrófica primária é uma síndrome rara, caracterizada pela presença de baqueteamento digital de mãos e pés, aumento das extremidades e de tecidos periarticulares secundários à proliferação óssea, fisionomia facial grosseira, dor e edema articular. A forma primária representa 3 a 5% de todos os casos de osteoartropatia hipertrófica e tem como fatores etiológicos influência genética, anormalidade da atividade fibroblástica e alteração no suprimento sanguíneo periférico. Apresenta evolução crônica e insidiosa, alternando fases de exacerbação com períodos assintomáticos. As manifestações clínicas são variáveis. A denominação síndrome completa é reservada aos casos com paquidermia (espessamento cutâneo da face e couro cabeludo, periostite e cutis vertici gyrata); síndrome incompleta, quando não há envolvimento do couro cabeludo; e frustra, quando se observa paquidermia com periostite mínima ou ausente. Os autores descrevem um homem branco de 39 anos diagnosticado com a forma primária da osteoartropatia hipertrófica. A partir do relato de caso, discutem-se características clínicas, radiológicas e a abordagem terapêutica dessa patologia.<br>Primary hypertrophic osteoarthropathy is a rare syndrome, consisting of clubbed hands, fingers, and feet digits; enlarged extremities secondary to periarticular and bone proliferation; thickened facial skin; painful and swollen joints. The idiopathic form represents 3 to 5 per cent of all cases of hypertrophic osteoarthropaty. Genetic influence, abnormal fibroblasts activity, and changes of the peripherical blood flow appear to be significant on the pathogenesis. Clinical manifestations are variable: the term complete syndrome is used for the patient with pachydermia, coarsening of the face skin and scalp, periostitis, and cutis verticis gyrata); the incomplete form, when there is no sparing of the scalp; and the frusted form for pachydermia with minimal or absent periostitis. The authors describe a 39-year-old white man diagnosed with primary hypertrophic osteoarthropathy. We also report the clinical and radiological carateristics of this syndrome and terapeutical approach of pachydermoperiostosis

Effect of Melatonin in Epithelial Mesenchymal Transition Markers and Invasive Properties of Breast Cancer Stem Cells of Canine and Human Cell Lines.

Cancer stem cells (CSCs) have been associated with metastasis and therapeutic resistance and can be generated via epithelial mesenchymal transition (EMT). Some studies suggest that the hormone melatonin acts in CSCs and may participate in the inhibition of the EMT. The objectives of this study were to evaluate the formation of mammospheres from the canine and human breast cancer cell lines, CMT-U229 and MCF-7, and the effects of melatonin treatment on the modulation of stem cell and EMT molecular markers: OCT4, E-cadherin, N-cadherin and vimentin, as well as on cell viability and invasiveness of the cells from mammospheres. The CMT-U229 and MCF-7 cell lines were subjected to three-dimensional culture in special medium for stem cells. The phenotype of mammospheres was first evaluated by flow cytometry (CD44(+)/CD24(low/-) marking). Cell viability was measured by MTT colorimetric assay and the expression of the proteins OCT4, E-cadherin, N-cadherin and vimentin was evaluated by immunofluorescence and quantified by optical densitometry. The analysis of cell migration and invasion was performed in Boyden Chamber. Flow cytometry proved the stem cell phenotype with CD44(+)/CD24(low/-) positive marking for both cell lines. Cell viability of CMT-U229 and MCF-7 cells was reduced after treatment with 1mM melatonin for 24 h (P<0.05). Immunofluorescence staining showed increased E-cadherin expression (P<0.05) and decreased expression of OCT4, N-cadherin and vimentin (P<0.05) in both cell lines after treatment with 1 mM melatonin for 24 hours. Moreover, treatment with melatonin was able to reduce cell migration and invasion in both cell lines when compared to control group (P<0.05). Our results demonstrate that melatonin shows an inhibitory role in the viability and invasiveness of breast cancer mammospheres as well as in modulating the expression of proteins related to EMT in breast CSCs, suggesting its potential anti-metastatic role in canine and human breast cancer cell lines

Narrar histórias para se constituir educador ambiental pela pesquisa

O Programa de Pós-Graduação em Educação Ambiental da FURGtem, ao longo dos últimos anos, investido na pesquisa como princípioeducativo (DEMO, 1997; GALIAZZI, 2003) da formação de educadoresambientais pesquisadores. Este texto apresenta-se nessa perspectiva e a ela seagrega a pesquisa narrativa como elemento transformador da realidade daformação de educadores ambientais. Nesse sentido, a narrativa está sendopensada como eixo articulador de diferentes atividades e como processo dereflexão sobre a própria constituição do educador ambiental, pretendendo-se estabelecer um programa de pesquisa em comunidades de aprendizagemcom os alunos, mestrandos e doutorandos do PPGEA.A primeira autora, ao ser convidada a coordenar um grupo dediscussão – GDP – sobre pesquisa na formação de educadores ambientaisno IV EPEA – Encontro de Pesquisa em Educação Ambiental, propôs oGDP como o desenvolvimento de um exercício de pesquisa narrativa.Por isso, o texto aqui apresentado é a aposta de uma possibilidadeda narrativa como constituidora de educadores ambientais. Para realizá-la,utilizamos especialmente o ambiente virtual como ferramenta de produçãode significados dessas histórias (www.ceamecim.furg.br/avatool2/2). Osrelatos foram produzidos pelos participantes no IV EPEA – GDPFormação de Educadores Ambientais, realizado em Rio Claro-SP, de 15 a18 de julho de 2007.Neste trabalho, apresentamos inicialmente os princípios dapesquisa narrativa permeados pelas intenções postas, assumidas pelo grupode participantes e desenvolvidas durante o GDP. Foi assim que o texto foise acrescentando de autores que produziram relatos, leram e interferiramnele. Todos os participantes contribuíram para que se alcançasse esteresultado, porém, aqueles que não se inscreveram no ambiente virtual nãopuderam ser nomeados

Immunofluorescence.

<p>Detection of (<b>A</b>) OCT4, (<b>B</b>) E-cadherin, (<b>C</b>) N-cadherin and (<b>D</b>) vimentin in CMT-U229 cells mammospheres after melatonin treatment compared to control groups. <b>E</b>. Statistical analysis of OCT4, E-cadherin, N-cadherin and vimentin proteins expression. Data are shown as mean ± standard deviation. The magnification was 40 X. *<i>P</i><0.05 Statistical significance compared to control group was determine by Student´s t- test.</p

Immunofluorescence.

<p>Detection of (<b>A</b>) OCT4, (<b>B</b>) E-cadherin, (<b>C</b>) N-cadherin and (<b>D</b>) Vimentin in MCF-7 cells after melatonin treatment compared to control groups. <b>E</b>. Statistical analysis of OCT4, E-cadherin, N-cadherin and Vimentin protein expression. Data are shown as mean ± standard deviation. The magnification was 40 X. *<i>P</i> < 0.05 Statistical significance compared to control group was determine by Student´s t- test.</p

Effect of melatonin on viability of cells from mammospheres.

<p>(<b>A</b>) CMT-U229 and (<b>B</b>) MCF-7 cells were treated with 1 mM of melatonin for 24 h and cell viability was measured by MTT assay. The white column corresponds to control group. Each column represents the mean ± standard error of triplicate experiments. *<i>P</i><0.05 Statistical significance compared to control group was determine by Student´s t- test.</p