Cannabinoid Agonists Inhibit Neuropathic Pain Induced by Brachial Plexus Avulsion in Mice by Affecting Glial Cells and MAP Kinases

Many studies have shown the antinociceptive effects of cannabinoid (CB) agonists in different models of pain. Herein, we have investigated their relevance in neuropathic pain induced by brachial plexus avulsion (BPA) in mice.Mice underwent BPA or sham surgery. The mRNA levels and protein expression of CB(1) and CB(2) receptors were assessed by RT-PCR and immunohistochemistry, respectively. The activation of glial cells, MAP kinases and transcription factors were evaluated by immunohistochemistry. The antinociceptive properties induced by cannabinoid agonists were assessed on the 5(th) and 30(th) days after surgery. We observed a marked increase in CB(1) and CB(2) receptor mRNA and protein expression in the spinal cord and dorsal root ganglion, either at the 5(th) or 30(th) day after surgery. BPA also induced a marked activation of p38 and JNK MAP kinases (on the 30(th) day), glial cells, such as microglia and astrocytes, and the transcription factors CREB and NF-κB (at the 5(th) and 30(th) days) in the spinal cord. Systemic treatment with cannabinoid agonists reduced mechanical allodynia on both the 5(th) and 30(th) days after surgery, but the greatest results were observed by using central routes of administration, especially at the 30(th) day. Treatment with WIN 55,212-2 prevented the activation of both glial cells and MAP kinases, associated with an enhancement of CREB and NF-κB activation.Our results indicate a relevant role for cannabinoid agonists in BPA, reinforcing their potential therapeutic relevance for the management of chronic pain states

Metformin attenuates the exacerbation of the allergic eosinophilic inflammation in high fat-diet-induced obesity in mice

A positive relationship between obesity and asthma has been well documented. The AMP-activated protein kinase (AMPK) activator metformin reverses obesity-associated insulin resistance (IR) and inhibits different types of inflammatory responses. This study aimed to evaluate the effects of metformin on the exacerbation of allergic eosinophilic inflammation in obese mice. Male C57BL6/J mice were fed for 10 weeks with high-fat diet (HFD) to induce obesity. The cell infiltration and inflammatory markers in bronchoalveolar lavage (BAL) fluid and lung tissue were evaluated at 48 h after ovalbumin (OVA) challenge. HFD obese mice displayed peripheral IR that was fully reversed by metformin (300 mg/kg/day, two weeks). OVA-challenge resulted in higher influx of total cell and eosinophils in lung tissue of obese mice compared with lean group. As opposed, the cell number in BAL fluid of obese mice was reduced compared with lean group. Metformin significantly reduced the tissue eosinophil infiltration and prevented the reduction of cell counts in BAL fluid. In obese mice, greater levels of eotaxin, TNF-α and NOx, together with increased iNOS protein expression were observed, all of which were normalized by metformin. In addition, metformin nearly abrogated the binding of NF-κB subunit p65 to the iNOS promoter gene in lung tissue of obese mice. Lower levels of phosphorylated AMPK and its downstream target acetyl CoA carboxylase (ACC) were found in lung tissue of obese mice, which were restored by metformin. In separate experiments, the selective iNOS inhibitor aminoguanidine (20 mg/kg, 3 weeks) and the anti-TNF-α mAb (2 mg/kg) significantly attenuated the aggravation of eosinophilic inflammation in obese mice. In conclusion, metformin inhibits the TNF-α-induced inflammatory signaling and NF-κB-mediated iNOS expression in lung tissue of obese mice. Metformin may be a good pharmacological strategy to control the asthma exacerbation in obese individuals.Fundação de Amparo à Pesquisa do Estado de São Paulo, 2012/14225-

T1rho and T2 relaxation times of the normal adult knee meniscus at 3T: analysis of zonal differences

Abstract Background Prior studies describe histological and immunohistochemical differences in collagen and proteoglycan content in different meniscal zones. The aim of this study is to evaluate horizontal and vertical zonal differentiation of T1rho and T2 relaxation times of the entire meniscus from volunteers without symptom and imaging abnormality. Methods Twenty volunteers age between 19 and 38 who have no knee-related clinical symptoms, and no history of prior knee surgeries were enrolled in this study. Two T1rho mapping (b-FFE T1rho and SPGR T1rho) and T2 mapping images were acquired with a 3.0-T MR scanner. Each meniscus was divided manually into superficial and deep zones for horizontal zonal analysis. The anterior and posterior horns of each meniscus were divided manually into white, red-white and red zones for vertical zonal analysis. Zonal differences of average relaxation times among each zone, and both inter- and intra-observer reproducibility were statistically analyzed. Results In horizontal zonal analysis, T1rho relaxation times of the superficial zone tended to be higher than those of the deep zone, and this difference was statistically significant in the medial meniscal segments (84.3 ms vs 76.0 ms on b-FFE, p 0.74) or good (0.60–0.74) in all meniscal segments on both horizontal and vertical zonal analysis, except for inter-class correlation coefficients of the lateral meniscus on SPGR. Compared with SPGR T1rho images, b-FFE T1rho images demonstrated more significant zonal differentiation with higher inter- and intra-observer reproducibility. Conclusions There are zonal differences in T1rho and T2 relaxation times of the normal meniscus

Micropropagação do abacaxizeiro ornamental Protocol for in vitromicropropagation of ornamental pineapple

O Ananas comosus var. erectifolius, cultivar de abacaxi ornamental, tem apresentado grande interesse para paisagistas e floricultores do Brasil e do exterior, por ser uma planta ornamental tropical, exótica e rústica. A produção de plantas ornamentais a partir de técnicas de cultura de tecidos apresenta-se como uma alternativa viável para a obtenção de um grande número de plantas com qualidade genética e fitossanitária, em um curto espaço de tempo, suprindo, assim, a necessidade do mercado na aquisição de mudas com qualidade comprovada. Estudou-se a influência das concentrações de BAP (0; 0,5; 1,0 e 1,5 mg L-1) e ANA (0,0; 0,12; 0,24; 0,48 mg L-1) no meio de cultura MS com 0; 2,5; 5,0; e 7,5 g L-1 de ágar, visando estabelecer um protocolo para multiplicação e enraizamento in vitro de brotos de abacaxizeiro ornamental. Brotações com 1,5 &plusmn; 0,5 cm, já estabelecidas in vitro, oriundas das gemas da coroa do fruto do abacaxizeiro ornamental, foram inoculados assepticamente nos frascos. Após inoculados, os explantes foram mantidos em sala de crescimento com luminosidade em torno de 35 ìmol m-2 s-1, 26&plusmn;1ºC e fotoperíodo de 16 horas. Após 45 dias observou-se que a multiplicação in vitrodo abacaxi ornamental é viável em meio MS líquido acrescido de BAP 1,5 mg L-1 e o enraizamento também em meio MS líquido, na ausência de reguladores de crescimento.<br>The Ananas comosus var. erectifoliusis an ornamental pineapple cultivar which greatly interests Brazilians and foreign landscapers and flower producers for being an exotic and rustic tropical ornamental plant. The market demand for high quality of cuttings requires efficient methods of propagation and in this context the tissue culture stands out as a viable alternative to obtain plants with genetic and phytossanitary quality in a short time. In the present work we studied the influence of concentrations of BAP (0; 0.5; 1.0; 1.5 mg L-1) and NAA (0; 0.12; 0.24; 0.48 mg L-1) in the MS medium culture within 0; 2.5; 5.0; 7.5 g L-1 of agar, in order to establish an invitro protocol for multiplication and rooting of ornamental pineapple. Plantlets with 1.5&plusmn;0.5 cm already established in vitro, extracted from buds of ornamental pineapple fruits crown were inoculated aseptically in flasks. After inoculation the plantlets were kept in a growth room at 26&plusmn;1ºC, 35 ìmol m-2 s-1 irradiance and a 16-hour photoperiod. After 45 days we observed that the multiplication of ornamental pineapple is viable at liquid MS medium with BAP 1.5 mg L-1 and the rooting is also increased in liquid MS medium in absence of growth regulators