1,800 research outputs found

    Real-time PCR quantitation of glucocorticoid receptor alpha isoform

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    BACKGROUND: The expression of glucocorticoid-receptor (GR) seems to be a key mechanism in the regulation of glucocorticoid (GC) sensitivity and is potentially involved in cases of GC resistance or hypersensitivity. The aim of this study is to describe a method for quantitation of GR alpha isoform (GRα) expression using real-time PCR (qrt-PCR) with analytical capabilities to monitor patients, offering standard-curve reproducibility as well as intra- and inter-assay precision. RESULTS: Standard-curves were constructed by employing standardized Jurkat cell culture procedures, both for GRα and BCR (breakpoint cluster region), as a normalizing gene. We evaluated standard-curves using five different sets of cell culture passages, RNA extraction, reverse transcription, and qrt-PCR quantification. Intra-assay precision was evaluated using 12 replicates of each gene, for 2 patients, in a single experiment. Inter-assay precision was evaluated on 8 experiments, using duplicate tests of each gene for two patients. Standard-curves were reproducible, with CV (coefficient of variation) of less than 11%, and Pearson correlation coefficients above 0,990 for most comparisons. Intra-assay and inter-assay were 2% and 7%, respectively. CONCLUSION: This is the first method for quantitation of GRα expression with technical characteristics that permit patient monitoring, in a fast, simple and robust way

    RELATIONSHIP BETWEEN THE CORONAL SUTURE AND THE CENTRAL LOBE How important is it and how can we use it in surgical planning?

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    The surgical treatment of the lesions located in the central lobe is a very difficult task for the neurosurgeon. The overall aim of this study is to verify the correlation of the coronal suture and the structures of the central lobe in 32 cadaver hemisphere brains and the importance of this information in surgical planning. The measurement of the nasion to the coronal suture ranged from 11.5 to 13.5 cm. The distance between the coronal suture in the midline to the central, precentral and paracentral sulcus ranged from 5.0 to 6.6, 2.5 to 4.5 and 1.3 to 4.0 cm respectively. Particularly in the normal cortex these measurements can be used to guide the surgical access. However, the identification of the central sulcus is not easy when the anatomical pattern is distorted or displaced by a lesion or edema. In cases such as these the use of other tools becomes crucial for good surgical planning and cortical mapping or awake craniotomy for a safer resection of the lesion as well.66486887

    Activation of Type 1 Cannabinoid Receptor (CB1R) promotes neurogenesis in murine subventricular zone cell cultures

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    The endocannabinoid system has been implicated in the modulation of adult neurogenesis. Here, we describe the effect of type 1 cannabinoid receptor (CB1R) activation on self-renewal, proliferation and neuronal differentiation in mouse neonatal subventricular zone (SVZ) stem/progenitor cell cultures. Expression of CB1R was detected in SVZ-derived immature cells (Nestin-positive), neurons and astrocytes. Stimulation of the CB1R by (R)-(+)-Methanandamide (R-m-AEA) increased self-renewal of SVZ cells, as assessed by counting the number of secondary neurospheres and the number of Sox2+/+ cell pairs, an effect blocked by Notch pathway inhibition. Moreover, R-m-AEA treatment for 48 h, increased proliferation as assessed by BrdU incorporation assay, an effect mediated by activation of MAPK-ERK and AKT pathways. Surprisingly, stimulation of CB1R by R-m-AEA also promoted neuronal differentiation (without affecting glial differentiation), at 7 days, as shown by counting the number of NeuN-positive neurons in the cultures. Moreover, by monitoring intracellular calcium concentrations ([Ca2+](i)) in single cells following KCl and histamine stimuli, a method that allows the functional evaluation of neuronal differentiation, we observed an increase in neuronal-like cells. This proneurogenic effect was blocked when SVZ cells were co-incubated with R-m-AEA and the CB1R antagonist AM 251, for 7 days, thus indicating that this effect involves CB1R activation. In accordance with an effect on neuronal differentiation and maturation, R-m-AEA also increased neurite growth, as evaluated by quantifying and measuring the number of MAP2-positive processes. Taken together, these results demonstrate that CB1R activation induces proliferation, self-renewal and neuronal differentiation from mouse neonatal SVZ cell cultures.Fundacao para a Ciencia e a Tecnologia - Portugal [POCTI/SAU-NEU/68465/2006, PTDC/SAU-NEU/104415/2008, PTDC/SAU-NEU/101783/2008, POCTI/SAU-NEU/110838/2009]; Fundacao Calouste Gulbenkian [96542]; Fundacao para a Ciencia e Tecnologiainfo:eu-repo/semantics/publishedVersio

    Two new species of Phalangopsis Serville, 1831 (Orthoptera: Grylloidea: Phalangopsidae) from Brazilian Amazon Forest

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    We describe here two new species of the genus Phalangopsis Serville, 1831 from the Brazilian Amazon Forest. The male genitalia and the female copulatory papilla were described, and a combination of diagnostic characteristics was given to separate both new species from the other described species. The principal morphological characteristics of this genus were discussed.Aqui foram descritas duas espécies novas do gênero Phalangopsis Serville, 1831 da Floresta Amazônica brasileira. A genitália masculina e a papila copulatória feminina são descritas, bem como uma combinação de características diagnósticas para separar ambas as novas espécies das outras espécies descritas. As principais características morfológicas foram discutidas

    Experimental and numerical investigations on the seismic behavior of bridge piers with vertical unbonded prestressing strands

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    In the performance-based seismic bridge design, piers are expected to undergo large inelastic deformations during severe earthquakes, which in turn can result in large residual drift and concrete crack in the bridge piers. In this paper, longitudinal unbonded prestressing strands are used to minimize residual drift and residual concrete crack width in reinforced concrete (RC) bridge piers. Seven pier specimens were designed and tested quasi-statically and the numerical simulations were carried out. The effectiveness of using vertical unbonded prestressing strands to mitigate the residual drift and concrete crack width of RC bridge piers are examined and discussed in detail. It is found that the residual drift and residual concrete crack width of the piers can be reduced significantly by using the prestressing strands. Moreover, the strands can increase the lateral strength of the piers while have little influence on the ductility capacity of the piers. The hysteretic curves, residual drifts and strand stress of the piers predicted by the numerical model agree well with the testing data and can be used to assess the cyclic behavior of the piers
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