Short-Term Striatal Gene Expression Responses to Brain-Derived Neurotrophic Factor Are Dependent on MEK and ERK Activation

BACKGROUND: Brain-derived neurotrophic factor (BDNF) is believed to be an important regulator of striatal neuron survival, differentiation, and plasticity. Moreover, reduction of BDNF delivery to the striatum has been implicated in the pathophysiology of Huntington's disease. Nevertheless, many essential aspects of BDNF responses in striatal neurons remain to be elucidated. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we assessed the relative contributions of multipartite intracellular signaling pathways to the short-term induction of striatal gene expression by BDNF. To identify genes regulated by BDNF in these GABAergic cells, we first used DNA microarrays to quantify their transcriptomic responses following 3 h of BDNF exposure. The signal transduction pathways underlying gene induction were subsequently dissected using pharmacological agents and quantitative real-time PCR. Gene expression responses to BDNF were abolished by inhibitors of TrkB (K252a) and calcium (chelator BAPTA-AM and transient receptor potential cation channel [TRPC] antagonist SKF-96365). Interestingly, inhibitors of mitogen-activated protein kinase kinases 1 and 2 (MEK1/2) and extracellular signal-regulated kinase ERK also blocked the BDNF-mediated induction of all tested BDNF-responsive genes. In contrast, inhibitors of nitric oxide synthase (NOS), phosphotidylinositol-3-kinase (PI3K), and CAMK exhibited less prevalent, gene-specific effects on BDNF-induced RNA expression. At the nuclear level, the activation of both Elk-1 and CREB showed MEK dependence. Importantly, MEK-dependent activation of transcription was shown to be required for BDNF-induced striatal neurite outgrowth, providing evidence for its contribution to striatal neuron plasticity. CONCLUSIONS: These results show that the MEK/ERK pathway is a major mediator of neuronal plasticity and other important BDNF-dependent striatal functions that are fulfilled through the positive regulation of gene expression

A new portable vibrator for plaster pouring: effect on the marginal fit at cylinder-abutment

Objective: The aim of this study was to test a new portable vibrator for plaster pouring (developed for this purpose), comparing the effect of its use on the accuracy of working cast of implant-supported restorations to the conventional vibrator. Material and methods: From a master cast with 2 implants, 30 transfer moldings were made randomly and divided into three groups: Group I (GI): pouring performed in an outsourced dental laboratory with conventional plaster vibrator (10 casts), Group II (GII): pouring performed in the laboratory of the Federal University of Santa Catarina (UFSC) with conventional plaster vibrator (10 casts) and Group III (GIII): pouring performed with the portable vibrator fabricated for this study (10 casts). The position of the analogue and marginal adaptation of the infrastructure were verified by testing the single screw on the master model and on the working model. The measurement of misfit was blindly performed with a precision microscope and analyzing unit, Quadra-Check 200. The data were statistically analyzed by analysis of variance (ANOVA) and the Holm-Sidak test (alpha=0.05). Results: Means +/- standard deviations were as follows: GI: 19.19 +/- 4.73 mu m; GII: 21.72 +/- 5.41 mu m; GIII: 13.5 +/- 2.39 mu m (P<0.05), with GIII significantly lower as compared to the other groups. Conclusion: Within the limitations of this study, it was concluded that a greater accuracy of working cast was achieved when a portable vibrator was used for casting molds.20551752

Influence of the casting technique and dynamic loading on screw detorque and misfit of single unit implant-supported prostheses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Objective. This study aimed to evaluate the influence of the casting procedure and cyclic loading of prosthetic frameworks on detorque of prosthetic screws and marginal misfit of single unit implant-supported prostheses. Materials and methods. Twenty specimens were obtained, each one consisting of a set of an implant (external hexagon 3.75 x 13 mm - Branemark type), a prosthetic abutment (entirely calcinable or overcasted UCLA) and a prosthetic screw. After the specimens were obtained, the prosthetic screws were tightened with 30 Ncm torque and released 24 h later in order to evaluate initial detorque. The screws were retightened and marginal gaps were assessed. All specimens were submitted to 10(6) loading cycles, performed with 2 Hz frequency and 130 N load. The specimens were re-evaluated for marginal misfit and detorque after the mechanical loading (final marginal misfit/ final detorque). The results were submitted to analysis of variance for repeated measurements, followed by Tukey HSD test (alpha = 0.05). Results. No statistically significant differences were found on detorque values of the prosthetics screws for all groups and intervals evaluated (p = 0.8922). The entirely calcinable abutments showed higher initial marginal misfit compared to the overcasted ones (p = 0.0438). There was no statistically significant difference on marginal misfit before and after mechanical loading for both groups (p > 0.05). Conclusions. It can be concluded that the overcasted abutments showed lower misfit values when compared to the entirely casted abutments. No difference was observed on detorque values of prosthetic screws. After mechanical loading there was no difference on marginal misfit and detorque between the groups.7141732404409Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)FAPESP [FAPESP 2009/18473-7

An Adeno-Associated Virus-Based Intracellular Sensor of Pathological Nuclear Factor-κB Activation for Disease-Inducible Gene Transfer

Stimulation of resident cells by NF-κB activating cytokines is a central element of inflammatory and degenerative disorders of the central nervous system (CNS). This disease-mediated NF-κB activation could be used to drive transgene expression selectively in affected cells, using adeno-associated virus (AAV)-mediated gene transfer. We have constructed a series of AAV vectors expressing GFP under the control of different promoters including NF-κB -responsive elements. As an initial screen, the vectors were tested in vitro in HEK-293T cells treated with TNF-α. The best profile of GFP induction was obtained with a promoter containing two blocks of four NF-κB -responsive sequences from the human JCV neurotropic polyoma virus promoter, fused to a new tight minimal CMV promoter, optimally distant from each other. A therapeutical gene, glial cell line-derived neurotrophic factor (GDNF) cDNA under the control of serotype 1-encapsidated NF-κB -responsive AAV vector (AAV-NF) was protective in senescent cultures of mouse cortical neurons. AAV-NF was then evaluated in vivo in the kainic acid (KA)-induced status epilepticus rat model for temporal lobe epilepsy, a major neurological disorder with a central pathophysiological role for NF-κB activation. We demonstrate that AAV-NF, injected in the hippocampus, responded to disease induction by mediating GFP expression, preferentially in CA1 and CA3 neurons and astrocytes, specifically in regions where inflammatory markers were also induced. Altogether, these data demonstrate the feasibility to use disease-activated transcription factor-responsive elements in order to drive transgene expression specifically in affected cells in inflammatory CNS disorders using AAV-mediated gene transfer. © 2013 Chtarto et al.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Interleukin-33 in health and disease

Interleukin-33 (IL-33) — a member of the IL-1 family — was originally described as an inducer of type 2 immune responses, activating T helper 2 (TH2) cells and mast cells. Now, evidence is accumulating that IL-33 also potently stimulates group 2 innate lymphoid cells (ILC2s), regulatory T (Treg) cells, TH1 cells, CD8+ T cells and natural killer (NK) cells. This pleiotropic nature is reflected in the role of IL-33 in tissue and metabolic homeostasis, infection, inflammation, cancer and diseases of the central nervous system. In this Review, we highlight the molecular and cellular characteristics of IL-33, together with its major role in health and disease and the potential therapeutic implications of these findings in humans