RESUMO
Familial amyloidotic polyneuropathy (FAP) is a neurodegenerative disorder characterized by extracellular deposition of amyloid fibrils composed by mutated transthyretin (TTR) mainly in the peripheral nervous system. At present, liver transplantation is still the standard treatment to halt the progression of clinical symptoms in FAP, but new therapeutic strategies are emerging, including the use of TTR stabilizers. Here we propose to establish a new gene therapy approach using adeno-associated virus (AAV) vectors to deliver the trans-suppressor TTR T119M variant to the liver of transgenic TTR V30M mice at different ages. This TTR variant is known for its ability to stabilize the tetrameric protein. Analysis of the gastrointestinal tract of AAV-treated animals revealed a significant reduction in deposition of TTR non-fibrillar aggregates in as much as 34% in stomach and 30% in colon, as well as decreased levels of biomarkers associated with TTR deposition, namely the endoplasmic reticulum stress marker BiP and the extracellular matrix protein MMP-9. Moreover, we showed with different studies that our approach leads to an increase in tetrameric and more stable forms of TTR, in favor of destabilized monomers. Altogether our data suggest the possibility to use this gene therapy approach in a prophylactic manner to prevent FAP pathology.
Assuntos
Neuropatias Amiloides Familiares/terapia , Terapia Genética/métodos , Pré-Albumina/genética , Neuropatias Amiloides Familiares/genética , Animais , Dependovirus/genética , Modelos Animais de Doenças , Eletroforese em Gel Bidimensional , Estresse do Retículo Endoplasmático/genética , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Regulação da Expressão Gênica , Técnicas de Transferência de Genes , Marcadores Genéticos , Vetores Genéticos , Fígado/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Transgênicos , Sistema Nervoso Periférico/metabolismo , Pré-Albumina/metabolismo , ProteômicaRESUMO
The aim of this study was to investigate the influence of gender on the EMG signal of the muscles of the quadriceps femoris and the physical performance in high-intensity, short-term exercise. Fourteen volunteers (7 men = 29.1 +/- 2.8 years and 7 women = 22.6 +/- 2.9 years) performed a Wingate Test (WT) with a load of 7.5% of body mass. The variables analyzed during the WT were the Relative Peak Power (W.Kg(-1)) (RPP), Relative Mean Power (W.Kg(-1)) (RMP), Fatigue Index (%) (FI) and Peak Power Instant (s) (PPI). EMG signals of the superficial muscles of the quadriceps femoris (QF) from the right leg: rectus femoris (RF), vastus lateralis (VL) and vastus medialis (VM) were analyzed through root mean square (RMS) values and the normalized median frequency (MNF) determined using the Fast Fourier Transform (FFT). The RPP and the RMP were significantly higher in men when compared to women (9.99 +/- 0.96 vs. 7.66 +/- 1.00 W.kg(-1); 7.23 +/- 0.49 vs. 5.65 +/- 0.61 W.kg(1), P < 0.05; respectively). No significant difference between genders was found on RMS and NMF during WT (P > 0.05). Although RPP and RMP were influenced by gender, the RMS and the NMF of the superficial muscles of the QF did not show the same behavior, suggesting that other mechanisms, not related to motor unit recruitment and speed of nervous stimuli in the muscle fiber may be associated to the lower performance of women in high-intensity, short-term exercise.
Assuntos
Eletromiografia , Exercício Físico/fisiologia , Fadiga Muscular/fisiologia , Força Muscular/fisiologia , Músculo Quadríceps/fisiologia , Fatores Sexuais , Adulto , Índice de Massa Corporal , Feminino , Humanos , Masculino , Recrutamento Neurofisiológico , Adulto JovemRESUMO
To study experimental Neospora caninum infection initiated at the gastrointestinal tract, Toll-like Receptor 4- and functional IL-12Rbeta2 chain-deficient C57BL/10 ScCr mice were challenged intragastrically with 5 x 10(6) N. caninum tachyzoites. All parasite-inoculated mice eventually died with disseminated infection. In contrast, immunocompetent BALB/c mice challenged with 1 x 10(7) N. caninum tachyzoites by the intragastric (i.g.) or the intraperitoneal (i.p.) route remained alive for at least 6 months. Expansion of splenic B- and T-cells, the latter displaying both activated and regulatory phenotypes, and increased levels of IFN-gamma and IL-10 mRNA were detected in both groups of infected BALB/c mice compared with non-infected controls, whereas in the Peyer's patches only IFN-gamma mRNA levels were found to be increased. Parasite-specific IgG1, IgG2a and IgA antibody levels were elevated in the sera of all infected mice, whereas increased N. caninum-specific IgA levels were detected in intestinal lavage fluids of i.g. challenged mice only. These results show that N. caninum infection can be successfully established in mice by i.g. administration of tachyzoites. They also show that the immune response elicited in i.g. or i.p. infected BALB/c mice, although conferring some degree of protection, was not sufficient for complete parasite clearance.