Ubiquitin System-Dependent Regulation of Growth Hormone Receptor Signal Transduction and Effects of Oxidative Stress

The ubiquitin-proteasome system is known to be involved in GHR endocytosis, where an active ubiquitin system is necessary for receptor ubiquitination and its consequent internalization. Endocytosis of the GHR has also been shown to be regulated by the proteasome. This thesis focuses, first of all, on the effect of the ubiquitin system on the GHR signal transduction and downregulation. In chapter 2, we studied the role of the ubiquitin system in the signal transduction of the GHR. By using specific proteasome inhibitors, we determined their effect on GH-induced activity of both GHR and JAK2 in both the wild-type GHR and in the internalization- deficient GHR mutant F327A. We suggest possible models for the role of the ubiquitin-proteasome system in GHR signal transduction. Furthermore, we asked the question involving the continuation of signal transduction of the GHR after its 37 Chapter 1 internalization, where binding of JAK2 as well as other activated proteins to the GHR, not only at the cell-surface but also intracellularly, was determined. Degradation products of the GHR were determined in both cell-surface and in endosomes, indicating that part of the receptor is down-regulated by the proteasome. In chapter 3, we focused on the role of JAK2-mediated signal transduction in GHR internalization and downregulation. Using a box-1 mutant of the GHR, activation of both GHR and JAK2 was determined, as well as receptor-bound GH internalization and GHR turnover and transport to the plasma membrane. We determined the role of GHR signal transduction, via JAK2, on GHR ubiquitination, endocytosis and degradation, and the role of the ubiquitin system on the regulation of both GHR internalization and signal transduction. Finally, chapter 4 discusses the role of stress in regulating GHR levels in the cell. We determined the effect of H2O2 on receptor's turnover and on receptor's downregulation from the cell-surface. We further determined the effect of oxidative stress on the amount of ligand binding sites at the cell-surface and related these results with the enhancement of shedding activity directed to the GHR

High energy ion beam irradiation of Co NiFe Co Cu multilayers Effects on the structural, transport and magnetic properties

The aim of this work is to investigate the effects of 593 MeVAu irradiation using two different projectile charges, namely Au30 and Au46.3 on the structural, transport and magnetization properties of Co NiFe Co Cu multilayers. X Ray diffraction and extended X ray absorption fine structures measurements show no significant structural change for as deposited and irradiated multilayers. On the other hand, the magnetoresistance amplitude decreases with the ion fluence but it is insensitive to the projectile charge state. The correlation between changes in the magnetoresistance and remanent magnetization suggests that the main effect responsible for the decrease of the magnetoresistance is the creation of ferromagnetic pinholes. These results are discussed on basis of the electronic thermal spike model and nuclear cascades theory and show similarities to the effects observed at low energy ion beam irradiatio

Trypanocidal Activity Of Brazilian Plants Against Epimastigote Forms From Y And Bolivia Strains Of Trypanosoma Cruzi

Chagas disease is one of the main public health problems in Latin America. Since the available treatments for this disease are not effective in providing cure, the screening of potential antiprotozoal agents is essential, mainly of those obtained from natural sources. This study aimed to provide an evaluation of the trypanocidal activity of 92 ethanol extracts from species belonging to the families Annonaceae, Apiaceae, Cucurbitaceae, Lamiaceae, Lauraceae, Moraceae, Nyctaginaceae, and Verbenaceae against the Y and Bolivia strains of Trypanosoma cruzi. Additionally, cytotoxic activity on LLCMK2 fibroblasts was evaluated. Both the trypanocidal activity and cytotoxicity were evaluated using the MTT method, in the following concentrations: 500, 350, 250, and 100 μg/mL. Benznidazole was used for positive control. The best results among the 92 samples evaluated were obtained with ethanol extracts of Ocotea paranapiacabensis (Am93) and Aegiphila lhotzkiana (Am160). Am93 showed trypanocidal activity against epimastigote forms of the Bolivia strain and was moderately toxic to LLCMK2 cells, its Selectivity Index (SI) being 14.56, while Am160 showed moderate trypanocidal activity against the Bolivia strain and moderate toxicicity, its SI being equal to 1.15. The screening of Brazilian plants has indicated the potential effect of ethanol extracts obtained from Ocotea paranapiacabensis and Aegiphila lhotzkiana against Chagas disease.223528533Bastos, J.K., Albuquerque, S., Silva, M.L.A., Evaluation of the trypanocidal activity of lignans isolated from the leaves of Zanthoxylum naranjillo (1999) Planta Med, 65, pp. 1-4Batista Jr., J.M., Lopes, A.A., Ambrósio, D.L., Regasini, L.O., Kato, M.J., Bolzani, V.S., Cicarelli, R.M., Furlan, M., Natural chromenes and chromene derivatives as potencial antitrypanosomal agents (2008) Biol Pharm Bull, 31, pp. 538-540Botsaris, A., Plants used traditionally to treat malaria in Brazil: The archives of Flora Medicinal (2007) J Ethnobiol Ethnomed, 1, p. 18Buainain, A., Giazzi, J.F., Belda Neto, F.M., Martini, A.S., Rosa, J.A., Pozetti, G.L., Estudo da atividade de extratos vegetais sobre o desenvolvimento de Trypanosoma cruzi em meio líquido de Warren (1992) Rev Cien Farm, 14, pp. 93-102Cabral, M.M., Barbosa-Filho, J.M., Maia, G.L., Chaves, M.C., Braga, M.V., de Souza, W., Neolignans from plants in northeastern Brazil (Lauraceae) with activity against (2010) Trypanosoma Cruzi. Exp Parasitol, 124, pp. 319-324Costa-Lotufo, L.V., Silveira, E.R., Barros, M.C., Lima, M.A., de Moraes, M.E., de Moraes, M.O., Pessoa, C., Antiproliferative effects of abietane diterpenes from aegiphilla lhotzkyana (2004) Planta Med, 70, pp. 180-182Cotinguiba, F., Regasini, L.O., Bolzani, V.S., Debonsi, H.M., Passerini, D.O., Cicarelli, R.M.B., Kato, M.J., Furlan, M., Piperamides and their derivatives as potential antitrypanosomal agents (2009) Med Chem Res, 18, pp. 703-711Coura, J.R., Castro, S.L., A critical review on Chagas disease chemotherapy (2002) Mem I Oswaldo Cruz, 97, pp. 3-24Coura, J.R., Present situation and new strategies for Chagas disease chemotherapy: A proposal (2009) Mem I Oswaldo Cruz, 104, pp. 549-554Fernandes, O., Souto, R.P., Castro, J.A., Pereira, J.B., Fernandes, N.C., Junqueira, A.C., Naiff, R.D., Coura, J.R., Brazilian isolates of Trypanosoma cruzi from humans and triatomines classified into two lineages using mini-exon and ribosomal RNA sequences (1998) Am J Trop Med Hyg, 58, pp. 807-811Fournet, A., Ferreira, M.E., Rojas de Arias, A., Guy, I., Guinaudeau, H., Heinzen, H., Phytochemical and antiprotozoal activity of (2007) Ocotea Lancifolia. Fitoterapia, 78, pp. 382-384Lopes, A.A., López, S.N., Regasini, L.O., Batista, J.M., Ambrósio, D.L., Kato, M.J., da Silva, B.V., Furlan, M., In vitro activity of compounds isolated from Piper crassinervium against Trypanosoma cruzi (2008) Nat Prod Res, 22, pp. 1040-1046Macedo, A.M., Oliveira, R.P., Pena, S.D.J., Chagas disease: Role of parasite genetic variation in pathogenesis (2002) Exp Mol Med, 4, pp. 1-16Muelas-Serrano, S., Nogal-Ruiz, J.J., Gómez-Barrio, A., Setting of a colorimetric method to determine the viability of Trypanosoma cruzi epimastigotes (2000) Parasitol Res, 86, pp. 999-1002Nwaka, S., Ridley, R.G., Virtual drug discovery and development for neglected diseases through publicprivate partnerships (2003) Nat Rev Drug Discov, 2, pp. 919-928Osorio, E., Arango, G.J., Jiménez, N., Alzate, F., Ruiz, G., Gutiérrez, D., Paco, M.A., Robledo, S., Antiprotozoal and cytotoxic activities in vitro of Colombian Annonaceae (2007) J Ethnopharmacol, 111, pp. 630-635Regasini, L.O., Cotinguiba, F., Passerini, G.D., Bolzani, V.S., Cicarelli, R.M.B., Kato, M.J., Furlan, M., Trypanocidal activity of Piper arboreum and Piper tuberculatum (Piperaceae) (2009) Rev Bras Farmacog, 19, pp. 199-203Saraiva, J., Vega, C., Rolon, M., da Silva, R., Silva, M.L., Donate, P.M., Bastos, J.K., de Albuquerque, S., In vitro and in vivo activity of lignan lactones derivatives against Trypanosoma cruzi (2007) Parasitol Res, 100, pp. 791-795Tibayrenc, M., Ayala, F.J., The clonal theory of parasitic protozoa: 12 years on (2002) Trends Parasitol, 18, pp. 405-410(2010), http://www.who.int/mediacentre/factsheets/fs340/en/index.html, World Health Organization 2010, accessed in Au