RESUMO
The effectiveness of the caspase-9-based artificial "death switch" as a safety measure for gene therapy based on the erythropoietin (Epo) hormone was tested in vitro and in vivo using the chemical inducer of dimerization, AP20187. Plasmids encoding the dimeric murine Epo, the tetracycline-controlled transactivator and inducible caspase 9 (ptet-mEpoD, ptet-tTAk and pSH1/Sn-E-Fv-Fvls-casp9-E, respectively) were used in this study. AP20187 induced apoptosis of iCasp9-modified C2C12 myoblasts. In vivo, two groups of male C57BI/6 mice, 8-12 weeks old, were injected intramuscularly with 5 µg/50 g ptet-mEpoD and 0.5 µg/50 g ptet-tTAk. There were 20 animals in group 1 and 36 animals in group 2. Animals from group 2 were also injected with the 6 µg/50 g iCasp9 plasmid. Seventy percent of the animals showed an increase in hematocrit of more than 65 percent for more than 15 weeks. AP20187 administration significantly reduced hematocrit and plasma Epo levels in 30 percent of the animals belonging to group 2. TUNEL-positive cells were detected in the muscle of at least 50 percent of the animals treated with AP20187. Doxycycline administration was efficient in controlling Epo secretion in both groups. We conclude that inducible caspase 9 did not interfere with gene transfer, gene expression or tetracycline control and may be used as a safety mechanism for gene therapy. However, more studies are necessary to improve the efficacy of this technique, for example, the use of lentivirus vector.
Assuntos
Animais , Masculino , Camundongos , Anemia/terapia , Caspase 9/genética , Dimerização , Eritropoetina , Expressão Gênica/genética , Terapia Genética/métodos , Tacrolimo/análogos & derivados , Caspase 9/administração & dosagem , Eritropoetina , Vetores Genéticos/genética , Hematócrito , Injeções Intramusculares , Lentivirus/genética , Plasmídeos/uso terapêutico , Tacrolimo/uso terapêuticoRESUMO
The effectiveness of the caspase-9-based artificial "death switch" as a safety measure for gene therapy based on the erythropoietin (Epo) hormone was tested in vitro and in vivo using the chemical inducer of dimerization, AP20187. Plasmids encoding the dimeric murine Epo, the tetracycline-controlled transactivator and inducible caspase 9 (ptet-mEpoD, ptet-tTAk and pSH1/Sn-E-Fv'-Fvls-casp9-E, respectively) were used in this study. AP20187 induced apoptosis of iCasp9-modified C2C12 myoblasts. In vivo, two groups of male C57BI/6 mice, 8-12 weeks old, were injected intramuscularly with 5 microg/50 g ptet-mEpoD and 0.5 microg/50 g ptet-tTAk. There were 20 animals in group 1 and 36 animals in group 2. Animals from group 2 were also injected with the 6 microg/50 g iCasp9 plasmid. Seventy percent of the animals showed an increase in hematocrit of more than 65% for more than 15 weeks. AP20187 administration significantly reduced hematocrit and plasma Epo levels in 30% of the animals belonging to group 2. TUNEL-positive cells were detected in the muscle of at least 50% of the animals treated with AP20187. Doxycycline administration was efficient in controlling Epo secretion in both groups. We conclude that inducible caspase 9 did not interfere with gene transfer, gene expression or tetracycline control and may be used as a safety mechanism for gene therapy. However, more studies are necessary to improve the efficacy of this technique, for example, the use of lentivirus vector.
Assuntos
Anemia/terapia , Caspase 9/genética , Dimerização , Eritropoetina/administração & dosagem , Expressão Gênica/genética , Terapia Genética/métodos , Tacrolimo/análogos & derivados , Animais , Caspase 9/administração & dosagem , Eritropoetina/genética , Vetores Genéticos/genética , Hematócrito , Injeções Intramusculares , Lentivirus/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Plasmídeos/uso terapêutico , Proteínas Recombinantes , Tacrolimo/uso terapêuticoRESUMO
We describe an infant with Dandy-Walker malformation and tetramelic postaxial polydactyly type 1A. Parental consanguinity reinforces previous suggestions for autosomal recessive inheritance.
Assuntos
Síndrome de Dandy-Walker/genética , Genes Recessivos , Polidactilia/genética , Encéfalo/diagnóstico por imagem , Consanguinidade , Síndrome de Dandy-Walker/diagnóstico por imagem , Humanos , Lactente , Masculino , Tomografia Computadorizada por Raios XRESUMO
The structure, manpower and tasks of the Tuberculosis Programme (ELAT) in Mozambique are explained. The activities are organised on three levels (ministerial, provincial and district). From 1985 to 1990 (1st half) 59,339 cases were detected. Of them, 32,978 were new smear-positive, 17,772 new smear-negative, 5,664 relapses and 2,425 extra-pulmonary cases. Results achieved by 3 regimens used by the Programme varied on average from 55% for the standard, 77% for the short-course and 72% for the retreatment regimen. The Programme is run by paramedical staff and no specific personnel exists in the districts. To upgrade the few personnel involved in the ELAT, supervision and training are considered as backbones of the Programme. The country suffers from a devastating war, which has aggravated the poor transport network, reduced accessibility to remote areas, hampered regular supervision and makes distribution of supplies irregular and very expensive, since drugs, reagents, forms and spare parts have to be sent by air or sea. In spite of all these terrible misfortunes, results achieved show that it is possible to diagnose and treat cases under such condition.
