RESUMO
A 75-year-old man was admitted for combined low-frequency repetitive transcranial magnetic stimulation (rTMS) and intensive occupational therapy. Five days after the initiation of rTMS, he developed hypotension and temporary exacerbation of the right hemiplegia with thunderclap headache. MRA showed segmental stenosis of the left middle cerebral artery, which findings were improved at 9 days after the onset of the headache. He was diagnosed as having the reversible cerebral vasoconstriction syndrome (RCVS). The rTMS was recognized as safe rehabilitation treatment. However, it is necessary to recognize that RCVS can become one of the precipitants. This is the first report of RCVS triggered by rTMS.
Assuntos
Transtornos da Cefaleia Primários/etiologia , Estimulação Magnética Transcraniana/efeitos adversos , Vasoespasmo Intracraniano/etiologia , Idoso , Arteriopatias Oclusivas , Infarto Cerebral/terapia , Transtornos da Cefaleia Primários/diagnóstico , Hemiplegia , Humanos , Hipotensão , Angiografia por Ressonância Magnética , Masculino , Artéria Cerebral Média/diagnóstico por imagem , Terapia Ocupacional , Síndrome , Vasoespasmo Intracraniano/diagnósticoRESUMO
We investigated 2'-O,4'-C-ethylene-bridged nucleic acids (ENA) antisense oligonucleotides (AONs) for vascular endothelial growth factor (VEGF) in human lung carcinoma A549 cells. An ENA/DNA gapmer AON with RNase H-mediated activity was virtually stable in rat plasma and exhibited more than 90% inhibition of VEGF mRNA production. Moreover, 22 genes that are likely to bind to the AON were found in the GenBank database by BLAST and CLUSTAL W searches. Three of these genes were actually inhibited by the ENA AON. In shorter ENA AONs with fewer matched sequences of these genes, inhibitiory activities were decreased and off-target effects were improved. These results indicate that ENA AONs act in a sequence-specific manner and could be used as effective antisense drugs.
Assuntos
Regulação da Expressão Gênica/genética , Oligonucleotídeos Antissenso/genética , Oligonucleotídeos Antissenso/metabolismo , RNA Mensageiro/genética , Fator A de Crescimento do Endotélio Vascular/genética , Animais , Sequência de Bases , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Masculino , Dados de Sequência Molecular , Estrutura Molecular , RNA Mensageiro/metabolismo , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Especificidade por Substrato , Células Tumorais CultivadasRESUMO
ENA antisense oligonucleotides for vascular endothelial growth factor (VEGF) mRNA were synthesized and evaluated in A549 lung cancer cells. It was found that the VEGF ENA-antisense inhibited not only the expression of VEGF, but also the expression of three genes, which were found in Genbank by BLAST and Clustal W search and considered likely to bind to the VEGF ENA-antisense. These results indicate that ENA-antisense oligonucleotides act in a sequence-specific manner, and could be used as effective antisense drugs.
Assuntos
Oligodesoxirribonucleotídeos Antissenso/síntese química , Oligodesoxirribonucleotídeos/química , Linhagem Celular , Etilenos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Indicadores e Reagentes , Neoplasias Pulmonares , Oligodesoxirribonucleotídeos Antissenso/farmacologia , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/genética , Células Tumorais Cultivadas , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
We studied the properties of 2'-O,4'-C-ethylene-bridged nucleic acid (ENA) oligonucleotides as antisense molecules. Compared to a phosphorothioate (PS) DNA and RNA heteroduplex, a duplex of an ENA/PS/ENA gapmer with RNA was a more effective substrate for RNase H-mediated cleavage. We designed ENA antisense oligonucleotides (AON) targeting human vascular endothelial growth factor (VEGF) mRNA. ENA AON with a cationic polymer considerably down-regulated VEGF mRNA expression, while no down-regulation by PS AON was observed. We also investigated the influence on the gene expression of genes other than VEGF by a gene database homology search and RT-PCR analysis. This method turned out to be an efficient approach to search for non-target genes sequence-specifically down-regulated by AON. In rat plasma, an ENA/PS/ENA gapmer was very stable after 24 hr, while a bridged nucleic acid (BNA) or locked nucleic acid (LNA) oligonucleotide and a PS oligonucleotide were half degraded in 4 hr. The high stability of ENA oligonucleotides in plasma would make ENA oligonucleotides ideal for in vivo studies.
