RESUMO
Reverse transcriptase-linked PCR was used to determine the prevalence of varicella-zoster virus (VZV) gene 21 transcription in latently infected human ganglia. Under conditions wherein reverse transcriptase-linked PCR detected > or = 1,000 transcripts, VZV gene 21 RNA, but not VZV gene 40 RNA, was found in ganglia but not other tissues from five of seven humans.
Assuntos
Gânglios/virologia , Herpesvirus Humano 3/genética , Transcrição Gênica , Proteínas Virais/genética , Latência Viral , Sequência de Bases , Primers do DNA , Humanos , Dados de Sequência Molecular , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
The entire varicella-zoster virus (VZV) genome appears to be present in latently infected human ganglia, but the extent of virus DNA transcription is unknown. Conventional methods to study virus gene transcripts by Northern (RNA) blotting are not feasible, since ganglia are small and VZV DNA is not abundant. To circumvent this problem, we prepared radiolabeled cDNA from ganglionic RNA, hybridized it to Southern blots containing VZV DNA, and demonstrated the presence of a transcript within the SalI C fragment of the virus genome (R. Cohrs, R. Mahalingam, A. N. Dueland, W. Wolf, M. Wellish, and D. H. Gilden, J. Infect. Dis. 166:S24-S29, 1992). To further map VZV transcripts, in the work described here we constructed a cDNA library from poly(A)+ RNA obtained from latently infected human ganglia. Phage DNA isolated from the library was used in PCR amplifications to detect VZV-specific inserts. The specificity of the PCRs was provided by selection of a primer specific for VZV gene 17, 18, 19, 20, or 21 and a second vector-specific primer. VZV gene 21-specific sequences were identified by PCR amplification. The PCR product contained the XhoI cloning site and poly(A)+ sequences between vector and VZV gene 21 sequences. The sequence motif at the 3' end of VZV gene 21, determined by cloning and sequencing of the PCR product, consisted of 49 to 51 nucleotide bases of 3'-untranslated DNA, the termination codon for the VZV gene 21 open reading frame, and DNA sequences reading into the VZV gene 21 open reading frame.
Assuntos
Expressão Gênica , Genes Virais , Herpesvirus Humano 3/fisiologia , Gânglio Trigeminal/virologia , Latência Viral , Idoso , Idoso de 80 Anos ou mais , Animais , Anticorpos Antivirais/sangue , Sequência de Bases , Linhagem Celular , Chlorocebus aethiops , Primers do DNA , DNA Complementar , DNA Viral/biossíntese , DNA Viral/isolamento & purificação , Feminino , Biblioteca Gênica , Vetores Genéticos , Herpesvirus Humano 3/genética , Herpesvirus Humano 3/isolamento & purificação , Humanos , Técnicas In Vitro , Rim , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Transcrição GênicaRESUMO
Eighteen patients with idiopathic hypertension were studied to assess the anti-hypertensive control, both at rest and during exercise of increasing doses of hydrochlorothiazide, metoprolol and a combination of hydrochlorothiazide and metoprolol. Each active treatment period was given for four weeks with an intervening four week placebo period, with assessments made at 2 to 4 hours and 22 to 24 hours following tablet intake. The results show that the resting systolic and diastolic values for all active treatment periods were significantly different from placebo. During exercise, the systolic blood pressure levels were significantly reduced by all active treatments with the greatest overall reduction being achieved by either metoprolol or the metoprolol-hydrochlorothiazide combination. However, the exercise induced increase in systolic blood pressure was reduced by approximately 25% by both metoprolol and the combination product, when compared to placebo, but not by the diuretic alone.
