For the purpose of discovering novel agents that inhibit HIV-1 replication at the transcriptional level, we have established cell lines reflecting the HIV-1 long terminal repeatdriven gene expression. Using these cell lines, we have screened approximately 10,000
Bioconversion using microorganisms and their enzymes is an important tool in many industrial fields. The discovery of useful new microbial enzymes contributes to the development of industries utilizing bioprocesses. Streptomyces sp. EAS-AB2608, isolated from a soil sample collected in Japan, can convert the tetrahydrobenzotriazole CPD-1 (a selective positive allosteric modulator of metabotropic glutamate receptor 5) to its hydroxylated form at the C4-(R) position. The current study was performed to identify the genes encoding the enzymes involved in CPD-1 bioconversion and to verify their function. To identify gene products responsible for the conversion of CPD-1, we used RNA sequencing to analyze EAS-AB2608; from its 8333 coding sequences, we selected two genes, one encoding cytochrome P450 (easab2608_00800) and the other encoding ferredoxin (easab2608_00799), as encoding desirable gene products involved in the bioconversion of CPD-1. The validity of this selection was tested by using a heterologous expression approach. A bioconversion assay using genetically engineered Streptomyces avermitilis SUKA24 Δsaverm3882 Δsaverm7246 co-expressing the two selected genes (strain ES_SUKA_63) confirmed that these gene products had hydroxylation activity with respect to CPD-1, indicating that they are responsible for the conversion of CPD-1. Strain ES_SUKA_63 also showed oxidative activity toward other compounds and therefore might be useful not only for bioconversion of CPD-1 but also as a tool for synthesis of drug metabolites and in optimization studies of various pharmaceutical lead compounds. We expect that this approach will be useful for bridging the gap between the latest enzyme optimization technologies and conventional enzyme screening using microorganisms.
Key points• Genes easab2608_00800 (cyp) and easab2608_00799 (fdx) were selected by RNA-Seq.• Selection validity was evaluated by an engineered S. avermitilis expression system.• Strain ES_SUKA_63 showed oxidative activity toward CPD-1 and other compounds.
The aim of this study was to investigate the values of three acoustic parameters (PPQ, APQ and NHR) in 88 normal elementary school children between the 1st and 3rd grades. The procedure called for the children to vocalize sustained phonation of the vowel [ɑ] for 5 seconds, until a constant pitch of voice was achieved; each child underwent 2 trials. Acoustic analysis was carried out using the Multi-Dimensional Voice Program 5105 of Kay Model 4500. The results revealed that the mean PPQ was higher in the 3rd grade children than in the 2nd year children, with the mean APQ significantly lower in the 2nd trial than in the 1st. Comparing these data to those obtained by Nishio et al. (2002) it is clear that there may exist parameters yielding higher or lower values in children than in adults, suggesting that the normal range in adults may differ from that of school children. The results also suggested that APQ values may change in elementary school children depending on learning effect.
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