Coxsackievirus B3 (CVB3) is a human pathogen that causes acute and chronic infections, but an antiviral drug to treat these diseases has not yet been developed for clinical use. Several intracellular pathways are altered to assist viral transcription, RNA replication, and progeny release. Among these, fatty acid synthase (FAS) expression is increased. In order to test the potential of FAS inhibition as an anti-CVB3 strategy, several experiments were performed, including studies on the correlation of CVB3 replication and FAS expression in human Raji cells and an analysis of the time and dose dependence of the antiviral effect of FAS inhibition due to treatment with amentoflavone. The results demonstrate that CVB3 infection induces an up-regulation of FAS expression already at 1 h postinfection (p.i.). Incubation with increasing concentrations of amentoflavone inhibited CVB3 replication significantly up to 8 h p.i. In addition, suppression of p38 MAP kinase activity by treatment with SB239063 decreased FAS expression as well as viral replication. These data provide evidence that FAS inhibition via amentoflavone administration might present a target for anti-CVB3 therapy.
A facile
approach for the construction of reagent-free electrochemical
dehydrogenase-based biosensors is presented. Enzymes and cofactors
(NAD+ and Fe(CN)6
3–) were immobilized by modification
of screen-printed carbon electrodes with graphene oxide (GO) and an
additional layer of cellulose acetate. The sensor system was exemplarily
optimized for an l-lactate electrode in terms of GO concentration,
working potential, and pH value. The biosensor exhibited best characteristics
at pH 7.5 in 100 mM potassium phosphate buffer at an applied potential
of +0.250 V versus an internal pseudo Ag reference electrode. Thereby,
sensor performance was characterized by a linear working range from
0.25 to 4 mM and a sensitivity of 0.14 μA mM–1. The detection principle was additionally evaluated with three other
dehydrogenases (d-lactate dehydrogenase, alcohol dehydrogenase,
and formate dehydrogenase, respectively). The developed reagentless
biosensor array enabled simultaneous and cross-talk free determination
of l-lactate, d-lactate, ethanol, and formate.
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