Strain dependent structural effects and in vivo efficacy of enterovirus-D68 inhibitors

Acute flaccid myelitis (AFM) leads to loss of limb control in young children and is likely due to Enterovirus-D68 (EV-D68), for which there is no current treatment. We have developed a lead isoxazole-3-carboxamide analog of pleconaril (11526092) which displayed potent inhibition of the pleconaril-resistant CVB3-Woodruff (IC50 6-20 nM), EV-D68 (IC50 58 nM), and other enteroviruses. A mouse respiratory model of EV-D68 infection, in which pleconaril is inactive, showed decreased viremia of 3 log units as well as statistically significant 1 log reduction in lung titer reduction at day 5 after treatment with 11526092. A cryo-electron microscopy (cryo-EM) structure of EV-D68 in complex with 11526092 suggests that the increased potency may be due to additional hydrophobic interactions. Cryo-EM structures of 11526092 and pleconaril demonstrate destabilization of EV-D68 (MO strain) compared to the previously described stabilization of EV-D68 (Fermon strain) with pleconaril, illustrating clear strain dependent mechanisms of this molecule. 11526092 represents a more potent inhibitor in vitro with in vivo efficacy providing a potential future treatment for EV-D68 and AFM, suggesting an improvement over pleconaril for further optimization.

The Effect of Incubation Temperature, Substrate and Initial pH Value on Plantaricin Activity and the Relative Transcription of pln Genes of Six Sourdough Derived Lactiplantibacillus plantarum Strains

The aim of the present study was to assess the effect of sourdough related parameters on the growth and plantaricin activity of six Lactiplantibacillus plantarum strains against a mixture of 5 Listeria monocytogenes strains and to analyze the transcriptomic response of their pln genes. Parameters included 3 substrates (MRS broth, mMRS broth, WFE), 3 temperatures (20, 30, 37 °C), 2 initial pH values (5.0, 6.0), 2 NaCl concentrations (0.0, 1.8%) and 12 time points (ranging from 0 to 33 h). The transcriptomic response of the plantaricin genes to the aforementioned parameters was assessed after 21 h of growth. In general, plantaricin activity was strain dependent with that of Lp. plantarum strains LQC 2422, 2441, 2485 and 2516, harboring four pln genes, namely, pln423 (plαΑ), plαΒ, plαC and plαD, reaching 2560 AU/mL. However, strains LQC 2320 and 2520, in which 18 pln genes were detected, namely, plNC8a, plNC8b, plNC8c, plnL, plnR, plnJ, plnK, plnE, plnF, plnH, plnS, plnY, plNC8-IF, plNC8-HK, plnD, plnI, plnM and plnG, exhibited plantaricin activity barely reaching 160 AU/mL. Substrate, temperature, initial pH value and strains significantly affected plantaricin activity, while NaCl had only a marginal effect. Similarly, growth substrate and temperature had a more pronounced effect than initial pH value on gene transcription. A strong correlation between the transcription of the genes belonging to the same locus was observed; however, only a weak correlation, if any, was observed between plantaricin activity and the transcription of the genes assessed.

Discovering the Influence of Microorganisms on Wine Color

Flavor, composition and quality of wine are influenced by microorganisms present on the grapevine surface which are transferred to the must during vinification. The microbiota is highly variable with a prevalence of non-Saccharomyces yeasts, whereas Saccharomyces cerevisiae is present at low number. For wine production an essential step is the fermentation carried out by different starter cultures of S. cerevisiae alone or in mixed fermentation with non-Saccharomyces species that produce wines with significant differences in chemical composition. During vinification wine color can be influenced by yeasts interacting with anthocyanin. Yeasts can influence wine phenolic composition in different manners: direct interactions—cell wall adsorption or enzyme activities—and/or indirectly—production of primary and secondary metabolites and fermentation products. Some of these characteristics are heritable trait in yeast and/or can be strain dependent. For this reason, the stability, aroma, and color of wines depend on strain/strains used during must fermentation. Saccharomyces cerevisiae or non-Saccharomyces can produce metabolites reacting with anthocyanins and favor the formation of vitisin A and B type pyranoanthocyanins, contributing to color stability. In addition, yeasts affect the intensity and tonality of wine color by the action of β-glycosidase on anthocyanins or anthocyanidase enzymes or by the pigments adsorption on the yeast cell wall. These activities are strain dependent and are characterized by a great inter-species variability. Therefore, they should be considered a target for yeast strain selection and considered during the development of tailored mixed fermentations to improve wine production. In addition, some lactic acid bacteria seem to influence the color of red wines affecting anthocyanins’ profile. In fact, the increase of the pH or the ability to degrade pyruvic acid and acetaldehyde, as well as anthocyanin adsorption by bacterial cells are responsible for color loss during malolactic fermentation. Lactic acid bacteria show different adsorption capacity probably because of the variable composition of the cell walls. The aim of this review is to offer a critical overview of the roles played by wine microorganisms in the definition of intensity and tonality of wines’ color.

Hormones Can Influence Antibiotic Susceptibilities Even in Mono- and Co-Culture Conditions

Pseudomonas aeruginosa and Staphylococcus aureus are known as important nosocomial infectious agents also their co-infections are commonly seen in some patient groups. It is well known that host factors such as hormones have roles in modulation of growth, pathogenesis and susceptibilities to antimicrobials. In our study, the influences of norepinephrine (NE) and melatonin (MEL) on antibiotic susceptibilities were examined in mono and co-culture conditions. Methicilin resistant Staphylococcus aureus (MRSA) ATCC 43300 and Pseudomonas aeruginosa ATCC 27853 were investigated to determine the minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of ciprofloxacin and gentamicin in the absence/presence of NE (0.0017 and 0.04μg/mL) and MEL (6 and 60 pg/mL) by microdilution method in mono and co-culture. It was found that hormones decreased (among 2-64 fold) MIC and MBC values of both antibiotics for MRSA. However, it was shown that hormones had no effect on MIC values of both antibiotics for P. aeruginosa. MIC and MBC values of both antibiotics for co-culture were found to be reduced compared to monoculture of MRSA; were found to be increased compared to monoculture of P. aeruginosa. Whereas, hormones decreased MIC values of both antibiotics in co-culture conditions. Our results suggest that both hormones decreased MIC values and it seems that hormones could influence antibiotic susceptibilities in a strain-dependent manner.