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Ceramides are a major lipid class known to play an essential role in maintaining skin function. Thus, efforts have been made to produce ceramides and ceramide precursors in large quantities for industrial applications. The yeast Wickerhamomyces ciferrii, a natural producer of the ceramide precursor tetraacetyl phytosphingosine (TAPS), has been isolated and engineered through various mutagenesis approaches aiming to enhance TAPS production. Herein, a high-throughput screening platform for isolating W. ciferrii mutants with improved TAPS production is described. A fluorescence-mediated reporter system that allows initial quantification of TAPS content in yeast cells based on BODIPY staining was developed. The optimal concentration of BODIPY for monitoring intracellular TAPS levels in W. ciferrii was 400 µg/L, as shown by a linear correlation between the actual TAPS levels and mean fluorescence intensities. Fluorescence-activated cell sorting was used for isolating high TAPS-producing strains from an ethyl methanesulfonate-induced mutant library. After several rounds of sorting, mutants exhibiting a high-TAPS phenotype were isolated, and the M40 strain with the highest TAPS titer was chosen for large-scale cultivation. The influence of different carbon sources for optimizing TAPS production was also evaluated using the M40 strain. A maximum production yield of 5.114 g/L of ceramide precursors, including TAPS and triacetyl phytosphingosine, was achieved with the supplementation of molasses. This novel platform enables rapid screening of high TAPS-producing strains using the common dye BODIPY and can be easily extended for the development of mutants with high productivity of ceramide precursors in yeast and other microorganisms. [ABSTRACT FROM AUTHOR]

Objective: Recently, Rodgersia podophylla has been reported to exhibit anti-inflammatory activity. However, little is known about the potential mechanisms about its anti-inflammatory activity. We elucidated the anti-inflammatory mechanisms of leaves extracts from Rodgersia podophylla (RP-L) in RAW264.7 cells. Materials and methods: LPS-induced NO was measured by Griess and mRNA of pro-inflammatory mediators was analyzed by RT-PCR. Cell viability was measured using MTT assay. The protein level was analyzed by Western blot. Results: RP-L significantly inhibited the production of the pro-inflammatory mediators such as NO, iNOS, IL-1β and IL-6 in LPS-stimulated RAW264.7 cells. RP-L increased HO-1 expression in RAW264.7 cells, and the inhibition of HO-1 by ZnPP reduced the inhibitory effect of RP-L against LPS-induced NO production in RAW264.7 cells. Inhibition of p38, ROS and GSK3β attenuated RP-L-mediated HO-1 expression. Inhibition of ROS inhibited p38 phosphorylation and GSK3β expression induced by RP-L. In addition, inhibition of GSK3β blocked RP-L-mediated p38 phosphorylation. RP-L induced nuclear accumulation of Nrf2, and inhibition of p38, ROS and GSK3β abolished RP-L-mediated nuclear accumulation of Nrf2. Furthermore, RP-L blocked LPS-induced degradation of IκB-α and nuclear accumulation of p65. RP-L also attenuated LPS-induced phosphorylation of ERK1/2 and p38. In GC/MS analysis of RP-L, pyrogallol was detected as bioactive compound for anti-inflammatory activity of RP-L. Pyrogallol was observed to activate HO-1 expression through ROS/GSK3β/p38/Nrf2/HO-1 signaling. Conclusions: Our results suggest that RP-L exerts potential anti-inflammatory activity by activating ROS/GSK3β/p38/Nrf2/HO-1 signaling and inhibiting NF-κB and MAPK signaling in RAW264.7 cells. These findings suggest that RP-L may have great potential for the development of anti-inflammatory drug. [ABSTRACT FROM AUTHOR]

Microalgae have been widely considered for the production of valuable products, such as lipid-based biofuel, value-added pigments, and anti-photo aging reagents. More recently, microalgae have been considered an alternative host for recombinant protein production because of their economic benefits and ecofriendly characteristics. Additionally, many microalgal strains identified to date are generally recognized as safe (GRAS); therefore, the use of microalgae-based technology is promising. However, basic studies on the genetic engineering of microalgae are rare, despite their importance. Particularly, inducible promoter systems that can be applied for strain engineering or recombinant protein production are rarely studied; hence, a number of challenging issues remain unsolved. Therefore, in this study, we focused on the development of a convenient and compact-inducible promoter system that can be used in microalgae. Based on previous success with plant systems, we employed the alcohol-inducible AlcR-PalcA system, which originates from the filamentous fungus, Aspergillus nidulans. This system comprises only two components, a regulatory protein, AlcR, and an inducible promoter, PalcA. Therefore, construction and transformation of the gene cassettes can be easily performed. Ethanol-dependent gene expression was observed in the transformants with no significant growth retardation or inducer consumption observed in the cells cultivated under optimized conditions. [ABSTRACT FROM AUTHOR]

Oxidative desulfurization (ODS) of organic compounds containing sulfur element from a model oil was performed using tungsten oxide catalysts supported on mesoporous silica with cubic Ia3d mesostructure, well-defined mesopores (7.2 nm), high surface area (719 m2/g), and three-dimensional pore network (WOx/KIT-6). The prepared WOx/KIT-6 catalysts (5-20 wt% WOx) were characterized by X-ray diffraction analysis, N2 sorption measurements, electron microscopy, H2-temperature programmed reduction, Raman spectroscopy, and thermogravimetric analysis. Among the mesoporous catalysts, 10 wt% WOx/KIT-6 exhibited the best catalytic performance. Sulfur-containing organic compounds, such as dibenzothiophene, 4,6-dimethyldibenzothiophene, and benzothiophene, were completely (100 %) removed from the model oil over 10 wt% WOx/KIT-6 catalyst in 2 h. In addition, the catalyst could be reused several times with only slight decrease in catalytic activity. [ABSTRACT FROM AUTHOR]

Objectives: To evaluate the differences between filtered back projection (FBP) and model-based iterative reconstruction (MBIR) algorithms on semi-automatic measurements in subsolid nodules (SSNs).Methods: Unenhanced CT scans of 73 SSNs obtained using the same protocol and reconstructed with both FBP and MBIR algorithms were evaluated by two radiologists. Diameter, mean attenuation, mass and volume of whole nodules and their solid components were measured. Intra- and interobserver variability and differences between FBP and MBIR were then evaluated using Bland-Altman method and Wilcoxon tests.Results: Longest diameter, volume and mass of nodules and those of their solid components were significantly higher using MBIR (p < 0.05) with mean differences of 1.1% (limits of agreement, -6.4 to 8.5%), 3.2% (-20.9 to 27.3%) and 2.9% (-16.9 to 22.7%) and 3.2% (-20.5 to 27%), 6.3% (-51.9 to 64.6%), 6.6% (-50.1 to 63.3%), respectively. The limits of agreement between FBP and MBIR were within the range of intra- and interobserver variability for both algorithms with respect to the diameter, volume and mass of nodules and their solid components. There were no significant differences in intra- or interobserver variability between FBP and MBIR (p > 0.05).Conclusion: Semi-automatic measurements of SSNs significantly differed between FBP and MBIR; however, the differences were within the range of measurement variability.Key Points: • Intra- and interobserver reproducibility of measurements did not differ between FBP and MBIR. • Differences in SSNs' semi-automatic measurement induced by reconstruction algorithms were not clinically significant. • Semi-automatic measurement may be conducted regardless of reconstruction algorithm. • SSNs' semi-automated classification agreement (pure vs. part-solid) did not significantly differ between algorithms. [ABSTRACT FROM AUTHOR]

Autophagy is a self-degradation system wherein cellular materials are recycled. Although autophagy has been extensively studied in yeast and mammalian systems, integrated stress responses in microalgae remain poorly understood. Accordingly, we carried out a comparative study on the oxidative stress responses of Chlamydomonas reinhardtii wild-type and a starchless (sta6) mutant previously shown to accumulate high lipid content under adverse conditions. To our surprise, the sta6 mutant exhibited significantly higher levels of lipid peroxidation in the same growth conditions compared to controls. The sta6 mutant was more sensitive to oxidative stress induced by H2O2, whereas the wild-type was relatively more resistant. In addition, significantly up-regulated autophagy-related factors including ATG1, ATG101, and ATG8 were maintained in the sta6 mutant regardless of nitrogen availability. Also, the sta6 mutant exhibited relatively higher ATG8 protein level compared to wild-type under non-stress condition, and quickly reached a saturation point of autophagy when H2O2 was applied. Our results indicate that, in addition to the impact of carbon allocation, the increased lipid phenotype of the sta6 mutant may result from alterations in the cellular oxidative state, which in turn activates autophagy to clean up oxidatively damaged components and fuel lipid production. [ABSTRACT FROM AUTHOR]