Importance of Kupffer Cells in the Development of Acute Liver Injuries in Mice

Kupffer cells reside within the liver sinusoid and serve as gatekeepers. They produce pro- and anti-inflammatory cytokines and other biologically important molecules upon the engagement of pattern recognition receptors such as Toll-like receptors. Kupffer cell-ablated mice established by in vivo treatment with clodronate liposomes have revealed many important features of Kupffer cells. In this paper, we review the importance of Kupffer cells in murine acute liver injuries and focus on the following two models: lipopolysaccharide (LPS)-induced liver injury, which is induced by priming with Propionibacterium acnes and subsequent challenge with LPS, and hypercoagulability-mediated acute liver failure such as that in concanavalin A (Con A)-induced hepatitis. Kupffer cells are required for LPS sensitization induced by P. acnes and are a major cellular source of interleukin-18, which induces acute liver injury following LPS challenge. Kupffer cells contribute to Con A-induced acute liver failure by initiating pathogenic, intrasinusoidal thrombosis in collaboration with sinusoidal endothelial cells. The mechanisms underlying these models may shed light on human liver injuries induced by various etiologies such as viral infection and/or abnormal metabolism.     

Lingual Epithelial Stem Cells and Organoid Culture of Them

As tongue cancer is one of the major malignant cancers in the world, understanding the mechanism of maintenance of lingual epithelial tissue, which is known to be the origin of tongue cancer, is unquestionably important. However, the actual stem cells that are responsible for the long-term maintenance of the lingual epithelium have not been identified. Moreover, a simple and convenient culture method for lingual epithelial stem cells has not yet been established. Recently, we have shown that Bmi1-positive cells, residing at the second or third layer of the epithelial cell layer at the base of the interpapillary pit (IPP), were slow-cycling and could supply keratinized epithelial cells for over one year, indicating that Bmi1-positive cells are long-term lingual epithelial stem cells. In addition, we have developed a novel lingual epithelium organoid culture system using a three-dimensional matrix and growth factors. Here, we discuss current progress in the identification of lingual stem cells and future applications of the lingual culture system for studying the regulatory mechanisms of the lingual epithelium and for regenerative medicine.     

Identification of the gene PaEMT1 for biosynthesis of mannosylerythritol lipids in the basidiomycetous yeast Pseudozyma antarctica

The yeast Pseudozyma antarctica produces a large amount of glycolipid biosurfactants known as mannosylerythritol lipids (MELs), which show not only excellent surface‐active properties but also versatile biochemical actions. To investigate the biosynthesis of MELs in the yeast, we recently reported expressed sequence tag (EST) analysis and estimated genes expressing under MEL production conditions. Among the genes, a contiguous sequence of 938 bp, PA_004, showed high sequence identity to the gene emt1, encoding an erythritol/mannose transferase of Ustilago maydis, which is essential for MEL biosynthesis. The predicted translation product of the extended PA_004 containing the two introns and a stop codon was aligned with Emt1 of U. maydis. The predicted amino acid sequence shared high identity (72%) with Emt1 of U. maydis, although the amino‐terminal was incomplete. To identify the gene as PaEMT1 encoding an erythritol/mannose transferase of P. antarctica, the gene‐disrupted strain was developed by the method for targeted gene disruption, using hygromycin B resistance as the selection marker. The obtained ΔPaEMT1 strain failed to produce MELs, while its growth was the same as that of the parental strain. The additional mannosylerythritol into culture allowed ΔPaEMT1 strain to form MELs regardless of the carbon source supplied, indicating a defect of the erythritol/mannose transferase activity. Furthermore, we found that MEL formation is associated with the morphology and low‐temperature tolerance of the yeast. Copyright © 2010 John Wiley & Sons, Ltd.     

Complete erasing of ghost images on computed radiography plates and role of deeply trapped electrons

Computed radiography (CR) plates made of europium-doped Ba(Sr)FBr(I) were simultaneously exposed to filtered ultraviolet light and visible light in order to erase ghost images, i.e., latent image that is unerasable with visible light (LIunVL) and reappearing one, which are particularly observed in plates irradiated with a high dose and/or cumulatively over-irradiated. CR samples showing LIunVLs were prepared by irradiating three different types of CR plates (Agfa ADC MD10, Kodak Directview Mammo EHRM2, and Fuji ST-VI) with 50 kV X-ray beams in the dose range 8.1 mGy—8.0 Gy. After the sixth round of simultaneous 6 h exposures to filtered ultraviolet light and visible light, all the LIunVLs in the three types of CR plates were erased to the same level as in an unirradiated plate and no latent images reappeared after storage at 0 °C for 14 days. With conventional exposure to visible light, LIunVLs consistently remained in all types of CR plates irradiated with higher doses of X-rays and latent images reappeared in the Agfa M10 plates after storage at 0 °C. Electrons trapped in deep centers cause LIunVLs and they can be erased by simultaneous exposures to filtered ultraviolet light and visible light. To study electrons in deep centers, the absorption spectra were examined in all types of irradiated CR plates by using polychromatic ultraviolet light from a deep-ultraviolet lamp. It was found that deep centers showed a dominant peak in the absorption spectra at around 324 nm for the Agfa M10 and Kodak EHRM2 plates, and at around 320 nm for the Fuji ST-VI plate, in each case followed by a few small peaks. The peak heights were dose-dependent for all types of CR samples, suggesting that the number of electrons trapped in deep centers increases with the irradiation dose.     

Simple Method for Measuring the Peroxide Value in a Colored Lipid

We developed a simple method for quantification of the peroxide value (PV) in colored lipids on the basis of the reaction between triphenylphosphine (TPP) and oxidized oil to afford triphenylphosphineoxide (TPPO). Diphenylphosphineoxide (DPPO) was employed as internal standard. The formed TPPO was analyzed by high-pressure liquid chromatography–UV spectroscopy with absorption at 260 nm. The conditions that gave the highest correlative calibration curve between the peak area on the chromatogram and peroxide value were identified: the optimum TPP–oxidized oil mix ratio, reaction temperature, and reaction time were found to be 2:1, 40 °C, and 30 min, respectively. Linear calibration curves, passing through the origin, were obtained for PV versus TPPO and TPPO versus DPPO. The quantification limit for this method was 2.01 pmol hydroperoxyl group, which corresponds to a PV value of 0.2 meq/kg in a 10-mg oil sample. This method was used to measure the PV in colored fats and oils or lipids extracted from dark meat and processed food containing a coloring agent. Though the official method could not measure the PVs in the colored lipids, the method proposed here, which uses an inexpensive chemical reagent and machine, could. The developed method could play an important role for food quality control.     

Breeding of high malate‐producing diploid sake yeast with a homozygous mutation in the VID24 gene

Malate is an important taste component of sake (a Japanese alcoholic beverage) that is produced by the yeast Saccharomyces cerevisiae during alcoholic fermentation. A variety of methods for generating high malate‐producing yeast strains have been developed to date. We recently reported that a high malate‐producing strain was isolated as a mutant sensitive to dimethyl succinate (DMS), and that a mutation in the vacuolar import and degradation protein (VID) 24 gene was responsible for high malate productivity and DMS sensitivity. In this work, the relationships between heterozygous and homozygous mutants of VID24 and malate productivity in diploid sake yeast were examined and a method was developed for breeding a higher malate‐producing strain. First a diploid yeast was generated with a homozygous VID24 mutation by genetic engineering. The homozygous integrants produced more malate during sake brewing and grew more slowly in DMS medium than wild‐type and heterozygous integrants. Thus, the genotype of the VID24 mutation influenced the level of malate production and sensitivity to DMS in diploid yeast. Then a homozygous mutant from a heterozygous mutant was obtained without genetic engineering by ultraviolet irradiation and culturing in DMS with nystatin enrichment. The non‐genetically modified sake yeast with a homozygous VID24 mutation exhibited a higher level of malate productivity than the parent heterozygous mutant strain. These findings provide a basis for controlling malate production in yeast, and thereby regulating malate levels in sake. Copyright © 2016 The Institute of Brewing & Distilling