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Perilla frutescens (L.) Britton var. frutescens (egoma in Japan) is a traditional oilseed that has several varieties with different photoperiod responses. Although egoma pomace, industrial waste produced during oil extraction, is a rich source of macro- and micro-nutrients such as protein, fiber, minerals, and polyphenols, it has not yet been used for purposes other than livestock feeding. To find out a better use of perilla pomace and its function, we selected four varieties of egoma originating from different regions with different photoperiod responses: two varieties were from Japan, which are broadly cultivated for oilseed and are highly sensitive to light and temperature. The other two varieties from Nepal, which are tolerant to low light and low temperature. Rosmarinic acid-3-O-glucoside, rosmarinic acid, and apigenin-7-O-glucoside were detected as the main polyphenolic constituents in every variety, while apigenin and luteolin were present only in perilla pomace from Japan. In IgE-sensitized RBL-2H3 cells, polyphenols derived from two varieties of Japan suppressed degranulation of mast cells, but those derived from the two varieties of Nepal did not, indicating that apigenin and luteolin may be in part responsible for the anti-allergic response. In addition, it was found that proteins involved in the degranulation signaling pathway, such as PLCγ2, Syk, and Akt, were less phosphorylated in cells treated with the egoma pomace extracts of Japanese origin. Taken together, pomace from egoma varieties derived from different regions may differently modulate allergic response in part due to the difference in polyphenol composition and may be applied to develop nutraceuticals and functional foods fortified with anti-allergic properties.

We previously reported the successful establishment of multiple immortalized cell lines that preserved the original nature of the primary cells via co-expression of R24C mutant cyclin-dependent kinase 4 (CDK4(R24C)), Cyclin D1, and telomerase reverse transcriptase (TERT). However, as these genes are kind of oncogenes, tools to control their expression levels are favorable. In this study, we describe a new polycistronic lentiviral vector expressing proliferation factors, CDK4(R24C) and Cyclin D1 along with enhanced green fluorescence protein (EGFP) under the control of doxycycline (Dox)-dependent transactivator (rtTA) and tetracycline response element (TRE). By introducing the Dox-inducible lentiviral vector into human airway epithelial cells, we established a novel human airway epithelial cell line harboring polycistronic Dox-inducible CDK4(R24C) and Cyclin D1, referred to as Tet-on K4D cells. We showed that the cell growth of Tet-on K4D cells could be controlled by Dox. Furthermore, expression of K4D genes and rtTA gene can be independently monitored by fluorescent imaging. Cultured airway epithelial cells are useful as a tool for studying the pathogenesis of lung disorders. Altogether, our established human airway epithelial cells could be used for a variety of studies such as lung pathology and biology underlying the differentiation process to form the complex pseudostratified multicellular layers. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10616-021-00477-0.

To obtain a large number of human mesenchymal stem cell (hMSCs) for allograft, nonwoven fabrics (NWF) were used as a cell culture scaffold. NWF are three-dimensional fiber aggregates formed by heat bonding and have a high surface area for cell adhesion and elongation. Inoculation hMSC was done to a center of NWF disc (diameter, 15.1 mm; depth, 0.1 mm). A cell suspension inoculum had a volume of 10 µL, which was close to the void volume of the disc, and resulted in a high initial (24 h) cell adhesion efficiency. Use of green fluorescent protein expressing rat MSCs and fluorescence microscopy revealed that adding an additional 10 μL of medium at 0–2 h after the cell inoculation made the initial horizontal distribution of cells in the NWF disc more uniform. Addition of 10 μL of the medium after 1 and 2 h of hMSC inoculation (0.15 × 10(3) cells/cm(2) NWF-fiber) markedly increased the final cell density (21 days) from 2.48 to 7.45 × 10(3) cells/cm(2) NWF-fiber and fold increase in cell density by 16–48-fold. In conclusion, the addition of an additional medium after inoculation made the initial cells distribution in NWF more uniform, which might result in higher final cell density.

Lysozyme is an anti-bacterial protein that is widely distributed in nature. Our previous studies revealed that lysozyme shows anti-inflammatory effect on hyperinflammatory macrophages in vitro. The effect of lysozyme on lipopolysaccharide-induced inflammation model mice was examined in this study. Oral administration of lysozyme at 2250 mg/kg body weight/day (high-dose group) significantly suppressed interleukin (IL)-6 and tumor necrosis factor-α levels in the serum. IL-6 level in the spleen was significantly suppressed by lysozyme at 450 mg/kg body weight/day (middle-dose group) and high-dose group due to the suppression of gene expression level. The gene expression levels of IL-1β and IL-12 were also decreased by lysozyme in the high-dose group. In addition, lysozyme significantly suppressed IL-6 level in the liver in the high-dose group. Our findings suggest that lysozyme mitigates inflammatory condition in vivo by suppressing inflammatory cytokine levels in serum and organs from LPS-induced inflammation model mice.