Summary Mesenchymal stromal cells (MSCs) have gained widespread popularity in cell therapy, but their development into clinical products has been impeded by the scarcity of cell‐specific markers. We previously explored transcriptome and membrane proteome of MSCs, from which fibroblast activation protein α (FAP) was recognized as a prime surface marker candidate. The present study showed that FAP was constitutively expressed on MSCs, but not on other cells. FAP immunoselection yielded homogeneous MSCs from cryopreserved bone marrow (BM). These results suggest that FAP serves as a surface protein marker that can singly define MSCs from BM and possibly from other sources.

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Lipopolysaccharide (LPS), which is a component of the cell walls of gram-negative bacteria, is one of the most potent macrophage activators. LPS induces the production of pro-inflammatory cytokines and small mediators, such as nitric oxide (NO) and PGE 2 in macrophages. Macrophage activation is known to play an important role in the inflammatory process.1) Increasing evidence also suggests that NO, a bioactive free radical, is involved in various pathophysiological processes including inflammation and septic shock. 2,3) Therefore, NO production might reflect the inflammation process and provide a measure for assessing the effects of drugs on this process. NO production is regulated by NO synthases (NOSs) and three NOS isoforms have been identified and cloned. The neural (nNOS) and the endothelial (eNOS) isoforms are constitutively expressed in selected tissues, and their enzymatic activity is regulated by changes in the intracellular free Ca 2ϩ concentration. A third member of the NOS family is type II NOS (inducible NOS, iNOS), which is produced in large quantities in response to inflammatory stimuli such as LPS and interferon-g (IFN-g). 4,5) Proinflammmatory cytokines (tumor necrosis factor-a (TNF-a), IL-1b and IL-6) have been suggested to induce tissue damage, and are considered to be an important initiator of the inflammatory responses. The expression of these cytokines depends on the activation of the transcriptional factor, nuclear factor (NF)-kB. NF-kB is a critical intracellular mediator of the inflammatory cascade because these proinflammatory molecules are regulated at the transcription level. [6][7][8][9][10] In a study of anti-inflammatory activities of chemically synthetic propenone compound, 1-furan-2-yl-3-pyridin-2-ylpropenone (FPP-3) ( Fig. 1) was found to inhibit LPS-stimulated NO and TNF-a production in RAW264.7 macrophages in a preliminary in vitro test. The aim of the present study was to evaluate the mechanism by which FPP-3 inhibits NO and TNF-a production in macrophages. The results clearly demonstrated that treatment with FPP-3 decreases LPS-stimulated iNOS protein expression and TNF-a production through the inhibition of NF-kB activation. MATERIALS AND METHODSMaterials 2-Acetylfuran and 2-pyridinecarboxaldehyde were purchased from Aldrich Chemical Co. (St. Louis, MO, U.S.A.) and were reagent grade. Thin-layer chromatography (TLC) and column chromatography were performed with Kieselgel 60 F 254 (Merck) and silica gel Kieselgel 60 (230-240 mesh, Merck), respectively. Compounds were visualized on TLC plates with UV light and r-anisaldehyde solution. Nuclear magnetic resonance (NMR) spectra were taken on a Bruker AMX 250 MHz, and tetramethylsilane was used as an internal standard. Chemical shifts (d) were recorded in ppm, and coupling constants (J) in Hz. RPMI 1640 and phosphatebuffer saline were obtained from GIBCO-BRL (Grand Island, NY, U.S.A.). Fetal bovine serum was purchased from Hyclone Laboratories (Logan, UT, U.S.A.). Rabbit polyclonal iNOS antibody, and anti-rabbit IgG peroxidase-co...

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The bioassay-guided fractionation of the methylene chloride soluble portion of a methanol extract of Machilus thunbergii bark led to the isolation of four known lignans, machilin A (1), meso-monomethyl dihydroguaiaretic acid (2), nectandrin A (3) and nectandrin B (4), which exhibited potent inhibitory activity on melanin biosynthesis in cultured B-16 mouse melanoma cells (IC(50): 39.9, 15.1, 19.4 and 37.8 microM, respectively).