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Intrahepatic bile ducts (IHBDs) are indispensable for transporting bile secreted from hepatocytes to the hepatic duct. The biliary epithelial cells (BECs) of the IHBD arise from bipotent hepatoblasts around the portal vein, suggesting the portal mesenchyme is essential for their development. However, except for Notch or Activin/ TGF-␤ signaling molecules, it is not known which molecules regulate IHBD development. Here, we found that FGF receptors and BMP4 are specifically expressed in the developing IHBD and the hepatic mesenchyme, respectively. Using a mesenchyme-free culture of liver bud, we showed that bFGF and FGF7 induce the hepatoblasts to differentiate into BECs, and that BMP4 enhances bFGF-induced BEC differentiation. The extracellular matrix (ECM) components in the hepatic mesenchyme induced BEC differentiation. Forced expression of a constitutively active form of the FGF receptor partially induced BEC differentiation markers in vivo. These data strongly suggest that bFGF and FGF7 promote BEC differentiation cooperatively with BMP4 and ECMs in vivo.

The chick embryo has been used widely for studying liver development. However, in the past 30 years, the usage has decreased markedly due to lack of appropriate marker genes for differentiation in the developing chick liver. To use the chick embryo for analyzing the molecular mechanism of liver development, we surveyed marker genes in the developing chick liver by examining the expression pattern of genes that are well-characterized in the developing mammalian liver. By whole-mount in situ hybridization, Fibrinogengamma (FIB) expression was first detected at stage 12, specifically in the anterior intestinal portal, and its liver-specific expression persisted in the later stages. Albumin (ALB) expression was first detected at stage 30, when the liver starts maturing. Cytokeratin 19 (CK19) was first detected at stage 37 in the ductal plate of the liver, and its expression continued in the intrahepatic bile ducts derived from the ductal plate. Hex, a transcription factor, is an additional marker of bile duct differentiation. Hence, FIB, ALB, and CK19 expression can be used to trace hepatic induction, maturation, and bile duct differentiation, respectively.

Background Resolvin E1 (RvE1) derived from the ω-3 polyunsaturated fatty acid eicosapentaenoic acid is known to be a potent pro-resolving lipid mediator that prevents chronic inflammation and osteoclastogenesis. We investigated the inhibitory effects of RvE1 on osteoclastogenesis and bone resorption to clarify its therapeutic potential for rheumatoid arthritis (RA). Methods Receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation was assessed with tartrate-resistant acid phosphatase staining. RANKL-induced bone resorption was assessed by the measurement of pit formation using calcium phosphate-labeled fluorescent polyanionic molecules in RAW264.7 cells as osteoclast precursors. The effects of RvE1 on the RANKL-induced mRNA expression of osteoclast-specific genes and transcriptional factors such as c-fos and nuclear factor of activated T cells c1 (NFATc1) in RAW264.7 cells were measured by quantitative real-time PCR. The distribution of NFATc1 induced by RANKL was evaluated by immunofluorescence staining in RAW264.7 cells. To analyze the mechanism of the inhibitory effect of RvE1 on osteoclastogenesis, we measured IL-17-induced RANKL mRNA expression in MC3T3-E1 osteoblast cells treated with RvE1 using quantitative real-time PCR and determined the level of prostaglandin E 2 (PGE 2 ) production by enzyme-linked immunosorbent assay. Results RvE1 significantly suppressed RANKL-in-duced osteoclast differentiation and bone resorption. RvE1 inhibited the RANKL-induced mRNA expression of osteoclast-specific genes along with the transcription factors NFATc1 and c-fos. Moreover, NFATc1 translocation from the cytoplasm to the nucleus of RAW264.7 cells was suppressed following RvE1 treatment. RvE1 also inhibited IL-17-induced RANKL mRNA expression and PGE 2 production in MC3T3-E1 cells. Conclusion RvE1 inhibited osteoclastogenesis and bone resorption by suppressing RANKL-induced NFATc1 and c-fos expression in osteoclasts and IL-17induced RANKL expression through the autocrine action of PGE 2 in osteoblasts. Our data suggest RvE1 as a new therapeutic target of RA.

Background:Although considerable efforts have been made to develop diagnostic tools for predicting the outcome of oral food challenges, tests for predicting the outgrowth of food allergies are lacking.Objective:The aim of this study was to assess the diagnostic value of the wheal size and skin index (SI) (the ratio of an allergen-induced wheal to a histamine-induced wheal diameter) of the skin-prick test based on the outcome of a controlled oral provocation test for cow's milk. Moreover, we assessed whether wheal size and/or SI were useful for predicting the outgrowth of cow's milk allergy (CMA).Methods:This study included 135 children with suspected CMA. Eighty-one patients were definitely diagnosed by oral provocation tests for cow's milk, and their wheal diameters, SIs, and cow milk's–specific serum immunoglobulin E concentrations were determined.Results:The wheal diameters were significantly larger and the SIs significantly higher in children with positive oral provocation test results than in those with negative test results. We found that 50% of the patients were expected to be able to drink cow's milk by age 5 years. In these patients, the wheal diameters were significantly smaller and the SIs significantly lower at the time of CMA outgrowth than at the time of diagnosis, whereas these values were apt to increase in patients who did not outgrow CMA, with no significant difference.Conclusions:The skin-prick test can be used to diagnose CMA and predict CMA outgrowth. A wheal diameter of 8 mm or/and an SI of 1.0 is informative, not only in diagnosing CMA but also in predicting a natural CMA outgrowth.