To establish a human hepatic cell line for a convenient reporter gene assay of arylhydrocarbon receptor (AhR) activators, the chimera plasmid containing xenobiotic responsible element (XRE), minimal SV40 promoter, and luciferase reporter gene and the expression vector pRC/CMV containing a neomycin-resistant gene were co-transfected into a human hepatoma cell line, HepG2. Then, antibiotic (G418)-resistant HepG2 cells were selected and cloned. A cell clone, HepG2-A10, showed the highest responsibility to 3-methylcholanthrene (MC)-mediated induction of luciferase among the clones obtained. Expression levels of luciferase activity in HepG2-A10 cells were increased in a dose-and time-dependent manner by treatment with either MC, an AhR-ligand type activator, or omeprazole (OME), a non-AhR ligand type activator. In addition, expression levels of cytochrome P4501A subfamily genes (CYP1A1 and CYP1A2) were also increased in a dose-and time-dependent manner by treatment with MC. The presentˆndings demonstrate that a newly established human HepG2-A10 is a useful cell line for a convenient reporter gene assay of AhR activators.

Background:The blood metabolome profiles depend on the meal intake time zone regardless of having the same meal. The serum albumin (Alb) level, which is important in managing geriatric patients with chronic diseases, is included in the metabolome analysis. In this study, we aimed to examine the relationship between Alb and the nutritional value of hospital meals consumed at breakfast, lunch, and dinner among geriatric patients. Chrononutrition was considered while drawing inferences. Methods:We retrospectively surveyed 52 geriatric patients with chronic diseases (aged 79.7 ± 8.7 years) admitted at a small-scale hospital providing combined healthcare measures and oral nutritional support. The dietary intake per kilogram of body weight of nutritional components for breakfast, lunch, and dinner was individually expressed as the ratio to the whole daily food intake. The dietary pattern was determined by principal component analysis. We also conducted linear regression analysis, with Alb as the dependent variable, and age, sex, and grade assigned in this study as well as the first, second, and third principal components of the dietary patterns as the independent variables.Results: Three principal components with an eigenvalue of > 1 were extracted. The second principal component was a significantly negative determinant factor for Alb (B = -0.108, P = 0.016). In patients with high Alb levels, the energy, protein, and fat ratios at lunch were positively correlated, while the energy and carbohydrate ratios at dinner were negatively correlated. Mealtimes were fixed. Conclusions:The results of this study showed that the dietary pattern predominantly observed in patients with high Alb levels may be positively associated with Alb synthesis.

Butylidenephthalide, the components of Angelica acutiloba var. sugiyamae HIKINO (Apiaceae) has the biological activities, such as centrally acting muscle relaxant 1) and antispasmodic effects 2) with characteristic odor. In our previous work,3) (Z)-butylidenephthalide was isolated as the potent insecticidal compound against adults and larvae of Drosophila melanogaster. Recently, Ahn et al. reported that butylidenephthalide was active constituent of Cnidium officinale against Dermatophagoides farinae and Tyrophagus putrescentiae, and more effective than acaricide, N,N-diethyl-mtoluamide (DEET). 4,5) Therefore, butylidenephthalide is the potential candidate for new natural pesticide. However, above report was unclear on stereochemistry of butylidenephthalide even though both (E)-and (Z)-butylidenephthalide were found from natural resources, and (E)-butylidenephthalide has not been tested against insects and mites.Geometrical stereoselectivity is often crucial on biological activities. Asahina et al. reported that synthetic fluoroquinolone derivatives were tested for antibacterial activity, and (Z)-isomers exhibited 2-to 32-fold more potent than corresponding (E)-isomers. 6) Against insects, the importance of geometry was discussed that binding affinity of juvenile hormone carrier protein from the hemolymph of Manduca sexta was decreased as (Z)-double bond are substituted for (E)-double bond in synthetic juvenile hormone analogues. 7)Therefore, evaluation of biological activity for geometrical isomers is required.Biological activity of (E)-butylidenephthalide against insects and mites has not been previously reported. In this paper, we investigated biological activities of (E)-butylidenephthalide against larvae and adults of D. melanogaster, adults of Ctenocephalides felis and two house dust mites, D. farinae and T. putrescentiae, and compared with (Z)-butylidenephthalide to investigate geometrical stereoselectivity. Rotenone (positive control for insecticidal activity) and DEET (positive control for acaricidal activity) were also tested. MATERIALS AND METHODSGC and GC-MS Analysis GC was performed on a Hewlett-Packard 5890A gas chromatograph equipped with a flame ionization detector (FID). The column was a fused silica capillary column (DB-5, 30 m length, 0.25 mm i.d.). Chromatographic conditions were as follows: oven temperature was programmed from 80 to 250°C at 4°C/min and held at 250°C for 15 min; injector and detector temperature were 270 and 280°C, respectively; carrier gas, helium at a flow rate of 40 cm/s. GC-MS was performed on a HewlettPackard 5972A mass selective detector interfaced with a Hewlett-Packard 5890A gas chromatograph fitted with a capillary column (HP-5MS, 30 m length, 0.25 i.d.). Chromatographic conditions were the same for the GC analysis.Chemicals (Z)-Butylidenephthalide was previously isolated from Angelica acutiloba.3) (E)-Butylidenephthalide was isolated from chloroform extract of A. acutiloba by SiO 2 (200 mesh) column chromatography (hexane : ethyl acetate ϭ93 : 7), repeatedly. The pu...

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Background Chronic kidney disease (CKD) is frequently associated with renal anemia. Erythropoiesis-stimulating agent-hyporesponsive anemia is often caused by iron deficiency in patients with CKD. We hypothesized that high accumulation of indoxyl sulfate, a uremic toxin, accelerates iron deficiency in patients with CKD. The aim of this study was to clarify whether the accumulation of indoxyl sulfate is a cause of iron deficiency in patients with CKD. Therefore, we investigated the association between serum indoxyl sulfate concentration and iron dynamics in patients with end-stage kidney disease (ESKD). Methods We performed a cross-sectional study on 37 non-dialyzed patients with ESKD, who were hospitalized to undergo maintenance hemodialysis treatment at Shirasagi Hospital. Serum indoxyl sulfate concentration, iron dynamics parameters and other laboratory data were measured immediately before the initiation of hemodialysis treatment. Clinical characteristics were obtained from electronic medical records. Results The estimated glomerular filtration rate (eGFR) of 37 patients with ESKD was 5.08 (3.78–7.97) mL/min/1.73 m2 (median [range]). Serum ferritin and transferrin saturation (TSAT) were 90 (10–419) ng/mL and 20 (8–59)% (median [range]), respectively. Serum indoxyl sulfate concentration was 62 (11–182) μM (median [range]). Serum indoxyl sulfate concentration was inversely correlated with serum ferritin level (ρ = − 0.422, p = 0.011), but not with TSAT, age, gender, eGFR and c-reactive protein (CRP) in 37 patients. In eight patients taking iron-containing agents, serum indoxyl sulfate concentration was strongly correlated with serum ferritin level (ρ = − 0.796, p = 0.037); however, in 29 patients not taking an iron-containing agent, this correlation was not observed (ρ = − 0.336, p = 0.076). In the multivariate analysis including age, gender, eGFR and CRP, the correlation between serum indoxyl sulfate concentration tended to be, but not significantly correlated with serum ferritin level in 37 patients (regression coefficient = − 54.343, p = 0.137). Conclusion Our study suggests that serum accumulation of indoxyl sulfate is one of causes not to increase serum ferritin level in patients with ESKD taking an iron-containing agent. Further clinical study is needed to reveal the appreciable relationship between serum ferritin and serum indoxyl sulfate.