Evaluation of Drug-Drug Interaction Potential of Enarodustat (JTZ-951) Using a Cytochrome P450 Probe Cocktail.

The drug interaction potential of enarodustat (doses: 25, 50 mg) on the activity of cytochrome P450 (CYP) 1A2, 2C9, 2C19, 2D6, and 3A4 was evaluated after once-daily administration for 15 days in a phase 1 multiple-ascending-dose study in healthy subjects. Probe substrates specific for the enzymes, i.e., caffeine (CYP1A2), tolbutamide (CYP2C9), omeprazole (CYP2C19), dextromethorphan (CYP2D6), and midazolam (CYP3A4), were administered orally as a cocktail with (day 15) and without (day -3) enarodustat. Drug interaction was based on geometric mean maximum plasma concentration (Cmax ) and area under the plasma concentration-time curve from the time of dosing to infinity (AUCinf ) ratios (day 15/day -3) for CYP1A2, 2C9, 2C19, 2D6, 3A4, and urinary excretion of dextromethorphan metabolite dextrorphan for CYP2D6. At the 2 enarodustat doses, for caffeine, the geometric mean ratios (range) for Cmax and AUCinf were 0.99-1.06 and 1.61-1.63, respectively. The ratios for peak concentrations and total exposures were 0.98-1.07 and 0.71-1.78 for tolbutamide and omeprazole, respectively. For dextrorphan the Cmax and AUCinf ratios were 0.83-0.90 and 1.02-1.04, respectively. The mean dextrorphan cumulative amount excreted into the urine from the time of dosing to 24 hours values on day -3 and day 15 were 8.25 mg and 8.20 mg at the lower dose, and 9.40 mg and 9.51 mg at the higher dose. The ratios for midazolam Cmax and AUCinf were 1.42-1.63. Overall, there was a lack of enarodustat dose dependency regarding the geometric mean ratios and 90% confidence intervals and urinary excretion of dextrorphan. There were some cases where the 90% confidence intervals at the 2 enarodustat doses were outside the 0.80-1.25 range, but changes in the geometric mean ratios were all <2-fold.

Efficacy of a mixture of Ginkgo biloba, sesame, and turmeric on cognitive function in healthy adults: Study protocol for a randomized, double-blind, placebo-controlled trial.

BACKGROUND AND PURPOSE: Ginkgo biloba extract (GBE) reportedly ameliorates cognitive function in patients with chronic cerebrovascular insufficiency. However, its efficacy in healthy adults is ambiguous. It was reported that concentrations of terpene lactones, active components of GBE that are present in very low concentrations in the brain, were significantly increased following administration of a mixture of GBE, sesame seed, and turmeric (GBE/MST) in mice. This study aims to investigate the effectiveness of GBE/MST on the cognitive function of healthy adults by comparing it with that of GBE alone. METHODS: Altogether, 159 participants providing informed consent will be recruited from a population of healthy adults aged 20-64 years. Normal cognitive function at baseline will be confirmed using the Japanese version of the Montreal Cognitive Assessment battery. Participants will be randomly assigned in a double-blind manner to the GBE/MST, GBE, and placebo groups in a 1:1:1 ratio. The Wechsler Memory Scale, Trail Making Test, and Stroop Color and Word Test will be used to assess the memory and executive functions at baseline and at the endpoint (24 weeks). For biological assessment, resting state functional magnetic resonance imaging (rs-fMRI) will be performed simultaneously with the neuropsychological tests. DISCUSSION: This study aims to obtain data that can help compare the profile changes in memory and executive functions among participants consuming GBE/MST, GBE alone, and placebo for 24 weeks. Alterations in the default mode network will be evaluated by comparing the rs-fMRI findings between baseline and 24 weeks in the aforementioned groups. Our results may clarify the impact of GBE on cognitive function and the functional mechanism behind altered cognitive function induced by GBE components. TRIAL REGISTRATION: This study was registered in the University Hospital Medical Information Network Clinical Trials Registry (UMIN-CTR; registration number: UMIN000043494). This information can be searched on the website of the International Clinical Trials Registry Platform Search Portal of the World Health Organization under the Japan Primary Registries Network.

Theoretical Study on the Adhesion Interaction between Epoxy Resin Including Curing Agent and Plated Gold Surface.

In this study, the adhesive interaction between gold and epoxy resin is theoretically investigated. These materials make up crucial components of a wide range of electronic devices. The objectives of the study are (1) to elucidate the bonding mechanism between epoxy resin and a realistic gold surface, and (2) to obtain a device-design guideline for superior adhesion, thus reducing the bonding breakage that may potentially cause device failure. Die pad surfaces used in chip attachment methods for microelectronics are usually fabricated using an electrolytic plating technique. This technique involves ionic gold solutions like K[Au(CN)2]. The combined theoretical and experimental studies previously carried out by the authors have revealed that the CN- counteranion of the gold cation has a high affinity for gold and is likely to remain on the realistic gold surface generated by plating. However, the cyano group content on the surface of the plated gold is still unknown. Therefore, gold surfaces embedded with cyano groups with various coverages are constructed. The effect of the varying coverage of the cyano groups on the adhesion strength is inspected using first-principles density functional theory calculations. As the number of cyano groups on the surface increases, the direct interaction between the gold surface and the epoxy resin is hindered, but the hydroxy and amino groups in the epoxy resin and hardener form more hydrogen bonds with the cyano groups adsorbed on the surface. It is found that the surface with intermediate cyano coverage (about 33%) yields the highest adhesive strength.

Proposal of a Parameter for OATP1B1 Inhibition Screening at the Early Drug Discovery Stage.

It is known that potent inhibition of organic-anion-transporting polypeptide (OATP)1B1 increases exposure to statins, leading to severe adverse effects. The aim of this study was to propose a parameter and its criteria in OATP1B1 inhibition assay at the early drug discovery stage to avoid compounds with the risk of statin-related adverse effects. According to drug label information, most compounds classified as "contraindicated" or "should be avoided" when administered concomitantly with statins increased their AUCs more than 4-fold. Generally, R values where R = 1 + plasma unbound fraction (fu) × maximum inhibitor concentration at the inlet to the liver/IC50 are used to evaluate the extent of clinical drug interaction. However, clinical doses and Cmax cannot be determined at the screening stage. Therefore, we estimated the correlations between change in AUC of statins concomitantly administered with OATP1B1 inhibitors and various parameters including fu/IC50. Cyclosporin A, rifampicin, and telaprevir increased the AUC of statins more than 4-fold and fu/IC50 of these compounds was >0.1 L/µmol. On the other hand, fu/IC50 of other compounds was ≤0.03 L/µmol. This study indicates that fu/IC50 is a useful parameter to avoid compounds that seriously affect statin potency through interaction with OATP1B1 at the screening stage.

Density-Functional Tight-Binding Study on the Effects of Interfacial Water in the Adhesion Force between Epoxy Resin and Alumina Surface.

Adhesion is one of the most interesting subjects in interface phenomena from the viewpoint of wide-range applications as well as basic science. Interfacial water has significant effects on coatings, adhesives, and fiber-reinforced polymer composites, often causing adhesion loss. The way of thinking based on quantum mechanics is essential for a better understanding of physical and chemical properties of adhesive interfaces. In this work, the molecular mechanism of the adhesion interaction between epoxy resin and hydroxylated alumina surface in the presence of interfacial water molecules is investigated by using density-functional tight-binding calculations. Periodic slab model calculations demonstrate that hydrogen bond is an important factor at the adhesion interface. Effects of interfacial water molecules located between epoxy resin and hydroxylated alumina surface are assessed by using a dry model without interfacial water and wet models with water layers of 3, 6, and 9 Å thicknesses. Interesting first- and second-layer structures are observed in the distribution of interfacial water molecules in the tight space between the adhesive and adherend. Energy plots with respect to the displacement of epoxy resin from the alumina surface are nicely approximated by the Morse potential. The adhesion force and stress are theoretically obtained by differentiating the potential curve with respect to the displacement of epoxy resin. Computational results show that the adhesion force and stress are significantly weakened with an increase in the thickness of interfacial water layer. Thus, interfacial water molecules have a clue as to the role of water in the loss of adhesion.

Studies on the Unusual 1ß-Hydroxylated Bile Acid Biosynthesis in Infants.

Unusual bile acids (1ß-hydroxylated bile acids), particularly 1ß-hydroxyl-cholic acid (CA-1ß-ol) and 1ß-hydroxyl-chenodeoxycholic acid (CDCA-1ß-ol), have been detected in the urine of infants. These acids are conjugated with amino acids, such as taurine, and are then excreted mainly via the urine. CA-1ß-ol and CDCA-1ß-ol are the predominant bile acids during infancy and are present in relatively large amounts in the urine. However, the biosynthetic pathway of 1ß-hydroxylated bile acids in infants remains unclear. To investigate the biosynthetic pathway of 1ß-hydroxylated bile acids during infancy, we performed a metabolic reaction using infant hepatocytes at 3 months after delivery. Glyco- and tauro-CA-1ß-ol were identified by LC/tandem mass spectrometry (MS/MS) analysis of the extracted culture medium incubated with cholic acids (CAs). Further, we identified that ketoconazole suppressed CA 1ß-hydroxylation and that the CYP3A subfamily was the primary group of enzymes responsible for CA-1ß-ol formation. The present study provides new information about the biosynthetic pathway of 1ß-hydroxylated bile acids during infancy.

Enhanced Aerosol Particle Filtration Efficiency of Nonwoven Porous Cellulose Triacetate Nanofiber Mats.

Aerosol particle filtration in most penetrating particle size (MPPS) region is of great challenge for conventional nonwoven filter mats. The present work, therefore, redesigns conventional filter mats by introducing porous structure. A combination of thermally induced phase separation and breath figure mechanism was employed to synthesize porous cellulose triacetate fibers, in conjunction with the volatile solvent methylene chloride. The ambient humidity, the concentration of the polyvinylpyrrolidone (PVP) secondary polymer, and the ethanol cosolvent were all adjusted to modify the Taylor cone formation, jet stability, and fiber porosity. After fiber formation, the PVP was removed to obtain a superhydrophobic material. To distinguish the effect of pores, the performance of porous and nonporous nanofibers having similar sizes was conducted. Tests were performed using various dust particle sizes, and the results show that the collection efficiency of the porous fibers, resulting from particle diffusion, inertial impaction, and interception, was improved. Interestingly, the efficiency of the porous fibers in the MPPS region was exceptionally enhanced (up to 95%), demonstrating that the presence of dynamic pores greatly contributes to particle capture.

Synthesis of Dual-Size Cellulose-Polyvinylpyrrolidone Nanofiber Composites via One-Step Electrospinning Method for High-Performance Air Filter.

Dual-size nanofibers consisting of a random mixture of nano- and submicron-size nanofibers are promising structures for specific applications such as air filters because of their increased specific surface area and low pressure drop. Synthesis of dual-size nanofibers using one-step electrospinning was reported here for the first time. The formation of well-mixed nano- and submicron-size cellulose-polyvinylpyrrolidone nanofiber composites was accomplished utilizing the physical properties of TEMPO-oxidized cellulose nanofibers (i.e., high thixotropy and high magnitude of zeta potential) and tuning the charge of the polymer jet, which influences the formation and shape of Taylor cone, and Coulombic explosion. The dual-size nanofibers were then spun on the surface of a HEPA filter to obtain a multilayer air filter. Aerosol filtration measurements show that this multilayer air filter has an incredibly high performance, shown by the high quality factor (Qf), 0.117 Pa-1, which is 10 times the Qf of commercial HEPA filters.

SAIDO learning as a cognitive intervention for dementia care: a preliminary study.

OBJECTIVES: The purpose of this study was to examine the beneficial effects on cognitive function by a cognitive intervention program designed for dementia care called Learning Therapy in Japan and SAIDO Learning in the United States (hereinafter "SAIDO Learning," as appropriate). SAIDO Learning is a working memory training program that uses systematized basic problems in arithmetic and language, including reading aloud, as well as writing. DESIGN, SETTING, PARTICIPANTS, AND INTERVENTION: Twenty-three nursing home residents with dementia were assigned as an intervention group, and another 24 people with dementia at another nursing home were assigned as a control group. Both nursing homes were operated by the same organization, and residents of both nursing homes received essentially the same nursing care. Thirteen and 6 subjects of the intervention and control groups, respectively, were clinically diagnosed as Alzheimer disease (AD). RESULTS: After the 6-month intervention, the participants with AD of the intervention group showed statistically significant improvement in cognitive function, as measured by the Mini-Mental State Examination (MMSE) compared with the control participants. In addition, post hoc analysis revealed that the Frontal Assessment Battery at Bedside (FAB) scores of the intervention group tended to improve after 6-month intervention. Based on MDS scores, improvements in total mood severity scores also were observed, but only in the intervention group of the participants with AD. CONCLUSION: These results suggest that SAIDO Learning is an effective cognitive intervention and is useful for dementia care. An additional outcome of this intervention, which has not yet been evaluated in detail, appears to be that it promotes greater positive engagement of a diversity of nursing home staff in the residents' individual progress and care needs.

Suppression of efflux transporters in the intestines of endotoxin-treated rats.

Infection and inflammation suppress the expression and activity of several drug transporters in the liver. In the intestine, P-glycoprotein (PGP/mdr1) and the multidrug resistance-associated protein 2 (MRP2) are important barriers to the absorption of many clinically important drugs. The protein expression and activity of these transporters were examined during inflammation induced by lipopolysaccharide (LPS). The transport of rhodamine123 (Rho123) and 5-carboxyfluorescein (5-CF) was determined in isolated ileal segments from endotoxin-treated or control rats in the presence or absence of inhibitors. The reverse transcription-polymerase chain reaction was used to measure mRNA levels. Compared with the controls, the mRNA levels of mdr1a and mrp2 were significantly decreased by approximately 50% in the ilea of the LPS-treated rats. Corresponding reductions in the basolateral-apical efflux of Rho123 and 5-CF were observed, resulting in significant increases in the apical-basolateral absorption of these compounds. Neither the permeability of fluorescein isothiocyanate labeled dextran 4000 (FD-4), a paracellular marker, nor membrane resistance was altered. These results indicate that endotoxin-induced inflammation reduces the intestinal expression and activity of PGP and MRP2 in rats, which eliciting corresponding changes in the intestinal transport of their substrates. Hence, infection and inflammatory diseases may induce variability in drug bioavailability through alterations in the intestinal expression and activity of drug transporters.

Effect of lipopolysaccharide on P-glycoprotein-mediated intestinal and biliary excretion of rhodamine123 in rats.

The effects of lipopolysaccharide (LPS) on the ileal and biliary excretion of rhodamine123 were investigated in rats at different times after intraperitoneal (i.p.) injection (1 mg/kg and 5 mg/kg of body weight). P-gp protein decreased 8h after injection of LPS and returned to the control level 24h after i.p. injection of LPS in the ileum. There was a marked decrease in the expression level of mdr1a mRNA in the ileum and liver 8h after i.p. injection of LPS when compared with the control condition. Also, the ileal and biliary clearance of rhodamine123 significantly decreased 8h after i.p. injection of LPS, but returned to the control levels 24h after i.p. injection of LPS. These results suggest that LPS-induced decreases in P-gp-mediated ileal and biliary excretion of rhodamine123 were probably due to impaired P-gp-mediated transport ability. The levels of iNOS and IL-1beta mRNA in the ileum and liver increased 2 and 8h after i.p. injection of LPS, respectively, and returned to the control levels 24h after injection of LPS. These findings suggest that LPS markedly decreases P-gp-mediated ileal and biliary excretion of rhodamine123, probably by partly decreasing the expression of P-gp protein levels, likely due to increased lipid peroxidation levels through iNOS mRNA and inflammatory mediators such as IL-1beta.

Preferential accumulation of CD103+ T cells in human livers; its association with extrathymic T cells.

BACKGROUND/AIMS: CD103, a mucosal integrin alphaEbeta7, binds to E-cadherin expressed on hepatocytes and bile duct epithelium in the liver. Although CD103+ T cells are enriched in intestinal intraepithelial lymphocytes, the localization of those cells in the liver is unknown. METHODS: We investigated whether CD103+ cells are present in human livers, and how they are associated with the intrahepatic development of T cells by flow cytometry and immunohistochemistry. RESULTS: Human livers contain significantly (P<0.001) higher percentages of CD103+ cells in CD4+ and CD8+ T cells (25.7+/-13.5 and 27.1+/-19.3%, respectively) than peripheral blood lymphocytes. Moreover, CD103+ cells in the liver, but not in peripheral blood, contained T cells with intermediate expression level of T cell receptor alphabeta. Those cells consist of mostly CD4+ and CD4-CD8- cells, and expressed low level of CD56 and interleukin-2 receptor beta chain in most of the population. These characteristics are distinct from natural killer T cells, which have been thought to be extrathymic T cells in human livers. Moreover, intrahepatic CD103+ cells expressed mRNA for recombination-activating gene-1, -2 and pre T cell receptor-alpha detected by reverse transcription-polymerase chain reaction. CONCLUSIONS: CD103+ T cells are preferentially accumulated in human livers, and those T cells show characteristics of extrathymic T cells.

Detection and analysis of intracytoplasmic cytokines in peripheral blood mononuclear cells in patients with drug-induced liver injury.

BACKGROUND/AIMS: Idiosyncratic immune response to drugs causes two types of liver injury, cholestasis or hepatitis. However, the underlying immune mechanisms of drug-induced liver injury are presently unclear. METHODS: We examined the cytokine production of peripheral blood mononuclear cells (PBMCs) from 17 patients with drug-induced liver injury and healthy controls during their incubation with and without the drug by flow cytometry. We also analyzed the cytokine production in PBMCs from eight patients after stimulation with the drug-pulsed HepG2 lysates to examine the possibility that the drug or its metabolites conjugated with a putative molecule derived from HepG2 cells might be more immunogenic. RESULTS: Among several cytokines produced by the drug or the drug-pulsed HepG2 lysates, interferon-gamma production from CD8+ cells was associated with hepatocellular injury, and tumor necrosis factor-alpha production from CD14+ cells was with cholestasis. Especially, the latter was apparent when the drug-pulsed HepG2 lysates were used as stimulants, suggesting that a complex consist of the drug, or its metabolite, and a putative molecule derived from HepG2 cells might be more immunogenic than the drug itself. CONCLUSIONS: The analysis of intracytoplasmic cytokine in PBMCs after stimulation with the drug or the drug-pulsed HepG2 lysates is useful to analyze the immune mechanism underlying drug-induced liver injury.

Phenotypic analysis of circulating and intrahepatic dendritic cell subsets in patients with chronic liver diseases.

BACKGROUND/AIMS: Dendritic cells (DCs) are the most potent professional antigen-presenting cells. Although two subsets of circulating DCs, lineage(-)CD11c(+)CD4(low) (CD11c(+)DCs) and lineage (-)CD11c(-)CD4(+)CD123(+) (CD123(+)DCs) are identified in humans, the role of each DC subset in the immunopathogenesis of liver diseases is unknown. METHODS: We examined the numbers and activation status of each DC subset in the circulation and in the inflamed livers in patients with chronic liver diseases by flow cytometry and immunohistochemistry. RESULTS: The numbers of circulating CD11c(+)DCs were inversely correlated with serum alanine aminotransferase (ALT) levels in patients with chronic viral hepatitis, and that the expression of costimulatory molecules on circulating CD11c(+)DCs in patients with chronic viral hepatitis was significantly up-regulated in patients with high serum levels of ALT. Both DCs are also identified in the livers by flow cytometry, and the expression of costimulatory molecule CD40 on those DCs was significantly higher in liver DCs than that in circulating DCs. Moreover, the ratios of CD11c(+)DCs/CD123(+)DCs were higher in liver DCs (mean+/-SD, 7.2+/-6.0) than those of circulating DCs (4.0+/-4.6). Immunohistochemically, CD11c(+) or CD123(+) cells and CD83(+) activated DCs were observed mostly in portal areas with mononuclear cell infiltration in various liver diseases. These overall data suggest that DCs, especially CD11c(+)DCs, could be associated with the necroinflammatory response in the liver of chronic viral liver diseases. CONCLUSIONS: DCs, especially CD11c(+)DCs, may be involved in the immunopathogenesis of chronic liver diseases.