Prognostic value or clinical implications of fluid status monitoring in liver cirrhosis are not fully elucidated. Tolvaptan, an orally available, selective vasopressin V2-receptor antagonist approved ...for hyponatremia in the United States and European Union. It is also used for cirrhotic ascites at a relatively low dose (3.75 mg to 7.5 mg) in Japan, exerts its diuretic function by excreting electrolyte-free water. We hypothesized that bioimpedance-defined dynamic changes in fluid status allow prediction of response of V2 antagonism and survival in cirrhotic patients.
In this prospective observational study, 30 patients with decompensated liver cirrhosis who were unresponsive to conventional diuretics were enrolled. Detailed serial changes of body composition that were assessed by using non-invasive bioimpedance analysis (BIA) devices, along with biochemical studies, were monitored at 5 time points.
Sixteen patients were classified as short-term responders (53%). Rapid and early decrease of BIA-defined intracellular water, as soon as 6 h after the first dose (ΔICW
%-6 h), significantly discriminated responders from non-responders (AUC = 0.97, P < 0.0001). ΔICW
%-6 h was highly correlated with the change of BIA-derived phase angle of trunk, e.g. reduced body reactance operated at 50 kHz after 24 h of the first dose of tolvaptan. Lower baseline blood urea nitrogen and lower serum aldosterone were predictive of a rapid and early decrease of ICW
. A rapid and early decrease of ICW
in response to tolvaptan was also predictive of a better transplant-free survival.
BIA-defined water compartment monitoring may help predict short-term efficacy and survival in decompensated cirrhotic patients treated with tolvaptan.
Disease-specific induced pluripotent stem cells (iPSCs) are widely used for disease modeling and drug discovery. We have established iPSCs from 7 types of familial Parkinson’s disease (PD). Notably, ...each form of familial PD has specific cellular functional abnormalities that induce extensive loss of dopaminergic neurons. To accelerate PD-iPSC research, we sought to establish a 96-well based high-throughput neural differentiation and phenotype detection system. iPSCs were differentiated into dopaminergic neurons on the 96-well plates by a neurosphere-based differentiation protocol (Stem Cell Reports, 2016). Subsequently, several PD-related phenotypes, including impaired mitophagy, which is considered a major phenotype in PARK2 (autosomal recessive juvenile PD, caused by mutations in PRKN gene) and PARK6 (autosomal recessive juvenile PD, caused by mutations in PINK1 gene), cell vulnerability, and α-synuclein accumulation commonly seen in PD, were quantified automatically by using In Cell Analyzer 2200 and In Cell Developer Toolbox (GE Healthcare). We successfully detected the phenotypes characterized in each form of familial PD as compared with healthy controls both quantitatively, and reproducibly. By using this high-throughput assay system, we screened a library of 320 compounds to evaluate phenotypic recovery. We identified several compounds that modify impaired mitophagy observed in PARK2 and PARK6 dopaminergic neurons. Furthermore, we established a small scale iPSC induction method, and completed iPSC induction from over 100 sporadic PD patients. By applying both the high-throughput neural differentiation method and phenotype detection system, we are trying to capture abnormal phenotypes in sporadic PD patients. Thus, this high-throughput phenotype detection system could comprehensively compare phenotypes of each form of familial PD, as well as perform drug screening. This approach is applicable for sporadic cases and other neurodegenerative diseases.
White bamboo is the swollen stem of Zizania latifolia parasitized by the smut fungus Ustilago esculenta. Five samples of galls were harvested at different stages of swelling, along with young ...seedlings of Z. latifolia and isolated colonies of U. esculenta. The inhibition capacity of boiling water or 50% ethanol extracts on NO release by RAW264 cells, which was stimulated with lipopolysaccharide, was evaluated as anti-inflammatory activity, while the NO induction capacity was evaluated as immunostimulatory activity. Total polyphenol (TPP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity were also measured. The anti-inflammatory effect, as well as TPP and DPPH, was comprehensively detected in the ethanol extracts. The immunostimulating activity was observed in the boiled extracts at different levels, depending on the swelling stage, and was especially high in the top (apical) part. These data may indicate that functional components could be dynamically induced by interactions between Z. latifolia and U. esculenta.
We study phases and structures of mixtures of two representative chromonic liquid crystal materials, sunset yellow FCF (SSY) and disodium cromoglycate (DSCG), in water. A variety of combinations of ...isotropic, nematic (N), and columnar (also called M) phases are observed depending on their concentrations, and a phase diagram is made. We find a tendency for DSCG-rich regions to show higher-order phases while SSY-rich regions show lower-order ones. We observe uniform mesophases only when one of the materials is sparse in the N phases. Their miscibility in M phases is so low that essentially complete phase separation occurs. X-ray scattering and spectroscopy studies confirm that SSY and DSCG molecules do not mix when they form chromonic aggregates and neither do their aggregates when they form M phases.
Parkinson's disease (PD) is a neurodegenerative disorder with the degeneration of midbrain dopaminergic neurons. However, few disease-modifying drugs for patients with PD have been identified. ...Familial Parkinson's diseases are numbered in the order in which the causative genes are identified, and one of them, PARK9, is caused by autophagy dysfunction and has mutation in the ATP13A2 gene which encodes a lysosomal type 5 P-type ATPase. To elucidate the pathogenesis and to screen effective drugs for PARK9, we generated iPSCs from the T cells from a PARK9 patient. Three independent clones were used for the analysis after quality verification. First, we confirmed reduced autophagy function in dopaminergic neurons derived from PARK9 iPSCs by examining the accumulation of LC3B, an autophagosome marker, due to impaired lysosomal functions. Then, we successfully quantified LC3B accumulation in the cell bodies of PARK9 dopaminergic neurons using IN Cell Analyzer 2200. Using these methods, we screened a compound library containing 320 chemical compounds using reduced LC3B accumulation in PARK9 dopaminergic neurons as an indicator. The first screening narrowed the list of candidate compounds to 70, followed by the second screening to exclude the compounds that reduce LC3B production. This two-step and 96 well-based high-throughput screening resulted in the identification of five novel disease modifying drug candidates that ameliorate impaired lysosomal dysfunction in PARK9-dopaminergic neurons.
How liver tolerance is disrupted in immune‐mediated liver injury is currently unclear. There is also insufficient information available regarding susceptibility, precipitation, escalation, and ...perpetuation of autoimmune hepatitis. To explore how dietary fiber influences hepatic damage, we applied the concanavalin A (ConA)‐induced acute immune‐mediated liver injury model in mice fed a diet supplemented with 6.8% inulin, a water‐soluble fermentable fiber. Twelve hours after ConA administration, inulin‐supplemented diet‐fed mice demonstrated significantly alleviated hepatic damage histologically and serologically, with down‐regulation of hepatic interferon‐γ and tumor necrosis factor and reduced myeloperoxidase (MPO)‐producing neutrophil infiltration. Preconditioning with an inulin‐supplemented diet for 2 weeks significantly reduced hepatic adenosine triphosphate (ATP) content; suramin, a purinergic P2 receptor antagonist, abolished the protective effect. Of note, the portal plasma derived from mice fed the inulin‐supplemented diet significantly alleviated ConA‐induced immune‐mediated liver injury. Mechanistically, increased portal short‐chain fatty acid (SCFA) levels, such as those of acetate and butyrate, by inulin supplementation leads to up‐regulation of hepatic γ‐type peroxisome proliferator‐activated receptor (Pparg) and uncoupling protein 2 (Ucp2), which uncouples mitochondrial ATP synthesis downstream of PPARγ. Pparg down‐regulating small interfering RNA cancelled the protective effect of inulin supplementation against MPO‐producing neutrophil infiltration and the subsequent immune‐mediated liver injury, suggesting that the SCFA–PPARγ–UCP2 axis plays a key role in the protective effect by inulin supplementation. Moreover, significant changes in the gut microbiota, including increased operational taxonomic units in genera Akkermansia and Allobaculum, also characterized the protective effect of the inulin‐supplemented diet. Conclusion: There is a possible unraveled etiopathophysiological link between the maintenance of liver tolerance and dietary fiber. The SCFA–PPARγ–UCP2 axis may provide therapeutic targets for immune‐mediated liver injury in the future.
In the cyanobacterium Synechococcus elongatus PCC 7942, KaiA, KaiB, and KaiC are essential proteins for the generation of a circadian rhythm. KaiC is proposed as a negative regulator of the circadian ...expression of all genes in the genome, and its phosphorylation is regulated positively by KaiA and negatively by KaiB and shows a circadian rhythm in vivo. To study the functions of KaiC phosphorylation in the circadian clock system, we identified two autophosphorylation sites, Ser-431 and Thr-432, by using mass spectrometry (MS). We generated Synechococcus mutants in which these residues were substituted for alanine by using site-directed mutagenesis. Phosphorylation of KaiC was reduced in the single mutants and was completely abolished in the double mutant, indicating that KaiC is also phosphorylated at these sites in vivo. These mutants lost circadian rhythm, indicating that phosphorylation at each of the two sites is essential for the control of the circadian oscillation. Although the nonphosphorylatable mutant KaiC was able to form a hexamer in vitro, it failed to form a clock protein complex with KaiA, KaiB, and SasA in the Synechococcus cells. When nonphosphorylatable KaiC was overexpressed, the kaiBC promoter activity was only transiently repressed. These results suggest that KaiC phosphorylation regulates its transcriptional repression activity by controlling its binding affinity for other clock proteins.
Binary and ternary blends with poly(lactic acid) (PLA), poly(butylene succinate) (PBS), and thermoplastic starch (TPS) were prepared by a melt process to produce biodegradable biomass plastics with ...both economical and good mechanical properties. The mechanical and structural properties of each blend were evaluated. Molecular dynamics (MD) simulations were also conducted to examine the mechanisms underlying the mechanical and structural properties. PLA/PBS/TPS blends showed improved mechanical properties compared with PLA/TPS blends. The PLA/PBS/TPS blends with a TPS ratio of 25–40 wt% showed higher impact strength than PLA/PBS blends. Morphology observations showed that in the PLA/PBS/TPS blends, a structure similar to that of core-shell particles with TPS as the embedding phase and PBS as the coating phase was formed, and that the trends in morphology and impact strength changes were consistent. The MD simulations suggested that PBS and TPS tightly adhered to each other in a stable structure at a specific intermolecular distance. From these results, it is clear that PLA/PBS/TPS blends are toughened by the formation of a core-shell structure in which the TPS core and the PBS shell adhered well together and stress concentration and energy absorption occurred in the vicinity of the core-shell structure.
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The rhizome of Kaempferia parviflora has been used in traditional Thai medicine. In this study, we identified and compared specific compounds from the hexane extract of K. parviflora with those from ...other Zingiberaceous plants by using gas chromatography-mass spectrometry. We identified 5,7-dimethoxyflavone (DMF), 5-hydroxy-3,7,3',4'-tetramethoxyflavone (TMF), estimated 3,5,7-trimethoxyflavone, 5-hydroxy-7,4'-dimethoxyflavone, 3,5,7,4'-tetramethoxyflavone, and investigated their anti-inflammatory effects in rat basophilic leukemia (RBL-2H3) cells stimulated with an IgE antigen or a calcium ionophore. We found that DMF and TMF more potently inhibited antigen-induced degranulation than did nobiletin, a well-known anti-inflammatory agent. In addition, compared to RBL-2H3 cells stimulated with a calcium ionophore, those treated with DMF and TMF showed more marked inhibition of the degranulation and the production and mRNA expression of inflammatory mediators. These results suggest that DMF and TMF inhibit an early step in the high-affinity IgE receptor signaling cascade rather than intracellular calcium release and protein kinase C activation.
Molecular insight into the phase-separated interface formed when biodegradable polyesters and thermoplastic starch (TPS) are melt-blended is valuable for the design of composites. In this study, ...eight different interfaces combining four major biodegradable polyesters (PLA, PBS, PHB and PBAT) and two TPSs unmodified TPS (nTPS) and citrate-modified TPS (cTPS) were investigated by using molecular dynamics (MD) simulations. According to the MD simulation results, PBS, PHB and PBAT diffuse readily into the TPS and form compatible interfaces, whereas PLA is less compatible with the TPS. The results of tensile simulations show that PBS and PBAT adhere well to TPS; in particular, PBS/cTPS and PBAT/cTPS exhibit high interfacial-fracture energy (G). Both PLA and PHB blended with TPS exhibit low G because PLA is less compatible with TPS and PHB and TPS have low electrostatic interaction. The reason for the high G of PBS/cTPS and PBAT/cTPS is thought to be a combination of three factors: (i) formation of a deep compatible interface, (ii) suppression of void growth by electrostatic interactions and (iii) absorption of strain energy by a change in the conformation of the molecular chains. These three interfacial adhesion mechanisms should be considered when designing biodegradable polyester/TPS blends with good mechanical properties.
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