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We evaluated whether modified Model for End-Stage Liver Disease (MELD) scores are useful for predicting the postdischarge prognosis in hospitalized patients with heart failure (HF) who are discharged alive. The MELD-XI and MELD-Na scores were calculated at discharge for a total of 1156 patients in the HIJ-HF II study. We also studied 3 groups on the basis of the left ventricular ejection fraction (LVEF): the HFrEF (LVEF < 40%), HFmrEF (LVEF 40-49%) and HFpEF (LVEF ≥ 50%) groups. The primary outcome was all-cause mortality, and the secondary outcome was rehospitalization due to worsening HF. The median MELD-XI and MELD-Na scores were 12 and 14, respectively. After a median follow-up of 19 months, there were significantly higher rates of all-cause mortality in patients with MELD-XI scores ≥ 12 than in those with MELD-XI scores < 12; there were also higher rates of all-cause mortality in patients with MELD-Na scores ≥ 14 than in those with MELD-Na scores < 14 (both log-rank p < 0.001). The cumulative incidence function based on a competing risks model showed a higher rate of rehospitalization due to worsening HF in patients with MELD-XI scores ≥ 12 than in those with MELD-XI scores < 12 and a higher rate of rehospitalization due to worsening HF in those with MELD-Na scores ≥ 14 than in those with MELD-Na scores < 14 (both Gray's test p < 0.001). The adjusted hazard ratios (HRs) of all-cause mortality for patients with MELD-XI scores ≥ 12 and those with MELD-Na scores ≥ 14 were 2.07 [95% confidence interval (CI) 1.25-3.44] and 2.79 [95% CI 1.63-4.79], respectively, in the HFrEF group; however, the HRs were not significant in the HFmrEF or HFpEF groups. Thus, MELD-XI and MELD-Na scores may be useful for predicting prognosis in hospitalized HF patients who are discharged alive, especially for those in the HFrEF group., (© 2022. Springer Japan KK, part of Springer Nature.)

Background: Opioids have been reported to be effective for refractory dyspnea in patients with advanced heart failure (HF) in the palliative care setting., Objective: The aim of this study was to evaluate the incidence of adverse drug reactions (ADRs) and their relationship with morphine dose/duration or renal insufficiency in patients with end-stage HF receiving continuous morphine infusion., Methods: We retrospectively studied 38 patients with end-stage HF receiving continuous intravenous or subcutaneous morphine infusion for the relief of breathlessness between 2014 and 2019 (mean age 78 years). The endpoints were nausea/vomiting, respiratory depression, and drowsiness, which are common morphine-related ADRs., Results: Of 38 patients with end-stage HF receiving continuous intravenous/subcutaneous morphine infusion, 14 (37%) experienced ADRs. The median estimated glomerular filtration rate (eGFR) was lower in patients with than in those without ADRs (16 [range 9-48] vs. 41 [range 8-133], respectively; p = 0.011). The ADRs with the highest incidence were drowsiness (n = 13), nausea/vomiting (n =5), and respiratory depression (n =3). There were no differences in the maintenance dose or duration of morphine administration between patients with and without ADRs. A baseline eGFR of 32 mL/min/1.73 m 2 was a good cutoff value for ADR prediction (sensitivity 86%, specificity 96%). A baseline eGFR < 32 mL/min/1.73 m 2 was significantly associated with the occurrence of morphine-related ADRs (odds ratio 6.63, 95% confidence interval 1.19-37.01)., Conclusions: Our results showed that 37% of patients with end-stage HF receiving continuous intravenous/subcutaneous morphine infusion experienced ADRs, especially drowsiness. Patients with eGFR < 32 mL/min/1.73 m 2 were likely to experience morphine-related ADRs., (© 2021. The Author(s).)

Administration of a compound can induce drug-metabolizing enzymes (DMEs) in the liver. DME induction can affect various parameters in toxicology studies. Therefore, evaluation of DME induction is important for interpreting test compound-induced biological responses. Several methods such as measurement of hepatic microsomal DME activity using substrates, electron microscopy, or immunohistochemistry have been used; however, these methods are limited in throughput and specificity or are not quantitative. Liquid chromatography mass spectrometry (LC/MS)-based protein analysis can detect and quantify multiple proteins simultaneously per assay. Studies have shown that formalin-fixed paraffin-embedded (FFPE) samples, which are routinely collected in toxicology studies, can be used for LC/MS-based protein analysis. To validate the utility of LC/MS using FFPE samples for quantitative evaluation of DME induction, we treated rats with a DME inducer, phenobarbital, and compared the protein expression levels of 13 phase-I and 11 phase-II DMEs between FFPE and fresh frozen hepatic samples using LC/MS. A good correlation between data from FFPE and frozen samples was obtained after analysis. In FFPE and frozen samples, the expression of 6 phase-I and 8 phase-II DMEs showed a similar significant increase and a prominent rise in Cyp2b2 and Cyp3a1 levels. In addition, LC/MS data were consistent with the measurement of microsomal DME activities. These results suggest that LC/MS-based protein expression analysis using FFPE samples is as effective as that using frozen samples for detecting DME induction., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Sigma C protein-coding sequences have been used to phylogenetically classify avian reovirus (ARV) strains. However, the relationship between serotype and phylogenetic cluster classification of the five prototype serotype strains of ARV in Japan has not been established. Thus, we used sigma C protein-coding sequences to characterize avian reoviruses (ARVs) isolated from chickens with tendonitis in Japan together with the five prototype serotype strains of ARV in Japan. Phylogenetic analysis of ARVs based on the sigma C protein-coding sequences revealed that the five prototype serotype strains of ARV were each classified into different, independent clusters. Two field isolates (JP/Tottori/2016 and JP/Nagasaki/2017) that were isolated from chickens with arthritis/tenosynovitis were classified into different clusters. JP/Tottori/2016 was classified into cluster VI with the CS-108 strain, and JP/Nagasaki/2017 was classified into cluster I with strain TS-142. Serologically, JP/Tottori/2016 was well-neutralized by antisera against the CS-108 strain, whereas JP/Nagasaki/2017 cross-reacted with antisera against both the CS-108 and TS-142 strains. Embryo lethality test revealed that the two field isolates induced 80% and 67% embryo mortality, respectively, whereas the five prototype strains induced 0%-33% embryo mortality. Our findings will contribute to understanding the characteristics of ARV strains in Japan.

Cell division cycle 7 (CDC7), a serine/threonine kinase, plays important roles in DNA replication. We developed a highly specific CDC7 inhibitor, TAK-931, as a clinical cancer therapeutic agent. This study aimed to identify the potential combination partners of TAK-931 for guiding its clinical development strategies. Unbiased high-throughput chemical screening revealed that the highest synergistic antiproliferative effects observed were the combinations of DNA-damaging agents with TAK-931. Functional phosphoproteomic analysis demonstrated that TAK-931 suppressed homologous recombination repair activity, delayed recovery from double-strand breaks, and led to accumulation of DNA damages in the combination. Whole-genome small interfering RNA library screening identified sensitivity-modulating molecules, which propose the experimentally predicted target cancer types for the combination, including pancreatic, esophageal, ovarian, and breast cancers. The efficacy of combination therapy in these cancer types was preclinically confirmed in the corresponding primary-derived xenograft models. Thus, our findings would be helpful to guide the future clinical strategies for TAK-931., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

Seventeen isolates of infectious bronchitis virus (IBV) were obtained from various prefectures of Japan during 2008-2019 and genetically analyzed. The IBV isolates were classified into six genetic groups, based on phylogenetic analysis of the S1 gene. The S1 genotypes were distinguishable by a newly developed restriction fragment length polymorphism (RFLP) method using three endonucleases, Hae II, Hpa I, and Fok I. Moreover, the isolates were classified into four genetic groups, based on phylogenetic analysis of the S2 gene. However, novel genetic groups based on a combination of S1 and S2 genotypes, which were undetected previously, were confirmed in this study, indicating that various recombinant IBV strains were prevalent in poultry in Japan.

Background: We investigated the efficacy and safety of apararenone (MT-3995), a non-steroidal compound with mineralocorticoid receptor agonist activity, in patients with stage 2 diabetic nephropathy (DN)., Methods: The study had two parts: a dose-response, parallel-group, randomized, double-blind, placebo-controlled, multicenter, phase 2, 24-week study and an open-label, uncontrolled, 28-week extension study. Primary and secondary endpoints were the 24-week percent change from baseline in urine albumin to creatine ratio (UACR) and 24- and 52-week UACR remission rates. Safety parameters were changes from baseline in estimated glomerular filtration rate (eGFR) and serum potassium at 24 and 52 weeks, and incidences of adverse events (AEs) and adverse drug reactions (ADRs)., Results: In the dose-response period, 73 patients received placebo and 73, 74, and 73 received apararenone 2.5 mg, 5 mg, and 10 mg, respectively. As a percentage of baseline, mean UACR decreased to 62.9%, 50.8%, and 46.5% in the 2.5 mg, 5 mg, and 10 mg apararenone groups, respectively, at week 24 (placebo: 113.7% at week 24; all P < 0.001 vs placebo). UACR remission rates at week 24 were 0.0%, 7.8%, 29.0%, and 28.1% in the placebo and apararenone 2.5 mg, 5 mg, and 10 mg groups, respectively. eGFR tended to decrease and serum potassium tended to increase, but these events were not clinically significant. AE incidence increased with dose while ADR incidence did not., Conclusion: The UACR-lowering effect of apararenone administered once daily for 24 weeks in patients with stage 2 DN was confirmed, and the 52-week administration was safe and tolerable., Clinical Trial Registration: NCT02517320 (dose-response study) and NCT02676401 (extension study).

Cerebrospinal fluid (CSF) metabolites reflect biochemical diffusion/export from the brain and possibly serve as biomarkers related to brain disease severity, pathophysiology, and therapeutic efficacy/toxicity. Metabolomic studies using blood matrices have demonstrated interindividual and preanalytical variation of blood metabolites, whereas those of CSF metabolites remain unclear. In this study, we aimed to delineate the postprandial effects on CSF metabolites because fasting of patients with brain-related disorders is challenging. We collected pre- and postprandial (1.5, 3, and 6 h) plasma and CSF from nine healthy subjects. Using a mass-spectrometry-based global metabolomics approach, 150 and 130 hydrophilic metabolites and 263 and 340 lipids were detected in CSF and plasma, respectively. Principal component analysis of CSF hydrophilic metabolites and lipids primarily classified individual subjects at any time point, suggesting that the postprandial effects had a lower impact than interindividual variations on CSF metabolites. Individually, less than 10% of the CSF metabolites were putatively altered by postprandial effects (with either significant differences or over 2-fold changes, but not both) at any time point. Thus, global CSF metabolite levels are not directly associated with food intake, and except for several putatively altered CSF metabolites, postprandial effects are not a major concern when applying CSF metabolomics to screen biomarkers.

Background The association between first-degree atrioventricular block (AVB) and life-threatening cardiac events in patients with hypertrophic cardiomyopathy (HCM) remains unclear. This study sought to investigate whether presence of first-degree AVB was associated with HCM-related death in patients with HCM. Methods and Results We included 414 patients with HCM (mean age, 51±16 years; 64.5% men). The P-R interval was measured at the time of the initial evaluation and patients were classified into those with and without first-degree AVB, which was defined as a P-R interval ≥200 ms. HCM-related death was defined as a combined end point of sudden death or potentially lethal arrhythmic events, heart failure-related death, and stroke-related death. First-degree AVB was noted in 96 patients (23.2%) at time of enrollment. Over a median (interquartile range) follow-up period of 8.8 (4.9-12.9) years, a total of 56 patients (13.5%) experienced HCM-related deaths, including 47 (11.4%) with a combined end point of sudden death or potentially lethal arrhythmic events. In a multivariable analysis that included first-degree AVB and risk factors for life-threatening events, first-degree AVB was independently associated with an HCM-related death (adjusted hazard ratio, 2.41; 95% CI, 1.27-4.58; P =0.007), and this trend also persisted for the combined end point of sudden death or potentially lethal arrhythmic events (adjusted hazard ratio, 2.60; 95% CI, 1.28-5.27; P =0.008). Conclusions In this cohort of patients with HCM, first-degree AVB may be associated with HCM-related death, including the combined end point of sudden death or potentially lethal arrhythmic events.

The Ryukyu ayu Plecoglossus altivelis ryukyuensis is an endangered amphidromous fish that inhabits rivers in the Ryukyu Archipelago (Japan). Populations of the species have declined dramatically. Consequently, the Ryukyu ayu has been registered as a natural monument in Japan and monitoring surveys with direct catching are restricted legally. This restriction, unfortunately, makes monitoring of population abundances difficult and creates a barrier to both advancing understanding of the species' status and the development of appropriate conservation plans. We developed a non-invasive monitoring methodology using eDNA analyses. We designed a specific quantitative PCR assay for the Ryukyu ayu using the mitochondrial ND4 region. Using this primer/probe set, we conducted eDNA analyses in three rivers on Amami-Ohshima Island. The DNA fragments were amplified from the eDNA extracted from natural water in each river. The numbers of DNA fragments detected were positively correlated with individual counts of fish obtained by visual snorkelling surveys. Our method does not contravene restrictions and facilitates abundance monitoring of this endangered fish species., (Yoshihisa Akamatsu, Gen Kume, Masuji Gotou, Takanori Kono, Takuma Fujii, Ryutei Inui, Yoshihisa Kurita.)

Replication stress (RS) is a cancer hallmark; chemotherapeutic drugs targeting RS are widely used as treatments for various cancers. To develop next-generation RS-inducing anticancer drugs, cell division cycle 7 (CDC7) has recently attracted attention as a target. We have developed an oral CDC7-selective inhibitor, TAK-931, as a candidate clinical anticancer drug. TAK-931 induced S phase delay and RS. TAK-931-induced RS caused mitotic aberrations through centrosome dysregulation and chromosome missegregation, resulting in irreversible antiproliferative effects in cancer cells. TAK-931 exhibited significant antiproliferative activity in preclinical animal models. Furthermore, in indication-seeking studies using large-scale cell panel data, TAK-931 exhibited higher antiproliferative activities in RAS -mutant versus RAS -wild-type cells; this finding was confirmed in pancreatic patient-derived xenografts. Comparison analysis of cell panel data also demonstrated a unique efficacy spectrum for TAK-931 compared with currently used chemotherapeutic drugs. Our findings help to elucidate the molecular mechanisms for TAK-931 and identify potential target indications.

Chromosome band 8q24 is the most frequently amplified locus in various types of cancers. MYC has been identified as the primary oncogene at the 8q24 locus, whereas a long noncoding gene, PVT1 , which lies adjacent to MYC , has recently emerged as another potential oncogenic regulator at this position. In this study, we established and characterized a novel cell line, AMU-ML2, from a patient with diffuse large B-cell lymphoma (DLBCL), displaying homogeneously staining regions at the 8q24 locus. Fluorescence in situ hybridization clearly detected an elevation in MYC copy numbers corresponding to the homogenously staining region. In addition, a comparative genomic hybridization analysis using high-resolution arrays revealed that the 8q24 amplicon size was 1.4 Mb, containing the entire MYC and PVT1 regions. We also demonstrated a loss of heterozygosity for TP53 at 17p13 in conjunction with a TP53 frameshift mutation. Notably, AMU-ML2 cells exhibited resistance to vincristine, and cell proliferation was markedly inhibited by MYC -shRNA-mediated knockdown. Furthermore, genes involved in cyclin D, mTOR, and Ras signaling were downregulated following MYC knockdown, suggesting that MYC expression was closely associated with tumor cell growth. In conclusion, AMU-ML2 cells are uniquely characterized by homogenously staining regions at the 8q24 locus, thus providing useful insights into the pathogenesis of DLBCL with 8q24 abnormalities.

Recent studies in RAS wild-type (WT) metastatic colorectal cancer (mCRC) suggest that the survival benefits of therapy using anti-epidermal growth factor receptor (anti-EGFR) and anti-vascular endothelial growth factor (anti-VEGF) antibodies combined with chemotherapy are maximized when the anti-EGFR antibody is given as first-line, followed by subsequent anti-VEGF antibody therapy. We report reverse-translational research using LIM1215 xenografts of RAS WT mCRC to elucidate the biologic mechanisms underlying this clinical observation. Sequential administration of panitumumab then bevacizumab (PB) demonstrated a stronger tendency to inhibit tumor growth than bevacizumab then panitumumab (BP). Cell proliferation was reduced significantly with PB (P < .01) but not with BP based on Ki-67 index. Phosphoproteomic analysis demonstrated reduced phosphorylation of EGFR and EPHA2 with PB and BP compared with control. Western blotting showed reduced EPHA2 expression and S897-phosphorylation with PB; RSK phosphorylation was largely unaffected by PB but increased significantly with BP. In quantitative real-time PCR analyses, PB significantly reduced the expression of both lipogenic (FASN, MVD) and hypoxia-related (CA9, TGFBI) genes versus control. These results suggest that numerous mechanisms at the levels of gene expression, protein expression, and protein phosphorylation may explain the improved clinical activity of PB over BP in patients with RAS WT mCRC., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

B-cell lymphoma 6 (BCL6) is the most frequently involved oncogene in diffuse large B-cell lymphomas (DLBCLs). BCL6 shows potent transcriptional repressor activity through interactions with its corepressors, such as BCL6 corepressor (BCOR). The inhibition of the protein-protein interaction (PPI) between BCL6 and its corepressors suppresses the growth of BCL6-dependent DLBCLs, thus making BCL6 an attractive drug target for lymphoma treatment. However, potent small-molecule PPI inhibitor identification remains challenging because of the lack of deep cavities at PPI interfaces. This article reports the discovery of a potent, cell-active small-molecule BCL6 inhibitor, BCL6-i (8), that operates through irreversible inhibition. First, we synthesized irreversible lead compound 4, which targets Cys53 in a cavity on the BCL6-BTB domain dimer by introducing an irreversible warhead to high-throughput screening hit compound 1. Further chemical optimization of 4 based on k inact /K I evaluation produced BCL6-i with a k inact /K I value of 1.9 × 10 4 M -1 s -1 , corresponding to a 670-fold improvement in potency compared to that of 4. By exploiting the property of irreversible inhibition, engagement of BCL6-i to intracellular BCL6 was confirmed. BCL6-i showed intracellular PPI inhibitory activity between BCL6 and its corepressors, thus resulting in BCL6-dependent DLBCL cell growth inhibition. BCL6-i is a cell-active chemical probe with the most potent BCL6 inhibitory activity reported to date. The discovery process of BCL6-i illustrates the utility of irreversible inhibition for identifying potent chemical probes for intractable target proteins.

Background: Large-scale clinical trials have analyzed risk factors for any ischemic stroke in patients with atrial fibrillation (AF). However, the risk factors for cardioembolic stroke (CES), specifically, have not been reported. To clarify the risk factors for CES and clinically significant cardioembolic infarction, we examined the incidence of CES and larger infarct volume (IV) (> 30 mL) CES, employing the Fushimi AF Registry, a community-based prospective cohort of AF patients in the Fushimi ward, Kyoto, Japan., Methods: A total of 4,182 Fushimi AF patients were enrolled from March 2011 to December 2014. The risk factors for CES were evaluated using multivariate analysis., Results: Of 4,182 patients enrolled, 3,749 patients were observed for ≥1 year. During the follow-up period (mean duration, 979 ± 7.7 days), 91/3,749 patients experienced a CES (2.43%). Significant risk factors associated with CES were older age (odds ratio [OR], 1.31; 95% confidence interval [CI], 1.01-1.72; p = 0.046), low body weight (OR, 1.30; 95% CI, 1.03-1.65; p = 0.033), sustained AF (OR, 1.67; 95% CI, 1.05-2.71; p = 0.034), and previous stroke or transient ischemic attack (TIA) (OR, 1.94; 95% CI, 1.22-3.06; p = 0.004). Predictors of a large IV were chronic kidney disease (CKD) (OR, 2.08; 95% CI, 1.09-4.05; p = 0.027) and previous stroke/TIA (OR, 2.27; 95% CI, 1.19-4.24; p = 0.011)., Conclusions: In this population-based cohort of Japanese patients with AF, in addition to previous stroke/TIA and older age, sustained AF and low body weight emerged as risk factors for CES, as opposed to any stroke, which may have a different risk profile. Patients with CKD or previous stroke/TIA who developed cardioembolic infarction exhibited more advanced severity. There is a need for direct oral anticoagulants that can be used safely in patients with comorbid AF and CKD., (The Author(s). Published by S. Karger AG, Basel.)

Aim: In the present study, we examined the relationship between cytokine levels in the cerebrospinal fluid (CSF) and 11 C-Pittsburgh compound B (PiB) retention in patients with mild cognitive impairment., Methods: A total of 33 participants (12 men and 21 women; mean age 76.5 years) with mild cognitive impairment underwent neuropsychological assessments, PiB positron emission tomography and analysis of cytokine levels in the CSF. The CSF levels of 48 cytokines and growth factors were measured using multiplex immunoassays. PiB retention was assessed based on a standardized uptake value ratio. Mild cognitive impairment participants were classified as PiB-positive and PiB-negative, with a cut-off level of 1.4. We compared the CSF cytokine levels and Alzheimer's disease biomarkers, including β-amyloid 1-42, total tau and tau phosphorylated at threonine 181, between the two subgroups, and evaluated the correlation between PiB retention or CSF Alzheimer's disease biomarkers and CSF cytokine levels., Results: Cytokine levels in the CSF did not differ between the two subgroups. Macrophage inflammatory protein-1β levels in the CSF significantly correlated with PiB retention only in the PiB-positive subgroup, whereas stem cell growth factor-β levels significantly correlated with PiB retention in the PiB-negative subgroup. Furthermore, stem cell growth factor-β levels significantly correlated with total tau and tau phosphorylated at threonine 181 levels in only the PiB-negative subgroup., Conclusion: The present findings suggest that macrophage inflammatory protein-1β and stem cell growth factor-β are associated with chronic inflammatory processes accompanied by amyloid deposition or AD pathophysiology. Geriatr Gerontol Int 2017; 17: 1907-1913., (© 2017 Japan Geriatrics Society.)

Protein function is mainly modulated by dynamic reversible or irreversible post-translational modifications. Among them, the identification of protein phosphorylation sites and changes in phosphorylation levels in vivo are of considerable interest for a better understanding of the protein function. Thus, effective strategies for the quantitative determination of phosphorylation degrees for low abundant proteins, for which antibodies are not available, are required in order to evaluate the functional regulation of proteins attributed to phosphorylation. In this study, we used the heart β1-adrenergic receptor (Adrb1) as a model protein and developed FLAG-Adrb1 knock-in mice, in which the FLAG tag was inserted at the N-terminus of Adrb1. The phosphorylation sites and levels of Adrb1 in the heart were elucidated by immuno-affinity purification followed by quantitative mass spectrometry analysis using ion intensity ratio of the phosphorylated peptide versus corresponding unphosphorylated peptide. The phosphorylation levels at Ser274 and Ser462 of Adrb1 were approximately 0.25 and 0.0023. This effective strategy should be useful for not only analyzing site-specific phosphorylation levels of target proteins, but also quantifying the expression levels of proteins of interest when appropriate antibodies are not available., Competing Interests: There are no conflicts of interest to declare.

In a high-throughput screening (HTS) process, the chemical reactivity of test samples should be carefully examined because such reactive compounds may lead to false-positive results and adverse effects in vivo. Among all natural amino acids, the thiol side chain in cysteine has the highest nucleophilicity; thus, assessment of intrinsic thiol group reactivity in the HTS processes is expected to accelerate drug discovery. In general, k (M chem (M -1 s -1 ), the secondary reaction rate constant of a compound to thiol, can be evaluated via time course measurements of thiol-compound adducts using liquid chromatography-mass spectroscopy; this requires time-consuming and labor-intensive procedures. To overcome this issue, we developed a fluorescence-based competitive endpoint assay that allows quantitative calculation of the reaction rate of test compounds in an HTS format. Our assay is based on the competitive reaction for a free thiol (e.g., glutathione) between the test compounds and a fluorescent probe, o -maleimide BODIPY. Our assay provides robust data with a satisfactory throughput at an affordable cost. Our k chem evaluation method has advantages over previous assays in terms of higher throughput and quantitativeness. Thus, it contributes to early elimination of reactive compounds as well as quantitative evaluation of the k chem values of covalent inhibitors.

Panitumumab is a monoclonal antibody developed against the human epidermal growth factor receptor (EGFR). TAS-102 is a novel chemotherapeutic agent containing trifluridine (FTD) as the active cytotoxic component. Both panitumumab and TAS-102 have been approved for the treatment of metastatic colorectal cancer. In this study, we revealed the mechanism underlying the anticancer effects of panitumumab/TAS-102 combination using preclinical models. Panitumumab/FTD cotreatment showed additive antiproliferative effects in LIM1215 and synergistic antiproliferative effects in SW48 colon cancer cells. Consistent with the in vitro effects, panitumumab/TAS-102 combination caused tumor regression in LIM1215 and COL-01-JCK colon cancer patient-derived xenograft models. In LIM1215 cells, FTD induced extracellular signal-regulated kinase (ERK)/protein kinase B (AKT)/signal transducer and activator of transcription 3 (STAT3) phosphorylation and subsequent serine/threonine phosphorylation of EGFR, while it had no effects on EGFR tyrosine phosphorylation. Panitumumab and the tyrosine kinase inhibitor erlotinib reduced the basal level of EGFR tyrosine phosphorylation and reversed FTD-induced ERK/AKT/STAT3 and EGFR serine/threonine phosphorylation. These results suggested that FTD in combination with the basal activity of EGFR tyrosine kinase induced downstream prosurvival signaling through ERK/AKT/STAT3 phosphorylation. Collectively, we propose that panitumumab interacts with FTD by targeting EGFR-mediated adaptive responses, thereby exerting anticancer effects when used in combination with TAS-102. These preclinical findings provide a compelling rationale for evaluating the combination of anti-EGFR antibodies with TAS-102 against metastatic colorectal cancer., (© 2017 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.)

β1-adrenergic receptor (Adrb1) belongs to the superfamily of G-protein-coupled receptors (GPCRs) and plays a critical role in the regulation of heart rate and myocardial contraction force. GPCRs are phosphorylated at multiple sites to regulate distinct signal transduction pathways in different tissues. However, little is known about the location and function of distinct phosphorylation sites of Adrb1 in vivo. To clarify the mechanisms underlying functional regulation associated with Adrb1 phosphorylation in vivo, we aimed to identify Adrb1 phosphorylation sites in the mouse heart using phosphoproteomics techniques with nano-flow liquid chromatography/tandem mass spectrometry (LC-MS/MS). We revealed the phosphorylation residues of Adrb1 to be Ser274 and Ser280 in the third intracellular loop and Ser412, Ser417, Ser450, Ser451, and Ser462 at the C-terminus. We also found that phosphorylation at Ser274, Ser280, and Ser462 was enhanced in response to stimulation with an Adrb1 agonist. This is the first study to identify Adrb1 phosphorylation sites in vivo. These findings will provide novel insights into the regulatory mechanisms mediated by Adrb1 phosphorylation., (Copyright © 2017 Elsevier Inc. All rights reserved.)

[Purpose] This study aimed to examine the health-related and subjective quality of life of community-dwelling elderly females in orthopedic outpatients, and also examined how such quality of life correlate with their daily life activities and instrumental activities of daily living. [Subjects and Methods] Subjects were 27 community-dwelling elderly females in orthopedic outpatients (mean age: 76.3 ± 7.4 years). Their health-related quality of life and subjective quality of life, life-space assessment, frenchay activities index were researched. [Results] For the relationships between the total subjective quality of life scores and health-related quality of life scores, significant positive correlations were observed for body pain, general health, vitality, social functions and mental health. The correlations were not statistically significant between the subjective quality of life scores and the life-space assessment and frenchay activities index scores. The correlations were statistically significant between some health-related quality of life scores and the life-space assessment and frenchay activities index scores. [Conclusion] The results suggest that supporting community-dwelling elderly females in orthopedic outpatients to improve their sense of physical and mental well-being, and prevent and reduce their depression and physical pain, is required in order to improve their QOL.

[Purpose] This study examined the quality of life of homebound elderly hemiparetic stroke patients with cognitive impairment and the factors affecting their quality of life. [Subjects and Methods] The subjects of the study were 17 home-based elderly hemiparetic stroke patients with cognitive impairment (8 males and 9 females, average age: 76.3 ± 10.5 years old). Their physical and psychological conditions, quality of life and other items were investigated. Nishimura's Mental State Scale for the Elderly was used for the cognitive impairment assessment. The Functional Independence Measure was used to assess activities of daily living, and the Japanese Quality of Life Inventory for the Elderly with Dementia was used to assess quality of life. [Results] The subjects' quality of life was affected by their cognitive impairment level and independence of activities of daily living. However, no correlations were observed between the quality of life of the homebound elderly hemiparetic stroke patients with cognitive impairment, age, gender or care-need level. [Conclusion] In order to improve the quality of life of homebound elderly hemiparetic stroke patients with cognitive impairment, assistance helping them to maintain their cognitive abilities and on-going rehabilitation for improving activities of daily living independence are required.

Amylase-producing myeloma exhibits refractoriness to chemotherapy and a dismal prognosis. In this study, we established a human myeloma cell line, 8226/AMY1, in which a lentivirally transfected AMY1 gene was stably expressed and explored its biological characteristics. 8226/AMY1 showed a survival advantage over mock control when treated with dexamethasone, bortezomib, and lenalidomide in vitro partly through inhibition of apoptosis induced by these reagents. In a xenograft murine model, 8226/AMY1 showed rapid tumor growth and reduced sensitivity to bortezomib compared with mock. A microarray gene expression analysis identified TCL1A, which functions as a coactivator of the cell survival kinase Akt, differentially up-regulated in 8226/AMY1. The expression of phosphorylated Akt was increased in the 8226/AMY1 cells following bortezomib treatment, but not in the mock cells. In addition, treatment with perifosine, an inhibitor of Akt phosphorylation, enhanced the anti-myeloma effect of bortezomib in the 8226/AMY1 cells. Our data suggest that amylase-producing myeloma reduced the sensitivity to bortezomib in vitro and in vivo, and the up-regulation of TCL1A may influence the drug susceptibility of 8226/AMY1 via the phosphorylation of Akt. These findings provide clues for developing treatment approaches for not only amylase-producing myeloma, but also relapsed and refractory myelomas.

[Purpose] This study aimed to examine the quality of life (QOL) of community-dwelling elderly women with musculoskeletal disorders and factors that affect it. [Subjects] The subjects were 27 community-dwelling elderly women with musculoskeletal disorders (mean age: 76.3 ± 7.4 years). Their physical and psychological conditions, QOL, and other characteristics were researched. [Methods] The Japanese version of Life-Space Assessment was used to assess the subjects' daily life activities; the Japanese version of Fall Efficacy Scale (FES), to assess their fear of falling; the Geriatric Depression Scale (GDS 15), to assess their depression status; and the Life Satisfaction Index K (LSIK), to assess their QOL. [Results] The results indicated that the number of family members living together, degree of pain, fear of falling, and depression affect the LSIK scores of the community-dwelling elderly women with musculoskeletal disorders. [Conclusion] The study results suggest that the LSIK scores of community-dwelling elderly women with musculoskeletal disorders can be improved by easing their pain, improving their physical abilities to prevent falls, and improving their mobility. The results also suggest that continuing rehabilitation treatment is required.

To develop centromere-associated protein-E (CENP-E) inhibitors for use as anticancer therapeutics, we designed novel imidazo[1,2-a]pyridines, utilizing previously discovered 5-bromo derivative 1a. By site-directed mutagenesis analysis, we confirmed the ligand binding site. A docking model revealed the structurally important molecular features for effective interaction with CENP-E and could explain the superiority of the inhibitor (S)-isomer in CENP-E inhibition vs the (R)-isomer based on the ligand conformation in the L5 loop region. Additionally, electrostatic potential map (EPM) analysis was employed as a ligand-based approach to optimize functional groups on the imidazo[1,2-a]pyridine scaffold. These efforts led to the identification of the 5-methoxy imidazo[1,2-a]pyridine derivative (+)-(S)-12, which showed potent CENP-E inhibition (IC50: 3.6 nM), cellular phosphorylated histone H3 (p-HH3) elevation (EC50: 180 nM), and growth inhibition (GI50: 130 nM) in HeLa cells. Furthermore, (+)-(S)-12 demonstrated antitumor activity (T/C: 40%, at 75 mg/kg) in a human colorectal cancer Colo205 xenograft model in mice.

[Purpose] This study explored the roles of physical therapists (PTs) in introducing assistive products, which are essential for living securely and stably at home, and examined how PTs can fulfill these roles more efficiently and effectively. [Subjects and Methods] A questionnaire on introducing assistive products was administered to PTs working at randomly selected hospitals, health-care facilities for the elderly requiring long-term care, home-visit nursing stations, and other such facilities and to PTs providing physical therapy directly to the home-bound elderly disabled. The subjects of the study were 77 PTs who returned valid responses. [Results] For awareness of systems for assistive product's introduction, PTs were more aware of the system based on the Long-Term Care (LTC) Insurance Act than the system based on the Act on Welfare for the Home-Bound Elderly Disabled. For PTs handling assistive product's introduction for the home-bound elderly disabled, approximately 91% of the respondents answered that they had handled some cases of assistive product's introduction, and PTs with longer clinical experience had handled more assistive product's introduction cases. [Conclusion] The results demonstrated that PTs understand the work involved in introducing assistive products work well and that they handle it. The results, however, also suggested that educational and operational improvements are urgently required for PTs introducing assistive products essential for the lives of the home-bound elderly disabled.

[Purpose] This study examined the quality of life (QOL) of homebound elderly hemiparetic stroke patients and factors that affect it. [Subjects] The subjects of the study were 21 homebound elderly hemiparetic stroke patients who were 65 years old or over and required care for daily living (12 males and 9 females, average age: 79.3 ± 8.4 years old). Their physical and psychological conditions, QOL, and other characteristics were researched. [Methods] The Functional Independence Measure (FIM) was used for the activities of daily living (ADL) assessment, and the MOS 36-Item Short-Form Health Survey (SF-36, Japanese version 1.2) was used for the QOL assessment. [Results] No correlations were observed between the QOL of homebound elderly hemiparetic stroke patients and their age and gender. However, the results showed that their QOL was affected by their independence in ADL, bedridden degree, and care-need level. [Conclusion] These results suggest that in order to improve the QOL of homebound elderly hemiparetic stroke patients, ongoing rehabilitation to improve independence in ADL and lower the bedridden degree and care-need level is required.

We examined the effects of diet nutrients on xenotransplanted leukemia cells, THP-1 or NB4. THP-1 tumors showed more growth when fed with high fat diet, while NB4 tumors grew more with high carbohydrate diet. Then, administration of 2-deoxyglucose (a glycolysis inhibitor) showed a significant antitumor effect on both tumors: NB4 tumor showed large necrotic areas, while THP-1 tumor did not, but had augmented expression of enzymes for fatty acid oxidation. 2-Deoxyglucose inhibited the growth of NB4 by cell death because main energy producing pathway (glycolysis) was abolished, while 2-deoxyglucose slowed the growth of THP-1 by shifting energy metabolism to fatty acid β-oxidation., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

[Purpose] We compared problematic behaviors of children according to the severity of their mental retardation (MR) of intellect as categorized by the Ryouiku Techou in this study, to investigate the influence of MR of intellect on children's problematic behaviors. [Subjects] The subjects were 86 mentally retarded children undergoing physical therapy at hospitals and other facilities. [Methods] The examiners were 13 physical therapists and 8 occupational therapists who worked at the hospital and knew the children well. The examiners individually assessed the subjects using the Japanese version of the Aberrant Behavior Checklist. The subjects were divided into two groups (A and non-A) according to the Ryouiku Techou standard. [Results] No significant differences were observed between the groups except in the items of stereotypy and lethargy. [Conclusion] Problematic behaviors other than stereotypy and lethargy were not influenced by the Ryouiku Techou standard.

[Purpose] This study explored the roles of physical therapists (PTs) in living environment maintenance, which is essential for living securely and stably at home, and examines how physical therapists can fulfill these roles more efficiently and effectively. [Subjects and Methods] A questionnaire on living environment maintenance was given to PTs working at randomly selected hospitals, health care facilities for the elderly requiring long-term care, home-visit nursing stations, and other such facilities and directly providing physical therapy to the home-bound elderly disabled. The subjects of the study were 77 PTs who returned valid responses. [Results] For awareness of systems for living environment maintenance, PTs were more aware of the system based on the Long-Term Care (LTC) Insurance Act than the system based on the Act on Welfare for the Home-Bound Elderly Disabled. PTs who have worked at two or more types of medical, welfare, and intermediate institutions were more aware of such systems than PTs who have worked at only one type. For PTs handling living environment maintenance for the home-bound elderly disabled, approximately 80% of respondents answered that they have handled some living environment maintenance, and PTs with longer clinical experience have handled more living environment maintenance cases. [Conclusion] The results demonstrated that PTs understand their living environment maintenance work well and handle the work. The results, however, also suggested that educational and operational improvements are urgently required for PTs handling living environment maintenance essential for the lives of the home-bound elderly disabled.

The shift in energy metabolism from oxidative phosphorylation to glycolysis can serve as a target for the inhibition of cancer growth. Here, we examined the metabolic changes induced by 2-deoxyglucose (2-DG), a glycolysis inhibitor, in leukemia cells by metabolome analysis. NB4 cells mainly utilized glucose as an energy source by glycolysis and oxidative phosphorylation in mitochondria, since metabolites in the glycolytic pathway and in the tricarboxylic acid (TCA) cycle were significantly decreased by 2-DG. In THP-1 cells, metabolites in the TCA cycle were not decreased to the same extent by 2-DG as in NB4 cells, which indicates that THP-1 utilizes energy sources other than glucose. TCA cycle metabolites in THP-1 cells may be derived from acetyl-CoA by fatty acid β-oxidation, which was supported by abundant detection of carnitine and acetylcarnitine in THP-1 cells. 2-DG treatment increased the levels of pentose phosphate pathway (PPP) metabolites and augmented the generation of NADPH by glucose-6-phosphate dehydrogenase. An increase in NADPH and upregulation of glutathione synthetase expression resulted in the increase in the reduced form of glutathione by 2-DG in NB4 cells. We demonstrated that a combination of 2-DG and inhibition of PPP by dehydroepiandrosterone (DHEA) effectively suppressed the growth of NB4 cells. The replenishment of the TCA cycle by fatty acid oxidation by carnitine palmitoyltransferase in THP-1 cells, treated by 2-DG, might be regulated by AMPK, as the combination of 2-DG and inhibition of AMPK by compound C potently suppressed the growth of THP-1 cells. Although 2-DG has been effective in preclinical and clinical studies, this treatment has not been fully explored due to concerns related to potential toxicities such as brain toxicity at high doses. We demonstrated that a combination of 2-DG and DHEA or compound C at a relatively low concentration effectively inhibits the growth of NB4 and THP-1 cells, respectively. These observations may aid in the identification of appropriate combinations of metabolic inhibitors at low concentrations which do not cause toxicities.

We report a very rare coexistence of posterior mediastinal hemangioma with extramedullary haematopoieisis. A 69-year-old woman was detected to have an abnormal mass on the chest radiograph during a routine medical examination. No disorders were found by hematologic exams before and after surgery. Chest computed tomography (CT) revealed a well circumscribed posterior mediastinal tumor measured 32×11 mm in diameter, located on paravertebral space. The most likely diagnosis was a benign neurogenic tumor and we were planning to remove this tumor by video-assisted thoracic surgery (VATS) via a right transthoracic approach. Thoracoscopic finding revealed that this tumor was brownish and soft, such as hemangioma. Initially, incisional biopsy was done to obtain definitive diagnosis. Since intraoperative microscopic examination of the biopsy specimen revealed extramedullary haematopoieisis, we obtained another larger samples and finished the operation. Interestingly, the postoperative final histological diagnosis was mediastinal hemangioma with extramedullary haematopoieis.

In this study, we established and analyzed a novel human myeloid leukemia cell line, AMU-AML1, from a patient with acute myeloid leukemia with multilineage dysplasia before the initiation of chemotherapy. AMU-AML1 cells were positive for CD13, CD33, CD117, and HLA-DR by flow cytometry analysis and showed a single chromosomal abnormality, 46, XY, t(12;22)(p13;q11.2), by G-banding and spectral karyotyping. Fluorescent in situ hybridization analysis indicated that the chromosomal breakpoint in band 12p13 was in the sequence from the 5' untranslated region to intron 1 of TEL and that the chromosomal breakpoint in band 22q11 was in the 3' untranslated region of MN1. The chimeric transcript and protein of MN1-TEL could not be detected by reverse-transcriptase polymerase chain reaction or Western blot analysis. However, the MN1 gene was amplified to three copies detected by array comparative genomic hybridization analysis, and the expression levels of the MN1 transcript and protein were high in AMU-AML1 cells when compared with other cell lines with t(12;22)(p13;q11-12). Our data showed that AMU-AML1 cells contain t(12;22)(p13;q11.2) without chimeric fusion of MN1 and TEL. The AMU-AML1 cells gained MN1 copies and had high expression levels of MN1. Thus, the AMU-AML1 cell line is useful for studying the biological consequences of t(12;22)(p13;q11.2) lacking chimeric MN1-TEL., (Copyright © 2011 Wiley Periodicals, Inc.)

A 67-year-old woman was admitted with progressive exertional dyspnea. Arterial blood gas analysis showed severe hypoxia. A chest radiograph and a computed tomography showed bilateral interstitial infiltrations and patchy ground-grass opacities. Initially, she was given antibiotics with no effect. Since transthoracic echocardiography revealed right ventricular overload, cardiac catheterization was performed, which showed pulmonary hypertension. Pulmonary angiography revealed irregular filling defects along with pulmonary peripheral sites. Open lung biopsy was performed to establish a definitive diagnosis and to treat the diffuse lung disease. Microscopically, multiple tumor thrombi of mucinous adenocarcinoma were found in small sized pulmonary artery. Primary sites of malignancy has not been found in spite of postoperative examinations.

For generation of energy, cancer cells utilize glycolysis more vigorously than oxidative phosphorylation in mitochondria (Warburg effect). We examined the energy metabolism of four leukemia cell lines by using glycolysis inhibitor, 2-deoxy-d-glucose (2-DG) and inhibitor of oxidative phosphorylation, oligomycin. NB4 was relatively sensitive to 2-DG (IC(50): 5.75 mM), consumed more glucose and produced more lactate (waste product of glycolysis) than the three other cell lines. Consequently, NB4 was considered as a "glycolytic" leukemia cell line. Dependency on glycolysis in NB4 was confirmed by the fact that glucose (+) FCS (-) medium showed more growth and survival than glucose (-) FCS (+) medium. Alternatively, THP-1, most resistant to 2-DG (IC(50): 16.14 mM), was most sensitive to oligomycin. Thus, THP-1 was recognized to be dependent on oxidative phosphorylation. In THP-1, glucose (-) FCS (+) medium showed more growth and survival than glucose (+) FCS (-) medium. The dependency of THP-1 on FCS was explained, at least partly, by fatty acid oxidation because inhibitor of fatty acid β-oxidation, etomoxir, augmented the growth suppression of THP-1 by 2-DG. We also examined the mechanisms by which THP-1 was resistant to, and NB4 was sensitive to 2-DG treatment. In THP-1, AMP kinase (AMPK), which is activated when ATP becomes limiting, was rapidly phosphorylated by 2-DG, and expression of Bcl-2 was augmented, which might result in resistance to 2-DG. On the other hand, AMPK phosphorylation and augmentation of Bcl-2 expression by 2-DG were not observed in NB4, which is 2-DG sensitive. These results will facilitate the future leukemia therapy targeting metabolic pathways.