In Japan, some 30,000 people every year die from liver carcinoma. Approximately 65% of hepatocellular carcinomas (HCC) are caused by hepatitis C. Therefore, the treatment of hepatitis C is important. As an antiviral therapy for hepatitis C virus (HCV), interferon (IFN) monotherapy was initiated in the 1990s. Thereafter, combination therapy of IFN with the oral antiviral drug ribavirin (RBV) became available in December 2001, followed by pegylated IFN (PEG-IFN) in December 2003. Combination therapy of PEG-IFN with RBV for HCV genotype (GT) 1 infection was initiated in December 2004, and this combination therapy had been the standard therapy until 2011. The development of direct acting antivirals (DAAs) represents a major breakthrough in the treatment of HCV-infected patients. Since the end of 2011, triple therapy of protease inhibitor combined with PEG-IFN and RBV has markedly improved treatment outcome in HCV-infected patients. In September 2014, IFN-free DAA therapies became available for patients with chronic hepatitis C and HCV-related cirrhosis of Child-Pugh class A. In these patients, DAA therapies achieve an HCV-RNA undetectable rate of approximately 95%. The final goal of treatment for chronic hepatitis C is viral eradication and prevention of progression to cirrhosis and HCC. The goal of hepatitis C eradication has almost been reached.
In humans, the incidence of embryo wastage and pregnancy loss is extraordinarily high; therefore, pregnancy loss commonly occurs in reproductive women. Furthermore, some women suffer from recurrent pregnancy loss (RPL). In over 50% of RPL cases, no specific cause is identified even after detailed examinations. Impaired decidualisation of uterine endometrium with aberrantly high uterine natural killer (uNK) cell density is associated with unexplained RPL. Glucocorticoid includes active cortisol and inert cortisone, which are mutually catalysed by 11β-hydroxysteroid dehydrogenase (11βHSD). 11βHSD isoform type 1 activates cortisone to cortisol, which binds glucocorticoid receptor (GR) and mineralocorticoid receptor (MR). During decidual transformation in human endometrial stromal cells (HESCs), 11βHSD1, but not type 2, is particularly induced by progesterone, leading to the local production of active cortisol and regulation of GR and MR gene networks. 11βHSD1/GR signalling is a significant pathway for the regulation of uNK cells and inflammatory reaction upon decidualisation of HESCs, whereas the MR gene network includes retinoid metabolism. Retinol (vitamin A), which is hydrolysed from retinyl ester, is transformed to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. RA binds to CRABP2 and activates retinoic acid receptor (RAR), leading to cell apoptosis and senescence, whereas RA-dependent activation of PPARβ/δ is mediated through FABP5, leading to cell differentiation. The decidualisation of HESCs decreases the expression of CRABP2 and FABP5 and down- and upregulates RAR and PPARβ/δ, respectively. In addition, decidualisation is associated with the upregulation of retinoid metabolism-related genes, including retinol-binding protein 4 (RBP4), cytochrome P450 26A1 (CYP26A1), RDH12 and DHRS3, suggesting that decidualisation suppresses RA signalling by decreasing key cytoplasmic-binding proteins and upregulating retinoid metabolism. Taken together, corticosteroid signalling via 11βHSD1 induction regulates GR-specific gene networks including inflammation and angiogenesis at the implantation site. Furthermore, retinoid metabolism signalling is controlled via 11βHSD1/MR signalling, leading to an optimal balance of cell differentiation and apoptosis, such as decidualisation of HESCs. Recently, for the treatment of unexplained RPL, the GR and MR gene networks are targeted via corticosteroid signalling, leading to modulation of uNK cells as well as retinoid metabolism signalling.
Apart from their direct antimicrobial activities against invading pathogens, antimicrobial peptides exhibit additional protective functions that have led to their being named host defense peptides (HDPs). These functions include the stimulation of the production of cytokines/chemokines, the promotion of chemotaxis and cell proliferation and the induction of angiogenesis and wound healing. AG-30/5C is a novel angiogenic HDP that in addition to its antimicrobial activity also activates fibroblasts and endothelial cells and promotes angiogenesis and wound healing. Given that mast cells are found primarily in the vicinity of vessels, where they are intimately involved in wound healing, we hypothesized that AG-30/5C may activate mast cells. We demonstrated that AG-30/5C activated LAD2 human mast cells to degranulate and produce lipid mediators including leukotriene C4, prostaglandin D2 and E2. Moreover, AG-30/5C increased mast cell chemotaxis and Induced the production of the cytokines GM-CSF and TNF-α and various chemokines, such as IL-8, MCP-1, MCP-3, MIP-1α and MIP-1β. The chemotaxis and cytokine/chemokine production induced by AG-30/5C were suppressed by both pertussis toxin and U-73122, suggesting the involvement of the G protein and phospholipase C pathways in AG-30/5C-induced mast cell activation. Furthermore, these pathways were activated downstream of the MAPK and NF-κB signaling molecules, as demonstrated by the inhibitory effects of ERK-, JNK-, p38- and NF-κB-specific inhibitors on cytokine/chemokine production. Interestingly, AG-30/5C caused the phosphorylation of MAPKs and IκB. We suggest that the angiogenic and antimicrobial peptide AG-30/5C plays a key role in the recruitment and activation of human mast cells at inflammation and wound sites.
Host defense (antimicrobial) peptides not only display antimicrobial activities against numerous pathogens but also exert a broader spectrum of immune-modulating functions. Innate defense regulators (IDRs) are a class of host defense peptides synthetically developed from natural or endogenous cationic host defense peptides. Of the IDRs developed to date, IDR-1018 is more efficient not only in killing bacteria but also in regulating the various functions of macrophages and neutrophils and accelerating the wound healing process. Because mast cells intimately participate in wound healing and a number of host defense peptides involved in wound healing are also known to activate mast cells, this study aimed to investigate the effects of IDR-1018 on mast cell activation. Here, we showed that IDR-1018 induced the degranulation of LAD2 human mast cells and caused their production of leukotrienes, prostaglandins and various cytokines and chemokines, including granulocyte-macrophage colony-stimulating factor, interleukin-8, monocyte chemoattractant protein-1 and -3, macrophage-inflammatory protein-1α and -1β, and tumor necrosis factor-α. Furthermore, IDR-1018 increased intracellular calcium mobilization and induced mast cell chemotaxis. The mast cell activation was markedly suppressed by pertussis toxin, U-73122, U0126, SB203580, JNK inhibitor II and NF-κB activation inhibitor II, suggesting the involvement of G-protein, phospholipase C, ERK, p38, JNK and NF-κB pathways, respectively, in IDR-1018-induced mast cell activation. Notably, we confirmed that IDR-1018 caused the phosphorylation of MAPKs and IκB. Altogether, the current study suggests a novel immunomodulatory role of IDR-1018 through its ability to recruit and activate human mast cells at the sites of inflammation and wounds.
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.
In order to extend the healthy life expectancy and shorten the years lived in poor health in Japan, the early treatment and prevention of motor organ disorders are urgently required. A concept named ‘locomotive syndrome’ is defined as being restricted in one’s ability to walk or lead a normal life owing to a dysfunction in one or more of the parts of the motor organ (locomotive) system. We developed the Locomonitor app using Apple’s ResearchKit and conducted the first cross-sectional nationwide observational study to evaluate the epidemiology of locomotive syndrome. The locomotive function was evaluated using the locomotive syndrome risk tests implemented in the app. The potential benefits of the smartphone-based Locomonitor study are 1) attracting participants from a wide area and different generations within a short period, 2) helping disseminate knowledge of locomotive syndrome, 3) enabling locomotive syndrome risk tests without a measurer, 4) gathering objective and continuous activity data without any burden on the participants, and 5) providing highly personalized feedback to participants. However, uncertainty with regard to accuracy and some possible selection bias are potential limitations. We predict that smartphone-based medical studies will be conducted with increasing frequency in the future.
The hippocampus plays important roles in learning and memory. In the hippocampus, the extracellular matrix (ECM) is known to function in synaptic plasticity. Chondroitin sulfate (CS) chain, one of the major ECM components in the brain, is a long unbranched polysaccharide consisting of repeating sulfated disaccharide units. After sugar extension, sulfation occurs at specific positions in the saccharides, and the pattern of CS sulfation influences several biological processes. The hippocampus is also known as a primary target for age-related conditions such as Alzheimer’s disease. Still, how the CS chains differ over time in the aged mouse hippocampus is still poorly understood.
In this study, we examined the immunoreactivity and disaccharide components of CS chains in young and aged mouse hippocampi.
We found that the immunoreactivity for specific CS epitopes recognized by the CS56 antibody exhibited two major patterns in the hippocampus; one was in the form of patches and the other was diffused pattern, especially in the CA1 region. Furthermore, the immunofluorescence intensity of both patterns significantly decreased with aging. Correspondingly, the specific disaccharide components of CS chains (C- and D-unit) in the hippocampus significantly decreased with aging.
Therefore, our study may help to understand the roles of chondroitin sulfate in the neurological decline due to aging.
Objective: Differentiation-inducing factor-1 (DIF-1) and -3 are novel lead anti-tumor agents that were originally isolated from the cellular slime mold Dictyostelium discoideum. We previously showed that the DIF derivatives Br-DIF-1, DIF-3(+2), and Bu-DIF-3 strongly suppress lysophosphatidic acid (LPA)-induced cell migration in mouse osteosarcoma LM8 cells in vitro. Here, we investigated the effects of these three DIF derivatives on the cell migration (wound healing) of LM8 cells and mouse 3T3-L1 fibroblast cells (a model of non-transformed cells).
Methods: A wound healing assay was performed to estimate the anti-cell migration activities of the compounds; cell growth was assessed by using a cell number indicator.
Results: After monolayers of LM8 cells or 3T3-L1 cells were wounded by scratching and incubated with 10% serum-containing media for 20 h or 8 h, respectively, cell-free areas (wounds) in the monolayer were occupied (healed) by neighboring cells to some extent even in the presence of Ara-C (an inhibitor of cell proliferation). This phenomenon is attributed to the migration of the neighboring cells into the wounds. Wound healing (cell migration) of LM8 cells was strongly suppressed in the presence of 10 μM Br-DIF-1, DIF-3(+2), or Bu-DIF-3. These DIF derivatives more effectively suppressed the cell migration of LM8 cells than of 3T3-L1 cells.
Conclusion: The results support our expectation that these DIF derivatives are promising lead anti-metastatic agents that may not affect normal cell migration.
Objective: Online hemodiafiltration (OLHDF) therapy has clinical benefits in patients requiring blood purification; however, it may lead to essential serum protein elimination. Previous studies suggest that total complement hemolytic activity (CH50) is related to nutritional status. This study aimed to evaluate the association of CH50 fluctuations and nutritional status after transition from hemodialysis (HD) to OLHDF.
Methods: Serum samples from 20 patients were analyzed during transition from HD to OLHDF therapy and at 3 and 9 months after the transition. Nutritional status was assessed by the Geriatric Nutritional Risk Index (GNRI). Subgroup analysis was performed according to CH50 increase or decrease.
Results: Variability rate of CH50 was positively correlated with that of serum albumin (Alb) and other serum nutritional indicators at 9 months (p<0.05). In patients with a decrease in CH50 at 9 months, serum Alb and other serum nutritional indicators also reduced (p<0.05). GNRI at 9 months was lower in the CH50-decrease than in the CH50-increase group (p<0.05).
Conclusions: Significant CH50 fluctuations were found to be as a nutritional parameter after transition from HD to OLHDF.
Objective: Regorafenib (BAY 73-4506) is an oral multikinase inhibitor that has been shown to inhibit various oncogenic stromal receptor tyrosine kinases and intracellular signaling kinase. The aim of the study is to confirm the safety and effectiveness of dose escalation therapy of regorafenib for the third- or fourth- treatments of patients with unresectable and recurrent colorectal cancer.
Design: This study is designed as a single-arm, prospective, non-randomized, multi-centered open label phase II trial in Japan. The study regimen consists of 28-day cycles with escalated doses of regorafenib (80-160 mg/body on days 1 to 21). This study is a dose-escalation study using a starting dose of 80 mg/day during the first seven days. In case of no adverse events related to the administration after the first seven days of the starting dose, the administration dose will be increased to 120 mg/day on days 8 to 14. The administration dose will then be increased to 160 mg/day on days 15 to 21. Primary end point is progression free survival (PFS); secondary end points include time to treatment failure, response rate, overall survival, the interval between the initial of drug administration and performance status 2, the transition rate to the following line of treatment, the cumulative dose and the incidence of adverse events of grade 3 or higher, and genetic alterations (KRAS mutation/BRAF mutation) in circulating cell-free DNA.
Conclusions: This study may contribute to determining the efficacy of the escalation of the administration dose in patients with metastatic colorectal cancer.
Objective: It has been reported that vaccination rates with pneumococcal vaccine are low in elderly persons in Japan. The primary aim of this study was to examine whether introduction of a vaccination status checkbox in electronic medical records increased the vaccination rates.
Materials and Methods: Subjects were inpatients and outpatients of the Department of General Medicine at Juntendo University Hospital between April 1, 2016 and March 31, 2018. The vaccination rates of individuals ≥65 years old were calculated based on a checkbox in the electronic medical records.
Results: A total of 30,580 people ≥65 years of age were evaluated. The vaccination rate (19.5%) for individuals ≥65 years old after introducing a regular vaccination schedule was significantly higher than the vaccination rate (8.4%) before introducing the schedule (p<0.01).
Conclusions: With the introduction of a regular vaccination schedule for the 23-valent pneumococcal polysaccharide vaccination targeting elderly individuals since October 2014 in Japan, the vaccination rate was shown to be improved by the introduction of a checkbox to indicate a patient’s vaccination status in the electronic medical records. Vaccination rates with pneumococcal vaccine in elderly persons might be a good quality indicator for primary care physicians.
Mutations in the PRRT2 gene have been identified as a major cause of benign infantile epilepsy (BIE) and/or paroxysmal kinesigenic dyskinesia (PKD). We identified the PRRT2 common mutation (c.640_641insC) in a 13-year-old boy who was initially suspected of focal epilepsy. He visited our hospital due to a one-year history of hyperkinetic movements, dystonic movements triggered by sudden voluntary movements, and a past history of BIE. He had experienced on-and-off attacks of involuntary dystonic movements that lasted from 10 s to 5 min, and involved the limbs, trunk, and occasionally the face. In our department, he was clinically diagnosed with PKD without family history, and a PRRT2 mutation was found. His condition was characterized by a longer duration of attacks and good efficacy of low dose carbamazepine. In this report, we describe how the correct diagnosis in this case had implications for treatment and prognosis.
Japan’s health system has received the world’s highest rating from the WHO in 2000, but that is the story so far. Japan is facing serious problems, which is the sharp increase in the elderly population. The population ratio of the elderly continues to increase until 2042, and aging population ratio will exceed 40% in 2065. Then it is the growth of medical expenditure that will cause enormous problems in the future. Extension of healthy life expectancy (HLE) is the key, and shortening the difference between lifespan and HLE is a present task in Japan. In order to extend HLE, it is necessary to shorten the period of reduction of degree of independence and bedridden, that is, requiring support and long-term care. And, the point is essential care prevention. There is a strong association between visual impairment and reasons for requiring long-term care / support. Measures against visual impairment will help to extend HLE.