Impact of oral health management by nurses and dental professionals on oral health status in inpatients eligible for the Nutrition Support Team: A longitudinal study.

OBJECTIVE: As the participation of dental professionals in multidisciplinary care is often limited, instructions on oral health management provided by dental professionals to other professionals are important to achieve transdisciplinary oral health management; however, the effectiveness of such instructions remains unclear. In this longitudinal study, we aimed to determine the impact of oral health management provided by dental professionals and nurses instructed on oral health management by dental professionals on the oral health of inpatients eligible for a Nurition Support Team (NST). METHODS: The study participants were 117 patients (66 men and 51 women, mean age: 71.9 ± 12.5 years) who received oral health management during the NST intervention period. The participants received oral health management from nurses (Ns group) or dental professionals (D group). The nurses who conducted the oral health management received instructions from dental professionals. Oral health was assessed at the beginning and end of the NST intervention using the Oral Health Assessment Tool (OHAT). RESULT: The Ns and D groups showed significant improvements in the total OHAT scores at the end of the NST intervention. Both groups showed significant improvements in the OHAT subitems of lip, tongue, gums and tissues, saliva, oral cleanliness and dental pain, while only the D group showed a significant improvement in the denture subitem. CONCLUSION: Effective oral health management provided by dental professionals or by nurses trained by them improved the oral health status of inpatients eligible for NST at an acute-care hospital.

Elucidation of the needs for telecritical care services in Japan: a qualitative study.

OBJECTIVE: To clarify the reasons for consultation, advice sought by frontline physicians and relationship between the patient's pathology and the type of advice provided to guide the future development of telecritical care services. DESIGN: Secondary analysis of transcripts of telephone calls originally recorded for quality control purposes was conducted using a thematic content analysis. The calls were conducted between December 2019 and April 2021 (total cases: 70; total time: ~15 hour). SETTINGS: Intensivists provided consultation services to frontline physicians at secondary care institutions in the Kansai and Chubu regions. PARTICIPANTS: Non-intensive care frontline physicians working in five secondary care institutions in the Kansai and Chubu regions and intensivists providing a consultation service (n=26). INTERVENTIONS: Not applicable. PRIMARY AND SECONDARY OUTCOME MEASURES: The main outcome was the themes emerging from the language used during telephone and video consultations, indicating the gap filled by the telecritical care service. FINDINGS: We analysed 70 cases and approximately 15 hours of anonymised audio data. We identified the following reasons for consultation: 'lack of competence in treatment and diagnostic testing' and 'lack of access to consultation in their own hospital'. Frontline physicians most often sought advice related to 'treatment', followed by 'patient triage and transfer', 'diagnosis' and 'diagnostic testing and evaluation'. Regarding the relationship between the patient's pathology and type of advice provided, the most commonly sought advice by frontline physicians varied based on the patient's pathology. CONCLUSION: This study explored the characteristics of 70 telecritical sessions and identified the reasons for and nature of the consultations. These findings can be used to guide the future provision and scale up of telecritical services.

Factors influencing the selection of oral healthcare providers in multidisciplinary Nutrition Support Team for malnourished inpatients: A cross-sectional study.

BACKGROUND: Oral health management involving dental professionals in a multidisciplinary Nutritional Support Team (NST) is effective collaborative care. However, the indicators for the triage of oral health management requirements and selection of oral healthcare providers remain unclear. OBJECTIVE: This cross-sectional study included inpatients with malnutrition and aimed to investigate the factors associated with determining the need for oral health management involving dental professionals and selecting primary oral healthcare providers when needed. METHODS: Participants included 255 inpatients (154 males and 101 females, mean age 69.7 ± 14.4 years) aged ≥20 years who underwent oral assessment by the NST between April 2016 and July 2019. Participants were assigned to the following groups: good oral health group, oral health management by nurses under the supervision of dental professionals group, and oral health management by dental professionals group. The comprehensive oral health status was investigated using the Oral Health Assessment Tool (OHAT). The total OHAT score ranges from 0 to 16, with a higher score indicating a poorer oral environment. RESULTS: The total OHAT score had a significant correlation with the need for oral health management by dental professionals. Inpatients with denture problems required oral health management from dental professionals. The optimal cutoff value of the total OHAT score for determining the need for oral health management was four. CONCLUSION: The need for oral health management by dental professionals increased with worsening oral health status, especially denture problems. The OHAT score could be used to triage inpatients who require oral health management collaborated with dental professionals.

Association between Grit and depressive symptoms at the timing of job start among medical residents during the COVID-19 pandemic in Japan: a cross-sectional study.

INTRODUCTION: The COVID-19 pandemic had an additional negative impact on the mental health of healthcare workers, including residents. Previous studies revealed that grit, which is an individual trait to achieve long-term goals unrelated to intelligence quotient, is not only positively associated with academic achievement and career success but also negatively correlated with depression. This study aimed to examine the association between grit and depressive symptoms among residents at the time of job start during the coronavirus disease 2019 (COVID-19) pandemic in Japan. MATERIALS AND METHODS: This cross-sectional study used data from all post-graduate year 1 residents from March 2020 to April 2022 at Tokyo Medical and Dental University, Tokyo, Japan. Grit was measured by the Japanese version of Grit-S. The resident's depressive symptoms were assessed using the Center for Epidemiologic Studies Depression Scale. The association of interest was examined using logistic regression analysis. RESULTS: Among 221 residents, 28 (12.7%) have depressive symptoms. One unit increase in Grit-S score after adjusting for age, sex, graduated university, and sleeping hours was associated with lower odds of having depressive symptoms by 63% (odds ratio [OR]: 0.37; 95% confidence interval [CI]: 0.19-0.74). Further, the perseverance of effort subscale score was associated with lower odds of having depressive symptoms after covariate adjustment (OR: 0.43; 95% CI: 0.22-0.84). CONCLUSIONS: Higher grit scores were associated with lower odds of having depressive symptoms among residents at the timing of job start during the COVID-19 pandemic. Regular mental health assessment is particularly important for residents with low grit scores at entry.

Factors associated with liver injury and prognosis in advanced cancer patients treated with immune checkpoint inhibitors.

AIM: The use of immune checkpoint inhibitors (ICIs) has increased remarkably, and immune-related adverse events (irAEs) have also increased. This study aimed to identify factors associated with immune-related liver injury (irLI), and the relationship between the grades of irLI and overall survival (OS) in patients treated with ICIs. METHODS: A total of 571 patients who had been treated for advanced malignancies with ICIs between January 2015 and March 2022 were retrospectively recruited. The presence of liver injury was determined by the aspartate aminotransferase and alanine aminotransferase elevation. The irLI grading was based on Common Terminology Criteria for Adverse Events version 5.0. RESULTS: A total of 50 (8.8%) patients had grade ≥2 irLI and 24 (4.2%) had grade ≥3 irLI. Treatment with anti-cytotoxic T-lymphocyte-associated protein-4 agents and baseline grade 1 aspartate aminotransferase/alanine aminotransferase elevation were independent predictive factors of grade ≥2 irLI. Treatment with anti-cytotoxic T-lymphocyte-associated protein-4 was the only independent predictive factor of grade ≥3 irLI. The median OS for patients who experienced any irAEs was significantly longer than of those without irAEs (hazard ratio 0.503, 95% CI 0.398-0.636, p < 0.001). The median OS in patients with grade ≥2 irLI was significantly longer (HR 0.570, 95% CI 0.387-0.838, p = 0.022). There was no significant difference between the median OS in patients with grade ≥3 irLI and the others (p = 0.11). CONCLUSION: The incidence of irLI was significantly higher in patients treated with anti-cytotoxic T-lymphocyte-associated protein-4 agents. Even in patients with pre-existing grade 1 aspartate aminotransferase/alanine aminotransferase elevation, appropriate follow-up and control of the irLI can improve the prognosis.

Changes in Nutrition-Intake Method and Oral Health through a Multidisciplinary Team Approach in Malnourished Older Patients Admitted to an Acute Care Hospital.

Malnourished older inpatients referred to a nutrition support team (NST) usually receive multidisciplinary oral health management during NST intervention. However, the effects of multidisciplinary oral health management on the nutrition-intake method and oral health in these patients remain unclear. This longitudinal study aimed to investigate the effects of NST-mediated multidisciplinary oral health management on the nutrition-intake methods, oral health, and the systemic and oral factors influencing the changes in the nutrition-intake method. A total of 117 inpatients (66 men, 51 women, mean age, 71.9 ± 12.5 years) who underwent NST-mediated multidisciplinary oral health management between April 2016 and July 2019 were enrolled. Demographic data and Functional Oral Intake Scale (FOIS), Dysphagia Severity Scale (DSS), and Oral Health Assessment Tool (OHAT) scores at the time of referral to the NST and completion of the NST intervention were evaluated. After multidisciplinary NST intervention, FOIS, DSS, and OHAT scores showed significant improvements (p < 0.001). Even after adjusting the results for systemic parameters, FOIS score improvement correlated positively with the length of NST intervention (p = 0.030) and DSS score improvement (p < 0.001) as well as OHAT score improvement (p = 0.047). NST interventions with multidisciplinary oral health management could improve the nutrition-intake method.

Facilitators and barriers to implementing shared decision making: A cross-sectional study of physicians in Japan.

OBJECTIVE: Shared decision making (SDM) is a collaborative process in which patients and healthcare providers jointly make a medical decision. This cross-sectional study aimed to identify the facilitators and barriers to self-reported implementation of SDM in Japan, and to explore if there is effect modification by hospital types. METHODS: A total of 129 physicians in Japan completed a questionnaire that asked about their perception of SDM based on SDM-Q-Doc and its facilitators and barriers, which corresponded to each construct of the integrated behavioral model (IBM). The association between facilitators and barriers and SDM-Q-doc score was assessed using linear regression analysis. Stratified analysis by hospital types was also performed. RESULTS: Significant facilitators included physicians' attitude, injunctive norm, intention and habit. Significant barriers included physicians' unfavorable attitude, lack of self-efficacy, knowledge, salience and experience. Moreover, experiential attitude (concerning patient characteristics), injunctive norm (concerning patient preferences), and physician's habit were significant facilitators for physicians working in university hospitals when compared to those working in municipal hospitals. CONCLUSION: The facilitators and barriers to implementing SDM in Japan were identified. PRACTICE IMPLICATIONS: More opportunities for training on SDM are needed to provide knowledge and skills, which will enhance salience and contribute their habitual practice.

Oral health status and its association with nutritional support in malnourished patients hospitalised in acute care.

OBJECTIVES: This cross-sectional study aimed to examine the oral health of malnourished acute-care hospital inpatients, who were the subjects of a nutritional support team (NST). We also aimed to elucidate the systemic and nutritional factors associated with the oral health of those patients. BACKGROUND: Interventions by NST are essential for inpatient nutrition management and require the active participation of dental professionals. However, information is limited regarding the state of oral health among acute-stage malnourished inpatients. MATERIALS AND METHODS: We enrolled 255 hospitalised patients (101 women, mean age: 69.7 ± 14.4 years) who were referred to an NST for nutrition management between April 2016 and July 2019. The main outcome was the Oral Health Assessment Tool (OHAT) scores. Moreover, we assessed participants' demographic characteristics, nutritional status, number of natural and functional teeth, posterior occlusal support, denture use, Dysphagia Severity Scale, whether oral health management was needed, and the methods of nutrition intake. RESULTS: Several participants presented with a deteriorated oral health. Consequently, oral health management was often regarded necessary in these patients. Approximately half were fed by parenteral or tube feeding. Multiple regression analysis revealed the OHAT score has a positive association with age (P = .008), and a negative association with body mass index (P = .009) and the method of nutrition intake (P = .028). CONCLUSION: Malnourished inpatients at an acute care hospital who were subject to an NST had a deteriorated oral health status. Additionally, poor oral health was associated with poor nutritional status and nutrition intake methods.

Factors Related to Oral Intake of Food by Hospitalized Patients with Malnutrition under the Care of a Nutrition Support Team.

This study aimed to evaluate the role of the general condition and oral health status in determining the primary nutritional route and suitable food form for oral ingestion among malnourished inpatients. This cross-sectional study included 255 inpatients referred to a nutrition support team (NST), which included dental professionals, at an acute care hospital. We assessed the participants' basic information, and Dysphagia Severity Scale (DSS) and Oral Health Assessment Tool (OHAT) scores. The nutritional intake mode was evaluated based on the Functional Oral Intake Scale scores at the initial NST consultation (FOIS-I), and then revised by the NST based on the participants' general condition and oral health (FOIS-R). There was a divergence between FOIS-I and FOIS-R, with FOIS-R being significantly higher than FOIS-I (p < 0.001). Logistic regression analysis of FOIS-R identified that consciousness level (odds ratio (OR): 0.448; 95% confidence interval (CI): 0.214-0.935) and DSS (OR: 3.521; 95% CI: 2.574-4.815) significantly affected the oral nutrition intake. Among participants who could ingest orally (FOIS-R ≥ 3; n = 126), FOIS score had significant negative and positive associations with the OHAT and DSS scores, respectively. These findings suggest that appropriate assessment of oral health status, including swallowing function, might contribute to high-quality nutrition management.

Facilitators and barriers to physicians' entrepreneurial ventures in major Japanese cities: A qualitative study.

BACKGROUND: The Japanese healthcare system currently faces numerous challenges, including a super-aging society and an excessive burden on medical workers; therefore, the need for innovative solutions from healthcare ventures to tackle these issues has increased. Meanwhile, as physicians play important roles in healthcare ventures, the need for Japanese physician entrepreneurs is more important than ever. Given the lack of research examining barriers to physicians starting ventures and what skills, knowledge, and surrounding environments act as facilitators, this study aimed to identify the facilitators and barriers faced by physicians to start ventures. METHODS: Between September and November 2019 and in May 2021, qualitative interviews were conducted with 33 participants, which included eight physician entrepreneurs; two administrative officers at the Ministry of Health, Labour and Welfare and the Ministry of Economy, Trade and Industry; three faculty members at Tokyo Medical and Dental University (in-depth interviews); and 20 medical students (focus group discussions). The interviews were deductively coded based on the social ecological model. The inductive approach was applied to coding any knowledge necessary to start a business. We conducted member checking with three physician entrepreneurs and seven medical students to improve our results' credibility. RESULTS: The factors influencing a physician's decision to launch a new business include their willingness to contribute to society, the unique environment in which an individual is placed while in medical school and afterward, negative aspects of the lack of diversity in physicians' careers, the financial stability provided by a medical license, and self-efficacy. CONCLUSIONS: Our study revealed facilitators and barriers to physicians' entrepreneurial ventures. Knowledge about these factors might be useful in supporting physicians to launch or become involved in healthcare ventures.

Mac-2 binding protein glycosylation isomer as a novel predictive biomarker for patient survival after hepatitis C virus eradication by DAAs.

BACKGROUND: It is crucial to identify risk factors for life prognosis after hepatitis C virus (HCV) eradication among patients with or without a high risk of liver cancer or complications. METHODS: This is a prospective, multicenter and observational study using the database of 1031 patients after HCV eradication by direct-acting antiviral agents (DAAs) to evaluate the development of hepatocellular carcinoma (HCC) and patients' survival after a sustained virological response (SVR). The Cox proportional hazards regression model was used to estimate hazard ratios associated with HCC development and survival. RESULTS: AFP at SVR was significantly associated with HCC recurrence in the adjusted model. Liver fibrosis, Mac-2 binding protein glycosylation isomer (M2BPGi) at SVR and smoking status before treatment were positively associated with the development of HCC and M2BPGi was positively associated with HCC recurrence, although not reaching statistical significance. Among patients without a history of HCC, M2BPGi and estimated glomerular filtration rate (eGFR) at SVR were significantly associated with death after viral eradication [M2BPGi (HR 4.07, 95% CI 1.22, 13.57), eGFR (HR 0.97, 95% CI 0.94, 0.99)]. Strikingly, of 16 patients who died, among participants without a history of HCC, only two died of liver cancer associated with HCV, whereas 11 died of non-HCV- related cancer or cardiovascular diseases. CONCLUSION: M2BPGi at SVR is a potential predictor for patients' survival and a candidate biomarker for detecting individuals who are at greater risk of death due to cancer-related and unrelated to HCV, as well as cardiovascular diseases, after viral eradication.

Vasoactive Intestinal Peptide Derived From Liver Mesenchymal Cells Mediates Tight Junction Assembly in Mouse Intrahepatic Bile Ducts.

Formation of intrahepatic bile ducts (IHBDs) proceeds in accordance with their microenvironment. Particularly, mesenchymal cells around portal veins regulate the differentiation and ductular morphogenesis of cholangiocytes in the developing liver; however, further studies are needed to fully understand the arrangement of IHBDs into a continuous hierarchical network. This study aims to clarify the interaction between biliary and liver mesenchymal cells during IHBD formation. To identify candidate factors contributing to this cell-cell interaction, mesenchymal cells were isolated from embryonic day 16.5 matrix metalloproteinase 14 (MMP14)-deficient (knockout [KO]) mice livers, in which IHBD formation is retarded, and compared with those of the wild type (WT). WT mesenchymal cells significantly facilitated the formation of luminal structures comprised of hepatoblast-derived cholangiocytes (cholangiocytic cysts), whereas MMP14-KO mesenchymal cells failed to promote cyst formation. Comprehensive analysis revealed that expression of vasoactive intestinal peptide (VIP) was significantly suppressed in MMP14-KO mesenchymal cells. VIP and VIP receptor 1 (VIPR1) were mainly expressed in periportal mesenchymal cells and cholangiocytic progenitors during IHBD development, respectively, in vivo. VIP/VIPR1 signaling significantly encouraged cholangiocytic cyst formation and up-regulated tight junction protein 1, cystic fibrosis transmembrane conductance regulator, and aquaporin 1, in vitro. VIP antagonist significantly suppressed the tight junction assembly and the up-regulation of ion/water transporters during IHBD development in vivo. In a cholestatic injury model of adult mice, exogenous VIP administration promoted the restoration of damaged tight junctions in bile ducts and improved hyperbilirubinemia. Conclusion: VIP is produced by periportal mesenchymal cells during the perinatal stage. It supports bile duct development by establishing tight junctions and up-regulating ion/water transporters in cholangiocytes. VIP contributes to prompt recovery from cholestatic damage through the establishment of tight junctions in the bile ducts.

Time course alterations of virus sequences and immunoglobulin titers in a chronic hepatitis E patient.

AIM: Hepatitis E virus (HEV) can cause chronic infection in immunocompromised hosts. However, the dynamics of HEV during persistent infection is not well understood. To elucidate time course alterations in virus sequences and anti-HEV antibodies during persistent infection, we analyzed the HEV sequences and titers of anti-HEV antibodies from a chronic hepatitis E patient. METHODS: Serum samples were obtained from a chronic hepatitis E patient under corticosteroid therapy for neurological disease. The titers of anti-HEV antibodies (immunoglobulin A, immunoglobulin M, and immunoglobulin G) in serum samples were detected by enzyme immunoassay. The full or near-full nucleotide sequences of HEV isolated from consecutive serum samples were identified and compared. Phylogenetic analysis was also performed. RESULTS: Alterations of anti-HEV antibodies from a chronic hepatitis E patient were different from those previously reported in acute hepatitis E patients. The virus sequence was unchanged in the period without treatment, but nucleotide mutations were observed after ribavirin treatment was started. In addition, the sequence of this strain had extremely high identity to that isolated from swine liver in Japan. CONCLUSIONS: Virus mutations in HEV emerged after ribavirin treatment was started. Sequence analysis may useful for deciding the treatment strategy for chronic hepatitis E patients who did not eliminate the virus with 3 months of RBV treatment and inferring the origin of the infection. This report provides insights into the chronicity of hepatitis E, and the impact of persistent infection and ribavirin treatment on the emergence of virus mutations.

Hypervirulent clade 2, ribotype 019/sequence type 67 Clostridioides difficile strain from Japan.

BACKGROUND: Clostridioides difficile ribotype (RT) 019/sequence type (ST) 67 strains belong to a hypervirulent lineage closely related to RT027/ST1; however, limited data are available for hypervirulent clade 2 lineages in Japan. Herein, we report the draft genome of a C. difficile strain B18-123 belonging to clade 2, RT019/ST67 for the first time in Japan. RESULTS: The pathogenicity locus carried by B18-123 (19.6 kb) showed higher homology (97.29% nucleotide identity) with strain R20291 (RT027/ST1) than the reference strain 630 (RT012/ST54), and B18-123 harbored 8-nucleotide substitutions in tcdC. However, it did not contain an 18-base pair (bp) deletion or a single-bp deletion at position 117 in tcdC, which was identified in the previous strain R20291. A cytotoxicity assay revealed similar cytotoxicity levels between strains B18-123 and ATCC BAA-1870 (RT027/ST1). The B18-123 strain was found to be susceptible to metronidazole and vancomycin. CONCLUSION: Our findings contribute to the further understanding of the characteristics of hypervirulent clade 2 including RT019/ST67 lineages.

A case of tracheal obstruction caused by reflux and aspiration of semi-solid nutrients via the nasogastric tube.

INTRODUCTION: Semi-solid nutrients have several advantages, including reduced cases of diarrhea and aspiration pneumonia, and are usually administered via percutaneous endoscopic gastrostomy owing to its high viscosity. Administering semi-solid nutrients via a nasogastric tube was recently introduced in clinical practice; however, its safety has not been well confirmed. PRESENTATION OF CASE: An 82-year-old man with a right occipital hemorrhage and severe diarrhea consulted the nutritional support team. Administrations of semi-solid nutrients (HINE E-GEL®) via the nasogastric tube was initiated, which gradually alleviated his symptoms. Fourteen days after initiation, he suddenly had pulmonary failure owing to a tracheal obstruction caused by the reflux and aspiration of semi-solid nutrients. Intubation and subsequent reflex cough expectorated sputum with gel-form particles, which quickly stabilized his pulmonary condition. After this, his hospital course was stable, and he was referred to another hospital for further rehabilitation. DISCUSSION: Semi-solid nutrients administered via the nasogastric tube have different ingredients compared with those administered via percutaneous endoscopic gastrostomy. HINE E-GEL®, for example, contains pectin and calcium phosphate that changes from liquid to semi-solid inside the stomach via chemical reactions under acidic conditions. Data on the viscosity of HINE E-GEL® in vivo are insufficient. Uncertainty regarding the form and viscosity of HINE E-GEL® inside the stomach complicates clinical practice. CONCLUSIONS: Although semi-solid nutrients have several advantages, including reduced diarrhea and gastroesophageal reflux, evidence on semi-solid nutrients via the nasogastric tube is insufficient. It should be noted that semi-solid nutrient reflux can be more fatal than liquid nutrients.

Comprehensive genetic analysis of cholangiolocellular carcinoma with a coexistent hepatocellular carcinoma-like area and metachronous hepatocellular carcinoma.

AIM: The genetic profile of cholangiolocellular carcinoma (CLC) and its origin in relation to hepatocellular carcinoma (HCC) remain unclear. To elucidate the genetic profile of CLC, a comprehensive analysis of genetic mutations was carried out in a case of CLC with an HCC-like focal area and metachronous HCC. METHOD: Liver tissue was obtained from CLC, a co-existent HCC-like area, and metachronously developed HCC by laser capture microdissection of formalin-fixed paraffin-embedded specimens obtained by hepatectomy. Gene mutational profiles were analyzed comprehensively by next-generation sequencing and digital PCR. Relationships among gene profiles, immunohistochemistry, and clinicopathological findings were investigated. RESULTS: Mutations in EGFR, PTEN, RB1, TP53, and ERBB2 were found in CLC, whereas mutations in KIT, BRAF, PTEN, TP53, and SMAD4 were found in the coexistent HCC-like area. Only the mutation in PTEN has a common Catalogue of Somatic Mutations in Cancer ID in the CLC and coexistent HCC-like area, and is related to the kinase-RAS module. In contrast, no cancer-related mutations were found in the metachronous HCC. No TERT mutations were found in any of the regions by digital PCR. Immunohistochemical staining for p53 was negative in CLC, although ≤10% positive in the coexistent HCC-like area. Immunostaining of C-kit, HER2, PTEN, and SMAD4 were negative. CONCLUSION: The genomic features of CLC and the focal area of an HCC-like region differ, but are related to the kinase-RAS module. The development of carcinogenesis in the CLC and HCC-like areas in this case might differ, following a common PTEN mutation, although alteration of the kinase-RAS module is the most common molecular event in CLC.

Impact of novel NS5A resistance-associated substitutions of hepatitis C virus detected in treatment-experienced patients.

Resistance-associated substitutions (RASs) of hepatitis C virus (HCV) in the NS5A region impair the efficacy of NS5A inhibitors. In this study, we evaluated the characteristics of the novel RASs observed in treatment-failure patients, A92K and a deletion at P32 (P32del), and the susceptibility of viruses with these RASs to various anti-HCV reagents by using JFH-1 based recombinant HCV with NS5A from a genotype 1b Con1 strain (JFH1/5ACon1). We introduced A92K or P32del solely or in combination with Q24K, L28M, R30Q or L31F into the NS5A of JFH1/5ACon1. Viruses harboring R30Q/A92K showed high extracellular core antigens and infectivity titers, whereas the other viruses with RASs showed low replication levels and infectivity titers. All the viruses with A92K or P32del were markedly resistant to ledipasvir, velpatasvir and elbasvir. Interestingly, viruses with R30Q/A92K were more susceptible to grazoprevir than viruses without RAS. All the viruses had a similar susceptibility to ribavirin and sofosbuvir. In conclusion, combination RASs R30Q/A92K enhanced virus production whereas other RASs impaired virus replication. Both A92K and P32del conferred severe resistance even to second generation NS5A inhibitors. However, these viruses were susceptible to grazoprevir, ribavirin and sofosbuvir. Thus, combination regimens with these reagents may eradicate viruses harboring A92K or P32del.

Loss of fibrocystin promotes interleukin-8-dependent proliferation and CTGF production of biliary epithelium.

BACKGROUND & AIMS: Congenital hepatic fibrosis (CHF) is a genetic liver disease resulting in abnormal proliferation of cholangiocytes and progressive hepatic fibrosis. CHF is caused by mutations in the PKHD1 gene and the subsequent dysfunction of the protein it encodes, fibrocystin. However, the underlying molecular mechanism of CHF, which is quite different from liver cirrhosis, remains unclear. This study investigated the molecular mechanism of CHF pathophysiology using a genetically engineered human induced pluripotent stem (iPS) cell model to aid the discovery of novel therapeutic agents for CHF. METHODS: PKHD1-knockout (PKHD1-KO) and heterozygously mutated PKHD1 iPS clones were established by RNA-guided genome editing using the CRISPR/Cas9 system. The iPS clones were differentiated into cholangiocyte-like cells in cysts (cholangiocytic cysts [CCs]) in a 3D-culture system. RESULTS: The CCs were composed of a monolayer of cholangiocyte-like cells. The proliferation of PKHD1-KO CCs was significantly increased by interleukin-8 (IL-8) secreted in an autocrine manner. IL-8 production was significantly elevated in PKHD1-KO CCs due to mitogen-activated protein kinase pathway activation caused by fibrocystin deficiency. The production of connective tissue growth factor (CTGF) was also increased in PKHD1-KO CCs in an IL-8-dependent manner. Furthermore, validation analysis demonstrated that both the serum IL-8 level and the expression of IL-8 and CTGF in the liver samples were significantly increased in patients with CHF, consistent with our in vitro human iPS-disease model of CHF. CONCLUSIONS: Loss of fibrocystin function promotes IL-8-dependent proliferation of, and CTGF production by, human cholangiocytes, suggesting that IL-8 and CTGF are essential for the pathogenesis of CHF. IL-8 and CTGF are candidate molecular targets for the treatment of CHF. LAY SUMMARY: Congenital hepatic fibrosis (CHF) is a genetic liver disease caused by mutations of the PKHD1 gene. Dysfunction of the protein it encodes, fibrocystin, is closely associated with CHF pathogenesis. Using an in vitro human induced pluripotent stem cell model and patient samples, we showed that the loss of fibrocystin function promotes proliferation of cholangiocytes and the production of connective tissue growth factor (CTGF) in an interleukin 8 (IL-8)-dependent manner. These results suggest that IL-8 and CTGF are essential for the pathogenesis of CHF.

LIM homeobox 2 promotes interaction between human iPS-derived hepatic progenitors and iPS-derived hepatic stellate-like cells.

Human induced pluripotent stem (iPS) cells can differentiate into hepatocyte lineages, although the phenotype of the differentiated cells is immature compared to adult hepatocytes. Improvement of cell-cell interactions between epithelium and mesenchyme is a potential approach to address this phenotype issue. In this study, we developed a model system for improving interactions between human iPS-derived hepatic progenitor cells (iPS-HPCs) and human iPS-derived hepatic stellate cell-like cells (iPS-HSCs). The phenotype of iPS-HSCs, including gene and protein expression profiles and vitamin A storage, resembled that of hepatic stellate cells. Direct co-culture of iPS-HSCs with iPS-HPCs significantly improved hepatocytic maturation in iPS-HPCs, such as their capacity for albumin production. Next, we generated iPS cell lines overexpressing LIM homeobox 2 (LHX2), which suppresses myofibroblastic changes in HSCs in mice. Hepatocytic maturation in iPS-HPCs was significantly increased in direct co-culture with iPS-HSCs overexpressing LHX2, but not in co-culture with a human hepatic stellate cell line (LX-2) overexpressing LHX2. LHX2 regulated the expression of extracellular matrices, such as laminin and collagen, in iPS-HSCs. In conclusion, this study provides an evidence that LHX2 upregulation in iPS-HSCs promotes hepatocytic maturation of iPS-HPCs, and indicates that genetically modified iPS-HSCs will be of value for research into cell-cell interactions.

Diabetic Retinopathy as a Risk Factor Associated with the Development of Hepatocellular Carcinoma in Nonalcoholic Fatty Liver Disease.

BACKGROUND: The risk factors associated with the development of hepatocellular carcinoma (HCC) in nonalcoholic fatty liver disease (NAFLD) are still unclear. The aim of the present study was to identify such risk factors in NAFLD patients who developed HCC. METHODS: Between April 2000 and -December 2016, a total of 182 patients with NAFLD were enrolled in this study; of these, only 22 patients had HCC. To identify risk factors, univariate and multivariate analyses were performed. To identify risk factors other than the degree of fibrosis, propensity matched analysis adjusted by the NAFLD fibrosis score (NFS) was carried out on 44 patients. Multivariate and survival analyses were also performed in HCC patients. RESULTS: In 182 patients, multivariate analysis highlighted the NFS (OR 2.275; p < 0.001) and hypertension (OR 5.868; p = 0.037) as independent factors that were significantly associated with the development of HCC. After adjustment for the NFS, multivariate analysis identified diabetic retinopathy (OR 8.654; p = 0.017) as an independent factor that was significantly associated with the development of HCC. For predicting the development of HCC, the area under the receiver operating characteristic curve of diabetic retinopathy was significantly higher than that of diabetes (0.731 vs. 0.615; p < 0.001). In patients with HCC, multivariate analysis indicated that the NFS were significantly associated with diabetic retinopathy. CONCLUSIONS: Diabetic retinopathy as well as liver fibrosis is a risk factor that associates with the development of HCC in NAFLD patients. Therefore, NAFLD patients with diabetic retinopathy should undergo careful screening for HCC.