Kobe Institute Of Computing

Abstract Chronic hepatitis B (HBV) and hepatitis D (HDV) viral infections pose serious global health challenges, and the lack of curative therapies calls for the development of new antiviral approaches. Recently, we have identified the Kinesin Family Member 4 (KIF4) as a crucial regulator of the surface transport for NTCP (HBV/HDV entry receptor). Our research has shown that the pan-RXR agonist, Bexarotene, suppresses KIF4 expression and inhibits HBV/HDV infections. However, the clinical use of Bexarotene is limited by poor physicochemical and pharmacokinetic (PK) properties and significant side effects. Here we investigated chemically diverse RXR modulators with tuned activity, and identified the RXR agonist S169 as a novel HBV/HDV entry inhibitor. S169 effectively blocks HBV and HDV infections in primary human hepatocyte (PXB) cultures without evident cytotoxicity. S169’s suppressive effect on HBV infection was confirmed by reduction of secreted HBsAg and HBeAg, as well as inhibition of HBV cccDNA, pregenomic RNA, and intracellular HBV DNA. Mechanistically, S169 suppressed FOXM1-mediated KIF4 expression, leading to decreased NTCP surface levels and a marked decrease in HBV preS1 peptide binding to the hepatocyte cell surface. S169 is reported to exhibit enhanced oral bioavailability, favorable PK parameters, lower toxicity compared to Bexarotene. Interestingly, S169 induced a modest inhibition of NTCP-mediated bile acid transport up to 10 μM, which remained unchanged at higher concentrations. Furthermore, S169 significantly impeded HBV spread in HBV infected long-time cultured PHHs (PXB). We speculate that S169 can be a promising seed for the development of novel oral HBV and HDV antiviral therapeutics.

Abstract Background The uncultured adipose-derived stromal vascular fraction (SVF), consisting of adipose-derived stromal cells (ADSCs), M2 macrophages (M2Φ) and others, has shown therapeutic potential against osteoarthritis (OA), however, the mechanisms underlying its therapeutic effects remain unclear. Therefore, this study investigated the effects of the SVF on OA in a human–immunodeficient rat xenotransplantation model. Methods OA model was induced in the knees of female immunodeficient rats by destabilization of the medial meniscus. Immediately after the surgery, human SVF (1 × 105), ADSCs (1 × 104), or phosphate buffered saline as a control group were transplanted into the knees. At 4 and 8 weeks postoperatively, OA progression and synovitis were analyzed by macroscopic and histological analyses, and the expression of collagen II, SOX9, MMP-13, ADAMTS-5, F4/80, CD86 (M1), CD163 (M2), and human nuclear antigen (hNA) were evaluated immunohistochemically. In vitro, flow cytometry was performed to collect CD163-positive cells as M2Φ from the SVF. Chondrocyte pellets (1×105) were co-cultured with SVF (1×105), M2Φ (1×104), and ADSCs (1×104) or alone as a control group, and the pellet size was compared. TGF-β, IL-10 and MMP-13 concentrations in the medium were evaluated using enzyme-linked immunosorbent assay. Results In comparison with the control and ADSC groups, the SVF group showed significantly slower OA progression and less synovitis with higher expression of collagen II and SOX9, lower expression of MMP-13 and ADAMTS-5, and lower F4/80 and M1/M2 ratio in the synovium. Only the SVF group showed partial expression of hNA-, CD163-, and F4/80-positive cells in the rat synovium. In vitro, the SVF, M2Φ, ADSC and control groups, in that order, showed larger pellet sizes, higher TGF-β and IL-10, and lower MMP-13 concentrations. Conclusions The M2Φ in the transplanted SVF directly affected recipient tissue, enhancing the secretion of growth factors and chondrocyte-protecting cytokines, and partially improving chondrocytes and joint homeostasis. These findings indicate that the SVF is as an effective option for regenerative therapy for OA, with mechanisms different from those of ADSCs.

Societal Impact StatementThe research explores the complex interaction between flowering plants and their pollinators, specifically focusing on the genus Arisaema. Unlike most plants, Arisaema has a distinctive trait in which killing pollinators can be beneficial. Traditionally, this interaction has been viewed as highly antagonistic because it appears to favor the plants at the expense of the pollinators. However, new evidence reveals that a pollinator uses the lethal floral trap of Arisaema thunbergii as a nursery. Remarkably, some individuals probably even escape from the trap after laying eggs. This finding challenges the prevailing notion that deceptive pollination is the sole outcome in Arisaema, a genus known for its intricate lethal pollination mechanisms.Summary While many flowering plants engage in mutualistic relationships with their pollinators, flower‐insect interactions are not always mutually beneficial. Some plants exploit their pollinators by deceptively promising rewards without delivering them. Among such plants, the genus Arisaema (Araceae) is notable for its antagonistic behavior towards pollinators, as female plants invariably entrap them with lethal effects. Here we tested the hypothesis that the succulent appendix of some Arisaema species, such as Arisaema thunbergii, may serve as a food source for pollinator larvae, leading to the possibility of nursery pollination. To investigate this, we retrieved adult insect corpses trapped within the spathes of A. thunbergii. Subsequently, we incubated the spathes after collecting the corpses to determine if conspecific insects emerged from them. We present novel evidence that one of the main pollinating fungus gnats, Leia ishitanii, uses the lethal floral trap of A. thunbergii inflorescences as a nursery. Astonishingly, some L. ishitanii individuals may even succeed in escaping from the female spathe after depositing their eggs. These findings challenge the conventional understanding that deceptive pollination is the norm within the genus Arisaema. The interaction between A. thunbergii and L. ishitanii, in which a significant portion of female adults die inside but still reproduction seems successful, suggests an intermediate stage between brood‐site deception and nursery pollination mutualism.

Adult T-cell leukemia/lymphoma (ATL) is caused by a retrovirus human T-cell leukemia virus type 1 (HTLV-1). Its oncogenes Tax and HBZ play essential roles in malignant initiation and progression of ATL However, in tumor microenvironment (TME), the HIF-1 and carbonic anhydrase IX (CA9) have been suggested to be rather important for finalization of its complicated oncogenic processes of ATL.This review aims to refine up the three-step oncogenesis model of ATL based on the HIF-1/CA9 axis and clarify the interaction of AKT and NF-B with the HIF/CA9 axis and other factors to finalize its three-step oncogenesis.In TME, HIF-1/HIF-1 has multiple functions and activates several signaling pathways, including AKT/mTOR, NF-B, CA9 and others.In turn, AKT and NF-B activate translation and transcription of HIF-1 mRNA, respectively.AP-1 and other factors also can contribute to finalization of the ATL three-step oncogenesis.To clarify detailed mechanisms, further studies are required.

Abstract Purpose: To investigate the detailed relationship between high wall shear stress (WSS) and plaque rupture (PR) in longitudinal and circumferential locations.Methods: Overall, 100 acute coronary syndrome (ACS) patients whose culprit lesions had PR documented by optical coherence tomography (OCT) were enrolled. Lesion-specific three-dimensional coronary artery models were created using OCT data. At the ruptured portion, tracing of the luminal edge of the residual fibrous cap was smoothly extrapolated to reconstruct the luminal contour before PR. Then, WSS was computed from computational fluid dynamics analysis. PR was classified into central-PR and lateral-PR according to the disrupted fibrous cap location.Results: In the longitudinal 3-mm segmental analysis, multivariate analysis demonstrated that higher WSS in the upstream segment was independently associated with Upstream-PR and a thinner fibrous cap was independently associated with Downstream-PR. In PR cross-sections, PR region had a significantly higher average WSS than the non-PR region. In the cross-sectional analysis, peak WSS was most frequently observed in the lateral region (66.7%) in lateral-PR, whereas that in central-PR was most frequently observed in the central region (70%). Multivariate analysis demonstrated that the presence of peak WSS at the lateral region, thinner broken fibrous cap, and larger lumen area were independently associated with lateral-PR, while the presence of peak WSS at the central region and thicker broken fibrous cap were independently associated with central-PR.Conclusions: A combined approach with computational fluid dynamics simulation and morphological plaque evaluation by OCT might help predict future PR-induced ACS events.

Abstract BackgroundRich vascularity of the induced membrane (IM) is important for Masquelet reconstruction surgery. The factors affecting IM vascularity are not completely understood. This study aimed to investigate these factors using histological samples of human IMs.MethodsWe retrospectively evaluated 36 patients whose bone defects were treated using the Masquelet technique. Two clinical pathologists analyzed histological sections of IM pieces (1-cm2). The number of blood vessels per 1 mm2 was counted and compared among men and women, femur or tibia, with and without free flap surgery, antibiotic impregnation to the cement, osteogenesis inside the membrane, smoking, and diabetes mellitus. The number of blood vessels within the same patient was compared among different time points. Correlation analysis was performed among blood vessel numbers and patient age, duration of cement spacer placement, and histological grading scales (inflammation, foreign body reaction, and fibrosis).ResultsIM formation with rich vascularity and some inflammation, foreign body reaction, and fibrosis was histologically confirmed in all patients. We found 37.4 ± 19.1 blood vessels per 1 mm2. Blood vessel numbers were significantly lower in patients with than in those without free flap surgery. Blood vessel numbers were higher in patients with osteogenesis inside the IM. No significant correlations were found in any of the analyses.ConclusionSex, patient age, smoking, diabetes mellitus, femur or tibia, duration of cement spacer placement, and antibiotic impregnation to the cement did not affect IM vascularization. IM vascularization was reduced in patients with than in those without free flap surgery.

Abstract Background: The gut microbiome plays an important role in the immune system and has attracted attention as a new biomarker of various diseases, including cancer. As such, in this study, we examined the relationship between the gut microbiome and lung cancer progression. In addition, we assessed the correlation between the gut microbiome and epidermal growth factor receptor (EGFR) mutation status.Methods: Female never-smokers diagnosed with lung adenocarcinoma were consecutively enrolled between May 2018 and August 2019. Fecal samples were collected within 1 month before or after diagnosis and before administration of any lung cancer treatment. Principal coordinate analyses were performed using Bray-Curtis distance matrices to investigate the effects of clinical variables (age, body mass index, Tumor-Node-Metastasis stage, T category, N category, M category, primary tumor size, performance status, and EGFR mutation status) on the gut microbial community. A correlation analysis was also performed to determine the strength of association between the dominant taxonomy (comprising ≥1% of the relative abundance of bacterial DNA sequences) and clinical variables.Results: A total of 37 patients were enrolled. T category and primary tumor size were significantly correlated with the gut microbial community (p=0.018 and 0.041, respectively). At the genus level, a significant positive correlation was observed between the relative abundance of Faecalibacterium and both T category (correlation coefficient, R=0.51, p=0.0013) and primary tumor size (R=0.37, p=0.024), whereas a significant negative correlation was observed between the relative abundances of Fusicatenibacter and Bacteroides and T category (R=−0.35, p=0.034 and R=−0.32, p=0.05, respectively) and primary tumor size (R=−0.36, p=0.029 and R=−0.41, p=0.012, respectively). EGFR mutation status had no statistically significant effect on the gut microbial community (p=0.11). However, the relative abundances of Bifidobacterium and Faecalibacterium were significantly higher in EGFR mutation–negative patients than EGFR mutation–positive patients (p=0.012 and 0.041), whereas the relative abundance of Blautia was significantly lower in EGFR mutation–negative patients (p=0.036).Conclusions: This is the first study identifying the gut microbiome as a promising biomarker of lung cancer progression. Further elucidation of the role of the gut microbiome in lung cancer progression could facilitate development of new treatments for lung cancer.

Abstract Background Frailty is associated with morbidity and mortality in patients in intensive care units (ICUs). However, the characteristics of frail patients with suspected infection, including sepsis, remain unclear. We aimed to investigate the characteristics and outcomes of frail patients with suspected infection in ICUs. Methods This is a secondary analysis of a multicenter cohort study conducted by 22 ICUs in Japan. Adult patients (≥ 16 years) with newly suspected infection from December 2017 to May 2018 were included. We compared baseline patient characteristics and outcomes between three frailty groups based on the Clinical Frailty Scale (CFS) score: fit (score 1–3), vulnerable (score 4), and frail (score 5–9). We conducted subgroup analysis of patients with sepsis defined based on Sepsis-3 criteria. We also produced Kaplan–Meier survival curves for 90-day survival. Results We enrolled 650 patients with suspected infection, including 599 (92.2%) patients with sepsis. Patients with a median rating of 3 (3–5) on the CFS were included: 337 (51.8%) were fit, 109 (16.8%) were vulnerable, and 204 were (31.4%) frail. The median age of the patients was 72 years (IQR 60 − 81). The Sequential Organ Failure Assessment scores for fit, vulnerable, and frail patients were 7 (4–10), 8 (5–11), and 7 (5–10), respectively (p = 0.59). The patients' median body temperatures were as follows: fit 37.5 (36.5–38.5)℃; vulnerable 37.5 (36.4–38.6)℃; and frail 37.0 (36.3–38.1)℃ (p < 0.01). C-reactive protein levels of fit, vulnerable, and frail patients were 13.6 (4.6–24.5), 12.1 (3.9–24.9), 10.5 (3.0–21.0) mg/dL, respectively (p < 0.01). In-hospital mortality did not statistically differ according to frailty (p = 0.19). Kaplan–Meier survival curves showed little difference in the mortality rate during the acute phase. However, more vulnerable and frail patients died after the acute phase than fit patients; this difference was not statistically significant (p = 0.25). Compared with fit or vulnerable patients, fewer frail patients were discharged. Conclusion Frail and vulnerable patients with suspected infection, including sepsis, tend to have poor disease outcomes after the acute phase of infection.