Search

Diffuse intrinsic pontine glioma (DIPG) remains a fatal brainstem tumor demanding innovative therapies. As B7-H3 (CD276) is expressed on central nervous system (CNS) tumors, we designed B7-H3–specific chimeric antigen receptor (CAR) T cells, confirmed their preclinical efficacy, and opened BrainChild-03 (NCT04185038), a first-in-human phase I trial administering repeated locoregional B7-H3 CAR T cells to children with recurrent/refractory CNS tumors and DIPG. Here, we report the results of the first three evaluable patients with DIPG (including two who enrolled after progression), who received 40 infusions with no dose-limiting toxicities. One patient had sustained clinical and radiographic improvement through 12 months on study. Patients exhibited correlative evidence of local immune activation and persistent cerebrospinal fluid (CSF) B7-H3 CAR T cells. Targeted mass spectrometry of CSF biospecimens revealed modulation of B7-H3 and critical immune analytes (CD14, CD163, CSF-1, CXCL13, and VCAM-1). Our data suggest the feasibility of repeated intracranial B7-H3 CAR T-cell dosing and that intracranial delivery may induce local immune activation.Significance:This is the first report of repeatedly dosed intracranial B7-H3 CAR T cells for patients with DIPG and includes preliminary tolerability, the detection of CAR T cells in the CSF, CSF cytokine elevations supporting locoregional immune activation, and the feasibility of serial mass spectrometry from both serum and CSF.This article is highlighted in the In This Issue feature, p. 1

Supplementary Data from Intraventricular B7-H3 CAR T Cells for Diffuse Intrinsic Pontine Glioma: Preliminary First-in-Human Bioactivity and Safety

Background Immune responses to the SARS-CoV-2 vaccination may be influenced by immunomodulatory drugs (IMDs). We investigated the immune responses and safety in fully vaccinated Japanese patients with IBD. Methods IBD patients and control subjects at 39 institutes were invited to participate in the study from March to October 2021. Blood sample collections to measure anti-SARS-CoV-2 spike IgG antibody titers were planned pre-1st vaccination, pre-2nd vaccination, and at 4 weeks, 3 months and 6 months post-2nd vaccination. Immune responses were compared between groups, considering baseline characteristics and IMD treatments. (UMIN000043545) The interim analyses presented here include mainly data from the 4-weeks post-2nd vaccination time-point. Results In total, 679 IBD patients and 203 controls were enrolled (Table 1). The IBD group received the BNT162b2 vaccine (86.2%) and the mRNA-1273 vaccine (12.5%), and the control group received the BNT162b2 vaccine (86.9%) and the mRNA-1273 vaccine (12.1%). Only 4 cases (0.7%) in the IBD group and 2 (1.0%) in the control group were infected with COVID-19. Adverse events of 2nd vaccination occurred in 48.4% of the IBD group and 35.1% of the control group. Comparison between administrated and non-administrated IBD patients for each IMD revealed an attenuated genomic mean titer (GMT [U/mL]) in those taking systemic steroids (18.85 vs 31.24), anti-TNF monotherapy (28.31 vs 42.99), anti-TNF therapy+ immunomodulator (IM) (12.86 vs 35.26), vedolizumab+IM (19.49 vs 30.39), ustekinumab+IM (20.44 vs 30.79), and tofacitinib (9.54 vs 32.08), but not in those taking oral 5-ASA (29.50 vs 32.40), or vedolizumab (41.85 vs 40.20) and ustekinumab (55.56 vs 39.26) monotherapies. Estimated least square means of the GMT by a multiple linear regression model are shown in Table 2. GMTs were significantly influenced by increasing age and allergy (51.2, 95%CI 42.1–62.3; p=0.0293), and tended to be influenced by COVID-19 infection (139.1, 41.0–472.2; p=0.0572). Sex, smoking, drinking, IBD, and adverse events of 2nd vaccination did not affect the GMT. The GMT was significantly higher for mRNA-1273 (90.3 [60.8–134.1]) than for BNT162b2 (39.6 [35.2–44.6], p= 0.0001). Systemic steroids (22.9 [13.9–37.7], p=0.0119), IM (24.2 [18.7–31.4], p Conclusion Aging and most IMD options attenuated immunogenicity in fully vaccinated IBD patients. Prioritization of a booster vaccination should be considered for IBD patients treated with IMDs.

Figure S1. ISX activated transcription of IDOs, AHR, CD86, and PD-L1 in hepatoma cells by binding to the promoter region. a, p < 0.001. A, ISX transcriptionally activated luciferase activity driven by IDO1, TDO2 B, PD-L1 and CD86 C, and AHR D, promoter in Huh7 and SK-Hep1 cells. Indicated deletion luciferase mutants were constructed as described in the Materials and Methods. E, ChIP analysis of ISX''s binding to the promoters of IDO1, TDO2 and AHR in SK-Hep1 cells as described in Supplemental Materials and Methods. a, p

Supplementary Information

Figure S5, A, Western blots analysis of IDOs, AHR, PD-L1 and CD86 protein expression in various hepatoma cells. CNL: Chang normal liver cells. B, The Kaplan-Meier survival curve analysis between HCC patients with low and high expression of CD80.

Figure S3, A, Western blotting analysis of ISX, IDO1, TDO2, KYNU, and CYP1B1proteins in hepatocytes of AhR-null mice with treated proinflammatory cytokine IL-6 (10 ng/ml) B, The levels of kynurenine in hepatocytes of AHR+/+, AHR+/â^'and AHR-/â^' mice were determined by ELISA. C, Western blotting analysis of ISX, IDO1 and TDO2 proteins in tumors of mice transplanted with AHR or ISX-knockdown cells. D, Hepatoma cell (Huh 7) transfected with ISX-GFP showed to increase the concentration of kynurenine and PD-L1 in the culture medium when co-cultured with cytotoxic CD8+ T cells (1x104). The opposite effect was observed when Huh 7 cells transfected with ISX-specific shRNAi. E, ISX expression in Huh 7 cells significantly suppressed the proliferation of cytotoxic CD8+ T cells induced by anti-CD3 antibody. F, Huh 7 cells (1x104) overexpressing ISX-GFP significantly abolished cell apoptotic activity of spleen cytotoxic CD8+ T cell and the opposite effects were observed when Huh 7 cells were treated with anti-PD-1 monoclonal antibody or IDO inhibitor (INCB-024360). G, Huh 7 cells (1x104) overexpressing ISX-GFP significantly abolished cell apoptotic activity induced by co-cultured spleen cytotoxic CD8+ T cell and the opposite effects were observed when Huh 7 cells were treated with anti-PD-1 monoclonal antibody or IDO inhibitor (INCB-024360). a, p

Figure S2, AHR signals positively activated ISX expression in hepatoma cells. A, Relative mRNA expression levels of CYP1B1 and ISX in Hep 3B and SK-Hep1 cells with AHR overexpression as detected by semi-quantitative RT-PCR. Data are presented as means {plus minus} S.D. a, p < 0.001. B, Relative mRNA expression levels of CYP1B1 and ISX in Huh 7 and SK-Hep1 cells with treated with AHR ligand, TCDD (5 nM) were detected by semi-quantitative RT-PCR. C, Western blotting analysis of ISX proteins in SK-Hep1 cells transfected with AHR, ARNT or AHR-ARNT complex D, AHR transcriptionally activated luciferase activity driven by promoters of ISX in SK-Hep1 cells. Relative luciferase activity was calculated as described in the Supplemental Materials and Methods. E, Western blotting analysis of ISX, IDOs, KYNU and downstream gene, CYP 1B1 proteins in Huh 7 and SK-Hep1 cells transfected with knockdown vector shRNAi against AHR. F, Schematic presentation of a series of luciferase deletions of ISX promoter. Relative luciferase activity in SK-Hep1 was calculated as described in the Materials and methods section. G, Chromatin was prepared and immunoprecipitated with anti-AHR antibody from SK-Hep1 and Huh7 cells treated with TCDD. H, The promoter regions of ISX with AHR-binding element (XRE) deletion were not pulled down by the anti-AHR antibody. I, Western blotting analysis of ISX, IDO1, IDO2, CYP1B1 and TDO2 expression in hepatocytes of AHR+/+, AHR+/â^'and AHR-/â^' mice.

We prepared three-dimensional (3-D) organoids of human stomach cancers and examined the correlation between the tumorigenicity and cytotoxicity of Helicobacter pylori (H. pylori). In addition, the effects of hepatoma-derived growth factor (HDGF) and tumor necrosis factor (TNFα) on the growth and invasion activity of H. pylori-infected gastric cancer organoids were examined. Cytotoxin-associated gene A (CagA)-green fluorescence protein (GFP)-labeled H. pylori was used to trace the infection in gastric organoids. The cytotoxicity of Cag encoded toxins from different species of H. pylori did not affect the proliferation of each H. pylori-infected cancer organoid. To clarify the role of HDGF and TNFα secreted from H. pylori-infected cancer organoids, we prepared recombinant HDGF and TNFα and measured the cytotoxicity and invasion of gastric cancer organoids. HDGF controlled the growth of each organoid in a species-specific manner of H. pylori, but TNFα decreased the cell viability in H. pylori-infected cancer organoids. Furthermore, HDGF controlled the invasion activity of H. pylori-infected cancer organoid in a species-dependent manner. However, TNFα decreased the invasion activities of most organoids. We found different signaling of cytotoxicity and invasion of human gastric organoids in response to HDGF and TNFα during infection by H. pylori. Recombinant HDGF and TNFα inhibited the development and invasion of H. pylori-infected gastric cancer differently. Thus, we propose that HDGF and TNFα are independent signals for development of H. pylori-infected gastric cancer. The signaling of growth factors in 3-D organoid culture systems is different from those in two-dimensional cancer cells.

The intestine-specific homeobox transcription factor intestine-specific homeobox (ISX) is an IL6-inducible proto-oncogene implicated in the development of hepatocellular carcinoma, but its mechanistic contributions to this process are undefined. In this study, we provide evidence that ISX mediates a positive feedback loop integrating inflammation, tryptophan catabolism, and immune suppression. We found that ISX-mediated IL6-induced expression of the tryptophan catabolic enzymes Indoleamine 2,3-dioxygenase 1 (IDO1) and tryptophan 2,3-dioxygenase in hepatocellular carcinoma cells, resulting in an ISX-dependent increase in the tryptophan catabolite kynurenine and its receptor aryl hydrocarbon receptor (AHR). Activation of this kynurenine/AHR signaling axis acted through a positive feedback mechanism to increase ISX expression and enhance cellular proliferation and tumorigenic potential. RNAi-mediated attenuation of ISX or AHR reversed these effects. In an IDO1-dependent manner, ectopic expression of ISX induced expression of genes encoding the critical immune modulators CD86 (B7-2) and programmed death ligand-1 (PD-L1), through which ISX conferred a significant suppressive effect on the CD8+ T-cell response. In hepatocellular carcinoma specimens, expression of IDO1, kynurenine, AHR, and PD-L1 correlated negatively with survival. Overall, our results identified a feed-forward mechanism of immune suppression in hepatocellular carcinoma organized by ISX, which involves kynurenine-AHR signaling and PD-L1, offering insights into immune escape by hepatocellular carcinoma, which may improve its therapeutic management. Cancer Res; 77(15); 4065–77. ©2017 AACR.

We show that the Priess-Crampe & Ribenboim fixed point theorem is provable in R C A 0 . Furthermore, we show that Caristi’s fixed point theorem for both Baire and Borel functions is equivalent to the transfinite leftmost path principle, which falls strictly between A T R 0 and Π 1 1 - C A 0 . We also exhibit several weakenings of Caristi’s theorem that are equivalent to W K L 0 and to A C A 0 . This article is part of the theme issue ‘Modern perspectives in Proof Theory’.

Koji mold, which belongs to the Aspergillus section Nigri, is used in the production of shochu. The section Nigri is composed of very morphologically similar members that in some cases produce mycotoxins, which rises concerns as to whether the presence of mycotoxin-producing fungi in shochu producing sites can compromise consumer safety. Thus, we examined the presence of mycotoxin-producing sec. Nigri fungi in six shochu factories (named A-F) in Japan. Airborne fungal levels in the factories were determined, and a traditional koji called "kona-koji" made from the mold naturally present in factory C (Aogashima village) was analyzed. Isolates of sec. Nigri fungi were identified morphologically and confirmed via cytochrome b gene analysis. In factory A (Nago city), airborne fungal levels of sec. Nigri were 4,000 and 100 cfu/m3 in the koji-making and fermentation rooms, respectively. In factories B, C, and D, the levels were 40, >104 cfu/m3, and 100 cfu/m3, respectively. In factory F (Iki city), there were high levels of airborne white-koji mold (a white mutant of Asp. luchuensis). The most dominant fungal species of sec. Nigri was isolated and identified as Asp. luchuensis via genetic analysis. This is likely to have originated from the commercial fermentation culture used. Asp. niger and Asp. luchuensis were isolated from kona-koji. Mycotoxin production (ochratoxin and fumonisin B2) by Asp. luchuensis (eight strains) and Asp. niger (three strains) was virtually inexistent; only one strain of Asp. niger was positive for fumonisin B2. This study clearly shows that mycotoxin-producing fungi are not dominant in the fungal flora present in the shochu factories examined and therefore, that the liquor can be safely fermented.Implications: In this study, we examined the presence of mycotoxin-producing Aspergillus sec. Nigri fungi in six shochu (Japanese distilled beverage) factories. The most dominant fungal species of sec. Nigri was isolated and identified as Aspergillus luchuensis (black-koji mold). The proportion of mycotoxin-producing Aspergillus niger and Aspergillus carbonarius was very small. In addition, the Asp. niger isolated from koji mold did not have the ability to produce ochratoxins or fumonisin B. This study clearly shows that shochu can be safely fermented.

It is postulated as a general concept of cancer stem cells (CSCs) that they can produce cancer cells overtly and repopulate cancer progenitor cells indefinitely. The CSC niche is part of a specialized cancer microenvironment that is important to keep the phenotypes of CSCs. Stem cell- and induced pluripotent stem cell (iPSC)-derived organoids with genetic manipulation are beneficial to the investigation of the regulation of the microenvironment of CSCs. It would be useful to assess the efficiency of the cancer microenvironment on initiation and progression of cancers. To identify CSCs in cancer tissues, normal cell organoids and gastric cancer organoids from the cancerous areas, as well as iPSCs, were established several years ago. However, many questions remain about the extent to which these cultures recapitulate the development of the gastrointestinal tract and the mechanism of Helicobacter pylori-induced cancer progression. To clarify the fidelity of human organoid models, we have noted several key issues for the cultivation of, and differences between, normal and cancerous organoids. We developed precise culture conditions for gastric organoids in vitro to improve the accuracy of the generation of organoid models for therapeutic and medical applications. In addition, the current knowledge on gastrointestinal CSC research, including the topic of CSC markers, cancer cell reprogramming, and application to target cancer cell plasticity through niches, should be reinforced. We discuss the progression of cancers derived from human gastric organoids and the identification of CSCs.

Locoregional delivery of chimeric antigen receptor (CAR) T cells has resulted in objective responses in adults with glioblastoma, but the feasibility and tolerability of this approach is yet to be evaluated for pediatric central nervous system (CNS) tumors. Here we show that engineering of a medium-length CAR spacer enhances the therapeutic efficacy of human erb-b2 receptor tyrosine kinase 2 (HER2)-specific CAR T cells in an orthotopic xenograft medulloblastoma model. We translated these findings into BrainChild-01 ( NCT03500991 ), an ongoing phase 1 clinical trial at Seattle Children’s evaluating repetitive locoregional dosing of these HER2-specific CAR T cells to children and young adults with recurrent/refractory CNS tumors, including diffuse midline glioma. Primary objectives are assessing feasibility, safety and tolerability; secondary objectives include assessing CAR T cell distribution and disease response. In the outpatient setting, patients receive infusions via CNS catheter into either the tumor cavity or the ventricular system. The initial three patients experienced no dose-limiting toxicity and exhibited clinical, as well as correlative laboratory, evidence of local CNS immune activation, including high concentrations of CXCL10 and CCL2 in the cerebrospinal fluid. This interim report supports the feasibility of generating HER2-specific CAR T cells for repeated dosing regimens and suggests that their repeated intra-CNS delivery might be well tolerated and activate a localized immune response in pediatric and young adult patients. In an interim analysis of a phase 1 trial, repeated intracranial infusions of HER2-specific CAR T cells were well tolerated with no observed dose-limiting toxicities in three young adult patients with CNS tumors.

Background The short-term effects of teduglutide (TED) for short bowel syndrome with chronic intestinal failure (SBS–IF) in patients with Crohn’s disease (CD) remain unknown and real-world data have not been scarce. This study aimed to investigate the short-term efficacy and safety of TED in CD patients on parenteral support (PS) for SBS–IF. Methods We retrospectively investigated the medical records of CD patients with SBS–IF who initiated TED between January 2020 and October 2022 in Hyogo Medical University hospital. The primary outcomes were the change in PS volume and proportion of patients with a reduction of PS volume by ≥20% at week 4. Secondary outcomes were the changes in body mass index (BMI), estimated glomerular filtration rate (eGFR), albumin, hematocrit (Ht), PS calorie requirements, withdrawal from PS, and adverse events during the observation period. Results Of 605 CD patients, 23 who underwent home PS for SBS–IF were included in this study. Twenty patients continued TED throughout the observation period. The median PS duration was 11.6 (range 1.0-20.0) years and the observation period after starting TED was 48.0 (7.0-60.0) weeks. TED significantly reduced the PS volume from 17989(570-49000) mL/week to 14257(0-42000) mL/week at week 4 (p = 0.0013), and the PS volume decreased by ≥20% in 8 patients (40.0%) at week 4 and in 16 patients (80.0%) during the observation period after TED administration. The BMI significantly increased from 17.9 (12.9–23.9) kg/m2 before TED administration and 18.9 (14.6–25.4) kg/m2 after TED administration (p = 0.014). Although there were no significant differences in albumin (p = 0.53), eGFR (p = 0.66), and Ht (p = 0.089), the PS calorie /week was significantly decreased from 6581 (244–14000) kcal to 3747(0– 8820) kcal after TED administration (p = 0.0001). Ten patients (50.0%) experienced gastrointestinal stoma complications, including stoma swelling and/or prolapse during the observation period. Abdominal pain occurred in 7 cases (35.0%), catheter-related infection in five patients (25.0%), oedema in 4 patients (20.0%), nausea in 2 patients (10.0%), abdominal distension in 3 patients (15.0%), and upper respiratory tract infection in 1 patient (5.0%). All the symptoms were transient and tolerable. Although 3 patients discontinued TED due to nausea and abdominal pain and were excluded from this analysis, these symptoms improved after the discontinuation. Conclusion TED reduced PS volume at week 4 in CD patients with SBS–IF, and the BMI and the PS calorie requirement were improved during the observational period without serious adverse events.

Photoexcited muon spin spectroscopy (photo- μSR) is used to study excess charge carrier lifetimes in silicon. Experiments are performed on silicon wafers with very high bulk lifetimes with the surface passivation conditions intentionally modified to control the effective lifetime. When the effective lifetime is low (10 ms) results in detectable degradation of the measured lifetime. Re-passivation of degraded samples with a temporary room temperature superacid-based passivation scheme demonstrates that degradation occurs in the silicon bulk. Deep-level transient spectroscopy measurements reveal the existence of several defect-related traps near the muon-exposed surface in concentrations of order 1010 cm−3 that are not present near the surface not exposed to muons. In contrast to the common perception of the μSR technique, our results demonstrate that muons are not inert probes and that beam-induced recombination activity modifies the bulk lifetime significantly in samples with high effective carrier lifetimes.

Diffuse intrinsic pontine glioma (DIPG) remains a fatal brainstem tumor demanding innovative therapies. As B7-H3 (CD276) is expressed on central nervous system (CNS) tumors, we designed B7-H3–specific chimeric antigen receptor (CAR) T cells, confirmed their preclinical efficacy, and opened BrainChild-03 (NCT04185038), a first-in-human phase I trial administering repeated locoregional B7-H3 CAR T cells to children with recurrent/refractory CNS tumors and DIPG. Here, we report the results of the first three evaluable patients with DIPG (including two who enrolled after progression), who received 40 infusions with no dose-limiting toxicities. One patient had sustained clinical and radiographic improvement through 12 months on study. Patients exhibited correlative evidence of local immune activation and persistent cerebrospinal fluid (CSF) B7-H3 CAR T cells. Targeted mass spectrometry of CSF biospecimens revealed modulation of B7-H3 and critical immune analytes (CD14, CD163, CSF-1, CXCL13, and VCAM-1). Our data suggest the feasibility of repeated intracranial B7-H3 CAR T-cell dosing and that intracranial delivery may induce local immune activation. Significance: This is the first report of repeatedly dosed intracranial B7-H3 CAR T cells for patients with DIPG and includes preliminary tolerability, the detection of CAR T cells in the CSF, CSF cytokine elevations supporting locoregional immune activation, and the feasibility of serial mass spectrometry from both serum and CSF. This article is highlighted in the In This Issue feature, p. 1

The aryl hydrocarbon receptor (AhR) is a ligand-binding protein that responds to environmental aromatic hydrocarbons and stimulates the transcription of downstream phase I enzyme–related genes by binding the cis element of dioxin-responsive elements (DREs)/xenobiotic-responsive elements. Dimethyl sulfoxide (DMSO) is a well-known organic solvent that is often used to dissolve phase I reagents in toxicology and oxidative stress research experiments. In the current study, we discovered that 0.1% DMSO significantly induced the activation of the AhR promoter via DREs and produced reactive oxygen species, which induced apoptosis in mouse embryonic fibroblasts (MEFs). Moreover, Jun dimerization protein 2 (Jdp2) was found to be required for activation of the AhR promoter in response to DMSO. Coimmunoprecipitation and chromatin immunoprecipitation studies demonstrated that the phase I–dependent transcription factors, AhR and the AhR nuclear translocator, and phase II–dependent transcription factors such as nuclear factor (erythroid-derived 2)–like 2 (Nrf2) integrated into DRE sites together with Jdp2 to form an activation complex to increase AhR promoter activity in response to DMSO in MEFs. Our findings provide evidence for the functional role of Jdp2 in controlling the AhR gene via Nrf2 and provide insights into how Jdp2 contributes to the regulation of ROS production and the cell spreading and apoptosis produced by the ligand DMSO in MEFs. Graphical abstract

Nine domestic wine samples collected from a Japanese winery were examined for the presence of fumonisin B1 (FB1), fumonisin B2 (FB2), and fumonisin B3 (FB3), as well as ochratoxin A (OTA) and ochratoxin B (OTB). Wine samples spiked with 13C-labeled internal standards (13C34-FB1 and 13C20-OTA) were diluted with phosphate buffered saline (PBS) buffer, loaded on immunoaffinity cartridges to purify of fumonisins and ochratoxins, and subjected to liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis. The data revealed that the domestic wine samples were possibly contaminated with FB1 and FB3, in addition to FB2, whereas none of the tested wine samples were contaminated with OTA and OTB. These results suggest that Fusarium fungi can be associated with the fumonisin contamination of Japanese domestic wine, whereas Aspergillus niger seems to be frequently reported as the major causal fungus of fumonisin contamination of wine in Europe. Analysis of the intermediate samples during the wine brewing indicated that fumonisin concentrations did not increase during wine production, suggesting that fumonisin contamination did not occur during the brewing process, but was derived from the raw materials of grape berries.

Background and study aims: This study evaluated the long- term outcomes of mainly endoscopic hemostatic therapy for gastrointestinal variceal bleeding and of the transition of hemostatic therapy. Patients and methods: Among 1,163 patients treated for gastrointestinal varices between April 2006 and June 2020, a total of 125 patients who underwent emergency hemostatic therapy were enrolled. Survival rates and secondary evaluation points were analyzed. Additionally, patients were classified into two groups: the previous and latter term. Patients’ background, therapeutic method, and treatment results were compared between the groups. Results: 94.4% had cirrhosis. The average Child-Pugh score was 8.90. Successful primary hemostasis rate was 98.4%, and 5.6% died within 2 weeks, all with a Child-Pugh score ≥9. The respective 1- and 5-year survival rates for Child-Pugh grade A/B were 81.3% and 55.4%, while those for Child-Pugh grade C were 58.1% and 17.8%. Child-Pugh grade C or hepatocellular carcinoma was significantly associated with poor prognosis. In total, 21.6% experienced variceal re-bleeding; 62.9% of these cases were triggered by continued alcohol consumption. There was no significant difference in survival between patients with and without variceal re-bleeding and in post-treatment survival between the previous and latter terms. In the latter term, the number of cases caused by continued alcohol consumption significantly increased. Conclusions: Multidisciplinary treatment and continuation of proper management after hemostatic therapy for variceal bleeding are crucial. Continued alcohol consumption leads to variceal bleeding and re-bleeding; its proper management, including alcohol abstinence, is one of the major challenges left in the post-direct- acting antivirals era.

V/A-ATPase is a motor protein that shares a common rotary catalytic mechanism with FoF1 ATP synthase. When powered by ATP hydrolysis, the V1 domain rotates the central rotor against the A3B3 hexamer, composed of three catalytic AB dimers adopting different conformations (ABopen, ABsemi, and ABclosed). Here, we report the atomic models of 18 catalytic intermediates of the V1 domain of V/A-ATPase under different reaction conditions, determined by single particle cryo-EM. The models reveal that the rotor does not rotate immediately after binding of ATP to the V1. Instead, three events proceed simultaneously with the 120˚ rotation of the shaft: hydrolysis of ATP in ABsemi, zipper movement in ABopen by the binding ATP, and unzipper movement in ABclosed with release of both ADP and Pi. This indicates the unidirectional rotation of V/A-ATPase by a ratchet-like mechanism owing to ATP hydrolysis in ABsemi, rather than the power stroke model proposed previously for F1-ATPase.

Stomach cancer has a high mortality, which is partially caused by an absence of suitable biomarkers to allow detection of the initiation stages of cancer progression. Thus, identification of critical biomarkers associated with gastric cancer (GC) is required to advance its clinical diagnoses and treatment. Recent studies using tracing models for lineage analysis of GC stem cells indicate that the cell fate decision of the gastric stem cells might be an important issue for stem cell plasticity. They include leucine-rich repeat-containing G-protein-coupled receptor 5 (Lgr5+), Cholecystokinin receptor 2 (Cckr2+), and axis inhibition protein 2 (Axin2+) as the stem cell markers in the antrum, Trefoil Factor 2 (TFF2+), Mist1+ stem cells, and Troy+ chief cells in the corpus. By contrast, Estrogen receptor 1 (eR1), Leucine-rich repeats and immunoglobulin-like domains 1 (Lrig1), SRY (sex determining region Y)-box 2 (Sox2), and B lymphoma Mo-MLV insertion region 1 homolog (Bmi1) are rich in both the antrum and corpus regions. These markers might help to identify the cell-lineage identity and analyze the plasticity of each stem cell population. Thus, identification of marker genes for the development of GC and its environment is critical for the clinical application of cancer stem cells in the prevention of stomach cancers.

There is considerable cellular diversity in the human stomach, which has helped to clarify cell plasticity in normal development and tumorigenesis. Thus, the stomach is an interesting model for understanding cellular plasticity and for developing prospective anticancer therapeutic agents. However, many questions remain regarding the development of cancers in vivo and in vitro in two- or three-dimensional (2D/3D) cultures, as well as the role of Helicobacter pylori (H. p.) infection. Here, we focus on the characteristics of cancer stem cells and their derived 3D organoids in culture, including the formation of stem cell niches. We define the conditions required for such organoid culture in vitro and examine the ability of such models for testing the use of anticancer agents. We also summarize the signaling cascades and the specific markers of stomach-cancer-derived organoids induced by H. p. infection, and their stem cell niches.

In the present work we demonstrate that C-doped Zr$_{5}$Pt$_{3}$ is an electron-phonon superconductor (with critical temperature T$_\mathrm{C}$ = 3.7\,K) with a nonsymmorphic topological Dirac nodal-line semimetal state, which we report here for the first time. The superconducting properties of Zr$_{5}$Pt$_{3}$C$_{0.5}$ have been investigated by means of magnetization and muon spin rotation and relaxation ($\mu$SR) measurements. We find that at low temperatures the depolarization rate is almost constant and can be well described by a single-band $s-$wave model with a superconducting gap of $2\Delta(0)/k_\mathrm{B}T_\mathrm{C}$ = 3.84, close to the value of BCS theory. From transverse field $\mu$SR analysis we estimate the London penetration depth $\lambda_{L}$ = 469 nm, superconducting carrier density $n_{s}$ = 2$\times$10$^{26}$ $m^{-3}$, and effective mass m$^{*}$ = 1.584 $m_{e}$. Zero field $\mu$SR confirms the absence of any spontaneous magnetic moment in the superconducting ground state. To gain additional insights into the electronic ground state of C-doped Zr$_5$Pt$_3$, we have also performed first-principles calculations within the framework of density functional theory (DFT). The observed homogenous electronic character of the Fermi surface as well as the mutual decrease of $T_\mathrm{C}$ and density of states at the Fermi level are consistent with the experimental findings. However, the band structure reveals the presence of robust, gapless fourfold-degenarate nodal lines protected by $6_{3}$ screw rotations and glide mirror planes. Therefore, Zr$_5$Pt$_3$ represents a novel, unprecedented condensed matter system to investigate the intricate interplay between superconductivity and topology., Comment: 9 pages, 5 figures

V/A-ATPase is a motor protein that shares a common rotary catalytic mechanism with F

Recently, there have been contrary claims of Kitaev spin-liquid behavior and ordered behavior in the honeycomb compound ${\mathrm{Ag}}_{3}\mathrm{Li}{\mathrm{Ir}}_{2}{\mathrm{O}}_{6}$ based on various experimental signatures. Our investigations on this system reveal a low-temperature ordered state with persistent dynamics down to the lowest temperatures. Magnetic order is confirmed by clear oscillations in the muon spin relaxation ($\ensuremath{\mu}\mathrm{SR}$) time spectrum below 9 K until 52 mK. Coincidentally in $^{7}\mathrm{Li}$ nuclear magnetic resonance, a wipeout of the signal is observed below $\ensuremath{\sim}10$ K, which again strongly indicates magnetic order in the low-temperature regime. This is supported by our density functional theory calculations which show an appreciable Heisenberg exchange term in the spin Hamiltonian that favors magnetic ordering. The $^{7}\mathrm{Li}$ shift and spin-lattice relaxation rate also show anomalies at $\ensuremath{\sim}50$ K. They are likely related to the onset of dynamic magnetic correlations, but their origin is not completely clear. Detailed analysis of our $\ensuremath{\mu}\mathrm{SR}$ data is consistent with a coexistence of incommensurate N\'eel and striped environments. A significant and undiminished dynamical relaxation rate ($\ensuremath{\sim}5$ MHz) as seen in $\ensuremath{\mu}\mathrm{SR}$ deep into the ordered phase indicates enhanced quantum fluctuations in the ordered state.

Simple Summary Tumor suppressor genes are critical in the control of many biological functions. They can be classified based on their roles in proliferation, cell-cycle progression, DNA repair/damage, and crucial signaling functions, including apoptosis, autophagy, and necrosis. The absence of functional tumor suppressor genes entails a higher risk of dysfunction of cell growth, differentiation, cell death, and cancer development. Loss of function or mutations of such genes has been identified in many types of cancer, such as breast, bladder, colorectal, head and neck, lung, ovarian, uterine, and pancreatic cancers. Familial cancer syndromes, such as Li–Fraumeni syndrome, are associated with loss of TP53 function. Extensive studies have been carried out to clarify the roles of the products of these genes, as well as their mechanistic link to cancers, to identify novel targets for specific cancer types. Here, we introduce the roles of tumor suppressor gene products in pancreatic cancer development and its therapeutics for tumorigenesis prevention. Abstract The high mortality of pancreatic cancer is attributed to the insidious progression of this disease, which results in a delayed diagnosis and advanced disease stage at diagnosis. More than 35% of patients with pancreatic cancer are in stage III, whereas 50% are in stage IV at diagnosis. Thus, understanding the aggressive features of pancreatic cancer will contribute to the resolution of problems, such as its early recurrence, metastasis, and resistance to chemotherapy and radiotherapy. Therefore, new therapeutic strategies targeting tumor suppressor gene products may help prevent the progression of pancreatic cancer. In this review, we discuss several recent clinical trials of pancreatic cancer and recent studies reporting safe and effective treatment modalities for patients with advanced pancreatic cancer.

Ni is one of the important metal resources. Because Ni-containing waste liquid is drained after several plating turns in the factories, an effective recycling technique should be developed. A unique magnetic separation technique using high temperature superconducting bulk magnet has succeeded in collecting Ni-sulfate crystals, which were fabricated from the Ni-plating waste liquid. Pulsed-field magnetizing method was employed to activate the bulk magnet up to 2.80 T, which produced a field space of 1.40 T on the surface of the waste channel. Green coarse crystals were attracted from the flowing stream of Ni-saturated liquid containing weakly-magnetic particles of Ni-compounds. The magnetically-collected particles were identified as paramagnetic NiSO4/6H2O crystals, and slight differences in Ni concentration and grain size were observed between the particles attracted and not-attracted to the 1.8 T magnetic pole. In both cases, the large grains were found to consist of a single phase. The compound can be used as a raw material in the Ni-recycle process. This preferential extraction suggests a novel recycling method of Ni resource.

Background The cerebellum is the sensitive region of the brain to developmental abnormalities related to the effects of oxidative stresses. Abnormal cerebellar lobe formation, found in Jun dimerization protein 2 (Jdp2)-knockout (KO) mice, is related to increased antioxidant formation and a reduction in apoptotic cell death in granule cell progenitors (GCPs). Here, we aim that Jdp2 plays a critical role of cerebellar development which is affected by the ROS regulation and redox control. Objective Jdp2-promoter-Cre transgenic mouse displayed a positive signal in the cerebellum, especially within granule cells. Jdp2-KO mice exhibited impaired development of the cerebellum compared with wild-type (WT) mice. The antioxidation controlled gene, such as cystine-glutamate transporter Slc7a11, might be critical to regulate the redox homeostasis and the development of the cerebellum. Methods We generated the Jdp2-promoter-Cre mice and Jdp2-KO mice to examine the levels of Slc7a11, ROS levels and the expressions of antioxidation related genes were examined in the mouse cerebellum using the immunohistochemistry. Results The cerebellum of Jdp2-KO mice displayed expression of the cystine-glutamate transporter Slc7a11, within the internal granule layer at postnatal day 6; in contrast, the WT cerebellum mainly displayed Sla7a11 expression in the external granule layer. Moreover, development of the cerebellar lobes in Jdp2-KO mice was altered compared with WT mice. Expression of Slc7a11, Nrf2, and p21Cip1 was higher in the cerebellum of Jdp2-KO mice than in WT mice. Conclusion Jdp2 is a critical regulator of Slc7a11 transporter during the antioxidation response, which might control the growth, apoptosis, and differentiation of GCPs in the cerebellar lobes. These observations are consistent with our previous study in vitro.

${\mathrm{Ba}}_{4}{\mathrm{Ru}}_{3}{\mathrm{O}}_{10}$, the quasi-one-dimensional spin-1 ($S$ = 1) compound, has rather intricate magnetic properties. The compound consists of structural Ru trimers, which together form the zig-zag chains where the Ru has two inequivalent crystallographic sites. While at high temperature both the inequivalent Ru ions stay in the $4+$ state with effective $S=$ 1 spin state, upon lowering the temperature, the magnetic moment of the central Ru atom is completely quenched accompanied by a change in the octahedral environment. This effectively gives rise to a bond-alternating chain and provides an opportunity to study the excitation of such a spin network in a real material. We have used microscopic tools such as neutron scattering and muon spin relaxation along with the density functional theory based calculations to address the spin state of the two inequivalent Ru ions in this material. From our neutron powder diffraction, on lowering of temperature, we find a large tetragonal distortion of the central ${\mathrm{RuO}}_{6}$ octahedra of the trimer. The splitting of the ${t}_{2g}$ level of the central Ru due to this distortion is found to be significant leading to the quenching of the moment underscoring the Hund's exchange. The nonmagnetic central Ru promotes a strong antiferromagnetic superexchange between the other two Ru ions in the trimer, which gives rise to a dimeric state. The presence of spin dimers is reflected by the manifestation of a gap in the spin excitation spectra. The spin-dimer formation in ${\mathrm{Ba}}_{4}{\mathrm{Ru}}_{3}{\mathrm{O}}_{10}$ is at par with the effective model proposed by Affleck and Haldane for the $S$ = 1 bond alternating chains in the light of valence bond solid formalism. Eventually, at a lower temperature, a long-range ordered antiferromagnetic state emerges from the gapped dimer state due to the significant interdimer interactions.

In the present work, we demonstrate that C-doped Zr

We propose that beyond its role in WNT secretion, WLS/GPR177 (wntless, WNT ligand secretion mediator) acts as an essential regulator controlling protein glycosylation, endoplasmic reticulum (ER) homeostasis, and dendritic cell (DC)-mediated immunity. WLS deficiency in bone marrow-derived DCs (BMDCs) resulted in poor growth and an inability to mount cytokine and T-cell responses in vitro, phenotypes that were irreversible by the addition of exogenous WNTs. In fact, WLS was discovered to integrate a protein complex in N-glycan-dependent and WLS domain-selective manners, comprising ER stress sensors and lectin chaperones. WLS deficiency in BMDCs led to increased ER stress response and macroautophagy/autophagy, decreased calcium efflux from the ER, and the loss of CALR (calreticulin)-CANX (calnexin) cycle, and hence protein hypo-glycosylation. Consequently, DC-specific wls-null mice were unable to develop both Th1-, Th2- and Th17-associated responses in the respective autoimmune and allergic disease models. These results suggest that WLS is a critical chaperone in maintaining ER homeostasis, glycoprotein quality control and calcium dynamics in DCs. Abbreviations: ATF6: activating transcription factor 6; ATG5: autophagy related 5; ATG12: autophagy related 12; ATG16L1: autophagy related 16 like 1; ATP2A1/SERCA1: ATPase sarcoplasmic/endoplasmic reticulum Ca2+ transporting 1; BALF: bronchoalveolar lavage fluid; BFA: brefeldin A; BMDC: bone marrow-derived dendritic cell; CALR: calreticulin; CANX: calnexin; CCL2/MCP-1: C-C motif chemokine ligand 2; CNS: central nervous system; CT: C-terminal domain; DTT: dithiothreitol; DNAJB9/ERDJ4: DnaJ heat shock protein family (Hsp40) member B9; EAE: experimental autoimmune encephalomyelitis; EIF2A/eIF2α: eukaryotic translation initiation factor 2A; EIF2AK3/PERK: eukaryotic translation initiation factor 2 alpha kinase 3; ERN1/IRE1: endoplasmic reticulum (ER) to nucleus signaling 1; GFP: green fluorescent protein; HSPA5/GRP78/BiP: heat shock protein A5; IFNA: interferon alpha; IFNAR1: interferon alpha and beta receptor subunit 1; IFNB: interferon beta; IFNG/INFγ: interferon gamma; IFNGR2: interferon gamma receptor 2; IL6: interleukin 6; IL10: interleukin 10; IL12A: interleukin 12A; IL23A: interleukin 23 subunit alpha; ITGAX/CD11c: integrin subunit alpha X; ITPR1/InsP3R1: inositol 1,4,5-trisphosphate receptor type 1; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; OVA: ovalbumin; PIK3C3/VPS34: phosphatidylinositol 3-kinase catalytic subunit type 3; PLF: predicted lipocalin fold; PPP1R15A/GADD34: protein phosphatase 1 regulatory subunit 15A; RYR1/RyanR1: ryanodine receptor 1, skeletal muscle; SD: signal domain; TGFB/TGF-β: transforming growth factor beta family; Th1: T helper cell type 1; Th17: T helper cell type 17; TM: tunicamycin; TNF/TNF-α: tumor necrosis factor; UPR: unfolded protein response; WLS/wntless: WNT ligand secretion mediator.

Induced pluripotent stem cells (iPSCs) are derived from various types of cell reprogramming methodologies. iPSCs are associated with a risk of tumorigenesis because they exhibit the capabilities of self-renewal, hyperproliferation, and plasticity. Moreover, pluripotency-inducing genes are expressed in not only the stem cells but also in a variety of cancer cells. Cancer initiation is mainly caused by the mutations in oncogenes and tumor-suppressor genes during the conversion of normal cells, somatic PSCs, and iPSCs to cancer cells or cancer stem cells (CSCs). Thus, the potential risks of tumorigenesis should be increased when CSCs-like cells are transplanted to the bodies of patients. Several methods and systems have been developed for the effective prevention of tumor formation from PSCs, cancer cells, and their derivatives. This chapter discusses the recent research progress in the undoubted correlation between reprogramming and cancer initiation from the standpoints of genetic, epigenetic, cellular, and microenvironmental alterations. Novel technologies aimed at the prevention of the risks of tumorigenesis in PSCs and cancer cells will shed new light on the development of anticancer drugs and regenerative medicine.

The use of biomarkers in cancer diagnosis, therapy, and prognosis has been highly effective over several decades. Studies of biomarkers in cancer patients pre- and post-treatment and during cancer progression have helped identify cancer stem cells (CSCs) and their related microenvironments. These analyses are critical for the therapeutic application of drugs and the efficient targeting and prevention of cancer progression, as well as the investigation of the mechanism of the cancer development. Biomarkers that characterize CSCs have thus been identified and correlated to diagnosis, therapy, and prognosis. However, CSCs demonstrate elevated levels of plasticity, which alters their functional phenotype and appearance by interacting with their microenvironments, in response to chemotherapy and radiotherapeutics. In turn, these changes induce different metabolic adaptations of CSCs. This article provides a review of the most frequently used CSCs and stem cell markers.