ZBP1 activation triggers hematopoietic stem and progenitor cell death resulting in bone marrow failure in mice.

Human bone marrow failure (BMF) syndromes result from the loss of hematopoietic stem and progenitor cells (HSPC), and this loss has been attributed to cell death; however, the cell death triggers, and mechanisms remain unknown. During BMF, tumor necrosis factor-α (TNFα) and interferon-γ (IFNγ) increase. These ligands are known to induce necroptosis, an inflammatory form of cell death mediated by RIPK1, RIPK3, and MLKL. We previously discovered that mice with a hematopoietic RIPK1 deficiency (Ripk1HEM KO) exhibit inflammation, HSPC loss, and BMF, which is partially ameliorated by a RIPK3 deficiency; however, whether RIPK3 exerts its effects through its function in mediating necroptosis or other forms of cell death remains unclear. Here, we demonstrate that similar to a RIPK3 deficiency, an MLKL deficiency significantly extends survival and like Ripk3 deficiency partially restores hematopoiesis in Ripk1HEM KO mice revealing that both necroptosis and apoptosis contribute to BMF in these mice. Using mouse models, we show that the nucleic acid sensor Z-DNA binding protein 1 (ZBP1) is up-regulated in mouse RIPK1-deficient bone marrow cells and that ZBP1's function in endogenous nucleic acid sensing is necessary for HSPC death and contributes to BMF. We also provide evidence that IFNγ mediates HSPC death in Ripk1HEM KO mice, as ablation of IFNγ but not TNFα receptor signaling significantly extends survival of these mice. Together, these data suggest that RIPK1 maintains hematopoietic homeostasis by preventing ZBP1 activation and induction of HSPC death.

Prognostic heterogeneity and clonal dynamics within distinct subgroups of myelodysplastic syndrome and acute myeloid leukemia with TP53 disruptions.

TP53 aberrations constitute the highest risk subset of myelodysplastic neoplasms (MDS) and acute myeloid leukemia (AML). The International Consensus Classification questions the blast threshold between MDS and AML. In this study, we assess the distinction between MDS and AML for 76 patients with TP53 aberrations. We observed no significant differences between MDS and AML regarding TP53 genomics. Median overall survival (OS) was 223 days for the entire group, but prognostic discrimination within subgroups showed the most inferior OS (46 days) for AML with multihit allelic state plus TP53 variant allele frequency (VAF) > 50%. In multivariate analysis, unadjusted Cox models revealed the following variables as independent risk factors for mortality: AML (vs. MDS) (hazard ratio [HR]: 2.50, confidence interval [CI]: 1.4-4.4, p = 0.001), complex karyotype (HR: 3.00, CI: 1.4-6.1, p = 0.003), multihit status (HR: 2.30, CI 1.3-4.2, p = 0.005), and absence of hematopoietic cell transplant (HCT) (HR: 3.90, CI: 1.8-8.9, p = 0.0009). Clonal dynamic modeling showed a significant reduction in TP53 VAF with front-line hypomethylating agents. These findings clarify the impact of specific covariates on outcomes of TP53-aberrant myeloid neoplasms, irrespective of the diagnosis of MDS versus AML, and may influence HCT decisions.

Thermal stress, hydration, and salivary and respiratory stress markers in curling players performing a match in the cold.

Curling is a target-based team sport played in a cold environment. The type of stress curling players face during a curling match remains to be determined. In the present study, 16 Japanese curling players performed a practice curling match (six ends lasting 90 min), wherein the following variables were documented: core and skin temperatures, heart rate, thermal sensation and comfort, urine-specific gravity, body fluid loss, salivary cortisol, α-amylase activity, salivary secretory immunoglobulin A (SIgA), and fractionated exhaled nitric oxide (FeNO, a respiratory stress marker). Pre-match resting core temperature was 37.24 ± 0.31°C, which increased up to 37.73 ± 0.41°C during the match (p < 0.001). Facial skin temperatures decreased after the match (all p ≤ 0.015), whereas finger skin temperatures remained unchanged (p ≥ 0.375). Thermal discomfort increased following the match but thermal sensation remained unchanged. Following the match, players lost 0.29 ± 0.15 L body fluid (sweat, respiratory evaporation, and urine), which was nearly compensated by fluid ingestion of 0.22 ± 0.13 L (p = 0.119). Nevertheless, urine-specific gravity increased from 1.021 ± 0.010 to 1.024 ± 0.008 after the match (p = 0.012), with 31% and 50% players being dehydrated at pre- and post-match, respectively. Salivary cortisol decreased (p < 0.001) after the match without changes in salivary SIgA, α-amylase activity, and FeNO (all p ≥ 0.113). Therefore, during a curling match, the core temperature and thermal discomfort increase, whereas the face skin temperature decreases. Additionally, players may undergo dehydration before the match, which could be exacerbated after the match.

Safety and tolerability of a multilineage-differentiating stress-enduring cell-based product in neonatal hypoxic-ischaemic encephalopathy with therapeutic hypothermia (SHIELD trial): a clinical trial protocol open-label, non-randomised, dose-escalation trial.

INTRODUCTION: Neonatal hypoxic-ischaemic encephalopathy (HIE) is an important illness associated with death or cerebral palsy. This study aims to assess the safety and tolerability of the allogenic human multilineage-differentiating stress-enduring cell (Muse cell)-based product (CL2020) cells in newborns with HIE. This is the first clinical trial of CL2020 cells in neonates. METHODS AND ANALYSIS: This is a single-centre, open-label, dose-escalation study enrolling up to 12 patients. Neonates with HIE who receive a course of therapeutic hypothermia therapy, which cools to a body temperature of 33°C-34°C for 72 hours, will be included in this study. A single intravenous injection of CL2020 cells will be administered between 5 and 14 days of age. Subjects in the low-dose and high-dose cohorts will receive 1.5 and 15 million cells per dose, respectively. The primary outcome is the occurrence of any adverse events within 12 weeks after administration. The main secondary outcome is the Bayley Scales of Infant and Toddler Development Third Edition score and the developmental quotient per the Kyoto Scale of Psychological Development 2001 at 78 weeks. ETHICS AND DISSEMINATION: This study will be conducted in accordance with the Declaration of Helsinki and Good Clinical Practice. The Nagoya University Hospital Institutional Review Board (No. 312005) approved this study on 13 November 2019. The results of this study will be published in peer-reviewed journal and reported in international conferences. TRIAL REGISTRATION NUMBERS: NCT04261335, jRCT2043190112.

The 10th Japan Bioanalysis Forum Symposium conference: a report.

The Japan Bioanalysis Forum Symposium was held on 12-14 February 2019 (Yokohama, Japan), in celebration of its 10th anniversary, and over 370 participants from pharmaceutical industries, contractors, academia and regulatory authorities from home and abroad came together in Yokohama. The 3-day symposium particularly aimed to foster collaboration with the scientists surrounding bioanalysts, according to the theme 'Open to the Public.' The symposium also included a broad range of pioneering programs, such as lectures by speakers from DMPK/metabolomics fields, discussions of future bioanalysis and poster presentations by publicly offered presenters as well as the regular ones we had organized. This report summarizes the major topics as a conference report.

Verification of recall criteria for masses detected on ultrasound breast cancer screening.

PURPOSE: Mammography is the only modality for breast cancer screening demonstrated to reduce the mortality rate. However, ultrasonographic screening is already being widely performed as opportunistic screening in Japan. The recall criteria for masses are very important as quality controls. The purpose of this study was to verify these criteria at multiple institutions. METHODS: Screening was performed by five institutions in various regions in Japan. The total number of cases screened at all five institutions was 10,519. RESULTS: The findings that could be concluded to be benign were a cystic pattern and three features of a solid pattern. The cystic pattern was noted in 6512 cases, typical fibroadenoma in 1483 cases, and typical complicated cyst in 70 cases. Only three of these 8065 cases were cancers, so the negative predictive value was 99.9%. The solid pattern with obvious malignant features, i.e., masses with an echogenic halo and/or interruption of the interface and masses with multiple echogenic foci, were noted in 33 cases. Twenty of the 33 cases were malignancy, resulting in a positive predictive value of 66.7%. CONCLUSION: Although some parts of the criteria should be considered further for verification and revision, the current recall criteria are mostly valid.

Impairment of fetal hematopoietic stem cell function in the absence of Fancd2.

Fanconi anemia (FA) results from mutations in the genes necessary for DNA damage repair and often leads to progressive bone marrow failure. Although the exhaustion of the bone marrow leads to cytopenias in FA patients as they age, evidence from human FA and mouse model fetal livers suggests that hematopoietic defects originate in utero, which may lead to deficient seeding of the bone marrow. To address this possibility, we examined the consequences of loss of Fancd2, a central component of the FA pathway. Examination of embryonic day 14.5 (E14.5) Fancd2 knockout (KO) fetal livers showed a decrease in total cellularity and specific declines in long-term and short-term hematopoietic stem cell (LT-HSC and ST-HSC, respectively) numbers. Fancd2 KO fetal liver cells display similar functional defects to Fancd2 adult bone marrow cells, including reduced colony-forming units, increased mitomycin C sensitivity, increased LT-HSC apoptosis, and heavily impaired competitive repopulation, implying that these defects are intrinsic to the fetal liver and are not dependent on the accumulation of DNA damage during aging. Telomere shortening, an aging-related mechanism proposed to contribute to HSC apoptosis and bone marrow failure in FA, was not observed in Fancd2 KO fetal livers. In summary, loss of Fancd2 yields significant defects to fetal liver hematopoiesis, particularly the HSC population, which mimics key phenotypes from adult Fancd2 KO bone marrow independently of aging-accrued DNA damage.

Crystal structures of a subunit of the formylglycinamide ribonucleotide amidotransferase, PurS, from Thermus thermophilus, Sulfolobus tokodaii and Methanocaldococcus jannaschii.

The crystal structures of a subunit of the formylglycinamide ribonucleotide amidotransferase, PurS, from Thermus thermophilus, Sulfolobus tokodaii and Methanocaldococcus jannaschii were determined and their structural characteristics were analyzed. For PurS from T. thermophilus, two structures were determined using two crystals that were grown in different conditions. The four structures in the dimeric form were almost identical to one another despite their relatively low sequence identities. This is also true for all PurS structures determined to date. A few residues were conserved among PurSs and these are located at the interaction site with PurL and PurQ, the other subunits of the formylglycinamide ribonucleotide amidotransferase. Molecular-dynamics simulations of the PurS dimer as well as a model of the complex of the PurS dimer, PurL and PurQ suggest that PurS plays some role in the catalysis of the enzyme by its bending motion.

Anodal transcranial direct current stimulation over the auditory cortex improved hearing impairment in a patient with brainstem encephalitis.

Transcranial direct current stimulation (tDCS) can alter cortical excitability, and has been effective in treating some neurological disorders. This case report describes the use of tDCS in a 13-year-old female who developed bilateral hearing impairment after brainstem encephalitis when she was 6 years old. Her auditory function was more impaired in her right ear than her left. Anodal stimulation (1 mA) was applied for 10 min to the left auditory cortex once per day for 4 consecutive days to improve her right ear speech discrimination score. Sustained and significant improvement in maximum speech discrimination was observed after the four tDCS treatments. To our knowledge, this is the first case report of improvement in speech discrimination after anodal stimulation of the auditory cortex. These results encourage further studies investigating the beneficial effects of tDCS in patients with hearing impairments.