Assessing seizure liability in vitro with voltage-sensitive dye imaging in mouse hippocampal slices

Non-clinical toxicology is a major cause of drug candidate attrition during development. In particular, drug-induced seizures are the most common finding in central nervous system (CNS) toxicity. Current safety pharmacology tests for assessing CNS functions are often inadequate in detecting seizure-inducing compounds early in drug development, leading to significant delays. This paper presents an in vitro seizure liability assay using voltage-sensitive dye (VSD) imaging techniques in hippocampal brain slices, offering a powerful alternative to traditional electrophysiological methods. Hippocampal slices were isolated from mice, and VSD optical responses evoked by stimulating the Schaffer collateral pathway were recorded and analyzed in the stratum radiatum (SR) and stratum pyramidale (SP). VSDs allow for the comprehensive visualization of neuronal action potentials and postsynaptic potentials on a millisecond timescale. By employing this approach, we investigated the in vitro drug-induced seizure liability of representative pro-convulsant compounds. Picrotoxin (PiTX; 1–100 μM), gabazine (GZ; 0.1–10 μM), and 4-aminopyridine (4AP; 10–100 μM) exhibited seizure-like responses in the hippocampus, but pilocarpine hydrochloride (Pilo; 10–100 μM) did not. Our findings demonstrate the potential of VSD-based assays in identifying seizurogenic compounds during early drug discovery, thereby reducing delays in drug development and providing insights into the mechanisms underlying seizure induction and the associated risks of pro-convulsant compounds

JWST Identification of Extremely Low C/N Galaxies with [N/O]≳0.5\gtrsim 0.5 at z∼6−10z\sim 6-10 Evidencing the Early CNO-Cycle Enrichment and a Connection with Globular Cluster Formation

We present chemical abundance ratios of 70 star-forming galaxies at $z\sim4$-10 observed by the JWST/NIRSpec ERO, GLASS, and CEERS programs. Among the 70 galaxies, we have pinpointed 2 galaxies, CEERS_01019 at $z=8.68$ and GLASS_150008 at $z=6.23$, with extremely low C/N ([C/N]$\lesssim -1$), evidenced with CIII]$\lambda\lambda$1907,1909, NIII]$\lambda$1750, and NIV]$\lambda\lambda$1483,1486, which show high N/O ratios ([N/O]$\gtrsim 0.5$) comparable with the one of GN-z11 regardless of whether stellar or AGN radiation is assumed. Such low C/N and high N/O ratios found in CEERS_01019 and GLASS_150008 (additionally identified in GN-z11) are largely biased towards the equilibrium of the CNO cycle, suggesting that these 3 galaxies are enriched by metals processed by the CNO cycle. On the C/N vs. O/H plane, these 3 galaxies do not coincide with Galactic HII regions, normal star-forming galaxies, and nitrogen-loud quasars with asymptotic giant branch stars, but globular-cluster (GC) stars, indicating a connection with GC formation. We compare C/O and N/O of these 3 galaxies with those of theoretical models, and find that these 3 galaxies are explained by scenarios with dominant CNO-cycle materials, i.e. Wolf-Rayet stars, supermassive ($10^{3}-10^{5}\ M_{\odot}$) stars, and tidal disruption events, interestingly with a requirement of frequent direct collapses. For all the 70 galaxies, we present measurements of Ne/O, S/O, and Ar/O, together with C/O and N/O. We identify 4 galaxies with very low Ne/O, $\log(\rm Ne/O)<-1.0$, indicating abundant massive ($\gtrsim30\ M_\odot$) stars.Comment: Accepted for publication in Ap

Inhibition of Hsp90 Leads to Cell Cycle Arrest and Apoptosis in Human Malignant Pleural Mesothelioma

IntroductionHeat shock protein 90 (Hsp90) is an abundant molecular chaperone that mediates the maturation and stability of a variety of proteins associated with the promotion of cell growth and survival. Inhibition of Hsp90 function leads to proteasomal degradation of its mis-folded client proteins. Recently, Hsp90 has emerged as being of prime importance to the growth and survival of cancer cells and its inhibitors have already been used in phase I and II clinical trials.MethodsWe investigated how 17-allylamino-17-demethoxygeldanamycin (17-AAG), a small molecule inhibitor of Hsp90, is implicated in human malignant pleural mesothelioma (MM).ResultsWe found that 17-AAG led to significant G1 or G2/M cell cycle arrest, inhibition of cell proliferation, and decrease of AKT, AKT1, and survivin expression in all human malignant pleural mesothelioma cell lines examined. We also observed significant apoptosis induction in all MM cell lines treated with 17-AAG. Furthermore, 17-AAG induced apoptosis in freshly cultured primary MM cells and caused signaling changes identical to those in 17-AAG treated MM cell lines.ConclusionThese results suggest that Hsp90 is strongly associated with the growth and survival of MM and that inhibition of Hsp90 may have therapeutic potential in the treatment of MM

EMPRESS. XIII. Chemical Enrichments of Young Galaxies Near and Far at z ~ 0 and 4-10: Fe/O, Ar/O, S/O, and N/O Measurements with Chemical Evolution Model Comparisons

We present gas-phase elemental abundance ratios of 7 local extremely metal-poor galaxies (EMPGs) including our new Keck/LRIS spectroscopy determinations together with 33 JWST $z\sim 4-10$ star-forming galaxies in the literature, and compare chemical evolution models. We develop chemical evolution models with the yields of core-collapse supernovae (CCSNe), Type Ia supernovae, hypernovae (HNe), and pair-instability supernovae (PISNe), and compare the EMPGs and high-$z$ galaxies in conjunction with dust depletion contributions. We find that high Fe/O values of EMPGs can (cannot) be explained by PISN metal enrichments (CCSN/HN enrichments even with the mixing-and-fallback mechanism enhancing iron abundance), while that the observed Ar/O and S/O values are much smaller than the predictions of the PISN models. The abundance ratios of the EMPGs can be explained by the combination of Type Ia SNe and CCSNe/HNe whose inner layers of argon and sulfur mostly fallback, which are comparable with Sculptor stellar chemical abundance distribution, suggesting that early chemical enrichment is taken place in the EMPGs. Comparing our chemical evolution models with the star-forming galaxies at $z\sim 4-10$, we find that the Ar/O and S/O ratios of the high-$z$ galaxies are comparable with those of the CCSNe/HNe models, while majority of the high-$z$ galaxies do not have constraints good enough to rule out contributions from PISNe. The high N/O ratio recently reported in GN-z11 cannot be explained even by rotating PISNe, but could be reproduced by the winds of rotating Wolf Rayet stars that end up as a direct collapse

Endoscopic Removal of Ingested Dentures and Dental Instruments: A Retrospective Analysis

Background. Dentures and dental instruments are frequently encountered ingested foreign bodies. The aim of the present study was to assess the safety and efficacy of endoscopically removing ingested dental objects. Methods. Twenty-nine consecutive patients with 29 dental objects who were treated at the Niigata University Medical and Dental Hospital from August 2009 to December 2015 were retrospectively reviewed. Characteristics of the patients and the ingested dental objects, the clinical features and findings of radiological imaging tests, and outcomes of endoscopic removal were analyzed. Results. Patients’ mean age was 62.9±21.0 years. The ingested dental objects included 23 dentures (13 crowns, 4 bridges, 4 partial dentures, and 2 other dentures) and 6 dental instruments. Twenty-seven upper gastrointestinal endoscopies and 2 colonoscopies were performed, and their success rates were 92.6% and 100%, respectively. There were 2 cases of removal failure; one case involved an impacted partial denture in the cervical esophagus, and this case required surgical removal. Conclusions. Endoscopic removal of ingested dentures and dental instruments is associated with a favorable success rate and acceptable complications. The immediate intervention and appropriate selection of devices are essential for managing ingested dental objects

DOCK2 is involved in the host genetics and biology of severe COVID-19

「コロナ制圧タスクフォース」COVID-19疾患感受性遺伝子DOCK2の重症化機序を解明 --アジア最大のバイオレポジトリーでCOVID-19の治療標的を発見--. 京都大学プレスリリース. 2022-08-10.Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge. Here we conducted a genome-wide association study (GWAS) involving 2, 393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3, 289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target