A rat model of human FENIB (familial encephalopathy with neuroserpin inclusion bodies)

金沢大学大学院医学系研究科脳細胞分子学FENIB (familial encephalopathy with neuroserpin inclusion bodies) is caused by intracellular accumulation/polymerization of mutant neuroserpins in the endoplasmic reticulum (ER). Transgenic rats overexpressing megsin (Tg meg), a newly identified serine protease inhibitor (serpin), demonstrated intraneuronal periodic-acid Schiff (PAS)-positive inclusions distributed throughout deeper layers of cerebral cortex, CA1 of the hippocampus, and substantia nigra. Hippocampal extracts from Tg meg rats showed increased expression of ER stress proteins, and activation of caspases-12 and -3, associated with decreased neuronal density. Enhanced ER stress was also observed in dopaminergic neurons in the substantia nigra, in parallel with decreased neuronal viability and motor coordination. In each case, PAS-positive inclusions were also positive for megsin. These data suggest that overexpression of megsin results in ER stress, eventuating in the formation of PAS-positive inclusions. Tg meg rats provide a novel model of FENIB, where accumulation of serpins in the ER induces selective dysfunction/loss of specific neuronal populations. © 2006 Elsevier Inc. All rights reserved

早期離床に対する看護師の認識と課題 －プロジェクトＦの活動を通じて－

早期離床を院内に定着させることを目標に，看護部でプロジェクトＦを発足させ活動を行った．まず，現状把握のため看護師538名に対し早期離床の認識についてのアンケート調査を行った．有効回答は428名（有効回答率79.6％）から得られた．その結果，離床援助技術の卒後の教育は不十分で，ほとんどの看護師が体位変換や移乗を困難であると感じていた．また，早期離床に関する医師の指示や離床に関する記録が統一されていないことがわかった．早期離床を妨げる要因として，記録などの多忙な業務や，痛みや循環変動，複数のラインなど患者の要因が挙げられた．看護師が行う早期離床の現実と理想との間に大きな乖離があり，現状に満足していない状況が示唆された．調査の結果から，「離床援助技術の教育」「早期離床フローチャートの策定と運用」「離床に関する記録整備」を課題とし，組織をあげて改善への取り組みを行った．その経緯についても加えて報告する．In order to establish early mobilization in the hospital, the "project F" was created in the Nursing Department and activities were started. A questionnaire was held regarding early wake-up and valid response were obtained from 428 out of 538 people. According to the results, factors such as busyness due to nursing records were one of the reasons that hindered this. As a result, post-graduate education for mobilization skills was inadequate, and most nurses felt it was difficult to change positions or transfer. It was also found that doctors' instructions regarding early mobilization, and records related to mobilization, were not unified. Factors that hindered early mobilization included diligent work such as nursing records, patient factors such as pain and circulatory fluctuations, and multiple lines. There was a big divergence between the reality, and the ideals of early mobilization performed by nurses, suggesting that they are not satisfied with the current situation. Based on the results of the survey, the issues were "education of mobilization assistance technology", "development and operation of early mobilization flowchart", and "maintenance of records concerning mobilization", and the entire organization worked on improvement. We will also report on the process

Comprehensive survey of p94/calpain 3 substrates by comparative proteomics – Possible regulation of protein synthesis by p94

Calpain represents a family of Ca2+-dependent cytosolic cysteine proteases found in almost all eukaryotes and some bacteria, and is involved in a variety of biological phenomena, including brain function. Several substrates of calpain are aggressively proteolyzed under pathological conditions, e.g., in neurodegenerating processes, fodrin is proteolyzed by calpain. Because very small amounts of substrate are proteolyzed by calpain under normal biological conditions, the molecular identities of calpain substrates are largely unknown. In this study, an extensive survey of the substrates of p94/calpain 3 in COS7 cells was executed using iTRAQ™ labeling and 2-D LC-MALDI analysis. p94 was used because: (i) several p94 splicing variants are expressed in brain tissue even though p94 itself is a skeletal-muscle-specific calpain, and (ii) it exhibits Ca2+-independent activity in COS cells, which makes it useful for evaluating the effects of p94 protease activity on proteins without perturbing the cells. Our approach revealed several novel protein substrates for p94, including the substrates of conventional calpains, components of the protein synthesis system, and enzymes of the glycolytic pathway. The results demonstrate the usefulness and sensitivity of this approach for mining calpain substrates. A combination of this method with other analytical methods would contribute to elucidation of the biological relevance of the calpain family

The whole blood transcriptional regulation landscape in 465 COVID-19 infected samples from Japan COVID-19 Task Force

「コロナ制圧タスクフォース」COVID-19患者由来の血液細胞における遺伝子発現の網羅的解析 --重症度に応じた遺伝子発現の変化には、ヒトゲノム配列の個人差が影響する--. 京都大学プレスリリース. 2022-08-23.Coronavirus disease 2019 (COVID-19) is a recently-emerged infectious disease that has caused millions of deaths, where comprehensive understanding of disease mechanisms is still unestablished. In particular, studies of gene expression dynamics and regulation landscape in COVID-19 infected individuals are limited. Here, we report on a thorough analysis of whole blood RNA-seq data from 465 genotyped samples from the Japan COVID-19 Task Force, including 359 severe and 106 non-severe COVID-19 cases. We discover 1169 putative causal expression quantitative trait loci (eQTLs) including 34 possible colocalizations with biobank fine-mapping results of hematopoietic traits in a Japanese population, 1549 putative causal splice QTLs (sQTLs; e.g. two independent sQTLs at TOR1AIP1), as well as biologically interpretable trans-eQTL examples (e.g., REST and STING1), all fine-mapped at single variant resolution. We perform differential gene expression analysis to elucidate 198 genes with increased expression in severe COVID-19 cases and enriched for innate immune-related functions. Finally, we evaluate the limited but non-zero effect of COVID-19 phenotype on eQTL discovery, and highlight the presence of COVID-19 severity-interaction eQTLs (ieQTLs; e.g., CLEC4C and MYBL2). Our study provides a comprehensive catalog of whole blood regulatory variants in Japanese, as well as a reference for transcriptional landscapes in response to COVID-19 infection

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

Synthesis of (zinc(II) phthalocyanine)-containing cellulose derivative using phthalocyanine-ring formation reaction

2, 3-Di-O-myristyl-6-O-(zinc(II) phthalocyaninyl) cellulose (5) was synthesized from cellulose (1) by five reaction steps via 6-O-(3′, 4′-dicyanophenyl)-2, 3-di-O-myristyl cellulose (4). The key reaction was phthalocyanine-ring formation on a cellulose backbone, that is, the reaction of compound 4 with o-phthalodinitrile in the presence of hexamethyldisilazane and zinc acetate in DMF afforded to compound 5 in 35.4 % yield. Consequently, the degree of substitution with phthalocyanine moieties of compound 5 was 0.38. The LB monolayer film of compound 5 on an indium tin oxide (ITO) electrode was found to show photocurrent generation performance at 680 nm

Protein kinase C-mediated inhibition of an inward rectifier potassium channel by substance P in nucleus basalis neurons

AbstractIn nucleus basalis neurons, substance P (SP) causes a slow excitation, mediated through a pertussis toxininsensitive G protein, by suppressing an inward rectifier K+ channel. Here we report that SP applied outside the patch pipette inhibited the single-channel activity, recorded on-cell, of the inward rectifier. The PKC inhibitors staurosporine and PKC(19–36) suppressed this effect in whole-cell mode and in on-cell single-channel mode. A diacylglycerol analog mimicked the SP effect, and PKC(19–36) suppressed this analog effect. SP irreversibly suppressed the inward rectifier in neurons treated with okadaic acid. These results indicate that a diffusible messenger mediates the SP effect, that its signal transduction involves phosphorylation by PKC, and that dephosphorylation by a serine/threonine protein phosphatase mediates its recovery

Facile synthesis of acyl chitosan isothiocyanates and their application to porphyrin-appended chitosan derivative.

Chitosan (1) was reacted with phenylisothiocyanate in 5% AcOH/H2O to give N-phenylthiocarbamoyl chitosan (2) with a degree of substitution (DS) of N-phenylthiocarbamoyl groups of 0.86 in 87.1% yield. The following acylation of compound 2 with hexanoyl chloride in the presence of pyridine afforded 3, 6-di-O-2, 3-hexanoyl chitosan isothiocyanate (4a) with a DS of the isothiocyanate groups of 0.70 in high yield, unexpectedly. Compound 4a exhibited high levels of reactivity toward various amines to give the corresponding N-thiocarbamoyl chitosan derivatives in high yields. Other acyl (decanoyl (4b), myristroyl (4c), stearoyl (4d), benzoyl (4e)) chitosan isothiocyanates were also prepared from chitosan (1) in high yields. To evaluate the potential applications of acyl chitosan isothiocyanates, N-(triphenylporphynyl)thiocarbamoyl chitosan derivative 6 with a DS of the triphenylporphynyl groups of 0.46 was prepared from compound 4b. The Langmuir-Blodgett monolayer film of compound 6 gave a good photon-to-electron conversion performance