Dynamic Regulation of Extracellular Signal-Regulated Kinase (ERK) by Protein Phosphatase 2A Regulatory Subunit B56γ1 in Nuclei Induces Cell Migration

Extracellular signal-regulated kinase (ERK) signalling plays a central role in various biological processes, including cell migration, but it remains unknown what factors directly regulate the strength and duration of ERK activation. We found that, among the B56 family of protein phosphatase 2A (PP2A) regulatory subunits, B56γ1 suppressed EGF-induced cell migration on collagen, bound to phosphorylated-ERK, and dephosphorylated ERK, whereas B56α1 and B56β1 did not. B56γ1 was immunolocalized in nuclei. The IER3 protein was immediately highly expressed in response to costimulation of cells with EGF and collagen. Knockdown of IER3 inhibited cell migration and enhanced dephosphorylation of ERK. Analysis of the time course of PP2A-B56γ1 activity following the costimulation showed an immediate loss of phosphatase activity, followed by a rapid increase in activity, and this activity then remained at a stable level that was lower than the original level. Our results indicate that the strength and duration of the nuclear ERK activation signal that is initially induced by ERK kinase (MEK) are determined at least in part by modulation of the phosphatase activity of PP2A-B56γ1 through two independent pathways. © 2013 Kawahara et al

Feasibility of Pilates for Late-Stage Frail Older Adults to Minimize Falls and Enhance Cognitive Functions

Globally, we are facing the tendency of aging, and demands for health enhancement among the older population have been steadily increasing. Among various exercise interventions, Pilates has been popularly utilized in rehabilitation; therefore, it is considered suitable for vulnerable populations. In this study, frail late-stage older adults (>75 years) participated in a modified Pilates program (30 min per session, once a week for eight weeks). Age-and condition-matched Controls were also involved as the benchmark to reveal the effect of Pilates. While only the Pilates group participated in the exercise intervention, both groups undertook the health assessments twice (before and after the intervention period). Assessments included: (i) falling risk based on 3D motion capture systems and (ii) overall cognitive functions utilizing Mini-Mental State Examination and executive function with the use of Trail Making Test-A (TMT-A). Two-dimensional mood state was also used to measure changes in mood due to Pilates intervention. An 8-week Pilates intervention was effective in achieving higher and symmetrical swing foot control. Dynamic balance at heel contact was also improved by extending the spatial margin in case of slipping. Despite the trend of positive Pilates effects on executive functions (29% improvement) confirmed by TMT-A, no significant effects were observed for cognitive functions. Positive mood changes were achieved by Pilates intervention, which may be the key for late-stage seniors to continue their participation in exercise programs. While further studies with a larger sample size are essential, Pilates appears to provide adequate exercise for the frail late-stage older population to minimize frailty

Upregulation of casein kinase 1ε in dorsal root ganglia and spinal cord after mouse spinal nerve injury contributes to neuropathic pain

<p>Abstract</p> <p>Background</p> <p>Neuropathic pain is a complex chronic pain generated by damage to, or pathological changes in the somatosensory nervous system. Characteristic features of neuropathic pain are allodynia, hyperalgesia and spontaneous pain. Such abnormalities associated with neuropathic pain state remain to be a significant clinical problem. However, the neuronal mechanisms underlying the pathogenesis of neuropathic pain are complex and still poorly understood. Casein kinase 1 is a serine/threonine protein kinase and has been implicated in a wide range of signaling activities such as cell differentiation, proliferation, apoptosis, circadian rhythms and membrane transport. In mammals, the CK1 family consists of seven members (α, β, γ1, γ2, γ3, δ, and ε) with a highly conserved kinase domain and divergent amino- and carboxy-termini.</p> <p>Results</p> <p>Preliminary cDNA microarray analysis revealed that the expression of the <it>casein kinase 1 epsilon </it>(<it>CK1ε</it>) mRNA in the spinal cord of the neuropathic pain-resistant N- type Ca²⁺channel deficient (<it>Ca</it>_<it>v</it><it>2.2</it>^-/-) mice was decreased by the spinal nerve injury. The same injury exerted no effects on the expression of <it>CK1ε </it>mRNA in the wild-type mice. Western blot analysis of the spinal cord identified the downregulation of CK1ε protein in the injured <it>Ca</it>_<it>v</it><it>2.2</it>^-/-mice, which is consistent with the data of microarray analysis. However, the expression of CK1ε protein was found to be up-regulated in the spinal cord of injured wild-type mice. Immunocytochemical analysis revealed that the spinal nerve injury changed the expression profiles of CK1ε protein in the dorsal root ganglion (DRG) and the spinal cord neurons. Both the percentage of CK1ε-positive neurons and the expression level of CK1ε protein were increased in DRG and the spinal cord of the neuropathic mice. These changes were reversed in the spinal cord of the injured <it>Ca</it>_<it>v</it><it>2.2</it>^-/-mice. Furthermore, intrathecal administration of a CK1 inhibitor IC261 produced marked anti-allodynic and anti-hyperalgesic effects on the neuropathic mice. In addition, primary afferent fiber-evoked spinal excitatory responses in the neuropathic mice were reduced by IC261.</p> <p>Conclusions</p> <p>These results suggest that CK1ε plays important physiological roles in neuropathic pain signaling. Therefore CK1ε is a useful target for analgesic drug development.</p

Hydrolyzed eggshell membrane immobilized on phosphorylcholine polymer supplies extracellular matrix environment for human dermal fibroblasts

We have found that a water-soluble alkaline-digested form of eggshell membrane (ASESM) can provide an extracellular matrix (ECM) environment for human dermal fibroblast cells (HDF) in vitro. Avian eggshell membrane (ESM) has a fibrous-meshwork structure and has long been utilized as a Chinese medicine for recovery from burn injuries and wounds in Asian countries. Therefore, ESM is expected to provide an excellent natural material for biomedical use. However, such applications have been hampered by the insolubility of ESM proteins. We have used a recently developed artificial cell membrane biointerface, 2-methacryloyloxyethyl phosphorylcholine polymer (PMBN) to immobilize ASESM proteins. The surface shows a fibrous structure under the atomic force microscope, and adhesion of HDF to ASESM is ASESM-dose-dependent. Quantitative mRNA analysis has revealed that the expression of type III collagen, matrix metalloproteinase-2, and decorin mRNAs is more than two-fold higher when HDF come into contact with a lower dose ASESM proteins immobilized on PMBN surface. A particle-exclusion assay with fixed erythrocytes has visualized secreted water-binding molecules around the cells. Thus, HDF seems to possess an ECM environment on the newly designed PMBN-ASESM surface, and future applications of the ASESM-PMBN system for biomedical use should be of great interest

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

A pristine record of outer Solar System materials from asteroid Ryugu’s returned sample

Volatile and organic-rich C-type asteroids may have been one of the main sources of Earth’s water. Our best insight into their chemistry is currently provided by carbonaceous chondritic meteorites, but the meteorite record is biased: only the strongest types survive atmospheric entry and are then modified by interaction with the terrestrial environment. Here we present the results of a detailed bulk and microanalytical study of pristine Ryugu particles, brought to Earth by the Hayabusa2 spacecraft. Ryugu particles display a close compositional match with the chemically unfractionated, but aqueously altered, CI (Ivuna-type) chondrites, which are widely used as a proxy for the bulk Solar System composition. The sample shows an intricate spatial relationship between aliphatic-rich organics and phyllosilicates and indicates maximum temperatures of ~30 °C during aqueous alteration. We find that heavy hydrogen and nitrogen abundances are consistent with an outer Solar System origin. Ryugu particles are the most uncontaminated and unfractionated extraterrestrial materials studied so far, and provide the best available match to the bulk Solar System composition

The Myth of Royal Authority and Shinbutsu-Shūgō (Kami-Buddha Amalgamation)

Cet article étudie deux étapes historiques du développement du mythe de l'autorité impériale au Japon. La première se situe à l'époque où le bouddhisme fut adopté dans la structure mythologique japonaise. Dans le Japon ancien, l 'autorité impériale gouvernait au moyen du système des codes (jp. ritsuryō). La mythologie et les cérémonies qui faisaient la démonstration de cette autorité mirent toutefois de côté l'impureté et l'exclurent du schéma mythologique général. L'ironie est qu'une telle exclusion rendit l'autorité impériale moins capable d'expliquer mythologiquement la création de l'univers par le truchement de la mythologie. De manière générale, les structures mythologiques nourrissent la violence. Celle-ci apparaît comme le premier élément dans les théories de la création de l 'univers. Dans le cas du Japon, la force violente fut représentée par les esprits courroucés et vengeurs des morts (jp. onryō). Mais, cet aspect fut redouté et volontairement négligé dans la mythologie impériale. C'est dans ce contexte que le bouddhisme fut adopté pour apaiser les esprits des morts assoiffés de vengeance et ainsi redonner à cette autorité sa puissance mythique dans toute sa plénitude. Bien que les enseignements du bouddhisme ne continssent pas à l'origine les éléments nécessaires aux rituels pour calmer les esprits vengeurs, lorsqu 'il fut intégré à la structure mythologique, il devait jouer un rôle crucial dans la mise au point de cérémonies pour la protection de l 'État (jp. chingo kokka) - aspect indispensable de la structure mythologique d'ensemble de la royauté. La seconde eut lieu lorsque divers changements sociaux firent passer le Japon à l'âge médiéval ; avec cette transition, la mythologie impériale, ses cérémonies et le rôle du bouddhisme furent aussi modifiés. Alors que le système des codes se détériorait, le système de répartition des terres cultivables (jp. handen-sei,) qui avait jusque-là prévalu, fut remplacé par le système dit des terres publiques (jp. kōryō) et des domaines privés (jp. shōen). Dans ce système, apparu ultérieurement, la distinction entre les terres appartenant à l'État et celles appartenant au privé cessa d'avoir la moindre fonction sociale. Les institutions de la société se modifièrent et leur soutien à l'État-nation gouverné par l'autorité impériale tendit à diminuer. Les paysans ne formaient plus un groupe homogène. Ils constituaient plutôt des groupes de statuts divers qui, de façon active et autonome, prenaient part à la production, à la distribution et à la consommation. Chaque groupe se spécialisa dans un « art » (jp. geinō au sens du latin artifex « métier »), que celui-ci fût d'ordre technique ou itinérant, et développa une histoire de ses origines fondée sur la mythologie impériale ; elle avait alors fusionné avec le bouddhisme (jp. shinbutsu shūgō) Cette nouvelle structure sociale contraignit la puissance impériale à refonder la mythologie ; un certain nombre d'essais en la matière virent le jour. L'exemple le plus évident est celui de Kitabatake Chikafusa dans son Jinnō shōtō ki (Histoire de la succession légitime des divins empereurs). Ceci dit, cette réinvention de la mythologie impériale fut incapable de fournir la démonstration complète de l'autorité de l'État-nation. Mais, paradoxalement, le mythe de l'autorité impériale se maintint dans les récits des origines développés au sein des divers groupes professionnels. Même si la structure de ces récits d'origine semble indiquer chez ces groupes une résistance à l'autorité impériale, ils manifestent également un désir de celle-ci.Sakurai Yoshiro, Fujieda Eri. The Myth of Royal Authority and Shinbutsu-Shūgō (Kami-Buddha Amalgamation). In: Cahiers d'Extrême-Asie, vol. 13, 2002. Moines, rois et marginaux. Études sur le bouddhisme médiéval japonais / Buddhist Priests, Kings and Marginals Studies on Medieval Japanese Buddhism pp. 85-99