2,067 research outputs found

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    Mitochondrial Hsp90s suppress calcium-mediated stress signals propagating from mitochondria to the ER in cancer cells

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    Background: Resistance to cell death in the presence of stressful stimuli is one of the hallmarks of cancer cells acquired during multistep tumorigenesis, and knowledge of the molecular mechanism of stress adaptation can be exploited to develop cancer-selective therapeutics. Mitochondria and the endoplasmic reticulum (ER) are physically interconnected organelles that can sense and exchange various stress signals. Although there have been many studies on stress propagation from the ER to mitochondria, reverse stress signals originating from mitochondria have not been well reported.Methods: After inactivation of the proteins by pharmacologic and genetic methods, the signal pathways were analyzed by fluorescence microscopy, flow cytometry, MTT assay, and western blotting. A mouse xenograft model was used to examine synergistic anticancer activity and the action mechanism of drugs in vivo.Results: We show in this study that mitochondrial heat shock protein 90 (Hsp90) suppresses mitochondria-initiated calcium-mediated stress signals propagating into the ER in cancer cells. Mitochondrial Hsp90 inhibition triggers the calcium signal by opening the mitochondrial permeability transition pore and, in turn, the ER ryanodine receptor, via calcium-induced calcium release. Subsequent depletion of ER calcium activates unfolded protein responses in the ER lumen, thereby increasing the expression of a pro-apoptotic transcription factor, CEBP homologous protein (CHOP). Combined treatment with the ER stressor thapsigargin and the mitochondrial Hsp90 inhibitor gamitrinib augmented interorganelle stress signaling by elevating CHOP expression, and showed synergistic cytotoxic activity exclusively in cancer cells in vitro and in vivo.Conclusions: Collectively, mitochondrial Hsp90s confer cell death resistance to cancer cells by suppressing the mitochondria-initiated calcium-mediated interorganelle stress response.open0

    From Agasa Cristie to Group Image Play-Analysis of Horror Survival Game Panic Room : Escaping from the Den on Emotional Elements Development

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    A maniac computer game genre called "Survival Horror Gamesā€Ÿ is aimed for making gamers feel cathartic feeling when they escaped from the designed horror successfully. The degree of gaming quality, however, is not easy to measure. In this paper, we apply Cailloisā€Ÿ game playing categories and other standards to measure how a game induces the feeling of fear and other emotional experience to players. Once dominated horror survival game series called Panic Room: Escaping from the Den was chosen to analyze and evaluate with those standards as well as its narratives and subsystems. Especially the 2nd version was most welcomed to users among 4 versions thus we focused on the difference between the version 1 and the version 2 in terms of game playing and fear elements in the game content and story structure. In result, version 2 showed much more Agon and Mimicry and all other fear elements than version 1. The group image playing structure and conference/collection subsystem that were newly provided to version 2 were attributed to its success

    ā€˜Evidence of an auxin signal pathway, microRNA167-ARF8-GH3, and its response to exogenous auxin in cultured rice cellsā€™

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    MicroRNA167 (miR167) was shown to cleave auxin responsive factor 8 (ARF8) mRNA in cultured rice cells. MiR167 level was found to be controlled by the presence of auxin in the growth medium. When cells grew in auxin-free medium, miR167 level decreased, resulting in an increase in the level of ARF8 mRNA. Cells growing in the normal growth medium containing auxin showed a reversed trend. It was also shown that expression of OsGH3-2, an rice IAA-conjugating enzyme, was positively regulated by ARF8. Delivery of synthesized miR167 into cells led to decrease of both ARF8 mRNA and OsGH3-2 mRNA. This study provides an evidence in which the exogeneous auxin signal is transduced to OsGH3-2 through miR167 and ARF8 in sequence. This proposed auxin signal transduction pathway, auxin-miR167-ARF8-OsGH3-2, could be, in conjunction with the other microRNA-mediated auxin signals, an important one for responding to exogeneous auxin and for determining the cellular free auxin level which guides appropriate auxin responses

    PSMB9 Codon 60 Polymorphisms Have No Impact on the Activity of the Immunoproteasome Catalytic Subunit B1i Expressed in Multiple Types of Solid Cancer

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    The proteasome is a key regulator of cellular protein homeostasis and is a clinically validated anticancer target. The immunoproteasome, a subtype of proteasome expressed mainly in hematopoietic cells, was initially recognized for its role in antigen presentation during the immune response. Recently, the immunoproteasome has been implicated in several disease conditions including cancer and autoimmune disorders, but many of the factors contributing to these pathological processes remain unknown. In particular, the codon 60 polymorphism of the PSMB9 gene encoding the Ī²1i immunoproteasome catalytic subunit has been investigated in the context of a variety of diseases. Despite this, previous studies have so far reported inconsistent findings regarding the impact of this polymorphism on proteasome activity. Thus, we set out to investigate the impact of the PSMB9 codon 60 polymorphism on the expression and activity of the Ī²1i immunoproteasome subunit in a panel of human cancer cell lines. The Ī²1i-selective fluorogenic substrate Acetyl-Pro-Ala-Leu-7-amino-4-methylcoumarin was used to specifically measure Ī²1i catalytic activity. Our results indicate that the codon 60 Arg/His polymorphism does not significantly alter the expression and activity of Ī²1i among the cell lines tested. Additionally, we also examined the expression of Ī²1i in clinical samples from colon and pancreatic cancer patients. Our immunohistochemical analyses showed that ā‰ˆ 70% of clinical colon cancer samples and ā‰ˆ 53% of pancreatic cancer samples have detectable Ī²1i expression. Taken together, our results indicate that the Ī²1i subunit of the immunoproteasome is frequently expressed in colon and pancreatic cancers but that the codon 60 genetic variants of Ī²1i display similar catalytic activities and are unlikely to contribute to the significant inter-cell-line and inter-individual variabilities in the immunoproteasome activity

    Circulating levels of DNA-histone complex and dsDNA are independent prognostic factors of disseminated intravascular coagulation

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    AbstractIntroductionNeutrophils can be induced to release DNA combined with histones. The resulting neutrophil extracellular trap (NET) provides a scaffold for growing hemostatic plug. Therefore, the NET formation may be inevitable in clinical conditions that are characterized by formation of vascular thrombi. Thus far, there have been no reports on the clinical significance of NET in disseminated intravascular coagulation (DIC). Therefore, we investigated circulating levels of NET in DIC and analyzed their potential values to assess coagulation severity and predict clinical outcome.MethodsThe plasma levels of DNA-histone complexes and double-stranded DNA (dsDNA), considered to be in vivo markers of NET, were measured in 199 patients suspected of having DIC and 20 healthy controls.ResultThe circulating levels of DNA-histone complexes and dsDNA were significantly elevated in overt-DIC. The increased levels of these two markers correlated with the severity of coagulopathy including DIC score and D-dimer. Multivariable Cox regression analysis, adjusted for the conventional DIC markers, revealed that elevated DNA-histone complexes and dsDNA are poor independent prognostic markers.ConclusionThe circulating levels of NET release reflect the coagulation activation and adverse clinical outcomes in patients with DIC, thereby providing potential clinical relevance for mortality prediction in DIC

    Eco-Innovation Indices as Tools for Measuring Eco-Innovation

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    Measuring eco-innovation helps us understand the overall trends and raises awareness in society. Measuring eco-innovation at the national level and making comparisons across countries may allow us to benchmark performance and foster policy learning. This paper assesses two indices developed in two different regions: The ASEM Eco-Innovation Index (ASEI) by the ASEM SMEs Eco-Innovation Center, based in Republic of Korea; and the Eco-Innovation Scoreboard (Eco-IS) developed by the Eco-Innovation Observatory, based in the European Union. This paper aims to examine and compare the features of both and attempts to obtain insights on their strengths and weaknesses. Towards this aim, our paper assesses those scoreboards against four criteria stemming from innovation analysis: (1) relevance of areas and stakeholders covered; (2) ability to indicate changes; (3) directions towards common goals; and (4) ability to facilitate further changes. We conclude both are promising, despite data shortages, and have great potential to contribute towards the sustainable development goals (SDGs), particularly with regard to the SDGs on sustainable industrialization and sustainable consumption and production. In comparison, the ASEI covers more countries than the Eco-IS. However, the ASEI has limitations on measuring indicators due to limited data availability in Asian countries. The Eco-IS is closely linked with the regional and national policies for eco-innovation in Europe, while the ASEIā€™s impact appears more limited, as of now. In conclusion, the research results give insights into key areas, goals and applications of eco-innovation indices, and can help upgrading eco-innovation indices. This research helps interpret the scores of two indices better and facilitate application of the scores in the multiple ways. It is expected that this research contributes to developing and modifying a global eco-innovation index and enhancing the ability of these indices to facilitate eco-innovation strategies at national levels and across relevant actors
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