In vitro differentiation of lineage-negative bone marrow cells into microglia-like cells

金沢大学医薬保健研究域医学系Microglia are believed to be the only resident immune cells in the CNS, originating from hematopoietic-derived myeloid cells and invading the CNS during development. However, the detailed mechanisms of differentiation and transformation of microglial cells are not fully understood. Here, we demonstrate that murine microglial cells show two morphological forms in vitro, namely, small round cells expressing CD11b, Iba1, triggering receptor expressing on myeloid cells-2 (TREM2), and weakly expressing major histocompatibility complex class II and large flat cells expressing only CD11b and Iba1. Moreover, lineage-negative bone marrow (LN) cells cultured with primary mixed glial culture cells could differentiate into only the small round microglia-like cells, despite the absence of CCR2 and Gr-1 expression. Addition of macrophage colony stimulating factor (M-CSF) to LN cell culture allowed the proliferation and expression of TREM2 in LN cells, and the addition of neutralizing anti-M-CSF antibodies suppressed the proliferation of LN cells despite the expression of TREM2. When LN cells were cultured with M-CSF, the number of small round cells in the culture was considerably low, indicating that the small round morphology of the immature cells is not maintained in the presence of only M-CSF. On the other hand, when LN cells were grown in the presence of astrocytes, the small round cells were maintained at a concentration of approximately 30% of the total population. Therefore, cell-cell contact with glial cells, especially astrocytes, may be necessary to maintain the small round shape of the immature cells expressing TREM2. © Federation of European Neuroscience Societies and Blackwell Publishing Ltd

Coagulation and fibrinolysis abnormalities in familial amyloid polyneuropathy

Objective: Familial amyloid polyneuropathy (FAP) is an autosomal dominant form of hereditary amyloidosis. Several studies reported coagulation factor X deficiency and excessive fibrinolysis in immunoglobulin light chain amyloidosis. However, few have investigated coagulation and fibrinolysis in FAP. The objective of this study was to determine abnormalities in plasma biomarkers of coagulation and fibrinolysis in FAP. Methods: We prospectively recruited eight FAP patients with transthyretin mutations and ten age-matched control patients with other neuropathies in our university. We examined plasma biomarkers of coagulation and fibrinolysis including prothrombin time, activated partial thromboplastin time, fibrinogen, fibrin/fibrinogen degradation products, D-dimer, α2-antiplasmin, antithrombin, plasminogen, thrombin-antithrombin complex, plasmin-α2-antiplasmin complex, prothrombin fragment 1+2, and coagulation factor X. The MannWhitney U test was performed for statistical comparisons between FAP and control groups. Results: FAP patients exhibited significantly decreased levels of coagulation factor X, plasminogen and α2-antiplasmin, and significantly increased levels of prothrombin fragment 1+2 compared to control patients. Conclusion: Our results indicate abnormalities of coagulation and fibrinolysis in FAP patients. © 2012 Informa UK, Ltd

Efficient Retrieval and Secure Distribution of Information about Nonwritten Cultural Materials

Our joint research is aimed at developing basic technology necessary for information about nonwritten cultural materials to be exchanged and shared among researchers or between researchers and non-experts, and at verifying the effectiveness of the technology by using a demonstration system targeting actual materials and researchers. We will offer a proposal for basic technology required to achieve these goals, and then develop a basic system for Tadami Townʼs mingu（folk implements） record cards. To improve the keyword search function of the Tadami Internet Eco-Museum, unseen relationships will be clarified by using the Web Ontology Language（OWL） for an ontology of nonwritten cultural materials. This will lead to the discovery of new relationships, and enable users to accurately retrieve information they really need. To design a user interface suitable to, for example, creating an ontology of nonwritten cultural materials and retrieving information about them, data will be made visually manageable by regarding each datum as swarm intelligence data that can self-organize, and controlling these data. The proposed management method can be applied to self-organizing methods according to the process of data management. We will propose two information retrieval methods that take the quality of the retrieval process into consideration: a method using ant colony optimization（ACO）, a swarm intelligence algorithm; and a method using a so-called “recommender system.” To create a system in which personal and confidential information are protected and copyright mediation is autonomously conducted when information about nonwritten cultural materials is retrieved and distributed, we will also propose a digital watermarking method for the copyright management of bilevel line images that embeds information in images, not pixel values, using Bézier curves. Our other proposals include : swarm intelligence that regards access control as “language games,” and adapts to changing environments by using a concept of swarm and a concept of heredity ; a value exchange system designed to facilitate the documentation, data processing and distribution of nonwritten cultural materials that can reflect a variety of values by taking advantage of a characteristic of community currencies ― “value can be set independently” ; and a model that uses game theory to satisfy each user under particular conditions

CSF tau protein is a useful marker for effective treatment of superficial siderosis of the central nervous system: Two case reports

金沢大学附属病院神経内科We report two cases of superficial siderosis (SS) of the central nervous system (CNS), which is caused by chronic haemorrhaging into the subarachnoid space with haemosiderin deposition in the superficial portion of the CNS. Patient 1 had fluid collection in the spinal canal, which was reported as the source of the chronic bleeding. Patient 2 was bleeding from thickened dura at the level of the sacral vertebrae. Both of the patients had xanthochromic cerebrospinal fluid. We surgically repaired the sources of bleeding. Subsequently the cerebrospinal fluid (CSF) cleared and their symptoms were not aggravated for about 1 year. We measured several CSF markers of SS before and after surgery. Total tau protein (CSF-t-tau), phosphorylated tau protein (CSF-p-tau), iron (CSF-iron) and ferritin (CSF-ferritin) in the CSF were highly elevated at diagnosis. After surgery, the levels of CSF-t-tau and CSF-p-tau were markedly reduced while CSF-iron and CSF-ferritin had not decreased. It is suggested that CSF-t-tau and CSF-p-tau reflected the neural damage in SS and were useful to evaluate the effectiveness of SS therapies. © 2009 Elsevier B.V. All rights reserved

Enhanced GATA4 expression in senescent systemic lupus erythematosus monocytes promotes high levels of IFNα production

Enhanced interferon α (IFNα) production has been implicated in the pathogenesis of systemic lupus erythematosus (SLE). We previously reported IFNα production by monocytes upon activation of the stimulator of IFN genes (STING) pathway was enhanced in patients with SLE. We investigated the mechanism of enhanced IFNα production in SLE monocytes. Monocytes enriched from the peripheral blood of SLE patients and healthy controls (HC) were stimulated with 2′3′-cyclic GAMP (2′3′-cGAMP), a ligand of STING. IFNα positive/negative cells were FACS-sorted for RNA-sequencing analysis. Gene expression in untreated and 2′3′-cGAMP-stimulated SLE and HC monocytes was quantified by real-time PCR. The effect of GATA binding protein 4 (GATA4) on IFNα production was investigated by overexpressing GATA4 in monocytic U937 cells by vector transfection. Chromatin immunoprecipitation was performed to identify GATA4 binding target genes in U937 cells stimulated with 2′3′-cGAMP. Differentially expressed gene analysis of cGAS-STING stimulated SLE and HC monocytes revealed the enrichment of gene sets related to cellular senescence in SLE. CDKN2A, a marker gene of cellular senescence, was upregulated in SLE monocytes at steady state, and its expression was further enhanced upon STING stimulation. GATA4 expression was upregulated in IFNα-positive SLE monocytes. Overexpression of GATA4 enhanced IFNα production in U937 cells. GATA4 bound to the enhancer region of IFIT family genes and promoted the expressions of IFIT1, IFIT2, and IFIT3, which promote type I IFN induction. SLE monocytes with accelerated cellular senescence produced high levels of IFNα related to GATA4 expression upon activation of the cGAS-STING pathway

Research on Basic Technologies, Including Data Mining and a User Interface, for the Internet Eco-Museum

第三期共同研究活動報

非文字資料の効率的な検索と安全な流通

共同研究非文字資料の効率的な検索と安全な流

Application of an RDF to an Ontology-Based Database for Nonwritten Materials

The need for intellectually processing massive nonwritten information such as smells, scenery and tactile sensations has been increasing. Our research team had conducted a study on the establishment of an ontology-based database for folk implements to offer a new perspective by proving that it is possible to create an ontology-based database for nonwritten materials. Yet, we did not come up with a definite solution for an inference engine needed to compile such a database. This time, we tried a Jena-based resource description framework （RDF） to derive new relationships between resources to show the significance of ontology for nonwritten materials.　An RDF is a semantic data format used as a semantic web technology to describe linked data and metadata. It functions as a framework to describe relationships between resources identified by a uniform resource identifier（URI） and provides readable metadata for computing machines to information resources on the web, based on RDF data. An RDF finds new information not directly included in data according to rules like ontology and schemas.　Jena is a framework to develop Java-based semantic web applications. The semantic web is a movement to convert the current web to a new type that is comprehensible to computers. Jena provides various functions to process and use RDF data or knowledge.　The following is how we used Jena for this research. We asked Jena to make inferences on two sets of RDF data. This data included Kiri or an awl ; Mitsumegiri as a local name of Kiri ; and Hole as a purpose of using Mitsumegiri. Jena then inferred that a purpose of using Kiri is Hole. In other words, it added a tag Kiri to Hole and another tag Hole to Mitsumegiri. These tags were not contained in the original RDF data. This means that Jena automatically added them while performing the inference task and that the significance and feasibility of ontology for nonwritten materials have been demonstrated.　Nevertheless, some issues and challenges have been revealed. It takes a long time to build a system. Furthermore, an inference system that can be easily used for information in Excel format and a system to derive new relationships while building an ontology are yet to be developed.論

Optical spinning rheometry test on viscosity curves of less viscous fluids at low shear rate range

We propose a method, named optical spinning rheometry (OSR), to acquire kinematic viscosity curves of Newtonian and non-Newtonian fluids in the same framework. The OSR is independent of torque measurements and utilizes velocity information measured by particle tracking velocimetry. This optical approach enables flexibility in velocity resolution, and benefits exploring the low shear rate region. In addition, the kinematic viscosity of less viscous fluids like water or dilute polymer solutions can be assessed as being free from the mechanical limitations of torque sensors. The applicable range of the OSR is discussed in detail, and its performance is verified in Newtonian fluids. Demonstrations in dilute xanthan gum solutions, concentrations of O(10 ppm), show the capability of measuring their shear-thinning behaviors and the kinematic viscosity curves even in the first Newtonian regime