Novel 3D Liquid Cell Culture Method for Anchorage-independent Cell Growth, Cell Imaging and Automated Drug Screening

Abstract Cells grown in three-dimensional (3D) cultures are more likely to have native cell-cell and cell-matrix interactions than in 2D cultures that impose mechanical constraints to cells. However, most 3D cultures utilise gel matrix which, while serving as a scaffold, limits application due to its solid and opaque nature and inconsistency in cell exposure to exogenous signals. In 3D culture without gel matrix, cells tend to adhere to each other and form clumps with necrotic zone at the centre, making them unsuitable for analyses. Here we report that addition of low-molecular-weight agar named LA717 to culture media allows cells to grow as dispersed clonal spheroids in 3D. LA717 maintains cells dispersed and settled to the bottom of the medium while keeping the medium clear with little additional viscosity, making it suitable for microscopic observation. Importantly, cancer spheroids formed in LA717-containing medium show higher sensitivity to anti-cancer drugs such as Trametinib and MK-2206 that are not as effective in 2D. Because of the small and consistent size of spheroids, cell viability and drug toxicity are readily detectable in automated imaging analysis. These results demonstrate that LA717 offers a novel 3D culture system with great in vivo reflection and practicality

Two rare cases of adult-onset phalangeal microgeodic syndrome with magnetic resonance imaging-proven bone edema transiently occurring in winter

We report two rare cases of adult-onset phalangeal microgeodic syndrome (PMS), which commonly develops in children. Both cases were Japanese women, with case 1: 60. years old and case 2: 58. years old. They developed swelling and stiffness in their fingers in winter, and their symptoms disappeared without any treatment in summer. Magnetic resonance imaging (MRI) of bilateral hands showed diffuse bone edema of the middle and proximal phalanges in winter, but the abnormal findings also disappeared in summer. The specific abnormalities observed on MRI were very useful for the diagnosis of PMS. Interestingly, they appeared in winter and disappeared in summer in parallel with the symptoms

Distinguishing the cerebrospinal fluid cytokine profile in neuropsychiatric systemic lupus erythematosus from other autoimmune neurological diseases

Neuropsychiatric systemic lupus erythematosus (NPSLE) is a serious complication in SLE. Although the mechanism of NPSLE remains unclear, cytokines and chemokines are considered to be involved in their pathogenesis. Here we used Bio-Plex Pro assays to examine 27 types of cytokines and chemokines in the cerebrospinal fluid (CSF) of 32 NPSLE patients. We used the CSF of 20 patients with multiple sclerosis (MS) and 22 patients with neuromyelitis optica (NMO) as a disease control group. Fourteen of 27 cytokines/chemokines were significantly higher in the NPSLE patients compared to the MS/NMO patients. We could identify six "minimum predictive markers" by using a weighted-voting algorithm that could distinguish NPSLE from MS and NMO: interleukin (IL)-17, IL-2, interferon (IFN)-γ, IL-5, basic fibroblast growth factor (FGF)-basic and IL-15. The determination of various types of CSF cytokine profiles may contribute to the diagnosis of NPSLE and may help elucidate the mechanisms underlying this disease

The linkage between medical student readiness for interprofessional learning and interest in community medicine

Objectives: The purpose of this study is to investigate the relationship between medical student readiness for interprofessional learning and interest in community medicine prior to incorporating community-oriented interprofessional education into the curriculum. Methods: A questionnaire was administered to students at Nagasaki University School of Medicine in Japan during each of three consecutive years (N=2244). The Readiness for Interprofessional Learning Scale (RIPLS) was administered in addition to a questionnaire to evaluate interest in community medicine. The Kruskal-Wallis and Steel-Dwass tests were used to determine differencesbetween school years. Correlation between the RIPLS score and interest in community medicine was evaluated with Spearman\u27s rank correlation coefficient. Relationships between RIPLS score and demographic parameters, and interest in community medicine were evaluated with multiple linear regression analysis. Results: Eighty-four percent (1891/2244) of students responded. The RIPLS score was highest in school year 1, followed by year 6, year 5,year 3, and years 4 and 2. Interest in community medicine correlated with the RIPLS score (rs = 0.332, p < 0.001),but less in year 1 (rs = 0.125, p = 0.002) than in other years. RIPLS score was significantly associated with gender, age, school year, interest in community medicine, but not the year that the survey was conducted. Conclusions: Community-oriented interprofessional education has the potential to improve attitudes towards interprofessional learning. When introducing this promising education into the curriculum from year 1, attracting students\u27 interest in community medicine should be considered

Anti-NXP2 autoantibodies in adult patients with idiopathic inflammatory myopathies: Possible association with malignancy

Objectives: Myositis-specific autoantibodies (MSAs) are useful tools for identifying clinically homogeneous subsets and predicting prognosis of patients with idiopathic inflammatory myopathies (IIM) including polymyositis (PM) and dermatomyositis (DM). Recent studies have shown that anti-NXP2 antibody (Ab) is a major MSA in juvenile dermatomyositis (JDM). In this study the frequencies and clinical associations of anti-NXP2 Ab were evaluated in adult patients with IIM. Methods: Clinical data and serum samples were collected from 507 adult Japanese patients with IIM (445 with DM and 62 with PM). Eleven patients with JDM, 108 with systemic lupus erythematosus, 433 with systemic sclerosis and 124 with idiopathic pulmonary fibrosis were assessed as disease controls. Serum was examined for anti-NXP2 Ab by immunoprecipitation and western blotting using polyclonal anti-NXP2 Ab. Results: Seven patients (1.6%) with adult DM and one (1.6%) with adult PM were positive for anti-NXP2 Ab. Except for two patients with JDM, none of the disease controls were positive for this autoantibody. Among eight adult patients with IIM, three had internal malignancies within 3 years of diagnosis of IIM. Another patient with DM also had a metastatic cancer at the diagnosis. All of the carcinomas were at an advanced stage (stage IIIb-IV). Conclusions: While less common than in juvenile IIM, anti-NXP2 Ab was found in adult IIM. Anti-NXP2 Ab may be associated with adult IIM with malignancy

Utility of a simplified ultrasonography scoring system among patients with rheumatoid arthritis: A multicenter cohort study

ABSTRACT: We aimed to evaluate the utility of a simplified ultrasonography (US) scoring system, which is desired in daily clinical practice, among patients with rheumatoid arthritis (RA) receiving biological/targeted synthetic disease-modifying antirheumatic drugs (DMARDs).A total of 289 Japanese patients with RA who were started on tumor necrosis factor inhibitors, abatacept, tocilizumab, or Janus kinase inhibitors between June 2013 and April 2019 at one of the 15 participating rheumatology centers were reviewed. We performed US assessment of articular synovia over 22 joints among bilateral wrist and finger joints, and the 22-joint (22j)-GS and 22-joint (22j)-PD scores were evaluated as an indicator of US activity using the sum of the GS and PD scores, respectively.The top 6 most affected joints included the bilateral wrist and second/third metacarpophalangeal joints. Therefore, 6-joint (6j)-GS and -PD scores were defined as the sum of the GS and PD scores from the 6 synovial sites over the aforementioned 6 joints, respectively. Although the 22j- or 6j-US scores were significantly correlated with DAS28-ESR or -CRP scores, the correlations were weak. Conversely, 6j-US scores were significantly and strongly correlated with 22j-US scores not only at baseline but also after therapy initiation.Using a multicenter cohort data, our results indicated that a simplified US scoring system could be adequately tolerated during any disease course among patients with RA receiving biological/targeted synthetic DMARDs

Influence of the Noh theatre on Paul Claudel’s ‘‘synthesis of arts’’

Pendant son séjour diplomatique au Japon de 1921 à 1927, Paul Claudel a rencontré le nô, une forme du théâtre traditionnel. Le dramaturge catholique, issu de la sphère symboliste, découvre dans le nô ce qu’il imaginait pour une nouvelle forme de la « synthèse des arts », c’est-à-dire l’union parfaite entre la Poésie, la Musique et la Danse, imprégnée de spiritualité. A l’aide d’une approche à la fois génétique, historique et comparatiste, notre thèse s’attache à étudier comment il interprète le nô et comment il s’en inspire dans sa propre création. Nous analysons d’abord la manière originale dont il comprend le nô, en adaptant une double perspective, histoire de la réception du nô en Occident depuis XVIe siècle et esthétique dramatique de l’auteur. Son originalité peut être dégagée par la comparaison avec la conception japonaise du nô authentique et par la confrontation avec la vision de ses prédécesseurs occidentaux ; contrairement à ces derniers, il passe sous silence le kyôgen, farce qui accompagne le nô ; son attitude vis-à-vis du bouddhisme est ambiguë ; son essai sur le nô a de grandes qualités littéraires. Ensuite, nous étudions le processus de l’assimilation de la philosophie orientale dans son univers, à la lumière d’un adage de Zeami, fondateur du nô, cité dans le Journal de Claudel. Enfin, nous démontrons qu’il utilise des apports du nô, d’une part dans ses pièces écrites au Japon, telle La Femme et son ombre (1922), et d’autre part, dans son « oratorio dramatique », genre inauguré en 1927 : le nô est sublimé dans son univers chrétien. Claudel cosmopolite, comprend le nô, puis le métamorphose en fonction de ses propres convictions esthétiques et spirituelles.France's Ambassador to Japan from 1921 to 1927, Paul Claudel discovered there the Noh theatre which stems from a medieval performing tradition. The Catholic dramatist and symbolist poet found in the Noh what he had imagined as a new form of “synthesis of arts’’, which was for him a perfect union of Poetry, Music and Dance, open to spirituality. Our method is genetic, historic and comparative in studying his understanding of the Noh, which is a source of inspiration for him. We have adopted a double perspective: first of all we study the history of the reception of the Noh in the Western world from the 16th century onward, and combine this type of investigation with literary considerations about the dramatist's aesthetics. The originality of our poet’s vision appears in comparison with the authentic Japanese conception of the Noh plays and the interpretation of his Western predecessors. For unlike these European specialists, his silence about the Kyôgen, a comic sketch played in a Noh performance, is surprising; his attitude to Buddhism is ambiguous; his essay about the Noh has a specific literary quality. The adage of Zeami, founder of the Noh, quoted in the Claudel's Journal, throws light on the process of assimilation by his Catholic vision of oriental philosophy. Finally, we show the impact of the Noh on his plays written in Japan, like La Femme et son ombre (1922), and on his “dramatic oratorio,” a genre inaugurated in 1927. The Noh is sublimated in his Christian world. Claudel, a cosmopolitan author who aims at universality, after having penetrated the world of the Noh, integrates it in his creative practice, according to his own aesthetic and spiritual conviction

Study on Availavility of Glass Ceramics ((CaO-P2O5-MgO-SiO2-CaF) for a Synthetic Bone Grafting Material

本論文の要旨は,第5回バイオラミックス研究会(平成元年6月19日),第7回バイオラミックス研究会(平成3年5月20日),第8回バイオラミックス研究会(平成4年4月20日)において発表した