Monitoring of a novel bacterium, Lactobacillus thermotolerans, in chicken intestine by fluorescence in situ hybridization (FISH)

We successfully established fluorescence in situ hybridization (FISH) method for specific detection and enumeration of a novel bacterium, Lactobacillus thermotolerans, in chicken feces. The specific FISH probes were designed based on the L. thermotolerans 16S rRNA gene sequences, and these sequences were compared to those of all available 16S rRNA genes in the GenBank database. The assay, targeting 16S rRNA gene, was evaluated using DNA from a pure culture of L. thermotolerans, DNA from the closely related bacteria Lactobacillus mucosae DSM 13345T and Lactobacillus fermentum JCM 1173T, and DNA from other lactic acid bacteria in quantitative experiments. The assay was then applied to two individual chicken trials. In trial 1, the cell population of L. thermotolerans ranged from 1.6 „e 106 to 3.4 „e 108 cells/g feces and from 2.6 „e 107 to 3.6 „e 108 cells/g cecal content. In trial 2, L. thermotolerans had also almost similar concentration (2.0 „e 106 to 3.4 „e 108 cells/g feces and 2.7 „e 107 to 2.9 „e 108 cells/g cecal content). We were not able to detect any bacterial cells at day one in both the trials. The results suggest that the newly developed FISH technique might be used for monitoring L. thermotolerans in the chicken intestine despite of its low sensitivity.Key words: Lactobacillus thermotolerans, fluorescence in situ hybridization (FISH), probiotic, chickens

PR3-ANCA in Wegener's granulomatosis prime human mononuclear cells for enhanced activation via TLRs and NOD1/2

<p>Abstract</p> <p>Background</p> <p>Anti-neutrophil cytoplasmic antibodies (ANCA) is autoantibodies characteristic of vasculitis diseases. A connection between ANCA and Wegener's granulomatosis was well established. The interaction of both ANCA phenotypes (PR3-ANCA and MPO-ANCA) with leukocytes provoked cell activation, which might be involved in the pathogenesis of ANCA-related Wegener's granulomatosis.</p> <p>Methods</p> <p>In this study, we examined whether PR3-ANCA sera and purified immunoglobulins from patients with Wegener's granulomatosis prime human monocytic cells for enhanced responses to microbial components in terms of production of proinflammatory cytokines.</p> <p>Results</p> <p>Flow cytometry demonstrated that stimulation with antibodies to proteinase 3 enhanced the expression of TLR2, 3, 4, 7, and 9, NOD1, and NOD2 in human mononuclear cells. The sera and purified immunoglobulins significantly primed human mononuclear cells to secrete interleukin-8 in response to microbial components via TLRs and NODs. Priming effects were also observed for the production of interleukin-6, monocyte chemoattractant protein-1, and tumor necrosis factor-α. On the other hand, PR3-ANCA-negative sera from patients with polyarteritis nodosa which possibly related to MPO-ANCA and aortitis syndrome as well as control sera from a healthy volunteer did not have any priming effects on PBMCs.</p> <p>Conclusion</p> <p>In conclusion, PR3-ANCA prime human mononuclear cells to produce cytokines upon stimulation with various microbial components by up-regulating the TLR and NOD signaling pathway, and these mechanisms <it>may partially </it>participate in the inflammatory process in Wegener's granulomatosis.</p

Development of a scale to measure learners\u27 perceived preferences and benefits of data-driven learning

In spite of researchers\u27 and practitioners\u27 increasing attention to data-driven learning (DDL) and increasing numbers of DDL studies, a multi-item scale to measure learners\u27 attitude toward DDL has not been developed thus far. In the present study, we developed and validated a psychometric scale to measure learners\u27 perceived preferences and benefits of DDL for research and pedagogical purposes. First, we created an item pool by referring to open-ended responses from learners; second, the items were pilot tested with target-level learners of English as a foreign language; and third, with item analyses and exploratory factor analysis, the revised version of the questionnaire was prepared. Finally, the questionnaire was administered, and its psychometric properties were examined with confirmatory factor analysis and fit indices. The final phase also included a measure of task values to explore the convergent evidence of the construct validity of the proposed scale. The results suggest that the scale is a valid measure of learners\u27 attitudes toward DDL, with the hypothesized model providing a good fit with the data. We propose that the scale can be used in future studies that utilize the same type of questionnaire research to facilitate further investigation of DDL. Suggestions for further research are also provided.This study was supported by JSPS KAKENHI Grant Numbers 26704006 and 25284108

Lesson Program for Spatial Composition of “House” with Different Structural Methods by Third Graders of Primary School

本年度は，３年生児童によるワークショップを実施し，「結ぶ」，「組む」という異なる二つの製作手法を用いた，「安全な家」の模型づくりの授業を構成した。製作模型では，児童によって多少の個人差はあるが，床や屋根の構造のつくり方と制作のプロセスの変化に一定の類型があることが明らかになった。また、製作前後で行ったアンケートの分析では、「結ぶ」手法から「組む」手法へと変わった際に、空間に対して求めるものが変化することが分かった。模型製作手法という外的要因が児童の安全に対する意識に影響を与えたことを明らかにしたことが最大の成果である。児童の生活環境との相関，床面の構成の分析などが，今後の空間構成に関する研究の上での課題として残された。In this year, we had the workshop by third grade students and make the lesson program for the making of a model as “secure house” with structural methods of “tying” and “wooden jointing”. In the making of models, though there are some individual differences, we found the definite types to make models: the floor or the roof structure, and the making process itself. In analysis of questionnaire before and after the workshop, what children desired for living space changed, when the method was changed from “tying” to “wooden jointing”. It is the important result that children’s conscious for secure was affected by external factor; the model making method. Theme relation of children’s living environment and analysis of structure of floor require further study

Neuromelanin‐Sensitive Magnetic Resonance Imaging Using DANTE Pulse

BACKGROUND: Neuromelanin-sensitive magnetic resonance imaging techniques have been developed but currently require relatively long scan times. The aim of this study was to assess the ability of black-blood delay alternating with nutation for tailored excitation-prepared T1-weighted variable flip angle turbo spin echo (DANTE T1-SPACE), which provides relatively high resolution with a short scan time, to visualize neuromelanin in the substantia nigra pars compacta (SNpc). METHODS: Participants comprised 49 healthy controls and 25 patients with Parkinson's disease (PD). Contrast ratios of SNpc and hyperintense SNpc areas, which show pixels brighter than thresholds, were assessed between DANTE T1-SPACE and T1-SPACE in healthy controls. To evaluate the diagnostic ability of DANTE T1-SPACE, the contrast ratios and hyperintense areas were compared between healthy and PD groups, and receiver operating characteristic analyses were performed. We also compared areas under the curve (AUCs) between DANTE T1-SPACE and the previously reported gradient echo neuromelanin (GRE-NM) imaging. Each analysis was performed using original images in native space and images transformed into Montreal Neurological Institute space. Values of P < 0.05 were considered significant. RESULTS: DANTE T1-SPACE showed significantly higher contrast ratios and larger hyperintense areas than T1-SPACE. On DANTE T1-SPACE, healthy controls showed significantly higher contrast ratios and larger hyperintense areas than patients with PD. Hyperintense areas in native space analysis achieved the best AUC (0.94). DANTE T1-SPACE showed AUCs as high as those of GRE-NM. CONCLUSIONS: DANTE T1-SPACE successfully visualized neuromelanin of the SNpc and showed potential for evaluating PD. © 2020 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society

Two-Minute Quantitative Susceptibility Mapping From Three-Dimensional Echo-Planar Imaging: Accuracy, Reliability, and Detection Performance in Patients With Cerebral Microbleeds

Objectives: The aim of this study was to assess the accuracy, reliability, and cerebral microbleed (CMB) detection performance of 2-minute quantitative susceptibility mapping (QSM) from 3-dimensional echo-planar imaging (3D-EPI). Materials and Methods: Gadolinium phantom study was conducted using 3D-EPI, single–echo time (TE), and multi-TE gradient-recalled echo (GRE) sequences on two 3-T magnetic resonance (MR) scanners to assess the accuracy between measured and theoretical susceptibility values. The institutional review board approved this prospective study, and 40 healthy volunteers were enrolled with written consent between April 2018 and October 2019. Each underwent 3D-EPI, single-TE, and multi-TE GRE sequences consecutively on one 3-T MR scanner, and QSMs were calculated to assess the reliability of 3D-EPI QSM. Intraclass correlation coefficient (ICC), linear regression, and Bland-Altman plots were calculated. Patients with CMB who underwent both 3D-EPI and GRE QSM scans were retrospectively enrolled. Two radiologists evaluated images independently, and Cohen κ coefficients were calculated to compare CMB detection performance. Results: Phantom study showed excellent validity of 3D-EPI QSM on both MR scanners: Skyra, R2 = 0.996, P < 0.001, ICC = 0.997, mean difference, −2 ppb (95% confidence interval [CI], −45 to 40 ppb); Prisma, R2 = 0.992, P < 0.001, ICC = 0.988, mean difference, 15 ppb (95% CI, −67 to 97 ppb). A human study of 40 healthy volunteers (59 ± 13 years, 25 women) showed excellent reliability with 3D-EPI QSM for both single-TE and multi-TE GRE (R2 = 0.981, P < 0.001, ICC = 0.988; R2 = 0.983, P < 0.001, ICC = 0.990, respectively), supported by a Bland-Altman mean difference of 4 ppb (95% CI, −15 to 23 ppb) for single-TE GRE and 3 ppb (95% CI, −15 to 20 ppb) for multi-TE GRE. The CMB detection performance evaluation from 38 patients (51 ± 20 years, 20 women) showed almost perfect agreement between 3D-EPI and GRE QSM for both raters (κ = 0.923 and 0.942, P < 0.001). Conclusions: Faster QSM from 3D-EPI demonstrated excellent accuracy, reliability, and CMB detection performance

A stem cell-derived gene (Sddr) negatively regulates differentiation of embryonic stem cells

金沢大学医薬保健研究域医学系Embryonic stem (ES) cells, derived from the inner cell mass of blastocysts, are pluripotent and continue to self-renew. To better understand the molecular mechanisms under-lying self-renewal, we have been searching for a gene(s) which is specifically expressed in self-renewing ES cells. Here we report the isolation and characterization of a novel gene, Sddr (stem cell-derived differentiation regulator). Sddr was highly expressed in undifferentiated ES cells, and its expression was downregulated upon differentiation. In addition to ES cells, Sddr expression was observed strongly in ovary, and weakly in lung. Immunostaining and cellular fractionation analyses suggested that Sddr is a cytoplasmic protein associated with the cytoskeleton. Sddr-null ES cells showed no remarkable abnormalities in their undifferentiated state. In contrast, in differentiating Sddr-null cells, induction of several differentiation-associated markers was enhanced, and downregulation of self-renewal marker genes was accelerated, as compared with wild-type cells. These results suggest that although it is dispensable for ES cell self-renewal, Sddr is a negative regulator of ES cell differentiation. © 2009 UBC Press

A comprehensive analysis of the correlations between resting-state oscillations in multiple-frequency bands and big five traits

Recently, the association between human personality traits and resting-state brain activity has gained interest in neuroimaging studies. However, it remains unclear if Big Five personality traits are represented in frequency bands (~0.25 Hz) of resting-state functional magnetic resonance imaging (fMRI) activity. Based on earlier neurophysiological studies, we investigated the correlation between the five personality traits assessed by the NEO Five-Factor Inventory (NEO-FFI), and the fractional amplitude of low-frequency fluctuation (fALFF) at four distinct frequency bands (slow-5 (0.01–0.027 Hz), slow-4 (0.027–0.073 Hz), slow-3 (0.073–0.198 Hz) and slow-2 (0.198–0.25 Hz)). We enrolled 835 young subjects and calculated the correlations of resting-state fMRI signals using a multiple regression analysis. We found a significant and consistent correlation between fALFF and the personality trait of extraversion at all frequency bands. Furthermore, significant correlations were detected in distinct brain regions for each frequency band. This finding supports the frequency-specific spatial representations of personality traits as previously suggested. In conclusion, our data highlight an association between human personality traits and fALFF at four distinct frequency bands

高齢者への長期ビタミンD投与

Daily １，０００ IU vitamin D was supplemented to ８７ years old female（Case １）and ６８ years old male（Case ２）subjects with sarcopenia and vitamin D insufficiency／deficiency for ２ years and ５ months and ２ years and ７ months, respectively. Before, １ year and ９ months, and ２ years and ５ months after vitamin D supplementation in Case １, serum ２５‐hydroxyvitamin D［２５（OH）D］level was ９ ng/ml, ２３．８ ng/ml and ２３．１ ng/ml, skeletal muscle mass was ８．９５ kg, １０．２ kg and １０．０ kg, handgrip strength was ７．３ kg, ８．９ kg and ９．９ kg, and Barthel index was ４０, ８５ and ９０, respectively. Before, and ２ years and ７ months after vitamin D supplementation in Case ２, serum ２５（OH）D level was ２６ ng/ml and ３４．４ ng/ml, skeletal muscle mass was ２４．６ kg and ２５．１ kg, handgrip strength was ３７．２ kg and ３８．３ kg, and Barthel index was １００ and １００, respectively. Thus, activity of daily living（ADL）markedly improved in Case １ and keep high in Case ２. Therefore, it is suggested that daily １，０００ IU vitamin D supplementation is effective to keep QOL of aged persons higher

Prediction of common hepatic artery catheter insertion based on celiac trunk morphology

PURPOSEThis study aimed to predict the ability to insert a 4–5 French (Fr) catheter insertion with a guidewire into the common hepatic artery (CHA) based on celiac trunk morphology.METHODSThis retrospective study included 64 patients who underwent balloon-occluded transcatheter arterial chemoembolization (n = 56), transcatheter arterial chemotherapy (n = 2), or were fitted with an implantable port system (n = 6) between June 2019 and December 2019 in our institution. The morphology of the celiac trunk was classified into three types (upward, horizontal, and downward) based on celiac angiography. The aortic–celiac trunk angle was measured on sagittal images of preprocedural contrast-enhanced computed tomography (CT). We reviewed whether a 4–5-Fr shepherd’s hook catheter could advance beyond the CHA using a 0.035-inch guidewire (Radifocus® Guidewire M; Terumo). Three patients were diagnosed with median arcuate ligament syndrome (MALS) based on the characteristic hook shape of the celiac artery on sagittal images of contrast-enhanced CT. The predictive ability of celiac angiography and preprocedural CT for CHA insertion success was evaluated. In unsuccessful cases, the balloon anchor technique (BAT) was attempted as follows: (1) a 2.7/2.8-Fr microballoon catheter (Attendant Delta; Terumo) was placed beyond the proper hepatic artery, and (2) the balloon was inflated as an anchor for parent catheter advancement.RESULTSUpward, horizontal, and downward celiac trunk types were noted in 42, 9, and 13 patients, respectively. The median CT angle was 122.83° (first quartile–third quartile, 102.88°–136.55°). Insertion in the CHA using the guidewire was successful in 56 of 64 patients (87.50%), and the success rate in the downward type was significantly lower than that in the upward type [42/42 (100%) vs. 7/13 (53.85%), P < 0.001]. The CT angle was significantly larger downward in the unsuccessful group than in the successful group (121.03° vs. 140.70°, P = 0.043). Celiac angiography had a significantly higher area under the curve (AUC) than preprocedural CT (AUC = 0.91 vs. AUC = 0.72, P = 0.040). All three cases of MALS showed unsuccessful CHA insertion. In all eight patients with unsuccessful insertion, the catheter could be advanced using the BAT [8/8 (100%)].CONCLUSIONCeliac angiography and preprocedural CT could predict CHA catheter insertion using a guidewire, and celiac angiography had high predictability. CT could detect MALS, a risk factor for unsuccessful CHA insertion