Simultaneous monitoring of independent gene expression patterns in two types of cocultured fibroblasts with different color-emitting luciferases

<p>Abstract</p> <p>Background</p> <p>Luciferase assay systems enable the real-time monitoring of gene expression in living cells. We have developed a dual-color luciferase assay system in which the expression of multiple genes can be tracked simultaneously using green- and red-emitting beetle luciferases. We have applied the system to monitoring independent gene expressions in two types of cocultured fibroblasts in real time.</p> <p>Results</p> <p>Two Rat-1 cell lines were established that stably express either green- or red-emitting luciferases under the control of the <it>mBmal1 </it>promoter, a canonical clock gene. We cocultured these cell lines, and gene expression profiles in both were monitored simultaneously. The circadian rhythms of these cell lines are independent, oscillating following their intrinsic circadian phases, even when cocultured. Furthermore, the independent rhythms were synchronized by medium change as an external stimulus.</p> <p>Conclusion</p> <p>Using this system, we successfully monitored independent gene expression patterns in two lines of cocultured fibroblasts.</p

Rhodium-catalyzed annulative coupling of isothiazoles with alkynes through n-s bond cleavage

A Rh(III)-catalyzed annulative coupling of 3,5-diarylisothiazoles and alkynes is reported. The N–S bond in the isothi-azole ring acts as an internal oxidant to regenerate the Rh(III) species in combination with an external Cu(II) oxidant, and the corre-sponding 1:2 coupling products are obtained. The remarkable difference in the reaction outcome between isothiazoles and the relevant isoxazoles has been investigated by DFT calculations, revealing that the relative stability of the enolate intermediates dictates the product selectivity.Mihara G., Noguchi T., Nishii Y., et al. Rhodium-catalyzed annulative coupling of isothiazoles with alkynes through n-s bond cleavage. Organic Letters. 22(2), 661-665, (2020), 17 January 2020; ©2020 American Chemical Society. https://doi.org/10.1021/acs.orglett.9b04437

A comparison of the usefulness of nuclear beta‐catenin in the diagnosis of desmoid‐type fibromatosis among commonly used anti‐beta‐catenin antibodies

Desmoid-type fibromatosis (DF) is a locally aggressive but non-metastatic (myo)fibroblastic neoplasm. A hallmark of the tumor is nuclear positivity for beta-catenin in immunohistochemistry due mostly to CTNNB1 mutations. However, a recent study has reported that even beta-catenin ‘nuclear-negative’ DFs can harbor CTNNB1 mutations and that the positive ratio of nuclear beta-catenin in DF is different among antibodies. Here, we reviewed soft tissue lesions for which the possibility of DF was considered and compared the sensitivity and specificity of nuclear beta-catenin for the diagnosis of DF among commonly used anti-beta-catenin antibodies, i.e., clone beta-catenin 1, 17C2 and 14. We analyzed 26 cases of DF, 28 cases of benign fibroblastic lesions, and 27 cases of other soft tissue tumors. The sensitivity and specificity of nuclear beta-catenin for the diagnosis of DF were different among antibodies; 54% and 98% in clone beta-catenin 1, 85% and 84% in 17C2, and 96% and 62% in 14. IHC of LEF1 showed comparable results with IHC of beta-catenin, with a sensitivity of 88% and specificity of 76%. Additionally, when beta-catenin 1 was used, DFs showed characteristic dotted cytoplasmic staining, often appearing as rings. Our results might be helpful for making a correct diagnosis of DF

Scattered X-rays in Obscured Active Galactic Nuclei and their Implications for Geometrical Structure and Evolution

We construct a new sample of 32 obscured active galactic nuclei (AGNs) selected from the Second XMM-Newton Serendipitous Source Catalogue to investigate their multiwavelength properties in relation to the "scattering fraction", the ratio of the soft X-ray flux to the absorption-corrected direct emission. The sample covers a broad range of the scattering fraction (0.1%-10%). A quarter of the 32 AGNs have a very low scattering fraction (smaller than 0.5%), which suggests that they are buried in a geometrically thick torus with a very small opening angle. We investigate correlations between the scattering fraction and multiwavelength properties. We find that AGNs with a small scattering fraction tend to have low [O III]lambda5007/X-ray luminosity ratios. This result agrees with the expectation that the extent of the narrow-line region is small because of the small opening angle of the torus. There is no significant correlation between scattering fraction and far-infrared luminosity. This implies that a scale height of the torus is not primarily determined by starburst activity. We also compare scattering fraction with black hole mass or Eddington ratio and find a weak anti-correlation between the Eddington ratio and scattering fraction. This implies that more rapidly growing supermassive black holes tend to have thicker tori.Comment: 13 pages, 10 figures, accepted for publication in Ap

Factors Influencing Medicine Use Behavior in Adolescents in Japan Using a Bayesian Network Analysis

Background: Medicine education in Japan was introduced to junior high schools in 2012. However, the effectiveness of existing education programs is limited. In order to develop more effective programs for high school students, the present study investigated the variables that directly influence medicine use behavior and the magnitude of their influence, using a Bayesian network analysis.Methods: A national cross-sectional survey was conducted in 2017. Eighty-three public high schools across Japan were randomly selected, and questionnaires were administered to 15–16 years old 10th grade students. The number of valid responses was 17,437 (effective response rate was 98.46%). Responses were analyzed to measure students’ behavior toward, attitudes regarding, and knowledge of medicines, and awareness of their prior medicine education.Results: Students’ “attitude score” and “awareness of a class” directly influenced their “behavior score.” The “score on attitude,” which had a large influence on “score on behavior,” was directly influenced by “score on knowledge of proper use” and “awareness of class.”Conclusion: The present study argues that acquiring knowledge of appropriate medicine use leads to the acquisition of favorable attitudes, which may result in behavioral change. Therefore, for medicine education, it is expected that incorporating content related to knowledge acquisition for changing attitudes will be important for promoting behavioral change

Effectiveness of distributing pocket cards in improving the behavior, attitude, and knowledge regarding proper medication use among junior high school students in Japan

ObjectiveThis study aimed to explore the effectiveness of distributing pocket cards with summaries of key information on appropriate medication usage after the implementation of a structured school-based medication education program for junior high school students in Japan.MethodsA total of 227 3rd-grade high school students participated in the intervention. Students who received the program without the provision of pocket cards in 2022 were included in the comparison group, and students who took the program with the provision of pocket cards in 2023 were included in the intervention group. After propensity score matching, the final sample of N = 116 comprised n = 58 comparison group participants and n = 58 intervention group participants. Questionnaires were administered at baseline, end-of-class, and 3-month follow-up to assess the changes in behavior, attitude, and knowledge scores.ResultsThe matched intervention group showed significantly lower scores at the 3-month follow-up than the matched comparison group. The results of the multiple linear regression analysis showed that for both groups, only the attitude scores were significantly correlated with the behavior scores. In addition, regardless of the baseline scores, the matched intervention group demonstrated smaller or negative changes in scores at the 3-month follow-up.ConclusionOverall, the results of this study did not support the effectiveness of distributing pocket cards after in-class intervention. However, the usefulness of medication education intervention was confirmed. These results emphasize the need to explore other supplemental teaching tools to further enhance the impact of structured medication education programs

The evolution of the Compton thick fraction and the nature of obscuration for AGN in the Chandra Deep Field South

(Abridged) We present the results from the X-ray spectral analysis of high-z AGN in the CDFS, making use of the new 4Ms data set and new X-ray spectral models from Brightman & Nandra, which account for Compton scattering and the geometry of the circumnuclear material. Our goals are to ascertain to what extent the torus paradigm of local AGN is applicable at earlier epochs and to evaluate the evolution of the Compton thick fraction (f_CT) with z, important for XRB synthesis models and understanding the accretion history of the universe. In addition to the torus models, we measure the fraction of scattered nuclear light, f_scatt known to be dependant on covering factor of the circumnuclear materal, and use this to aid in our understanding of its geometry. We find that the covering factor of the circumnuclear material is correlated with NH, and as such the most heavily obscured AGN are in fact also the most geometrically buried. We come to these conclusions from the result that f_scatt decreases as NH increases and from the prevalence of the torus model with the smallest opening angle as best fit model in the fits to the most obscured AGN. We find that a significant fraction of sources (~ 20%) in the CDFS are likely to be buried in material with close to 4 pi coverage having been best fit by the torus model with a 0\degree opening angle. Furthermore, we find 41 CTAGN in the CDFS using the new torus models, 29 of which we report here for the first time. We bin our sample by z in order to investigate the evolution of f_CT. Once we have accounted for biases and incompleteness we find a significant increase in the intrinsic f_CT, normalised to LX= 10^43.5 erg/s, from \approx 20% in the local universe to \approx 40% at z=1-4.Comment: Accepted for publication in MNRA

Usefulness of medium-Energy Collimator for Sentinel Node Lymphoscintigraphy Imaging in Breast Cancer Patients

金沢大学大学院医学系研究科小野口, 昌久This study was performed to investigate the usefulness of a general-purpose medium-energy (ME) collimator for the accurate localization of the sentinel lymph node (SLN) in breast cancer patients. Methods: We compared phantom images and lymphoscintigraphy images obtained under different conditions for a patient with breast cancer. Comparisons were performed between 2 cameras, between a low-energy high-resolution (LEHR) collimator and a general-purpose ME collimator, and between energy windows centered at 141 keV and at 146 keV. Profile curves and image contrast were evaluated along with the visual interpretation of images. The most suitable imaging time was selected from the relationship between contrast and the data acquisition time. Results: The images obtained with the general-purpose ME collimator and the energy window centered at 141 keV were of poorer quality than those obtained with the LEHR collimator and the same energy window. However, the quality of the images obtained with the general-purpose ME collimator improved when the energy window was centered at 146 keV. The method involving the general-purpose ME collimator and the energy window centered at 146 keV showed excellent image quality similar to that obtained with the LEHR collimator. The enhancement of contrast was confirmed at more than 3 cm away from the center of the injection site. Stable contrast was obtained with a data acquisition time of 5 min, with the general-purpose ME collimator, and with the energy window centered at 146 keV. Conclusion: The method involving the general-purpose ME collimator and the energy window centered at 146 keV has the merit of the lymph node not being concealed by a lead shield. This new method is expected to improve the rate of detection of SLN and has the potential for shortening the acquisition time