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Nagasaki-shi, Japan

Nagasaki International University is a private university in Nagasaki, Nagasaki, Japan, established in 2000. Wikipedia.

Chromatoid body (CB) was identified as granules stained by basic dye 130 years ago and called by various names. Electron microscopy revealed that the CB belonged to nuage (cloud in French) specific for germ cells. We described the localization of several proteins, including RNA helicases, in the nuage compartments classified into six types and in several spermatogenic cell-specific structures. All the proteins examined were detected in the nuage, including the CB with different staining intensities. Several proteins were localized to non-nuage structures, suggesting that these nuage proteins structures are related to nuage function. © Springer-Verlag 2012. Source

Yokota S.,Nagasaki International University
Methods in Molecular Biology

Colloidal gold probes, including protein A-, IgG-F(ab') 2-, and streptavidin-labeled gold particles, are useful tools for localization of antigens in cells and tissues by immunoelectron microscopy (IEM). This chapter describes different methods for the preparation of colloidal gold and conjugation of colloidal gold to protein A, IgG, and streptavidin. © Springer Science+Business Media, LLC 2010. Source

Otera H.,Kyushu University | Wang C.,U.S. National Institutes of Health | Cleland M.M.,U.S. National Institutes of Health | Setoguchi K.,Kyushu University | And 3 more authors.
Journal of Cell Biology

The cytoplasmic dynamin-related guanosine triphosphatase Drp1 is recruited to mitochondria and mediates mitochondrial fission. Although the mitochondrial outer membrane (MOM) protein Fis1 is thought to be a Drp1 receptor, this has not been confirmed. To analyze the mechanism of Drp1 recruitment, we manipulated the expression of mitochondrial fission and fusion proteins and demonstrated that (a) mitochondrial fission factor (Mff) knockdown released the Drp1 foci from the MOM accompanied by network extension, whereas Mff overexpression stimulated mitochondrial recruitment of Drp1 accompanied by mitochondrial fission; (b) Mff-dependent mitochondrial fission proceeded independent of Fis1; (c) a Mff mutant with the plasma membrane - targeted CAAX motif directed Drp1 to the target membrane; (d) Mff and Drp1 physically interacted in vitro and in vivo; (e) exogenous stimuli - induced mitochondrial fission and apoptosis were compromised by knockdown of Drp1 and Mff but not Fis1; and (f) conditional knockout of Fis1 in colon carcinoma cells revealed that it is dispensable for mitochondrial fission. Thus, Mff functions as an essential factor in mitochondrial recruitment of Drp1. © 2010 Otera et al. Source

Kamiya S.,Nagasaki International University
Yakugaku Zasshi

The importance of nanoparticle formulation is increasingly recognized in supporting pharmaceutical development. Thus, maintaining nanoparticles in a constant state is a major issue. A method involving lyophilization with the addition of saccharides can be used to maintain the steady state of nanoparticles. In this study, trisaccharides, tetrasaccharides, and pentasaccharides were added to nanoparticle suspensions, followed by rehydration of the samples, which had been either dried normally or freeze-dried. The particle size after rehydration was measured. In addition, each powder was measured using a powder X-ray diffractometer and thermal analysis device to investigate the correlation between the nanoparticles' aggregation and the crystal form of saccharides. The diameter of the nanoparticles was maintained when it was freeze-dried, while particle aggregation occurred when normally dried samples were used. In addition, crystalline saccharide was not observed in the freeze-dried group, but did appear in the normally dried group. © 2015 The Pharmaceutical Society of Japan. Source

BACKGROUND:: Dabigatran (DT) is a direct thrombin inhibitor used to prevent venous and arterial thromboembolism due to atrial fibrillation. DT is the active form of the commercially available prodrug dabigatran etexilate. Although DT has many clinical advantages over warfarin, it increases the incidence of bleeding in patients with renal dysfunction. Circulating levels of DT are increased in such patients because it is mainly eliminated by renal excretion. Therapeutic drug monitoring may therefore help to prevent adverse DT effects, but no method for measuring circulating DT levels has been reported, except for an analysis by liquid chromatography-tandem mass spectrometry. The present study sought to develop a novel enzyme-linked immunosorbent assay (ELISA) to measure DT concentrations.METHODS:: Mice were immunized with a DT-keyhole limpet hemocyanin conjugate to generate an anti-DT antibody. Immunized mouse splenocytes and myeloma cells (SP2/0) were fused to obtain an anti-DT monoclonal antibody (DT-mAb). DT-mAb and DT solutions were added to microplate wells coated with a DT-human serum albumin conjugate. DT concentrations were determined based on the principles of ELISA.RESULTS:: DT-mAb was successfully purified from a hybridoma, and the competitive ELISA developed using this DT-mAb could evaluate DT concentrations ranging from 7.8 to 125 ng/mL. The ELISA signal was not linear using DT-spiked serum; however, it was linear when serum ultrafiltrate was used. Weak cross-reactivity with dabigatran etexilate was detected, but no cross-reactivity was observed with other structurally related drugs, or drugs commonly used for the treatment of atrial fibrillation.CONCLUSIONS:: The developed competitive ELISA is a valuable and specific tool to analyze free DT in serum ultrafiltrate for therapeutic drug monitoring and pharmacokinetic studies. © 2015 by Lippincott Williams & Wilkins Source

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