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The Cytomegalovirus Enhancer Induces an Immediate Response to the Myosin Light Chain 2v Promoter during P19CL6 Cell Differentiation

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American Journal of Molecular Biology 2017年第4期7卷 190-203页

作者： Takanari Wakayama kazuaki ohashi Yasuyuki Fujimoto Masatomo Maeda Laboratory of Biochemistry and Molecular Biology Graduate School of Pharmaceutical Sciences Osaka University Osaka Japan Department of Medical Biochemistry School of Pharmacy Iwate Medical University Shiwa-Gun Japan Department of Molecular Biology School of Pharmacy Iwate Medical University Shiwa-Gun Japan Laboratory of Gene Therapy Faculty of Pharmaceutical Sciences Niigata University of Pharmacy and Applied Life Sciences Niigata Japan

The P19CL6 mouse embryonic carcinoma cells efficiently differentiate into cardiac muscle cells in the presence of DMSO. A reporter plasmid for cardiac muscle differentiation was constructed by connecting the CMV enhancer and a 250 bp MLC-2v promoter in front of the GFP gene to further evaluate the role of the CMV enhancer. This plasmid (pCBVenh/MLC-2vpro/EGFP) was stably introduced into P19CL6 cells, and the transfectant differentiated into cardiomyocytes with DMSO. Upon DMSO addition, GFP was immediately transcribed (within 2 days) and the amount of the transcript increased with cultivation. Concomitantly, GFP fluorescence was detected in the cells under a microscope. However, native MLC-2v was transcribed later on day 4. This expression time course is different from that of GFP. Clearly the CMV enhancer responded immediately to DMSO. Since GATA DNA-binding proteins play crucial roles in the initiation of cardiomyocyte differentiation, such a response could be ascribed to the presence of multiple GATA motifs in the enhancer sequence but not in the native MLC-2v promoter. Thus the CMV enhancer may be not only useful for gene therapy and monitoring cell differentiation but also the study of the role of GATA transcription factors expressed in P19CL6 cells.

关键词： Cytomegalovirus Enhancer Differentiation GATA Transcription Factor Gene Expression Heart Muscle MLC-2v P19CL6 Cells Promoter

Recombinant Human REG I<i>α</i>Aggregates <i>Staphylococcus aureus</i>—Exhibits a Lectin-Like Function

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Advances in Bioscience and Biotechnology 2017年第3期8卷 79-90页

作者： Nausheen Jamal Yuichiro Kezuka Takamasa Nonaka kazuaki ohashi Koji Nata Department of Medical Biochemistry School of Pharmacy Iwate Medical University Yahaba Japan Department of Structural Biology School of Pharmacy Iwate Medical University Yahaba Japan

Staphylococcus aureus is pathogenic to humans with worldwide health care concern due to its ability to evade the immune system and develop resistance to multiple drugs. Reg family proteins are C-type lectins with antimicrobial properties. Bacterial aggregation through binding to microbial cell surface sugar and/or lipid moieties is key mechanism employed in the process. In the present study we have analysed the antimicrobial effect of human REG Iα on S. aureus. Aggregation of mid-log phase culture of S. aureus was observed in presence of purified recombinant REG Iα. Therefore REG Iα can be applied in eliminating S. aureus infections in humans.

关键词： Reg Proteins Lectin-Like Activity Bacterial Aggregation

Increased Electrophoretic Mobility of Long-Type GATA-6 Transcription Factor upon Substitution of Its PEST Sequence

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Advances in Bioscience and Biotechnology 2014年第13期5卷 1032-1042页

作者： Kanako Obayashi Kayoko Takada kazuaki ohashi Ayako ohashi-Kobayashi Mayumi Nakanishi-Matsui Makoto Araki Masatomo Maeda Laboratory of Biochemistry and Molecular Biology Graduate School of Pharmaceutical Sciences Osaka University Suita Japan Department of Medical Biochemistry School of Pharmacy Iwate Medical University Shiwa-Gun Japan Department of Immunobiology School of Pharmacy Iwate Medical University Shiwa-Gun Japan Department of Biochemistry School of Pharmacy Iwate Medical University Shiwa-Gun Japan Department of Molecular Biology School of Pharmacy Iwate Medical University Shiwa-Gun Japan

The transcriptional factor GATA-6 gene produces two translational isoforms from a single mRNA through ribosomal leaky scanning. L-type GATA-6 has an extension of 146 amino acid residues at its amino terminus. In the extension, there is a unique PEST sequence (Glu31-Cys46), which is composed of an amino terminal Pro-rich segment and a carboxyl terminal Ser-cluster. Substitution of either half of the PEST sequence with Ala residues by cassette mutagenesis reduced the apparent molecular size of L-type GATA-6 on SDS-polyacrylamide gel-electrophoresis. However, the effect of substitution of the Pro-rich segment was much more significant;the mobility increase of the Pro-rich segment on the gel was 13% while that of the Ser-cluster was 8%. Substitution of each amino acid residue demonstrated that the effect of Pro substitution is greater than that of the Ser and Thr residues. Such increased mobility of L-type GATA-6 in the presence of a detergent may apparently correlate with the decrease in transcription activity in vivo as determined by means of luciferase reporter gene assay. The activity of ΔAla (with Ala residues instead of the PEST sequence) was reduced to one fifth of that of ΔA (with the PEST sequence). These results suggest that the PEST sequence of L-type GATA-6 does not function as a constitutive protein degradation signal, but rather plays structural and functional roles in the activation of gene expression on the GATA responsive promoter.

关键词： Cassette Mutagenesis Long-Type GATA-6 Mobility on Gel-Electrophoresis PEST Sequence Proline-Rich Segment Transcription Factor

A Case of Warthin’s Tumor Together with IgG4-Related Sialadenitis

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Open Journal of Pathology 2024年第3期14卷 81-89页

作者： Ryogo Aoki Risa Ito Asuka ohashi Riyoko Niwa Masashi Matsuyama Sachiko Oka Naomi Kawaguchi Toshimasa Sakakima Daichi Kodama Naoki Watanabe Daisuke Okamoto Takafumi Naiki Takuji Tanaka Department of Diagnostic Pathology (DDP) & Research Center of Diagnostic Pathology (RC-DiP) Gifu Municipal Hospital Gifu Japan Department of Central Clinical Laboratory Gifu Municipal Hospital Gifu Japan Department of Hematology Gifu Municipal Hospital Gifu Japan

Warthin’s tumor is the second most frequent neoplasm next to pleomorphic adenoma in the salivary gland. The tumor contains the epithelial oncocyte cells with the presence of rich-mitochondria and their surrounding abundant lymphocytes. A relatively new disease entity of IgG4-related disease frequently occurs in the salivary gland. However, the coexistence of Warthin’s tumor and IgG4-related disease is scarcely observed. We have recently experienced a rare case of Warthin’s tumor with IgG4-related sialadenitis. A 51-year-old man presented to our hospital, complaining of a mass with right submandibular tenderness and spontaneous pain. A computed tomography scan of the cervical region revealed a suspicion of lymph node proliferative disease, including malignant lymphoma. Elevated serum levels of IL-2R: 1843 U/ml (reference value 122 - 496 U/ml), IgG: 3430 mg/dl (reference value 861 - 1747 mg/dl), and IgG4: 3140 mg/dl (reference value 11 - 121 mg/dl) were observed. Other laboratory data showed within normal ranges. The cervical tumor was diagnosed as Warthin’s tumor by the findings of fine-needle aspiration cytology and biopsy examination. Immunohistochemistry revealed numerous IgG4- and IgG-positive cells with fibrosis surrounding the epithelial component of Warthin’s tumor, suggesting IgG4-rerated sialadenitis. Finally, we diagnosed the cervical tumor as Warthin tumor with IgG4-related sialadenitis. This is the second report describing a case of Warthin’s tumor with possible involvement of IgG4-related sialadenitis.

关键词： Warthin’s Tumor Sialadenitis IgG4-Related Disease Immunohistochemistry

Retinal ganglion cell protection by hop-flower extract as a novel neuroprotective strategy for glaucoma

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Neural Regeneration Research 2022年第6期17卷 1267-1268页

作者： Tomoko Hasegawa Hanako ohashi Ikeda Department of Ophthalmology and Visual Sciences Kyoto University Graduate School of MedicineKyotoJapan Research Fellow of Japan Society for the Promotion of Science TokyoJapan

In glaucoma,a leading cause of blindness,retinal ganglion cells are progressively *** pressure reduction is the only established treatment for glaucoma(Collaborative Normal-Tension Glaucoma Study Group,1998;Vass et al.,2007).However,in some cases,visual field loss progresses despite sufficiently reduced intraocular pressure(Collaborative Normal-Tension Glaucoma Study Group,1998;Killer and Pircher,2018).While intraocular pressure and age are known risk factors for glaucoma(Ernest et al.,2013),the underlying mechanisms of glaucoma progression are not fully understood.

关键词： glaucoma al. protective

Adenosine triphosphate maintenance by branched chain amino acids as a novel neuroprotective strategy for retinal neurodegenerative diseases

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Neural Regeneration Research 2019年第1期14卷 82-84页

作者： Tomoko Hasegawa Hanako ohashi Ikeda Department of Ophthalmology and Visual Sciences Kyoto University Graduate School of Medicine Department of Experimental Therapeutics Institute for Advancement of Clinical and Translational Science Kyoto University Hospital

Retinal neuronal cell death is caused in many incurable eye diseases such as retinitis pigmentosa(RP)and glaucoma,which are leading causes of adult *** RP,progressive loss of photoreceptor cells leads to visual *** established treatments are available to date for this condition,although potential treatments,including regenerative medicine,gene therapy,and neurotropic factor therapy are being *** glaucoma。

关键词：眼睛疾病视网膜氨基酸基因治疗腺苷分叉维护光敏电阻器

Modulation of valosin-containing protein by Kyoto University Substances(KUS) as a novel therapeutic strategy for ischemic neuronal diseases

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Neural Regeneration Research 2017年第8期12卷 1252-1255页

作者： Masayuki Hata Hanako ohashi Ikeda Department of Ophthalmology and Visual Sciences Kyoto University Graduate School of Medicine Neuroprotective Treatment Project for Ocular Diseases Institute for Advancement of Clinical and Translational Science Kyoto University Hospital

Retinal ischemia causes several vision-threatening diseases, including diabetic retinopathy, retinal artery occlusion, and retinal vein occlusion. Intracellular adenosine triphosphate（ATP） depletion and subsequent induced endoplasmic reticulum（ER） stress are proposed to be the underlying mechanisms of ischemic retinal cell death. Recently, we found that a naphthalene derivative can inhibit ATPase activity of valosin-containing protein, universally expressed within various types of cells, including retinal neural cells, with strong cytoprotective activity. Based on the chemical structure, we developed novel valosin-containing protein modulators, Kyoto University Substances（KUSs）, that not only inhibit intracellular ATP depletion, but also ameliorate ER stress. Suppressing ER stress by KUSs is associated with neural cell survival in animal models of several neurodegenerative diseases, such as glaucoma and retinal degeneration. Given that a major pathology of ischemic retinal diseases, other than intracellular ATP depletion, is ER stress-induced cell death, KUSs may provide a novel strategy for cell protection in ischemic conditions. Hence, we investigated the efficacy of KUS121 in a rat model of retinal ischemic injury. Intravitreal injections of KUS121, which is clinically preferable route of drug administration in retinal diseases, significantly suppressed inner retinal thinning and retinal cell death, and maintained visual functions. Valosin-containing protein modulation by KUS is a promising novel therapeutic strategy for ischemic retinal diseases.

关键词： adenosine triphosphatase C/EBP homologous protein central retinal artery occlusion endoplasmicreticulum stress neuroprotective therapy retinal ganglion cell

Cryopreserved fat: our clinical experience and applications

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Plastic and Aesthetic Research 2020年第5期7卷 47-56页

作者： Masanori ohashi The Clinic Tokyo Minato-kuTokyo 106-0031Japan

Aim:Cryopreservation of fat is an effective method for repeat fat grafting,but there are few reports about the clinical use of cryopreserved *** aim of this study was to determine the effectiveness and safety of cryopreserved fat for clinical ***:Between Aug 2015 and Dec 2018,we investigated 590 patients who underwent fat harvesting at our *** harvested fat was cryopreserved at a temperature of-196°C at a cell processing center and injections were performed in our ***:Of the 590 patients studied,216(312 cases)have undergone fat injections so *** augmentations using harvested fat,such as facial and breast augmentations,were performed on 180 *** 84 patients,harvested fat was utilized only for revitalization/fertilization purposes,such as to improve skin *** were no severe complications in any ***,volume maintenance was rarely *** rejuvenation effects were comparable to that in cases using fresh ***:The clinical use of cryopreserved fat is thought to be safe and effective.

关键词： Cryopreserved fat frozen fat fat grafting skin rejuvenation

Gorenstein Q-homology projective planes

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Science China Mathematics 2015年第3期58卷 501-512页

作者： HWANG DongSeon KEUM JongHae ohashi Hisanori Department of Mathematics Ajou University School of Mathematics Korea Institute For Advanced Study Department of Mathematics Faculty of Science and Technology Tokyo University of Science

We present the complete list of all singularity types on Gorenstein Q-homology projective planes,i.e.,normal projective surfaces of second Betti number one with at worst rational double *** list consists of 58 possibl... 详细信息

关键词： Q-homology projective plane Enriques surface rational double point