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Biomedical Research

Ryoichi Yoshimura, Misaki Nakagawa, Yasuhisa Endo
The mechanism of peripheral blood circulation failure by low temperatures, known as Raynaud's phenomenon, is not well understood at present. We examined the possibility that the efferent fibers of peripheral sympathetic neurons may release transmitters directly by cold sensation. PC12 cells, a culture model of sympathetic neurons, were placed at 25°C for 30 min, fixed with the tannic acid-glutaraldehyde-osmium method and observed by electron microscopy. The number of omega-shaped exocytotic vesicle figures increased significantly compared with that in the control (37°C), although it was lower than that of the positive control (high potassium stimulation at 25°C and 37°C)...
2016: Biomedical Research
Yuji Shimoda, Takeshi Nagashima, Kenichi Urakami, Tomoe Tanabe, Junko Saito, Akane Naruoka, Masakuni Serizawa, Tohru Mochizuki, Keiichi Ohshima, Sumiko Ohnami, Shumpei Ohnami, Masatoshi Kusuhara, Ken Yamaguchi
The use of next-generation sequencing (NGS) techniques to analyze the genomes of cancer cells has identified numerous genomic alterations, including single-base substitutions, small insertions and deletions, amplification, recombination, and epigenetic modifications. NGS contributes to the clinical management of patients as well as new discoveries that identify the mechanisms of tumorigenesis. Moreover, analysis of gene panels targeting actionable mutations enhances efforts to optimize the selection of chemotherapeutic regimens...
2016: Biomedical Research
Takeshi Nagashima, Yuji Shimoda, Tomoe Tanabe, Akane Naruoka, Junko Saito, Masakuni Serizawa, Keiichi Ohshima, Kenichi Urakami, Sumiko Ohnami, Shumpei Ohnami, Tohru Mochizuki, Masatoshi Kusuhara, Ken Yamaguchi
Identification of causal genomic alterations is an indispensable step in the implementation of personalized cancer medicine. Analytical methods play a central role in identifying such changes because of the vast amount of data produced by next generation sequencer. Most analytical techniques are designed for the Illumina platform and are therefore suboptimal for analyzing datasets generated by whole exome sequencing (WES) using the Ion Proton System. Accurate identification of somatic mutations requires the characterization of platform-dependent error profiles and genomic properties that affect the accuracy of sequence data as well as platform-oriented optimization of the pipeline...
2016: Biomedical Research
Shunsuke Kimura, Junko Nio-Kobayashi, Ayuko Kishimoto, Toshihiko Iwanaga
GP2, a GPI-anchored glycoprotein that is a useful marker for M cells of Peyer's patches, is functionally related to the uptake of pathogenic bacteria in the gut lumen. Our immunostaining throughout the whole body of mice detected a broader localization than previously found of GP2 in various mucous glands and secretory cells. In the oral cavity, the palatine gland and lingual gland intensely expressed GP2 with immunolabeling along the basolateral membrane of acini and in luminal secretions of ducts. Secretory portions of the duodenal gland as well as the pancreas were immunoreactive for GP2 in the digestive tract...
2016: Biomedical Research
Kensuke Nakajima, Shigeru Oiso, Takuhiro Uto, Osamu Morinaga, Yukihiro Shoyama, Hiroko Kariyazono
Ghrelin is an appetite-stimulating peptide hormone with an octanoyl modification at serine 3 that is essential for its orexigenic effect. Ghrelin O-acyltransferase (GOAT) is the enzyme that catalyzes ghrelin acylation using fatty acyl-coenzyme A as a substrate. We previously developed an assay system based on the AGS-GHRL8 cell line that produces octanoylated ghrelin in the presence of octanoic acid, and demonstrated that some fatty acids suppressed octanoylated ghrelin production. Recent studies have reported that triterpenes have anti-obesity effect...
2016: Biomedical Research
Satoshi Tsuzuki, Takahiko Amitsuka, Tatsuya Okahashi, Yuki Kozai, Masayuki Yamasaki, Kazuo Inoue, Tohru Fushiki
Cluster of differentiation 36 (CD36) is a broadly expressed transmembrane protein that has multiple ligands, including oxidized low-density lipoproteins. We found recently that CD36 is expressed in olfactory sensory neurons and postulated that it plays a role in the detection of distinct odorants in the nasal cavity. To date, however, there have been few examples of attempts to identify CD36-recognizable odorants. In this study, by an in vitro assay using a peptide mimic of the receptor, we provided evidence that CD36 recognizes (Z,Z)-4,7-tridecadienal, an odor-active volatile compound that is known to occur in Katsuobushi (dried, fermented, and smoked skipjack tuna commonly used in Japanese cuisine as a seasoning) and in the preorbital secretion of male oribi...
2016: Biomedical Research
Kaori Murata, Ayumi Takahashi, Takao Morita, Akihiro Nezu, Satoshi Fukumoto, Masato Saitoh, Akihiro Tanimura
Genetically encoded calcium indicators (GECIs) are suitable for long-term imaging studies. In this study, we employed a highly sensitive GECI, G-GECO, and achieved efficient gene delivery with an adenoviral vector. The adenoviral vector allowed us to express G-GECO in more than 80% of cells. More than 80% of G-GECO-expressing cells showed an ATP-induced increase in fluorescence intensity due to Ca(2+) release from intracellular stores and subsequent Ca(2+) entry. The fluorescence intensity of these cells was increased more than 2-fold by stimulation with 10 μM ATP...
2016: Biomedical Research
Masako Yajima, Shin-Ichiro Karaki, Takeshi Tsuruta, Shunsuke Kimura, Junko Nio-Kobayashi, Atsukazu Kuwahara, Takaji Yajima
Non-neuronal and atropine-sensitive ileal contractile responses to short chain fatty acids (SCFAs) are detected in the neonatal stage, and change with age or inflammatory conditions. However, the roles of luminal SCFAs in developmental changes have not yet been elucidated. We examined ileal contractile responses to SCFAs in mice colonized with different SCFA-producing intestinal microbiota under normal and inflammatory conditions. Using conventional (Conv), germ-free (GF), and gnotobiotic mice infected with Bifidobacterium (GB-bif), Propionibacterium (GB-prop), or Lactobacillus (GB-lact), ileal contractions were measured in 1-day-old neonates and 7-week-old mice using an isotonic transducer...
2016: Biomedical Research
Dae Seong Kim, Myoung Woo Lee, Young Jong Ko, Hyun Jin Park, Yoo Jin Park, Dong-Ik Kim, Hye Lim Jung, Ki Woong Sung, Hong Hoe Koo, Keon Hee Yoo
Human mesenchymal stem cell (MSC) heterogeneity and problems associated with the ex vivo expansion of MSC are linked with the failure of MSC clinical trials. In this study, we compared the effect of MSCs cultured in different oxygen concentrations on GVHD in mice to elucidate whether hypoxia improves the immunosuppressive capacity of MSCs. Hypoxia increased the proliferative activity and the expression of several stemness and chemokine genes, such as KLF4, OCT4, C-MYC, CCL2, and CXCL10. Mice that received MSCs cultured in normoxia or hypoxia showed alleviated symptoms of GVHD and increased survival times...
2016: Biomedical Research
Tomoyuki Shibata, Hiromi Yamashita, Tomohiko Kawamura, Yasutaka Jodai, Takafumi Omori, Kazuya Sumi, Yuichiro Ichikawa, Masaaki Okubo, Takamitsu Ishizuka, Tomomitsu Tahara, Mitsuo Nagasaka, Yoshihito Nakagawa, Ichiro Hirata, Naoki Ohmiya, Makoto Nakao
The aim of this study was to investigate the effect of consuming small amounts of beer or a nonalcoholic beer taste beverage (non-beer) on gastric emptying and the polymorphisms in alcohol metabolism-related enzyme-encoding genes. Twenty male healthy volunteers were questioned regarding their alcohol consumption status, and body measurement was performed. The genetic polymorphisms in ADH1B (rs1229984, Arg47His) and ALDH2 (rs671 Glu487Lys) were analyzed. The subjects consumed 150 mL of beer or non-beer once per week, followed by the ingestion of 200 kcal of the test nutrient containing (13)C-acetate 15 min later, after which the subjects' exhalations were collected up to 120 min...
2016: Biomedical Research
Kazuhiro Shiina, Ken-Ichiro Hayashida, Kazuo Ishikawa, Masahito Kawatani
Overactive bladder is one of the major health problem especially in elderly people. Adenosine triphosphate (ATP) is released from urinary bladder cells and acts as a smooth muscle contraction and sensory signal in micturition but little is known about the role of ATP release in the pathophysiology of overactive bladder. To assess the relationship between ATP and overactive bladder, we used a partial bladder outlet obstruction (pBOO) model in rats. The bladder caused several changes by pBOO: An increase in bladder weight, hypertrophy of sub-urothelium and sub-serosal area, and frequent non-voiding bladder contraction during urine storage...
2016: Biomedical Research
Kenji Matsunaga, Chika Ito, Kaichi Nakakogawa, Akina Sugiuchi, Ryo Sako, Masahiro Furusawa, Takashi Muramatsu
The objective of this study is to investigate the responses of human cementoblasts to light compressive force in vitro. A human cementoblast cell line (HCEM) was loaded for 12 h by mounting coverslips (0.25 gf/cm(2)). The coverslips were removed and the cells were cultured for up to 21 days. Cells without glass loading were used as controls. Cell growth, morphological changes, and the mRNA expression of RUNX2, ALP, WNT5A and SPON1 were investigated. No significant differences were observed in cell numbers between the compressed group and control group...
2016: Biomedical Research
Norihiko Sumiyoshi, Hiroyuki Ishitobi, Shigeru Miyaki, Kenji Miyado, Nobuo Adachi, Mitsuo Ochi
Although osteoarthritis (OA) is the most prevalent aging-related joint disease, the understanding of mechanisms of OA pathogenesis remains limited. Key features include the progressive degradation of articular cartilage, synovial hyperplasia, and angiogenesis in joint tissues. CD9, a member of the tetraspanin family, is localized in the cell membranes and partly in the endosomes of all mammalian cell types. CD9 is associated with inflammation and angiogenesis through cell adhesion, migration, and signal transduction...
2016: Biomedical Research
Hideko Azuma, Eri Watanabe, Yohei Otsuka, Yasuyuki Negishi, Sadayuki Ohkura, Eiji Shinya, Hidemi Takahashi
Langerhans cells (LCs), a subset of dendritic cells (DCs), reside in body surface presenting antigens from various pathogens and activate immune system after migrating to vicinal lymph nodes. We recently demonstrated that the E-cadherin interaction allowed peripheral blood (PB) CD14(+) cells to differentiate into LC-like cells that closely resemble primary LCs. Here, with a combination of GM-CSF, TGF-β, and TNF-α, we induced LC-like cells from umbilical cord blood (UCB)derived CD34(+) cells and compared them with those induced from PB CD14(+) cells...
2016: Biomedical Research
Miao Zheng, Shinhye Lee, Satoshi Tsuzuki, Kazuo Inoue, Daisaku Masuda, Shizuya Yamashita, Toshihiko Iwanaga
The sebaceous glands secrete sebum to protect the epidermis and hairs by the oily products. The glands express several transporters and binding proteins for the production of fatty acids and uptake of their sources. The present immunohistochemical study examined the expression and localization of CD36, MCT1, FATP4, and E-FABP in the sebaceous glands, including the meibomian and preputial glands of mice. CD36 and MCT1 in sebaceous glands were largely co-localized along the plasma membrane of secretory cells, while they were separately expressed in the glandular portion of meibomian and preputial glands...
2016: Biomedical Research
Ken Yamaguchi, Kenichi Urakami, Takeshi Nagashima, Yuji Shimoda, Shumpei Ohnami, Sumiko Ohnami, Keiichi Ohshima, Tohru Mochizuki, Keiichi Hatakeyama, Masakuni Serizawa, Yasuto Akiyama, Kouji Maruyama, Hirohisa Katagiri, Yuji Ishida, Kaoru Takahashi, Seiichiro Nishimura, Masanori Terashima, Taiichi Kawamura, Yusuke Kinugasa, Yushi Yamakawa, Tetsuro Onitsuka, Yasuhisa Ohde, Takashi Sugino, Ichiro Ito, Hiroyuki Matsubayashi, Yasue Horiuchi, Maki Mizuguchi, Mutsumi Yamazaki, Kengo Inoue, Kimiko Wakamatsu, Misato Sugiyama, Katsuhiko Uesaka, Masatoshi Kusuhara
Using whole exome sequencing data obtained from 1,685 Japanese cancer patients, we examined genetic variations of germline TP53 and found 10 types of non-synonymous single nucleotide variants. In the present study, we focused on 6 patients with germline D49H mutation located in the transactivation domain 2 of p53 protein, since the mutation seemed to be prevalent in cancer patients and to be pathogenic. According to the initial survey for family history of the proband with the germline TP53 D49H mutation, one osteosarcoma patient and his pedigree fulfill the criteria for Li-Fraumeni-like syndrome and the 2009 Chompret criteria for germline TP53 mutation screening...
2016: Biomedical Research
Jumpei Washio, Tamaki Ogawa, Keisuke Suzuki, Yosuke Tsukiboshi, Motohiro Watanabe, Nobuhiro Takahashi
Dental plaque metabolizes both carbohydrates and amino acids. The former can be degraded to acids mainly, while the latter can be degraded to various metabolites, including ammonia, acids and amines, and associated with acid-neutralization, oral malodor and tissue inflammation. However, amino acid metabolism in dental plaque is still unclear. This study aimed to elucidate what kinds of amino acids are available as metabolic substrates and how the amino acids are metabolized in supragingival plaque, by a metabolome analysis...
2016: Biomedical Research
Kenkichi Tsuruga, Toshikazu Hashimoto, Ryoko Kato, Rui Kato, Yousuke Uchida, Tetsutaro Hase, Yuji Morimoto
Brain-derived neurotrophic factor (BDNF) is released from activated microglia during neuropathic pain and is hypothesized to downregulate the expression of the potassium chloride cotransporter 2 (KCC2) via the TrkB receptor. Previous studies reported that KCC2 is downregulated 5 min after the plantar injection of formalin in rats; however, the mechanism behind this decrease in KCC2 expression during acute inflammatory pain remains unknown. In this study, we determined whether the TrkB receptor contributes to the expression of KCC2 during the acute pain...
2016: Biomedical Research
Yasuto Akiyama, Ryota Kondou, Akira Iizuka, Keiichi Ohshima, Kenichi Urakami, Takeshi Nagashima, Yuji Shimoda, Tomoe Tanabe, Sumiko Ohnami, Shumpei Ohnami, Masatoshi Kusuhara, Tohru Mochizuki, Ken Yamaguchi
Project HOPE (High-tech Omics-based Patient Evaluation) has been progressing since its implementation in 2014 using whole-exome sequencing (WES) and gene expression profiling (GEP). With the aim of evaluating immune status in cancer patients, a gene panel consisting of 164 immune response-associated genes (56 antigen-presenting cell and T-cell-associated genes, 34 cytokine- and metabolism-associated genes, 47 TNF and TNF receptor superfamily genes, and 27 regulatory T-cell-associated genes) was established, and its expression and mutation status were investigated using 1,000 cancer patient-derived tumors...
2016: Biomedical Research
Shoko Furuta, Rika Kuwahara, Eri Hiraki, Koichiro Ohnuki, Shinobu Yasuo, Kuniyoshi Shimizu
Hericium erinaceus (HE), an edible mushroom, has been used as a herbal medicine in several Asian countries since ancient times. HE has potential as a medicine for the treatment and prevention of dementia, a disorder closely linked with circadian rhythm. This study investigated the effects of the intake of HE extracts on behavioral rhythm, photosensitivity of the circadian clock, and clock gene mRNA expression in the suprachiasmatic nucleus (SCN), a central clock, in mice. Although the HE ethanol extract only affected the offset time of activity, the HE water extract advanced the sleep-wake cycle without affecting the free-running period, photosensitivity, or the clock gene mRNA expression in SCN...
2016: Biomedical Research
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