Papers - OHTAKE Kazuo
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L-NAME誘発性高血圧ラットの内皮機能不全と心臓リモデリングに対する魚油摂取の効果 Reviewed
薗田邦博, 河野有華, 大竹一男, 竹之内康広, 柴祥子, 小林順, 加園恵三
金城学院大学消費生活科学研究所紀要 28 ( 1 ) 13 - 28 2024.03
Language：Japanese Publishing type：Research paper (bulletin of university, research institution)
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異なるジューサーを用いて作製した野菜果物ジュースの硝酸塩含有量と官能検査の比較 Reviewed
薗田 邦博、河野 有華、清水 彩子、大竹 一男、柴 祥子、加園 恵三、小林 順
金城学院大学消費生活科学研究所紀要 27 ( 1 ) 11 - 20 2023.03
Language：Japanese Publishing type：Research paper (bulletin of university, research institution)
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非アルコール性脂肪肝炎モデル動物の肝炎と肝線維化に対するSodium nitriteとCaptoprilの併用投与による影響 Reviewed
河野 有華、 薗田 邦博、大竹 一男、 柴 祥子、北森 一哉、小林 順
金城学院大学消費生活科学研究所紀要 27 ( 1 ) 1 - 10 2023.03
Language：Japanese
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Nitric oxide bioavailability for red blood cell deformability in the microcirculation: A review of recent progress. Reviewed International journal
Jun Kobayashi, Kazuo Ohtake, Isamu Murata, Kunihiro Sonoda
Nitric oxide : biology and chemistry 129 25 - 29 2022.12
Authorship：Second author Language：English Publishing type：Research paper (scientific journal)
The rheological properties of red blood cells (RBCs) play an important role in their microcirculation. RBCs can elastically deform in response to mechanical forces to pass through narrow vessels for effective gas exchange in peripheral tissues. Decreased RBC deformability is observed in lifestyle-related diseases such as diabetes mellitus, hypercholesterolemia, and hypertension, which are pathological conditions linked to increased oxidative stress and decreased nitric oxide (NO) bioavailability. Redox-sensitive cysteine residues on RBC cytoskeletal proteins, such as α- and β-spectrins, responsible for membrane flexibility, are affected by prolonged oxidative stress, leading to reversible and irreversible oxidative modifications and decreased RBC deformability. However, endogenously, and exogenously generated NO protects RBC membrane flexibility from further oxidative modification by shielding redox-sensitive cysteine residues with a glutathione cap. Recent studies have shown that nitrate-rich diets and moderate exercise can enhance NO production to increase RBC deformability by increasing the interplay between RBCs and vascular endothelium-mediated NO bioavailability for microcirculation. This review focuses on the molecular mechanism of RBC- and non-RBC-mediated NO generation, and how diet- and exercise-derived NO exert prophylactic effects against decreased RBC deformability in lifestyle-related diseases with vascular endothelial dysfunction.
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Effects of dietary palmitoleic acid on vascular function in aorta of diabetic mice. Reviewed International journal
Yasuhiro Takenouchi, Yoshie Seki, Sachiko Shiba, Kazuo Ohtake, Koji Nobe, Keizo Kasono
BMC endocrine disorders 22 ( 1 ) 103 - 103 2022.04
Language：English Publishing type：Research paper (scientific journal)
BACKGROUND: Chronic hyperglycemia in diabetes causes atherosclerosis and progresses to diabetic macroangiopathy, and can lead to coronary heart disease, myocardial infarction and cerebrovascular disease. Palmitoleic acid (POA) is a product of endogenous lipogenesis and is present in fish and vegetable oil. In human and animal studies, POA is reported as a beneficial fatty acid related to insulin sensitivity and glucose tolerance. However, few studies have reported its effects on aortic function in diabetes. Here, we investigated the effects of POA administration on vascular function in KKAy mice, a model of type 2 diabetes. METHODS: Male C57BL/6 J (control) and KKAy (experimental) mice at the age of 14 weeks were used in the present study. For each mouse strain, one group was fed with reference diet and a second group was fed POA-containing diet for 2 weeks. The vascular reactivities of prepared aortic rings were then measured in an organ bath to determine if POA administration changed vascular function in these mice. RESULTS: KKAy mice treated with POA exhibited decreased plasma glucose levels compared with mice treated with reference diet. However, endothelium-dependent vasorelaxant responses to acetylcholine and protease-activated receptor 2 activating protein, which are attenuated in the aorta of KKAy mice compared to C57BL/6 J mice under a reference diet, were not affected by a 2-week POA treatment. In addition, assessment of vasoconstriction revealed that the phenylephrine-induced vasoconstrictive response was enhanced in KKAy mice compared to C57BL/6 J mice under a reference diet, but no effect was observed in KKAy mice fed a POA-containing diet. In contrast, there was an increase in vasoconstriction in C57BL/6 J mice fed the POA-containing diet compared to mice fed a reference diet. Furthermore, the vasoconstriction in aorta in both C57BL/6 J and KKAy mice fed a POA-containing diet were further enhanced under hyperglycemic conditions compared to normal glucose conditions in vitro. In the hyperinsulinemic, and hyperinsulinemic combined with hyperglycemic conditions, vasoconstriction was increased in KKAy mice fed with POA. CONCLUSION: These results suggest that POA intake enhances vasoconstriction under hyperglycemic and hyperinsulinemic conditions, which are characteristics of type 2 diabetes, and may contribute to increased vascular complications in diabetes.
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Beneficial Effects of Dietary Nitrite on a Model of Nonalcoholic Steatohepatitis Induced by High-Fat/High-Cholesterol Diets in SHRSP5/Dmcr Rats: A Preliminary Study. Reviewed International journal
Kunihiro Sonoda, Yuka Kono, Kazuya Kitamori, Kazuo Ohtake, Sachiko Shiba, Keizo Kasono, Jun Kobayashi
International journal of molecular sciences 23 ( 6 ) 2022.03
Language：English Publishing type：Research paper (scientific journal)
Nonalcoholic steatohepatitis (NASH) is a chronic liver disease that leads to liver cirrhosis and hepatocellular carcinoma. Endothelial dysfunction caused by hepatic lipotoxicity is an underlying NASH pathology observed in the liver and the cardiovascular system. Here, we evaluated the effect of dietary nitrite on a rat NASH model. Stroke-prone, spontaneously hypertensive 5/Dmcr rats were fed a high-fat/high-cholesterol diet to develop the NASH model, with nitrite or captopril (100 mg/L, each) supplementation in drinking water for 8 weeks. The effects of nitrite and captopril were evaluated using immunohistochemical analyses of the liver and heart tissues. Dietary nitrite suppressed liver fibrosis in the rats by reducing oxidative stress, as measured using the protein levels of nicotinamide adenine dinucleotide phosphate oxidase components and inflammatory cell accumulation in the liver. Nitrite lowered the blood pressure in hypertensive NASH rats and suppressed left ventricular chamber enlargement. Similar therapeutic effects were observed in a captopril-treated rat NASH model, suggesting the possibility of a common signaling pathway through which nitrite and captopril improve NASH pathology. In conclusion, dietary nitrite attenuates the development of NASH with cardiovascular involvement in rats and provides an alternative NASH therapeutic strategy.
DOI： 10.3390/ijms23062931
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Contribution of Pyk2 pathway and reactive oxygen species (ROS) to the anti-cancer effects of eicosapentaenoic acid (EPA) in PC3 prostate cancer cells. Reviewed International journal
Keiichi Oono, Kazuo Ohtake, Chie Watanabe, Sachiko Shiba, Takashi Sekiya, Keizo Kasono
Lipids in health and disease 19 ( 1 ) 15 - 15 2020.01
Authorship：Second author Language：English Publishing type：Research paper (scientific journal)
BACKGROUND: n-3 polyunsaturated fatty acids (n-3 PUFAs), including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are thought to exert protective effects in cardiovascular diseases. In addition, n-3 PUFAs have demonstrated anti-cancer effects in vitro and in vivo. OBJECTIVE: We investigated the anti-cancer effects and mechanism of action of EPA on PC3 prostate cancer cells in vitro. METHODS: PC3 cells were treated with various concentrations of EPA, and cell survival and the abilities of migration and invasion were evaluated. The time course of the growth inhibitory effect of EPA on PC3 cells was also assessed. The mechanism underlying the anti-cancer effects of EPA was investigated by human phosphokinase and human apoptosis antibody arrays, and confirmed by western blot analysis. We also examined the contribution of reactive oxygen species (ROS) to the effects of EPA using the ROS inhibitor N-acetyl cysteine. RESULTS: EPA decreased the survival of PC3 cells in a dose-dependent manner within 3 h of application, with an effective concentration of 500 μmol/L. EPA inhibited proline-rich tyrosine kinase (Pyk)2 and extracellular signal-regulated kinase 1/2 phosphorylation as determined by western blotting and the antibody arrays. The growth of PC3 cells was inhibited by EPA, which was dependent on ROS induction, while EPA inhibited Pyk2 phosphorylation independent of ROS production. CONCLUSIONS: Inhibition of Pyk2 phosphorylation and ROS production contribute to the anticancer effects of EPA on PC3 cells.
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食事性亜硝酸塩摂取の炎症および免疫機能への影響と心血管系の健康とのかかわり Invited Reviewed
大竹一男 薗田邦博 小林 順
栄養学レビュー 28 ( 3 ) 175 - 193 2020
Authorship：Lead author Language：Japanese Publishing type：Research paper (scientific journal)
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Chronic Treatment with α-Lipoic Acid Improves Endothelium-Dependent Vasorelaxation of Aortas in High-Fat Diet-Fed Mice. Reviewed
Yasuhiro Takenouchi, Kazuhito Tsuboi, Kenji Ohsuka, Koji Nobe, Kazuo Ohtake, Yasuo Okamoto, Keizo Kasono
Biological & pharmaceutical bulletin 42 ( 9 ) 1456 - 1463 2019
Language：English Publishing type：Research paper (scientific journal)
α-Lipoic acid (ALA) is used as a dietary supplement and known as an anti-oxidant. The present study aimed to examine whether ALA improves endothelial dysfunction in high-fat diet-fed obese mice. After feeding a high-fat diet to Institute of Cancer Research (ICR) mice for 4 weeks, the mice were maintained with a high-fat diet (group HF) or a high-fat diet containing ALA (25 mg/d, group HF + ALA) for an additional 20 weeks. Age-matched normal diet-fed mice were also used (group Normal). Chronic oral treatment with ALA did not affect various plasma parameters or body weights. As compared with the aortas of Normal mice, those from HF mice showed impaired endothelium-dependent relaxation in response to clonidine. However, such an impairment was not observed in the aortas from HF + ALA mice. The plasma levels of thiobarbituric acid reactive substances, an indicator of oxidative stress, were significantly decreased in HF + ALA mice compared with HF mice, confirming the anti-oxidative effects of ALA. In addition, when the impaired clonidine-induced vasorelaxation of aortas from normal mice under high glucose conditions was used as a model of acute oxidative stress, the vasorelaxation responses were improved in the presence of ALA at 100 µM. Our results suggested that the chronic oral administration of ALA improves endothelial dysfunction in high-fat diet-fed obese mice possibly through the reduction in oxidative stress in vivo.
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Eicosapentaenoic acid ethyl ester improves endothelial dysfunction in type 2 diabetic mice. Reviewed International journal
Yasuhiro Takenouchi, Kazuo Ohtake, Koji Nobe, Keizo Kasono
Lipids in health and disease 17 ( 1 ) 118 - 118 2018.05
Language：English Publishing type：Research paper (scientific journal)
BACKGROUND: Eicosapentaenoic acid (EPA) is thought to have many beneficial effects, such as anti-atherosclerogenic and anti-inflammatory properties. However, few studies have reported its effects of endothelial dysfunction in diabetes and its direct effects on the aorta. Here, we investigated the effects of EPA treatment on impaired endothelium-dependent relaxation of the aorta in KKAy mice, a model of type 2 diabetes. METHODS: Male KKAy mice were fed a high-fat (HF) diet for 8 weeks to induce diabetes, after which they were divided into two groups. One group was fed a HF diet, and the other group was fed a HF diet containing EPA ethyl ester (EPA-E, 10 mg/day) for 4 weeks. Then, the vascular reactivities of prepared aortic rings were measured in an organ bath to determine if EPA-E administration changed vascular function in these diabetic mice. In addition, we examined effect of EPA-E and its metabolites to vascular action using aorta separated from C57BL/6 J mice. RESULTS: Although EPA-E administration did not change the plasma glucose and insulin levels in diabetic mice, total cholesterol levels were significantly decreased. The aorta extracted from EPA-E untreated diabetic mice showed impaired endothelium-dependent relaxation in response to acetylcholine (ACh). However, EPA-E administration improved the relaxation response to ACh to the control levels observed in non-diabetic C57BL/6 J mice. On the other hand, endothelium-independent relaxation in response to sodium nitroprusside did not significantly differ among these three groups. The enhanced contractile response by phenylephrine in diabetic mice was not altered by the administration of EPA-E. In addition, the direct administration of EPA-E metabolites such as EPA, docosahexaenoic acid, and docosapentaenoic acid led to vasodilation in the aortic rings of C57BL/6 J mice. CONCLUSION: These results showed that chronic EPA-E administration prevented the development of endothelial dysfunction in KKAy mice, partly via the direct action of EPA-E metabolites on the aorta.
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L-NAME誘発性高血圧ラットの血圧に対する亜硝酸塩とニフェジピンの相互作用 Reviewed
金城学院大学消費生活科学研究所紀要 22 ( 1 ) 7 - 16 2018.03
Language：Japanese
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Dietary Nitrite Attenuates Elastase-Induced Pulmonary Emphysema in a Mouse Model. Reviewed
Kunihiro Sonoda, Kazuo Ohtake, Maya Tagiri, Miku Hirata, Hazuki Tamada, Hiroyuki Uchida, Junta Ito, Jun Kobayashi
Biological & pharmaceutical bulletin 41 ( 12 ) 1818 - 1823 2018
Authorship：Second author Language：English Publishing type：Research paper (scientific journal)
Pulmonary emphysema (PE) is a major pathological feature of chronic obstructive pulmonary disease (COPD) and is characterized by proteolytic destruction of the alveolar structure and subsequent inflammation of the respiratory tract. We hypothesized that nitrite attenuates the development of PE via anti-inflammatory actions. PE was induced by intratracheal instillation of porcine pancreas elastase (PPE) in mice. Dietary nitrite dose-dependently (50 and 150 mg/L in drinking water) attenuated emphysematous development and macrophage accumulation in the alveolar parenchyma 21 d after PPE treatment. The present study shows that dietary nitrite might be a possible nutritional strategy in preventing the development of PE in mice.
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Dietary nitrite supplementation attenuates cardiac remodeling in L-NAME-induced hypertensive rats Reviewed International journal
Kunihiro Sonoda, Kazuo Ohtake, Hiroyuki Uchida, Junta Ito, Masaki Uchida, Hideshi Natsume, Hazuki Tamada, Jun Kobayashi
NITRIC OXIDE-BIOLOGY AND CHEMISTRY 67 1 - 9 2017.07
Authorship：Second author Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE
Loss of nitric oxide (NO) bioavailability underlies the development of hypertensive heart disease. We investigated the effects of dietary nitrite on N-G-nitro-L-arginine methyl ester (L-NAME)-induced hypertension. Sprague-Dawley rats were divided into five groups: an untreated control group, an L-NAME-treated group, and three other L-NAME-treated groups supplemented with 10 mg/L or 100 mg/L of nitrite or 100 mg/L of captopril in drinking water. After the 8-week experimental period, mean arterial blood pressure was measured, followed by sampling of blood and heart tissue for assessment of nitrite/nitrate levels in the plasma and heart, the plasma level of angiotensin II (AT II), and the heart transcriptional levels of AT II type 1 receptor (AT(1)R), transforming growth factor-beta (TGF-beta 1), and connective tissue proteins such as type 1 collagen and fibronectin. Heart tissue was analyzed by histopathological morphomety, including assessments of ventricular and coronary vascular hypertrophy and fibrosis, as well as immunohistochemistry analyses of myocardial expression of AT(1)R. L-NAME treatment reduced the plasma nitrate level and led to the development of hypertension, with increased plasma levels of AT II and increased heart transcriptional levels of AT(1)R and TGF-beta 1-mediated connective tissue proteins, showing myocardial and coronary arteriolar hypertrophy and fibrosis. However, dietary nitrite supplementation inhibited TGF-beta 1-mediated cardiac remodeling by suppressing AT II and AT(1)R. These results suggest that dietary nitrite levels achievable via a daily high-vegetable diet could improve hypertensive heart disease by inhibiting AT II-AT(1)R-mediated cardiac remodeling. (C) 2017 Elsevier Inc. All rights reserved.
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Dietary nitrite reverses features of postmenopausal metabolic syndrome induced by high-fat diet and ovariectomy in mice. Reviewed International journal
Kazuo Ohtake, Nobuyuki Ehara, Hiroshige Chiba, Genya Nakano, Kunihiro Sonoda, Junta Ito, Hiroyuki Uchida, Jun Kobayashi
American journal of physiology. Endocrinology and metabolism 312 ( 4 ) E300-E308 2017.04
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
Menopausal women are at greater risk of developing metabolic syndrome with reduced endothelial nitric oxide synthase (eNOS) activity. Hormone replacement therapy increases eNOS activity and normalizes some characteristics of metabolic syndrome. We hypothesized that nitric oxide (NO) supplementation should have a therapeutic effect on this syndrome. We examined the effect of dietary nitrite in a mouse model with postmenopausal metabolic syndrome induced by ovariectomy (OVX) and a high fat diet (HF). C57BL/6 female mice were divided into five groups, sham+normal fat diet (NF), sham+ HF, OVX+HF with or without sodium nitrite (50 mg and 150 mg/l) in the drinking water. Daily food intake and weekly body weight were monitored for 18 wk. OVX and HF significantly reduced plasma levels of nitrate/nitrite (NOx), and mice developed obesity with visceral hypertrophic adipocytes and increased transcriptional levels of monocyte chemoattractant protein-1, TNF-α, and IL-6 in visceral fat tissues. The proinflammatory state in the adipocytes provoked severe hepatosteatosis and insulin resistance in OVX+HF group compared with sham+NF group. However, dietary nitrite significantly suppressed adipocyte hypertrophy and transcriptions of proinflammatory cytokines in visceral fat in a dose-dependent manner. The improvement of visceral inflammatory state consequently reversed the hepatosteatosis and insulin resistance observed in OVX+HF mice. These results suggest that an endogenous NO defect might underlie postmenopausal metabolic syndrome and that dietary nitrite provides an alternative source of NO, subsequently compensating for metabolic impairments of this syndrome.
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NO-Rich Diet for Lifestyle-Related Diseases. Reviewed International journal
Jun Kobayashi, Kazuo Ohtake, Hiroyuki Uchida
Nutrients 7 ( 6 ) 4911 - 37 2015.06
Authorship：Second author Language：English Publishing type：Research paper (scientific journal)
Decreased nitric oxide (NO) availability due to obesity and endothelial dysfunction might be causally related to the development of lifestyle-related diseases such as insulin resistance, ischemic heart disease, and hypertension. In such situations, instead of impaired NO synthase (NOS)-dependent NO generation, the entero-salivary nitrate-nitrite-NO pathway might serve as a backup system for NO generation by transmitting NO activities in the various molecular forms including NO and protein S-nitrosothiols. Recently accumulated evidence has demonstrated that dietary intake of fruits and vegetables rich in nitrate/nitrite is an inexpensive and easily-practicable way to prevent insulin resistance and vascular endothelial dysfunction by increasing the NO availability; a NO-rich diet may also prevent other lifestyle-related diseases, including osteoporosis, chronic obstructive pulmonary disease (COPD), and cancer. This review provides an overview of our current knowledge of NO generation through the entero-salivary pathway and discusses its safety and preventive effects on lifestyle-related diseases.
DOI： 10.3390/nu7064911
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Dietary nitrite supplementation improves insulin resistance in type 2 diabetic KKA(y) mice. Reviewed International journal
Ohtake K, Nakano G, Ehara N, Sonoda K, Ito J, Uchida H, Kobayashi J
Nitric oxide : biology and chemistry 44 31 - 38 2015.01
Language：English Publishing type：Research paper (scientific journal)
BACKGROUND: Because insulin signaling is essential for endothelial nitric oxide synthase (eNOS)-derived nitric oxide (NO) production, the loss of bioavailable NO might be a common molecular mechanism underlying the development of insulin resistance and endothelial dysfunction. Although dietary nitrite acts as a substrate for systemic NO generation, thereby serving as a physiological alternative source of NO for signaling, it is not precisely known how dietary nitrite affects type 2 diabetes mellitus. Here we report the therapeutic effects of dietary nitrite on the metabolic and histological features of KKA(y) diabetic mice. METHODS: KKA(y) mice were divided into three groups (without nitrite, and with 50 mg/L and 150 mg/L nitrite in drinking water), and two groups of C57BL/6J mice served as controls (without nitrite and with 150 mg/L nitrite in drinking water). After 10 weeks, blood samples, visceral adipose tissues, and gastrocnemius muscles were collected after a 16-hour fast to assess the homeostasis model assessment of insulin resistance (HOMA-IR) levels, the histology of the adipose tissue, insulin-stimulated sequential signaling to glucose transporter 4 (GLUT4), and nitrite and nitrate contents in the muscle using an HPLC system. RESULTS: KKA(y) mice developed obesity with enhanced fasting plasma levels of glucose and insulin and exhibited increased HOMA-IR scores compared with the C57BL/6J control mice. Dietary nitrite dose-dependently reduced the size of the hypertrophic adipocytes and TNF-α transcription in the adipose tissue of KKA(y) diabetic mice, which also restored the insulin-mediated signal transduction, including p85 and Akt phosphorylation, and subsequently restored the GLUT4 expression in the skeletal muscles. CONCLUSIONS: These results suggest that dietary nitrite provides an alternative source of NO, and subsequently improves the insulin-mediated signaling and the metabolic and histological features in KKA(y) diabetic mice.
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A Mechanism Enhancing Macromolecule Transport Through Paracellular Spaces Induced by Poly-L-Arginine: Poly-L-Arginine Induces the Internalization of Tight Junction Proteins via Clathrin-Mediated Endocytosis Reviewed International journal
Tsutomu Yamaki, Yusuke Kamiya, Kazuo Ohtake, Masaki Uchida, Toshinobu Seki, Hideo Ueda, Jun Kobayashi, Yasunori Morimoto, Hideshi Natsume
PHARMACEUTICAL RESEARCH 31 ( 9 ) 2287 - 2296 2014.09
Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER/PLENUM PUBLISHERS
Poly-L-arginine (PLA) enhances the paracellular permeability of the Caco-2 cell monolayer to hydrophilic macromolecules by disappearance of tight junction (TJ) proteins from cell-cell junctions. However, the mechanism of the disappearance of TJ proteins in response to PLA has been unclear. In this study, we investigated the mechanism of disappearance of TJ proteins from cell-cell junctions after the application of PLA to Caco-2 cell monolayers.
The membrane conductance (G(t)), FITC-dextran (FD-4) permeability, and localization of TJ proteins were examined after the treatment of Caco-2 cell monolayers with PLA in the presence of various endocytosis inhibitors. In addition, the localization of endosome marker proteins was also observed.
Clathrin-mediated endocytosis inhibitors suppressed the increase in G(t) and P-app of FD-4 induced by PLA, and also significantly suppressed the disappearance of TJ proteins induced by PLA. Furthermore, occludin, one of the TJ proteins, colocalized with early endosome and recycling endosomes after the internalization of occludin induced by PLA, and then was recycled to the cell-cell junctions.
PLA induced the transient internalization of TJ proteins in cell-cell junctions via clathrin-mediated endocytosis, subsequently increasing the permeability of the Caco-2 cell monolayer to FD-4 via a paracellular route. -
Aldehyde dehydrogenase 2 partly mediates hypotensive effect of nitrite on L-NAME-induced hypertension in normoxic rat. Reviewed International journal
Kunihiro Sonoda, Kazuo Ohtake, Yoshinori Kubo, Hiroyuki Uchida, Masaki Uchida, Hideshi Natsume, Miya Kobayashi, Jun Kobayashi
Clinical and experimental hypertension (New York, N.Y. : 1993) 36 ( 6 ) 410 - 8 2014
Authorship：Second author Language：English Publishing type：Research paper (scientific journal)
Nitrite has become a topic of interest in the field of medical research because of its potential therapeutic role as an alternative source of nitric oxide (NO). While the bioconversion of nitrite to NO occurs via either nonenzymatic or enzymatic reduction under acidic or hypoxic conditions, little is known about its conversion to NO under normoxic conditions. Because of a recent report of aldehyde dehydrogenase 2 (ALDH2)-catalyzed glyceryl trinitrate (GTN) vasorelaxation by denitration of GTN to 1,2-glyceryl dinitrate (1,2-GDN) and nitrite, we therefore investigated a catalytic activity of ALDH2 for nitrite reduction and subsequent effect on N(ω)-nitro-l-arginine methyl ester (l-NAME)-induced hypertension in normoxic rat. Male Sprague-Dawley rats treated with l-NAME in drinking water for 3 weeks developed hypertension with significantly reduced plasma levels of nitrite and nitrate. The intravenous injection of sodium nitrite lowered the arterial pressure in a dose-dependent manner (17, 50 and 150 μmol/kg). Pretreatment with ALDH2 inhibitors (cyanamide and chloral hydrate) partially inhibited the hypotensive responses to sodium nitrite. In addition, cyanamide significantly delayed the nitrite clearance from plasma and most of the organs examined during the experimental period. These results suggest that ALDH2 may be at least in part involved in nitrite-mediated hypotensive effects and nitrite catalysis in many organs of normoxic rats.
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Tsutomu Yamaki, Kazuo Ohtake, Keiko Ichikawa, Masaki Uchida, Hiroyuki Uchida, Shinji Oshima, Kazuhiko Juni, Jun Kobayashi, Yasunori Morimoto, Hideshi Natsume
Biological and Pharmaceutical Bulletin 36 ( 3 ) 432 - 441 2013.03
Authorship：Second author Language：English Publishing type：Research paper (scientific journal)
We investigated whether poly-l-arginine (PLA) enhances the paracellular permeability of the Caco-2 monolayer to hydrophilic macromolecules and clarified the disposition of tight junction (TJ) proteins. The transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC)-dextran (FD-4) permeation were determined after treatment with PLA. TJ proteins were visualized using immunofluorescence microscopy after PLA exposure and depletion, and their expression levels were determined. The barrier function of TJs was also evaluated by measuring the alterations in the TEER and in the localization of TJ proteins. PLA induced an increase in hydrophilic macromolecule, FD-4, permeation through Caco-2 cell monolayers and a decrease in the TEER in a concentration-dependent manner, without any significant impact on the cell viability. This increased paracellular permeability induced by PLA was found to be internalized of claudin-4, ZO-1, tricellulin and mainly occludin from cell-cell junction to the subcellular space. ZO-1 appeared to play an important role in the reconstitution of TJ strand structures following PLA depletion. These results indicate that the PLA led to the internalization of TJ proteins to the subcellular space, subsequently increasing the permeability of the Caco-2 cell monolayer to FD-4 via a paracellular route. © 2013 The Pharmaceutical Society of Japan.
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Effect of poly-L-arginine on intestinal absorption of hydrophilic macromolecules in rats. Reviewed
Tsutomu Yamaki, Masaki Uchida, Yusuke Kuwahara, Yohei Shimazaki, Kazuo Ohtake, Mitsutoshi Kimura, Hiroyuki Uchida, Jun Kobayashi, Masahiko Ogihara, Yasunori Morimoto, Hideshi Natsume
Biological & pharmaceutical bulletin 36 ( 3 ) 496 - 500 2013
Language：English Publishing type：Research paper (scientific journal)
We have already reported that poly-L-arginine (PLA) remarkably enhanced the in vivo nasal absorption of hydrophilic macromolecules without producing any significant epithelial damage in rats. In the present study, we examined whether PLA could enhance the absorption of a model hydrophilic macromolecule, fluorescein isothiocyanate-dextran (FD-4), across the intestinal mucosa, as well as the nasal mucosa, by an in situ closed-loop method using the rat intestine. PLA was found to enhance the intestinal absorption of FD-4 in a concentration-dependent manner within the concentrations investigated in this study, but segment-specific differences were found to be associated with this effect (ileum>jejunum>duodenum≧colon). The factors responsible for the segment-specific differences were also investigated by intestinal absorption studies using aprotinin, a trypsin inhibitor, and an analysis of the expression of occludin, a tight junction protein. In the small intestine, the differences in the effect of PLA on the absorption of FD-4 may be related to the enzymatic degradation of PLA. In the colon, the reduced effect of PLA on the absorption of FD-4 may be related to the smaller surface area for absorption and the higher expression of occludin compared with other segments.
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Oral nitrite ameliorates dextran sulfate sodium-induced acute experimental colitis in mice. Reviewed International journal
Ohtake K, Koga M, Uchida H, Sonoda K, Ito J, Uchida M, Natsume H, Kobayashi J
Nitric oxide : biology and chemistry 23 ( 1 ) 65 - 73 2010.08
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
Inflammatory bowel diseases (IBDs) such as Crohn's disease and ulcerative colitis are chronic inflammatory disorders of the intestinal tract with excessive production of cytokines, adhesion molecules, and reactive oxygen species. Although nitric oxide (NO) is reported to be involved in the onset and progression of IBDs, it remains controversial as to whether NO is toxic or protective in experimental colitis. We investigated the effects of oral nitrite as a NO donor on dextran sulfate sodium (DSS)-induced acute colitis in mice. Mice were fed DSS in their drinking water with or without nitrite for up to 7days. The severity of colitis was assessed by disease activity index (DAI) observed over the experimental period, as well as by the other parameters, including colon lengths, hematocrit levels, and histological scores at day 7. DSS treatment induced severe colitis by day 7 with exacerbation in DAI and histological scores. We first observed a significant decrease in colonic nitrite levels and increase in colonic TNF-alpha expression at day 3 after DSS treatment, followed by increased colonic myeloperoxidase (MPO) activity and increased colonic expressions of both inducible NO synthase (iNOS) and heme oxygenase-1 (HO-1) at day 7. Oral nitrite supplementation to colitis mice reversed colonic nitrite levels and TNF-alpha expression to that of normal control mice at day 3, resulting in the reduction of MPO activity as well as iNOS and HO-1 expressions in colonic tissues with clinical and histological improvements at day 7. These results suggest that oral nitrite inhibits inflammatory process of DSS-induced experimental colitis by supplying nitrite-derived NO instead of impaired colonic NOS activity.
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Proteomic approach for identification of protein S-nitrosation in mouse gastric mucosa treated with S-nitrosoglutathione. Reviewed International journal
Kazuo Ohtake, Norihisa Shimada, Hiroyuki Uchida, Jun Kobayashi
Journal of proteomics 72 ( 5 ) 750 - 60 2009.07
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
Nitric oxide, endogenously generated and exogenously supplied to the stomach, plays an important role in gastric physiological and pathological processes, including epithelial secretion, barrier function, immune response, and carcinogenesis. One of the mechanisms by which NO and related nitroso-compounds transmit their signals is S-nitrosation-mediated protein modification. To screen and identify gastric mucosal proteins that are uniquely sensitive to S-nitrosation, we reacted mouse gastric mucosal lysates with S-nitrosoglutathione, a physiologically relevant nitrosating agent, then performed proteomic analysis including the biotin-switch assay. The results showed that more than sixty protein spots on 2-DE were detected, and thirty-two of these were identified by MALDI-TOF MS and PMF. Eight of these identified proteins were unique S-nitrosated proteins not previously reported. Using Western blot technique, we further confirmed S-nitrosation especially in four proteins such as carbonic anhydrase-2, nucleoside diphosphate kinase B, Raf kinase inhibitor protein, and galectin-2, all known to be closely related to gastric mucosal protection and integrity, cell migration, and tumor metastasis. In addition, ex vivo study closer to in vivo situation also demonstrated these four proteins significantly S-nitrosated with S-nitrosoglutathione in mouse gastric mucosa. These findings will provide useful information regarding the linkage of protein S-nitrosation to gastric physiology and pathophysiology.
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Dietary nitrite inhibits early glomerular injury in streptozotocin-induced diabetic nephropathy in rats. Reviewed International journal
Kazuo Ohtake, Yuichi Ishiyama, Hiroyuki Uchida, Etsuko Muraki, Jun Kobayashi
Nitric oxide : biology and chemistry 17 ( 2 ) 75 - 81 2007.09
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
Increased production of reactive oxygen species (ROS) is a key event leading to microvascular complications, including nephropathy, in diabetes mellitus (DM). Excessive ROS and oxidative stress in DM have been reported to be associated with subsequent impaired nitric oxide (NO) bioavailability. The aim of this study is to examine the beneficial function of dietary nitrite supplementation as an interventional NO donor to attenuate early progression of diabetic nephropathy. To test this hypothesis, male Sprague-Dawley rats were randomly divided into four groups: non-diabetic rats given water with or without nitrite (nitrite-treated or untreated, respectively), and streptozotocin-induced diabetic rats given water with or without nitrite (nitrite-treated or untreated, respectively). After a 4 week experimental period, untreated diabetic rats exhibited significantly higher malondialdehyde (MDA) levels in the kidney compared with untreated non-diabetic rats, accompanied by a reduction in levels of endogenous NO synthase-derived nitrite. However, dietary nitrite supplementation to diabetic rats not only decreased MDA levels but also increased nitrite levels in the kidney to the same levels as in the non-diabetic kidney. These improvements accompanied an improvement in the parameters of glomerular injury, including urinary protein and albumin excretion, histopathological glomerular hypertrophy, and mesangial matrix accumulation. These results indicate that dietary nitrite is effective in the prevention of early diabetic glomerular injury in which NO bioavailability is impaired.
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Poly-L-arginine enhances paracellular permeability via serine/threonine phosphorylation of ZO-1 and tyrosine dephosphorylation of occludin in rabbit nasal epithelium. Reviewed International journal
Kazuo Ohtake, Takuya Maeno, Hideo Ueda, Masahiko Ogihara, Hideshi Natsume, Yasunori Morimoto
Pharmaceutical research 20 ( 11 ) 1838 - 45 2003.11
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
PURPOSE: The purpose of the present study is to explore whether a poly-L-arginine (poly-L-Arg)-induced increase in tight junctions (TJ) permeability of fluorescein isothiocyanate-labeled dextran (MW 4.4 kDa, FD-4) is associated with the Ca2+-dependent signaling and occurs following the phosphorylation/dephosphorylation of TJ proteins. METHODS: Excised rabbit nasal epithelium was mounted in an Ussing-type chamber for measurement of FD-4 transport and membrane conductance (Gt) in the presence of various inhibitors that are involved in the Ca2+-dependent pathway and the phosphorylation/dephosphorylation of TJ proteins. The resultant distribution of TJ proteins was observed using confocal laser scanning microscopy (CLSM) in an immunostaining. RESULTS: The increase in TJ permeability of FD4 induced by 0.2 mg/ml poly-L-Arg was not altered by treatment with inhibitors (of possible Ca2+ mobilization pathways followed by exposure of poly-L-Arg, suggesting that the promoting effect of poly-L-Arg is independent of Ca2+-related signaling. On the other hand, the protein kinase C (PKC) and tyrosine phosphatase inhibitors suppress the increase in TJ permeability by poly-L-Arg, indicating that serine/threonine phosphorylation by way of Ca2+-independent PKC and tyrosine dephosphorylation of junction proteins may have occurred. Furthermore, immunofluorescent monitoring of ZO-1, a TJ associated protein, and occludin, an integral membrane protein localizing at TJ, after preincubation with PKC and tyrosine phosphatase inhibitors followed by poly-L-Arg treatment has shown that the internalization of ZO-1 and occludin occurred by way of serine/threonine phosphorylation by PKC activation and by way of tyrosine dephosphorylation, respectively, providing TJ disassembly. CONCLUSIONS: We conclude that poly-L-Arg enhances the paracellular permeability of FD-4 (i.e., macromolecules), at least, by way of both serine/threonine phosphorylation of ZO-1 and tyrosine dephosphorylation of occludin in rabbit nasal epithelium.
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Poly-L-arginine predominantly increases the paracellular permeability of hydrophilic macromolecules across rabbit nasal epithelium in vitro. Reviewed International journal
Kazuo Ohtake, Takaya Maeno, Hideo Ueda, Hideshi Natsume, Yasunori Morimoto
Pharmaceutical research 20 ( 2 ) 153 - 60 2003.02
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
PURPOSE: The purpose of this work was to characterize the main transport pathway of hydrophilic macromolecules induced by poly-L-arginine (poly-L-Arg; molecular weight 42.4 kDa) across the excised rabbit nasal epithelium. METHODS: Excised rabbit nasal epithelium was mounted in an Ussing-type chamber for measurement of fluorescein isothiocyanate-labeled dextran (FD-4; molecular weight 4.4 kDa) transport and transepithelial electrical resistance (TEER). The main transport pathway of FD-4 enhanced by poly-L-Arg was evaluated using confocal laser scanning microscopy. Immunolocalization of junction proteins (ZO-1, occludin, and E-cadherin) after treatment with poly-L-Arg was also observed. RESULTS: After apical application of a poly-L-Arg (0.05, 0.5, and 5 mg/mL), the permeability coefficient of FD-4 increased by 1.6-, 2.9-, and 5.2-fold, respectively, compared with the control of 5.2 +/- 1.3 x 10(-7) cm/s. Consistent with the increase in transport, there was a concurrent reduction in TEER. At a concentration of 0.05 mg/mL poly-L-Arg. both FD-4 transport and TEER returned to the control level. A good correlation was obtained between the FD-4 permeability coefficient and 1/TEER. Basolateral application of poly-L-Arg at 5 mg/mL, however, did not increase FD-4 transport. Marked FD-4 fluorescence was located in the paracellular spaces after treatment with apical poly-L-Arg compared with that in the absence of poly-L-Arg. Immunofluorescence of ZO-1, occludin, and E-cadherin in cell-to-cell junctions was reduced and distributed into the cytoplasm by apical application of poly-L-Arg, suggesting that poly-L-Arg regulates the junction proteins to enhance paracellular permeability across the nasal epithelium. After pretreatment with either 2,4-dinitrophenol or ouabain, the enhancing effect of apical poly-L-Arg was abolished, indicating the contribution of metabolic energy (cell viability) to the poly-L-Arg-mediated enhancing effect. CONCLUSION: In the nasal epithelium, apical poly-L-Arg appears to increase predominantly the paracellular transport of hydrophilic macromolecules via disorganization of tight- and adherens-junction proteins. The regulatory mechanism of the poly-L-Arg effect is likely to be dependent on energy-requiring cellular processes.
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Analysis of transient and reversible effects of poly-L-arginine on the in vivo nasal absorption of FITC-dextran in rats. Reviewed
Ohtake K, Natsume H, Ueda H, Morimoto Y.
J Control Release 21 263 - 275 2002.04
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
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市販ビートルート粉末に含まれる硝酸塩量と抗酸化能の比較
薗田邦博 河野有華 大竹一男 清水彩子 柴 祥子 飯野汐里 加園恵三 小林 順
金城学院大学論集(自然科学編) 21 ( (1) ) 1 - 6 2024.09
Language：Japanese Publishing type：Research paper (bulletin of university, research institution)
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英国と日本国内のビートルートジュースに含まれる硝酸塩と抗酸化能(ORAC値)の比較
河野 有華, 薗田 邦博, 清水 彩子, 大竹 一男, 飯野 汐里, 柴 祥子, 小林 順
金城学院大学論集(自然科学編) 20 ( 1 ) 26 - 33 2023.09
Language：Japanese Publisher：金城学院大学
英国で販売されているビートルートジュースと国内で購入できるビートルートジュースの硝酸塩、亜硝酸塩、抗酸化能(ORAC値)等を比較した。比較したのは製品A:英国販売ビートルートジュース(70mL)、製品B:国内販売ビートルートジュース(200mL)、製品C:国内販売ビートルートジュース(100mL)であった。亜硝酸イオン含有量は製品Aと製品Cは同程度であったが、製品Bと比較して有意に多かった。硝酸イオン含有量は製品A(328.3±19.9mg)、製品B(54.6±3.2mg)、製品C(87.5±2.1mg)で、製品Aに最も多く含まれていた。抗酸化能であるORAC値は100mLあたりの値では製品Bと製品Cと比べて製品Aが有意に値が高かったが、1製品当たりに換算すると製品Cと同程度となり、製品BのORAC値が高かった。ビートルートジュース製品であっても英国販売と国内販売のもので成分含有量が大幅に異なること、国内販売のものにも含有量に違いがあることが示された。
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硝酸塩由来一酸化窒素補給を期待した市販野菜果物ジュース中の硝酸塩含有量と抗酸化能の比較
河野 有華, 薗田 邦博, 大竹 一男, 清水 彩子, 齋藤 百花, 柴 祥子, 加園 恵三, 小林 順
金城学院大学論集(自然科学編) 19 ( 1 ) 1 - 8 2022.09
Language：Japanese Publisher：金城学院大学
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河野 有華, 薗田 邦博, 北森 一哉, 大竹 一男, 小林 順
日本内分泌学会雑誌 97 ( 5 ) 1503 - 1503 2022.03
Language：Japanese Publisher：(一社)日本内分泌学会
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福祉・医療の現場から 高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果の検討(第2報) Reviewed
加園 恵三, 糸井 郁美, 本松 茂, 柴 祥子, 大竹 一男, 加藤 勇太
地域ケアリング 24 ( 1 ) 42 - 47 2022.01
Language：Japanese Publisher：(株)北隆館
日本は、2007年に65歳以上の高齢者人口が全体の21%を超え、超高齢社会に突入した。高齢者の増加に伴い、認知症患者数も増加の一途をたどっている。増加する認知症への対策として、認知症の前駆段階である軽度認知障害、あるいはそれ以前の段階で感知し、かつ、認知障害が進行しないための支援方法の確立は重要課題である。今回私どもは認知症の早期発見に、ADAS-Jcogが有用であること、また、高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果が期待できることを報告する。(著者抄録)
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福祉・医療の現場から 高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果の検討(第2報) Reviewed
加園 恵三, 糸井 郁美, 本松 茂, 柴 祥子, 大竹 一男, 加藤 勇太
地域ケアリング 23 ( 6 ) 56 - 61 2021.06
Language：Japanese Publisher：(株)北隆館
日本は、2007年に65歳以上の高齢者人口が全体の21%を超え、超高齢社会に突入した。高齢者の増加に伴い、認知症患者数も増加の一途をたどっている。増加する認知症への対策として、認知症の前駆段階である軽度認知障害、あるいはそれ以前の段階で感知し、かつ、認知障害が進行しないための支援方法の確立は重要課題である。今回私どもは認知症の早期発見に、ADAS-Jcogが有用であること、また、高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果が期待できることを報告する。(著者抄録)
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NO不足妊娠高血圧ラットの胎仔発育不全に対する亜硝酸塩補給の影響
薗田 邦博, 大竹 一男, 河野 有華, 小林 順
金城学院大学論集(自然科学編) 17 ( 2 ) 3 - 9 2021.03
Language：Japanese Publisher：金城学院大学
一酸化窒素(NO)合成阻害剤であるN-omega-nitro-L-arginine methyl ester(L-NAME)を用いたNO不足妊娠高血圧モデル動物を作成し、胎児発育不全(FGR)と胎盤構造に対する食事性亜硝酸塩の補給による影響を調査した。動物を、体重で4匹ずつControl(蒸留水)群、L-NAME(1g/L)群、L-NAME(1g/L)と亜硝酸ナトリウム(100mg/L)を混合した亜硝酸塩群の3群に無作為に分けた。L-NAMEや亜硝酸塩摂取による母体の体重、摂食量、摂水量、尿量について測定したが実験開始前と開始後で、各グループ間に有意な差はなかった。また、解剖時に母体重量、胎仔数、胎盤重量を測定した。その結果、各グループで有意な差は認められなかった。血圧においては、Control群に比べL-NAME群と亜硝酸塩群で有意に上昇したが、亜硝酸塩群はL-NAME群に比べ血圧の上昇を抑制した。各群における胎仔重量を比較したところ、Control群に比べL-NAME群で胎児体重の有意な減少が認められた。一方で、亜硝酸塩を補給したグループではControl群と有意な差は認められなかった。組織標本による胎盤の組織構造の変化を確認したところ、L-NAME群において母体と胎仔の栄養交換の場であるラビリンスゾーンの構造に空洞が多く確認された。一方で、亜硝酸塩群では構造の壊れている部分(空洞)が認められたもののL-NAME群に比べ少なかった。酸化ストレスマーカーである8-OHdGの尿中濃度は、Control群やL-NAME群に比べ亜硝酸塩群で有意に増加した。
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福祉の現場から 高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果の検討(第2報) Reviewed
加園 恵三, 糸井 郁美, 本松 茂, 柴 祥子, 大竹 一男, 加藤 勇太
地域ケアリング 23 ( 2 ) 60 - 65 2021.02
Language：Japanese Publisher：(株)北隆館
日本は、2007年に65歳以上の高齢者人口が全体の21%を超え、超高齢社会に突入した。高齢者の増加に伴い、認知症患者数も増加の一途をたどっている。増加する認知症への対策として、認知症の前駆段階である軽度認知障害、あるいはそれ以前の段階で感知し、かつ、認知障害が進行しないための支援方法の確立は重要課題である。今回私どもは認知症の早期発見に、ADAS-Jcogが有用であること、また、高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果が期待できることを報告する。(著者抄録)
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低酸素環境下トレーニングでの魚油濃縮物摂取によるヘモレオロジー的検討 Reviewed
櫛部 静二, 小林 悟, 新井 尚之, 浜崎 景, 野部 浩司, 竹之内 康広, 柴 祥子, 大竹 一男, 白幡 晶, 加園 恵三
脂質栄養学 29 ( 2 ) 127 - 135 2020.09
Language：Japanese Publisher：日本脂質栄養学会
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高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果の検討 亜鉛と認知症の関連 Reviewed
加園 恵三, 柴 祥子, 大竹 一男, 加藤 勇太
細胞 52 ( 3 ) 130 - 134 2020.03
Language：Japanese Publisher：(株)ニュー・サイエンス社
現在、高齢化社会を迎えた日本では、認知症患者数は増加の一途をたどっている。増加する認知症への対策として、認知症の前駆段階である軽度認知障害、あるいはそれ以前の段階で感知し、かつ、認知障害が進行しないための支援方法の確立は重要課題である。筆者らは、これまでに高齢者では複数の栄養素が不足する可能性が高いことを報告した。今回筆者らは認知症の早期発見に、認知機能検査のADAS-Jcogが有用であること、また、高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果が期待できることを報告するとともに、亜鉛欠乏と認知機能障害との関連についても考察する。(著者抄録)
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高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果の検討 Reviewed
加園恵三 糸井郁美 本松茂 高村亜矢乃 柴祥子 大竹一男 加藤勇太
別冊地域ケアリング 22 ( 1 ) 67 - 71 2020.01
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：北隆館
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福祉の現場から 高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果の検討 Reviewed
加園 恵三, 糸井 郁美, 本松 茂, 高村 亜矢乃, 柴 祥子, 大竹 一男, 加藤 勇太
地域ケアリング 22 ( 1 ) 67 - 71 2020.01
Language：Japanese Publisher：(株)北隆館
現在、高齢化社会を迎えた日本では、認知症患者数は増加の一途をたどっている。増加する認知症への対策として、認知症の前駆段階である軽度認知障害、あるいはそれ以前の段階で感知し、かつ、認知障害が進行しないための支援方法の確立は重要課題である。今回私どもは認知症の早期発見に、ADAS-Jcogが有用であること、また、高齢者に不足しがちな微量栄養素の補充による認知機能低下予防効果が期待できることを報告する。(著者抄録)
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絶食による廃用性腸管粘膜萎縮に及ぼす経口グルタチオン(GSH)投与の効果
内田 博之, 中島 由加里, 大竹 一男, 伊東 順太, 森田 匡彦, 神村 彩子, 小林 順
アミノ酸研究 12 ( 1 ) 59 - 60 2019.02
Language：Japanese Publisher：日本アミノ酸学会
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Enhancement Effect of Poly-L-ornithine on the Nasal Absorption of Water-Soluble Macromolecules in Rats. Reviewed
Shigehiro Omori, Yusuke Kamiya, Tsutomu Yamaki, Masaki Uchida, Kazuo Ohtake, Mitsutoshi Kimura, Hideshi Natsume
Biological & pharmaceutical bulletin 42 ( 1 ) 144 - 148 2019
Language：English Publishing type：Research paper (scientific journal)
The transnasal route for the delivery of water-soluble macromolecules, such as bioactive peptides and proteins, has attracted interest, although the use of permeation enhancers is required due to the poor permeabilities of these macromolecules across the nasal mucosa. With polycationic compounds, such as poly-L-arginine and chitosan, the nasal absorption of hydrophilic macromolecules is molecular weight- and concentration-dependently enhanced without causing cytotoxicity. In the present study, we evaluated the effect of various molecular weights and concentrations of poly-L-ornithine (PLO), a polycationic compound, on the nasal absorption and the damage to the nasal mucosa in vivo. PLO enhanced the nasal absorption of fluorescein isothiocyanate-dextran (FD-4), used as a model drug, and the bioavailability of FD-4 increased with the concentration of PLO. The enhancement effect was also dependent on the molecular weight. The administration of PLO at a concentration that sufficed for enhancing the nasal absorption had no effect on the activity of lactic dehydrogenase and the protein leakage in the nasal fluid, as indices of nasal mucosa damage. These findings suggest that a transnasal delivery system using PLO is a useful strategy for improving the nasal absorption of water-soluble macromolecules without toxicity to the nasal mucosa.
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ベイズ型age-period-cohort分析を用いた日本の予測平均余命の性差(2023～2047年) Reviewed
内田 博之, 三藤 瑠莉, 瓶子 英朗, 斎藤 雅文, 小田切 陽一, 大竹 一男, 八巻 努, 内田 昌希, 夏目 秀視, 小林 順
日本衛生学雑誌 73 ( 3 ) 338 - 353 2018.09
Language：Japanese Publisher：(一社)日本衛生学会
日本の1958～2012年までの全死因および主要死因別の性別年齢階級別死亡数は人口動態統計、1958～2012年までの性別年齢階級別人口は国勢調査報告および10月1日現在推計人口、2013～2047年までの日本の性別年齢階級別将来推計人口は国立社会保障・人口問題研究所より得た。1958～1962年から2008～2012年にかけて男女ともすべての年齢階級別死亡率は低下傾向を示し、特に15歳未満でその傾向が大きかった。出生年次別の死亡率は、男女ともすべての年齢階級で出生年の推移に伴い低下傾向を示し、特に20歳未満の傾きが大きかった。全死因の死亡率の年次推移に対する年齢効果、時代効果およびコホート効果の推計値の比較より、死亡率の年次推移は、男女ともに年齢効果が一番大きく影響し、男性でコホート効果の次に時代効果、女性で時代効果の次にコホート効果が影響した。平均余命の将来予測では、2008～2012年から2043～2047年までの期間に、0～4歳の平均余命は2023～2027年に一端短縮するものの男性が1.64年、女性が0.70年延伸し、他の年齢階級においても同様に平均余命の延伸が予測された。
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Improved Intranasal Retentivity and Transnasal Absorption Enhancement by PEGylated Poly-l-ornithine. Reviewed International journal
Yusuke Kamiya, Tsutomu Yamaki, Shigehiro Omori, Masaki Uchida, Kazuo Ohtake, Mitsutoshi Kimura, Hiroshi Yamazaki, Hideshi Natsume
Pharmaceuticals (Basel, Switzerland) 11 ( 1 ) 2018.01
Language：English Publishing type：Research paper (scientific journal)
We reported that the introduction of polyethylene glycol (PEG) to poly-l-ornithine (PLO), which is an homopolymeric basic amino acid having absorption-enhancement ability, prolonged retention time in an in vitro inclined plate test, probably due to an increase in viscosity caused by PEGylation. The aim of the present study is to investigate whether the introduction of PEG chains to PLO improves intranasal retention and transnasal absorption in vivo. We performed intranasal administration experiments using PLO and PEG-PLO with a model drug, fluorescein isothiocyanate dextran (FD-4), in rats under closed and open systems. In the open system, transition of plasma FD-4 concentration after co-administration with unmodified PLO was low, and the area under the plasma concentration-time curve (AUC) decreased to about 60% of that in the closed system. In contrast, the AUC after co-administration with PEG-PLO in the open system was about 90% of that in the closed system, and the transition of plasma FD-4 concentration and FD-4 absorption profile were similar to those of the closed system. These findings indicate that introducing PEG chains to homopolymeric basic amino acids (HPBAAs) is a very useful method for developing a functional absorption enhancer that can exhibit an efficient in vivo absorption-enhancing effect.
DOI： 10.3390/ph11010009
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Hiroyuki Uchida, Ruri Mito, Hideaki Heishi, Masafumi Saito, Youichi Odagiri, Kazuo Ohtake, Tutomu Yamaki, Masaki Uchida, Hideshi Natsume, Jun Kobayashi
Nihon eiseigaku zasshi. Japanese journal of hygiene 73 ( 3 ) 338 - 353 2018
Language：Japanese Publishing type：Research paper (scientific journal)
OBJECTIVES: In this study, we aimed to (1) determine the effects of age, period, and cohort on mortality rate trends between 1958 and 2012 in Japan and (2) assess gender differences in projected life expectancy (LE) for the 2023-2047 period. METHODS: A time trend study was conducted using age-period-cohort (APC) analysis. A Bayesian APC model was fitted to describe mortality rate trends for the 1958-2012 period and to project mortality rates for 2023-2047. LE was predicted by Chiang's method using projected mortality rates. RESULTS: Age, period, and cohort effects showed similar patterns between males and females. As time passes, gender differences in projected LE were larger among individuals over 65 years than among those under 65 years. Time series change rates of the extension of projected LE after excluding specific causes of death showed the following: smaller extension of projected LE in males in terms of mortality risk from malignant neoplasms, heart diseases, pneumonia, and accidents (under 65 years) and in females in terms of mortality risk from heart diseases, cerebrovascular diseases, and suicide (over 65 years). CONCLUSIONS: Gender differences in projected LE are expected to be smaller before middle age and to be larger among seniors. These projected gender differences stem in part from the lower mortality risk among men than among women from malignant neoplasms, heart diseases, pneumonia, and accidents (under 65 years), and among women compared to men from heart disease, cerebrovascular disease, and suicide (over 65 years).
DOI： 10.1265/jjh.73.338
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Protective effects of oral glutathione on fasting-induced intestinal atrophy through oxidative stress Reviewed International journal
Hiroyuki Uchida, Yukari Nakajima, Kazuo Ohtake, Junta Ito, Masahiko Morita, Ayako Kamimura, Jun Kobayashi
WORLD JOURNAL OF GASTROENTEROLOGY 23 ( 36 ) 6650 - 6664 2017.09
Language：English Publishing type：Research paper (scientific journal) Publisher：BAISHIDENG PUBLISHING GROUP INC
AIM
To determine whether oral glutathione (GSH) administration can alleviate the effects of fasting-induced intestinal atrophy in the small intestinal mucosa.
METHODS
Rats were divided into eight groups. One group was fed ad libitum, another was fed ad libitum and received oral GSH, and six groups were administrated saline (SA) or GSH orally during fasting. Mucosal height, apoptosis, and cell proliferation in the jejunum were histologically evaluated. iNOS protein expression (by immunohistochemistry), nitrite levels (by high performance liquid chromatography, as a measure of NO production), 8-hydroxydeoxyguanosine formation (by ELISA, indicating ROS levels), glutathione/ oxidized glutathione (GSH/GSSG) ratio (by enzymatic colorimetric detection), and gamma-glutamyl transpeptidase (Ggt1) mRNA levels in the jejunum (by semi-quantitative RT-PCR) were also estimated.
RESULTS
Oral GSH administration was demonstrated to drastically reduce fasting-induced intestinal atrophy in the jejunum. In particular, jejunal mucosal height was enhanced in GSH-treated animals compared to SA-treated animals [527.2 +/- 6.9 for 50 mg/kg GSH, 567.6 +/- 5.4 for 500 mg/kg GSH vs 483.1 +/- 4.9 (mu m), P < 0.01 at 72 h]. This effect was consistent with decreasing changes in GSH-treated animals compared to SA-treated animals for iNOS protein staining [0.337 +/- 0.016 for 50 mg/kg GSH, 0.317 +/- 0.017 for 500 mg/kg GSH vs 0.430 +/- 0.023 (area of staining part/area of tissue), P < 0.01 at 72 h] and NO [2.99 +/- 0.29 for 50 mg/kg GSH, 2.88 +/- 0.19 for 500 mg/kg GSH vs 5.34 +/- 0.35 (nmol/g tissue), P < 0.01 at 72 h] and ROS [3.92 +/- 0.46 for 50 mg/kg GSH, 4.58 +/- 0.29 for 500 mg/kg GSH vs 6.42 +/- 0.52 (8-OHdG pg/mu g DNA), P < 0.01, P < 0.05 at 72 h, respectively] levels as apoptosis mediators in the jejunum. Furthermore, oral GSH administration attenuated cell proliferation decreases in the fasting jejunum [182.5 +/- 1.9 for 500 mg/kg GSH vs 155.8 +/- 3.4 (5-BrdU positive cells/10 crypts), P < 0.01 at 72 h]. Notably, both GSH concentration and Ggt1 mRNA expression in the jejunum were also attenuated in rats following oral administration of GSH during fasting as compared with fasting alone [0.45 +/- 0.12 vs 0.97 +/- 0.06 (nmol/mg tissue), P < 0.01; 1.01 +/- 0.11 vs 2.79 +/- 0.39 (Ggt1 mRNA/Gapdh mRNA), P < 0.01 for 500 mg/kg GSH at 48 h, respectively].
CONCLUSION
Oral GSH administration during fasting enhances jejunal regenerative potential to minimize intestinal mucosal atrophy by diminishing fasting-mediated ROS generation and enterocyte apoptosis and enhancing cell proliferation. -
Inducible and neuronal nitric oxide synthases exert contrasting effects during rat intestinal recovery following fasting Reviewed International journal
Junta Ito, Hiroyuki Uchida, Naomi Machida, Kazuo Ohtake, Yuki Saito, Jun Kobayashi
EXPERIMENTAL BIOLOGY AND MEDICINE 242 ( 7 ) 762 - 772 2017.04
Language：English Publishing type：Research paper (scientific journal) Publisher：SAGE PUBLICATIONS LTD
We investigated the effects of endogenous inducible (iNOS) and neuronal nitric oxide synthase on recovery from intestinal mucosal atrophy caused by fasting-induced apoptosis and decreased cell proliferation during refeeding in rats. Rats were divided into five groups, one of which was fed ad libitum, and four of which underwent 72h of fasting, followed by refeeding for 0, 6, 24, and 48 h, respectively. iNOS and neuronal nitric oxide synthase mRNA and protein levels in jejunal tissues were measured, and mucosal height was histologically evaluated. Apoptotic indices, interferon-gamma (IFN-gamma) transcription levels, nitrite levels (as a measure of nitric oxide [NO] production),8-hydroxydeoxyguanosine formation (indicating reactive oxygen species [ROS] levels), crypt cell proliferation, and the motility indices (MI) were also estimated. Associations between mucosal height and NOS protein levels were determined using Spearman's rank correlation test. Notably, we observed significant increases in mucosal height and in neuronal nitric oxide synthase mRNA and protein expression as refeeding time increased. Indeed, there was a significant positive correlation between neuronal nitric oxide synthase protein level and mucosal height during the 48-h refeeding period (r=0.725, P<0.01). Conversely, iNOS mRNA and protein expression decreased according to refeeding time, with a significant negative correlation between iNOS protein level and mucosal height being recorded during the 48-h refeeding period (r=-0.898, P<0.01). We also noted a significant negative correlation between jejunal neuronal nitric oxide synthase and iNOS protein concentrations over this same period (r=-0.734, P<0.01). Refeeding also restored the decreased jejunal MI caused by fasting. Our finding suggests that refeeding likely repairs fasting-induced jejunal atrophy by suppressing iNOS expression and subsequently inhibiting NO, ROS, and IFN- as apoptosis mediators, and by promoting neuronal nitric oxide synthase production and inducing crypt cell proliferation via mechanical stimulation.
Impact statement
Besides providing new data confirming the involvement of iNOS and nNOS in intestinal mucosal atrophy caused by fasting, this study details their expression and function during recovery from this condition following refeeding. We demonstrate a significant negative correlation between iNOS and nNOS levels during refeeding, and associate this with cell proliferation and apoptosis in crypts and villi. These novel findings elucidate the relationship between these NOS isoforms and its impact on recovery from intestinal injury. A mechanism is proposed comprising the up-regulation of nNOS activity by mechanical stimulation due to the presence of food in the intestine, restricting iNOS-associated apoptosis and promoting cell proliferation and gut motility. Our investigation sheds light on the molecular basis behind the repercussions of total parenteral nutrition on intestinal mucosal integrity, and more importantly, the beneficial effects of early enteral feeding. -
Identification of the cysteine residue responsible for oxidative inactivation of mouse galectin-2. Reviewed International journal
Mayumi Tamura, Akari Sasai, Rika Ozawa, Masanori Saito, Kaori Yamamoto, Tomoharu Takeuchi, Kazuo Ohtake, Hiroaki Tateno, Jun Hirabayashi, Jun Kobayashi, Yoichiro Arata
Journal of biochemistry 160 ( 4 ) 233 - 241 2016.10
Language：English Publishing type：Research paper (scientific journal)
Galectins are a group of animal lectins characterized by their specificity for β-galactosides. Mouse galectin-2 (mGal-2) is predominantly expressed in the gastrointestinal tract and has been identified as one of the main gastric mucosal proteins that are uniquely sensitive to S-nitrosylation. We have previously reported that oxidation of mGal-2 by hydrogen peroxide (H2O2) resulted in the loss of sugar-binding ability, whereas pre-treatment of mGal-2 with S-nitrosocysteine prevented H2O2-induced inactivation. In this study, we used point-mutated recombinant mGal-2 proteins to study which of the two highly conserved Cys residues in mGal-2 must be S-nitrosylated for protection against oxidative inactivation. Mutation of Cys57 to a Met residue (C57M) did not result in lectin inactivation following H2O2 treatment, whereas Cys75 mutation to Ser (C75S) led to significantly reduced lectin activity, as is the case for wild-type mGal-2. However, pre-treatment of the C75S mutant with S-nitrosocysteine protected the protein from H2O2-induced inactivation. Therefore, Cys57 is suggested to be responsible for oxidative inactivation of the mGal-2 protein, and protection of the sulfhydryl group of the Cys57 in mGal-2 by S-nitrosylation is likely important for maintaining mGal-2 protein function in an oxidative environment such as the gastrointestinal tract.
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Development of a Transnasal Delivery System for Recombinant Human Growth Hormone (rhGH): Effects of the Concentration and Molecular Weight of Poly-L-arginine on the Nasal Absorption of rhGH in Rats Reviewed
Ryo Kawashima, Masaki Uchida, Tsutomu Yamaki, Kazuo Ohtake, Tomomi Hatanaka, Hiroyuki Uchida, Hideo Ueda, Jun Kobayashi, Yasunori Morimoto, Hideshi Natsumea
BIOLOGICAL & PHARMACEUTICAL BULLETIN 39 ( 3 ) 329 - 335 2016.03
Language：English Publishing type：Research paper (scientific journal) Publisher：PHARMACEUTICAL SOC JAPAN
A novel system for delivering recombinant human growth hormone (rhGH) that is noninvasive and has a simple method of administration is strongly desired to improve the compliance of children. The aim of this study was to investigate the potential for the intranasal (i.n.) co-administration of rhGH with poly-L-arginine (PLA) as a novel delivery system by evaluating the effects of the concentration and molecular weight of PLA on the nasal absorption of rhGH. The influence of the formation of insoluble aggregates and a soluble complex in the dosage formulation on nasal rhGH absorption was also evaluated by size-exclusion chromatography and ultrafiltration. PLA enhanced the nasal absorption of rhGH at each concentration and molecular weight examined. Nasal rhGH absorption increased dramatically when the PLA concentration was 1.0 % (w/v) due to the improved solubility of rhGH in the formulation. A delay in rhGH absorption was observed when the molecular weight of PLA was increased. This appeared to be because the increase in molecular weight caused the formation of a soluble complex. It seems that the PLA concentration affects the absorption-enhancing effect on rhGH, while the molecular weight of PLA affects the time when the maximum plasma rhGH concentration was reached (T-max) of rhGH after i.n. administration, mainly because of the interactions among rhGH, PLA, and additives. Therefore, the transnasal rhGH delivery system using PLA is considered to be a promising alternative to subcutaneous (s.c.) injection if these interactions are sufficiently controlled.
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大学男子駅伝選手の常圧下低酸素環境を利用したトレーニングにおける魚油製剤摂取の効果 Reviewed
櫛部 静二, 小林 悟, 竹之内 康広, 金 賢珠, 新井 尚之, 野部 浩司, 大竹 一男, 白幡 晶, 加園 恵三
脂質栄養学 25 ( 1 ) 61 - 74 2016.03
Language：Japanese Publisher：日本脂質栄養学会
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Identification of the cysteine residue responsible for oxidative inactivation of mouse galectin-2. Reviewed International journal
Tamura M, Sasai A, Ozawa R, Saito M, Yamamoto K, Takeuchi T, Ohtake K, Tateno H, Hirabayashi J, Kobayashi J, Arata Y.
J Biochem. 160 233 - 241 2016
Language：English Publishing type：Research paper (scientific journal)
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S-nitrosylation of mouse galectin-2 prevents oxidative inactivation by hydrogen peroxide. Reviewed International journal
Mayumi Tamura, Masanori Saito, Kaori Yamamoto, Tomoharu Takeuchi, Kazuo Ohtake, Hiroaki Tateno, Jun Hirabayashi, Jun Kobayashi, Yoichiro Arata
Biochemical and biophysical research communications 457 ( 4 ) 712 - 7 2015.02
Language：English Publishing type：Research paper (scientific journal)
Galectins are a group of animal lectins characterized by their specificity for β-galactosides. Galectin-2 (Gal-2) is predominantly expressed in the gastrointestinal tract. A proteomic analysis identified Gal-2 as a protein that was S-nitrosylated when mouse gastric mucosal lysates were reacted with S-nitrosoglutathione, a physiologically relevant S-nitrosylating agent. In the present study, recombinant mouse (m)Gal-2 was S-nitrosylated using nitrosocysteine (CysNO), which had no effect on the sugar-binding specificity and dimerization capacity of the protein. On the other hand, mGal-2 oxidation by H2O2 resulted in the loss of sugar-binding ability, while S-nitrosylation prevented H2O2-inducted inactivation, presumably by protecting the Cys residue(s) in the protein. These results suggest that S-nitrosylation by nitric oxides protect Gal-2 from oxidative stress in the gastrointestinal tract.
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都道府県別の平均要介護期間と損失生存可能年数の地域格差と医療・福祉資源の関連について 医薬品情報に着目した地域相関研究 Reviewed
内田 博之, 中村 拓也, 金子 彩野, 大竹 一男, 内田 昌希, 小田切 陽一, 夏目 秀視, 小林 順
厚生の指標 61 ( 3 ) 15 - 24 2014.03
Language：Japanese Publisher：(一財)厚生労働統計協会
目的 平均要介護期間と年齢調整YPLL(years of potential life lost)率に着目し,各指標の都道府県別の地域格差と医療・福祉資源との関連を明らかにし,医薬品情報を含んだ関連要因の抽出を目的として地域相関研究を行った。方法 2008年の厚生労働省,総務省の各種統計資料のデータを使用し,都道府県別の平均要介護期間と年齢調整YPLL率を算出した。また,都道府県別の医療・福祉資源に関する要因のデータも得た。2つの指標と各種要因との間の相関係数を算出し,統計学的に有意な相関を示す要因を抽出した。相関マトリクスを作成し,多重共線性を配慮して候補要因を絞り,2つの指標を目的変数とした重回帰分析を行った。結果 平均要介護期間との相関が認められた要因のうち医薬品情報に関する要因として,男性では「電算処理済み処方箋1枚当たりの報酬別内訳の技術料」および「特定保険医療材料料」が抽出されたが,重回帰分析の結果からは,「糖尿病内科医師数」と「居宅介護サービスの通所介護利用者数」が関連の大きい説明変数として得られた。女性では医薬品情報に関する要因として,「電算処理済み処方箋1枚当たりの報酬別内訳の特定保険医療材料料」が抽出されたが,重回帰分析の結果からは,「リウマチ科医師数」と「居宅介護サービスの訪問介護利用者数」が関連の大きい説明変数として得られた。年齢調整YPLL率との相関が認められた要因のうち医薬品情報に関する要因として,男性では「薬剤師数」「薬局総数」「調剤の電算化率」および「後発医薬品調剤率」が抽出されたが,重回帰分析の結果からは,「後発医薬品調剤率」「特別養護老人ホームの定員」および「薬局総数」が関連の大きい説明変数として得られた。女性では医薬品情報に関する要因として,「薬局総数」「内服薬処方箋1枚当たりの薬剤料の3要素分解(1種類1日当たり薬剤料)」が抽出されたが,重回帰分析の結果からは,「呼吸器内科医師数」が説明変数として得られた。結論 相関分析の結果より,男女ともに平均要介護期間および年齢調整YPLL率に影響を与えている要因には,医薬品情報に関連した要因が説明変数の候補として抽出されたが,重回帰分析の結果より,医薬品情報と関連した要因として,男性において「後発医薬品調剤率」と「薬局総数」が年齢調整YPLL率との関連の大きい要因として把握された。(著者抄録)
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[Age, period, and birth cohort-specific effects on cervical cancer mortality rates in Japanese women and projections for mortality rates over 20-year period (2012-2031)]. Reviewed
Hiroyuki Uchida, Mizuki Kobayashi, Ami Hosobuchi, Ayano Ohta, Kazuo Ohtake, Tutomu Yamaki, Masaki Uchida, Youichi Odagiri, Hideshi Natsume, Jun Kobayashi
Nihon eiseigaku zasshi. Japanese journal of hygiene 69 ( 3 ) 215 - 24 2014
Language：Japanese Publishing type：Research paper (scientific journal)
OBJECTIVES: We aimed to determine the effects of age, period, and birth cohort on cervical cancer mortality rate trends in Japanese women, by age-period-cohort (APC) analysis. Additionally, we analyzed projected mortality rates. METHODS: We obtained data on the number of cervical cancer deaths in Japanese women from 1975-2011 from the national vital statistics and census population data. A cohort table of mortality rate data was analyzed on the basis of a Bayesian APC model. We also projected the mortality rates for the 2012-2031 period. RESULTS: The period effect was relatively limited, compared with the age and cohort effects. The age effect increased suddenly from 25-29 to 45-49 years of age and gently increased thereafter. An analysis of the cohort effect on mortality rate trends revealed a steep decreasing slope for birth cohorts born from 1908-1940 and a subsequent sudden increase after 1945. The mortality rate projections indicated increasing trends from 40 to 74 years of age until the year 2031. CONCLUSIONS: The age effect increased from 25-29 years of age. This could be attributable to the high human papilloma virus (HPV) infection risk and the low cervical cancer screening rate. The cohort effect changed from decreasing to increasing after the early 1940s. This might be attributable to the spread of cervical cancer screening and treatment before 1940 and the high HPV infection risk and reduced cervical cancer screening rate after 1945. The projected mortality rate indicated an increasing trend until the year 2031.
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Acute lethal crush-injured rats can be successfully rescued by a single injection of high-dose dexamethasone through a pathway involving PI3K-Akt-eNOS signaling. Reviewed International journal
Isamu Murata, Kazuya Ooi, Shingo Shoji, Yohei Motohashi, Miwa Kan, Kazuo Ohtake, Soichiro Kimura, Hideo Ueda, Genya Nakano, Kunihiro Sonoda, Yutaka Inoue, Hiroyuki Uchida, Ikuo Kanamoto, Yasunori Morimoto, Jun Kobayashi
The journal of trauma and acute care surgery 75 ( 2 ) 241 - 9 2013.08
Language：English Publishing type：Research paper (scientific journal)
BACKGROUND: Crush syndrome (CS) is characterized by ischemia/reperfusion-induced rhabdomyolysis and the subsequent onset of systemic inflammation. CS is associated with a high mortality, even when patients are treated with conventional therapy. We hypothesized that treatment of lethal CS rat model with dexamethasone (DEX) have therapeutic effects on the laboratory findings and clinical course and outcome. METHODS: To create a CS model, anesthetized rats were subjected to bilateral hind limb compression with rubber tourniquets for 5 hours and randomly divided into three groups as follows: saline-treated CS group, CS groups treated with low (0.1 mg/kg) and high doses (5.0 mg/kg) of DEX. Saline for the CS group or DEX for the DEX-treated CS groups was intravenously administered immediately before reperfusion. Under continuous monitoring and recording of arterial blood pressures, blood and tissue samples were collected for histologic and biochemical analysis at designated period before and after reperfusion. RESULTS: Ischemic compression of rat hind limbs reduced the nitrite content in the crushed muscle, and the subsequent reperfusion induced reactive oxygen species-mediated circulatory collapse and systemic inflammation, finally resulting in a mortality rate of 76% by 48 hours after reperfusion. A single injection of high-dose DEX immediately before reperfusion activated endothelial nitric oxide synthase (eNOS) by sequential phosphorylation through the nongenomic phosphoinositide 3-kinase (PI3K)-Akt-eNOS signaling pathway. DEX also exhibited anti-inflammatory effects by modulating proinflammatory and anti-inflammatory mediators, consequently suppressing myeloperoxidase activities and subsequent systemic inflammation, showing a complete recovery of the rats from lethal CS. CONCLUSION: These results indicate that high-dose DEX reduces systemic inflammation and contributes to the improved survival rate in a rat CS model.
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肥満を伴うインスリン抵抗性マウスに及ぼす亜硝酸塩摂取の影響に関する研究 Invited
大竹一男
日本食品化学研究振興財団研究成果報告書 ( 19 ) 2013
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デキストラン硫酸ナトリウム誘導大腸炎モデルマウスに与えるセルロース粉末の影響 Reviewed
清水 純, 紙谷 ひとみ, 大竹 一男, 内田 博之, 小林 順, 真野 博
ルミナコイド研究 16 ( 2 ) 71 - 79 2012.12
Language：Japanese Publisher：(一社)日本食物繊維学会
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Nitrite reduces ischemia/reperfusion-induced muscle damage and improves survival rates in rat crush injury model Reviewed International journal
Isamu Murata, Ryo Nozaki, Kazuya Ooi, Kazuo Ohtake, Soichiro Kimura, Hideo Ueda, Genya Nakano, Kunihiro Sonoda, Yutaka Inoue, Hiroyuki Uchida, Ikuo Kanamoto, Yasunori Morimoto, Jun Kobayashi
JOURNAL OF TRAUMA AND ACUTE CARE SURGERY 72 ( 6 ) 1548 - 1554 2012.06
Language：English Publishing type：Research paper (scientific journal) Publisher：LIPPINCOTT WILLIAMS & WILKINS
BACKGROUND: Nitrite is an intrinsic signaling molecule with potential therapeutic implications in mammalian ischemia/reperfusion (I/R) injury of the heart, liver, and kidney. Although limb muscle compression and subsequent reperfusion are the causative factors in developing crush syndrome (CS), there has been no report evaluating the therapeutic effects of nitrite on CS. We therefore tested whether nitrite could be a therapeutic agent for the treatment of CS.
METHODS: To create a CS model, anesthetized rats were subjected to bilateral hind limb compression with rubber tourniquets for 5 hours, followed by reperfusion for 0 hour to 6 hours while monitoring blood pressure. Saline for the CS group or sodium nitrite (NaNO2-100, 200, and 500 mu mol/kg) for the nitrite-treated CS groups was intravenously administered immediately before reperfusion. Blood and tissue samples were collected for biochemical analysis.
RESULTS: Tissue nitrite levels in injured muscles were significantly reduced in the CS group compared with the sham group during I/R injury. Nitrite administration to CS rats restored nitric oxide bioavailability by enhancing nitrite levels of the muscle, resulting in a reduction of rhabdomyolysis markers such as potassium, lactate dehydrogenase, and creatine phosphokinase. Nitrite treatment also reduced plasma levels of interleukin-6 and myeloperoxidase activities in muscle and lung tissues, finally resulting in a dose-dependent improvement of survival rate from 24% (CS group) to 36% (NaNO2-100 group) and 64% (NaNO2-200 and 500 groups).
CONCLUSION: These results indicate that nitrite reduces I/R-induced muscle damage through its cytoprotective action and contributes to improved survival rate in a rat CS model. (J Trauma Acute Care Surg. 2012;72: 1548-1554. Copyright (C) 2012 by Lippincott Williams & Wilkins). -
I 型糖尿病モデルラットにおける造影剤腎症に対する N-acetylcysteine の予防効果 Reviewed
澁谷真紀、小原由香、小島俊彦、内田昌希、大竹一男、内田博之、小林順、横田千津子、夏目秀視
Progres in Medicine. 32 119 - 125 2012.04
Language：Japanese Publishing type：Research paper (scientific journal)
Other Link： http://libir.josai.ac.jp/il/meta_pub/G0000284repository_JOS-02873648-32-659
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肥満を伴うインスリン抵抗性マウスに及ぼす亜硝酸塩摂取の影響に関する研究 Invited
大竹一男
日本食品化学研究振興財団研究成果報告書 ( 18 ) 2012
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Characterization of systemic and histologic injury after crush syndrome and intervals of reperfusion in a small animal model. Reviewed International journal
Isamu Murata, Kazuya Ooi, Hiromi Sasaki, Soichiro Kimura, Kazuo Ohtake, Hideo Ueda, Hiroyuki Uchida, Norikiyo Yasui, Yasuhiro Tsutsui, Naoya Yoshizawa, Ichiro Hirotsu, Yasunori Morimoto, Jun Kobayashi
The Journal of trauma 70 ( 6 ) 1453 - 63 2011.06
Language：English Publishing type：Research paper (scientific journal)
BACKGROUND: Prolonged compression of limb muscles and subsequent decompression are important in the development of crush syndrome (CS). We applied a simple rubber tourniquet to rat hind limbs to create a CS model. METHODS: Anesthetized rats were subjected to bilateral hind limb compression for 5 hours followed by decompression and reperfusion for 0 hour, 1 hour, 3 hours, and 24 hours under monitoring of arterial blood pressure and electrocardiography. Blood and tissue samples were collected for histology, biochemical analysis, and tissue myeloperoxidase activity assessment. RESULTS: The survival rates of the CS-model groups remained at 100% until 3 hours, however, dropped to 25% at 24 hours after reperfusion mainly because of hyperkalemia and consequent hypotension observed at 1 hour and deteriorated at 3 hours after reperfusion. Rhabdomyolysis evaluated by circulating and histologic markers of injury was found as early as 1 hour and more marked at 3 hours, resulting in impaired renal function 24 hours after reperfusion. Myeloperoxidase activities increased with incremental periods after reperfusion not only in injured limb muscles but also in kidney and lung, suggesting an abnormal interaction between the vascular endothelium and circulating leukocytes after rhabdomyolysis, possibly causing subsequent multiple organ dysfunction frequently encountered in CS. CONCLUSION: The findings from this study demonstrate the feasibility of a novel small animal model of extremity crush injury. By using this model, the impact of incremental periods of reperfusion on mortality and remote organ dysfunctions can be characterized. Future studies are necessary to better define a threshold for this injury pattern and the impact of other factors underlying this syndrome.
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[Age-, period-, and birth-cohort-specific effects on the male proportion in Japanese newborns and projections for male proportion for 20 periods (2008-2027)]. Reviewed
Hiroyuki Uchida, Mayo Watanabe, Maho Naiki, Junta Ito, Kazuo Ohtake, Youichi Odagiri, Jun Kobayashi
Nihon eiseigaku zasshi. Japanese journal of hygiene 66 ( 3 ) 582 - 8 2011.05
Language：Japanese Publishing type：Research paper (scientific journal)
OBJECTIVES: To determine the age-, period-, and cohort-specific effects on the male proportion in Japanese newborns, we performed an age-period-cohort (APC) analysis in this study. In addition, projections for the male proportion were analyzed. METHODS: We obtained data on live births of newborns for Japanese women in 1947-2007 from the National Vital Statistics. Cohort tables containing data on the male proportion were analyzed using a Bayesian APC model. Projections of the male proportion (2008-2027) were calculated. RESULTS: The age effect decreased when the mothers were 40-44 years old; however, the effect was relatively limited as compared with the period and cohort effects. The period effect increased from 1947 to 1969 and decreased thereafter. Analysis of the cohort effect on male proportion trends revealed a decreasing slope for birth cohorts born between 1905 and 1945 and a subsequent increase after 1958. The projections for male proportion indicated that the male proportion in 2027 would be similar to that in the 1970s. CONCLUSIONS: The age of the mother hardly affected the male proportion. The period effect started decreasing from the latter half of the 1960s. This may be attributable to the high economic growth since 1965 that promoted industrial development that led to environmental pollution, which in turn may have lead to the deterioration of the intrauterine environment. Cohort effects changed from 1958 and exhibited trends toward increase in male proportion; this may be due to improvements in obstetric care. Our results suggest that the male proportion in Japanese newborns will increase in the future.
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肥満を伴うインスリン抵抗性マウスに及ぼす亜硝酸塩摂取の影響に関する研究 Invited
大竹一男
日本食品化学研究振興財団研究成果報告書 ( 17 ) 2011
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Intranasal administration of milnacipran in rats: evaluation of the transport of drugs to the systemic circulation and central nervous system and the pharmacological effect. Reviewed
Masaki Uchida, Takuya Katoh, Mutsuhiro Mori, Takuya Maeno, Kazuo Ohtake, Jun Kobayashi, Yasunori Morimoto, Hideshi Natsume
Biological & pharmaceutical bulletin 34 ( 5 ) 740 - 7 2011
Language：English Publishing type：Research paper (scientific journal)
Recently, transnasal drug delivery has attracted a great deal of attention as an administration route to deliver drugs directly to the central nervous systems (CNS) and drug targeting of the CNS is expected to increase. In the present study, we investigated the possibility of using a transnasal delivery system for milnacipran, a serotonin-noradrenaline reuptake inhibitor (SNRI), by evaluating the transport to the systemic circulation and cerebrospinal fluid (CSF) and the pharmacological effect after intranasal (i.n.) administration. Moreover, the effect of chitosan as a bioadhesive material on the transport to the systemic circulation and CSF and the pharmacological effect after i.n. administration were evaluated. As a result, i.n. administration of milnacipran was found to produce a higher direct delivery to the CNS as well as to the systemic circulation, suggesting that this is a promising route of administration and an alternative to peroral (p.o.) administration. Furthermore, the i.n. co-administration with chitosan led to increased plasma and CSF concentrations and an enhanced pharmacological effect, evaluated by means of the forced swimming test. The results suggested that chitosan produced a long residence time of milnacipran in the nasal cavity due to its bioadhesive effect, leading to the enhanced transport of milnacipran from the systemic circulation to the CNS via the blood-brain barrier by an increase in systemic absorption as well as direct transport to the CNS, resulting in a higher antidepressant effect compared to that with p.o. administration.
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Fasting-induced intestinal apoptosis is mediated by inducible nitric oxide synthase and interferon-{gamma} in rat. Reviewed International journal
Junta Ito, Hiroyuki Uchida, Takayuki Yokote, Kazuo Ohtake, Jun Kobayashi
American journal of physiology. Gastrointestinal and liver physiology 298 ( 6 ) G916-26 2010.06
Language：English Publishing type：Research paper (scientific journal)
Nitric oxide (NO) is associated with intestinal apoptosis in health and disease. This study aimed to investigate the role of intestinal NO in the regulation of apoptosis during fasting in rats. Male Wistar rats were divided into two groups and subcutaneously injected with saline (SA) or aminoguanidine (AG), followed by fasting for 24, 48, 60, and 72 h. At each time point, the jejunum was subjected to histological evaluation for enterocyte apoptosis by histomorphometric assessment and TUNEL analysis. We performed immunohistochemistry for inducible NO synthase (iNOS) expression in the jejunum and measured tissue nitrite levels using HPLC and 8-hydroxydeoxyguanosine adduct using ELISA, indicative of endogenous NO production and reactive oxygen species (ROS) production, respectively. Jejunal transcriptional levels of iNOS, neuronal NO synthase (nNOS), and interferon-gamma (IFN-gamma) were also determined by RT-PCR. Fasting caused significant jejunal mucosal atrophy due to attenuated cell proliferation and enhanced apoptosis with increase in iNOS transcription, its protein expression in intestinal epithelial cells (IEC), and jejunal nitrite levels. However, AG treatment histologically reduced apoptosis with inhibition of fasting-induced iNOS transcription, protein expression, and nitrite production. We also observed fasting-induced ROS production and subsequent IFN-gamma transcription, which were all inhibited by AG treatment. Furthermore, we observed reduced transcriptional levels of nNOS, known to suppress iNOS activation physiologically. These results suggest that fasting-induced iNOS activation in IEC may induce apoptosis mediators such as IFN-gamma via a ROS-mediated mechanism and also a possible role of nNOS in the regulation of iNOS activity in fasting-induced apoptosis.
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The role of neuronal and inducible NOS in the regulation of intestinal apoptosis in fasting and refeeding rat. Reviewed
Ito J, Uchida H, Ohtake K, Kobayashi J.
Nitric Oxide. 22 S72 - S72 2010.04
Language：English Publishing type：Research paper (international conference proceedings)
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Oral nitrite ameliorates dextran sulfate sodium-induced acute experimental colitis in mice. Reviewed
Ohtake K, Koga M, Uchida H, Sonoda K, Ito J, Uchida M, Natsume H, Kobayashi J.
Nitric Oxide 23 ( 1 ) 65 - 73 2010.04
Authorship：Lead author Language：English Publishing type：Research paper (international conference proceedings)
Other Link： http://libir.josai.ac.jp/il/meta_pub/G0000284repository_JOS-j.niox.2010.04.004
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カルセイン含有デンプン溶液の経鼻投与後の鼻腔滞留性とカルセイン血中動態との関係 Reviewed
入江 博美, 大竹 一男, 前野 拓也, 内田 昌希, 内田 博之, 小林 順, 夏目 秀視
薬剤学: 生命とくすり 69 ( 3 ) 219 - 227 2009.05
Language：Japanese Publisher：(公社)日本薬剤学会
ラットを用いた閉鎖実験系から得られる経鼻投与後の動態パラメータと、ラットを斜めに固定し、鼻腔に投与した薬液が傾斜により物理的に鼻腔から流出できるようにした開放実験系から得られる動態パラメータとを比較することで滞留性と吸収性を解析した。モデル薬物は、分子量約600のカルセインを選択した。また、粘膜付着性物質は、potatoおよびtapiocaの2種類のデンプンを用いた。デンプンの粘度が増加すると鼻粘膜に対する付着効果が高まり、除去クリアランスに対する抵抗が増して、粘性溶液製剤の滞留性が高まった。デンプンの添加濃度の低いタピオカデンプンの方が添加濃度が低くても滞留効果が高くなる傾向を認めた。閉鎖系と開放系から得た薬動学的パラメータを比較することで、カルセイン含有デンプン粘性溶液の付着・滞留効果とカルセインの吸収に対する効果を評価できた。
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Dietary nitrite ameliorates dextran sodium-induced acute experimental colitis in mice. Reviewed
Kobayashi J, Ohtake K, Koga M, Uchida H.
Nitric Oxide. 20 S38 - S38 2009.04
Language：English Publishing type：Research paper (international conference proceedings)
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Effect of mitochondrial Aldehyde dehydrogenase on pharmacokinetic profile of nitrite in plasma and organs. Reviewed
Nakanishi K, Kubo Y, Ohtake K, Uchida H, Kotake F, Natsume H, Kobayashi J.
Nitric Oxide. 20 S40 - S40 2009.04
Language：English Publishing type：Research paper (international conference proceedings)
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Proteomic approach for identification of protein S-nitrosylation in mouse gastric mucosa. Reviewed
Ohtake K, Shimada N, Uchida H, Kobayashi J.
Nitric Oxide. 20 S40 - S41 2009.04
Language：English Publishing type：Research paper (international conference proceedings)
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[Age, period and birth-cohort effects on marriage rates in Japanese women between 1985 and 2005, and comparison of trends of effects between marriage and birth rates]. Reviewed
Hiroyuki Uchida, Youichi Odagiri, Kazuo Ohtake, Jun Kobayashi
[Nihon koshu eisei zasshi] Japanese journal of public health 55 ( 7 ) 440 - 8 2008.07
Language：Japanese Publishing type：Research paper (scientific journal)
OBJECTIVE: An age-period-cohort (APC) analysis was performed to provide information about age-, period-, and cohort-specific effects on marriage trends in Japanese women. In addition, the relationships of the trends of age-, period-, and cohort-specific effects between marriage and birth were analyzed. METHODS: We obtained data regarding marriages of Japanese women aged between 19 and 38 years for the period of 1985 to 2005 from the National Vital Statistics. Population data used were for an estimated population, obtained from the Population Estimates Annual Reports. Standard cohort tables comprising marriage and population data were analyzed using a Bayesian APC model to identify age-, period-, and cohort-specific effects on marriage rate trends. Previously obtained data for a similar APC-analysis of birth trends were used to compare the trends in the effects of age, period, and cohort on marriage and birth patterns. For this purpose, the estimated values for each effect were normalized. RESULTS: With regard to the marriage trends in Japanese women, the effect of age was the greatest, peaking at the age of 25 years. The period effect increased after 1997; however, its effect was relatively limited as compared to the other effects. The cohort effect, which was greater than the period effect and less than the age effect, on marriage trends showed a decreasing slope for birth cohorts born after 1966 and subsequent increase after 1982. Comparison of age, period and cohort effects between the trends in marriage and birth rates showed that the age effect distinctly peaked at 25 and 28 years for marriage and births, respectively. The period effect on marriage and birth showed a decreasing trend until 1991 and subsequent increased in 1992 and 1997 for births and marriage, respectively. With regard to the cohort effect on birth rates, a decreasing trend was observed for the birth cohorts after 1961, with increase after 1977. However, with regard to the cohort effect on marriage rates, the decreasing trend observed for birth cohorts after 1966 showed an increase after 1982. CONCLUSION: Among age, period, and birth cohort, age is the most influential factor affecting marriage rates. Period effects appear relatively small, but they increased after 1997. Cohort effects reduced for birth cohorts born after 1966 and subsequently increased after 1982. Results of the comparison study showed that changing patterns of age, period and cohort effects had very similar influences on the trends for marriage and birth rates. However, a 3-year difference was observed between the peaks of the age effect on the two rates. A time lag of 5 years was observed between the turning point in the trend of period effects for marriage and birth rates. The changing patterns of cohort effects on marriage and birth rates were similar, but the turning point for the marriage pattern occurred in a 5-year younger cohort compared with the birth pattern.
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ヨード造影剤による腎障害に対するN-アセチルシステインの経口投与の有効性 Reviewed
岡本 浩, 宮内 雅弘, 西山 雅巳, 三宅 隆之, 桜田 真己, 大竹 一男, 小林 順, 荻原 政彦, 木村 光利, 夏目 秀視
心臓 38 ( 9 ) 915 - 923 2006.09
Language：Japanese Publisher：(公財)日本心臓財団
石心会狭山病院循環器科でのカルテ調査から、造影剤を用いた心カテーテル検査および経皮的冠動脈形成術(PTCA)で発症する腎障害に対するN-アセチルシステイン(ムコフィリン)の経口投与の有効性について評価した。2004年の1年間のカルテ調査(1,128名)から、中程度に腎機能が低下した患者(血清クレアチニン値1.2～3.0mg/dL)で臨床検査データが既知の患者を抽出した結果、ムコフィリン投与群24名、ムコフィリン未投与群(コントロール群)29名であった。腎機能にかかわる検査データ、造影剤腎症の発症率、入院期間などから評価した結果、ムコフィリン投与群の患者のほうが、造影剤投与による腎障害を予防できることが示唆された。リスク疾患別に評価すると、症例数が少なかったが糖尿病および、糖尿病と高血圧を併発している患者でムコフィリンの効果が高かった。また、造影剤腎症の独立予測因子である年齢について評価すると、70歳以上の高齢患者でもムコフィリンの経口投与は有効であった。以上の結果より、造影剤を用いた心カテーテル検査およびPTCAにおいて、ムコフィリンの経口投与は中程度の腎機能の低下を有する患者に対して有効であると考えられた。(著者抄録)
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食餌由来亜硝酸塩の摂取による生活習慣病予防に関する研究～糖尿病性腎症について～
大竹一男、石山裕一、内田博之、小林順
城西大学生命科学センター報告 6 15 - 23 2006.04
Authorship：Lead author, Corresponding author Language：Japanese Publishing type：Research paper (bulletin of university, research institution)
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Improvement of transnasal absorption of macromolecular drugs using tight-junction regulation Reviewed
Hideshi Natsume, Takuya Maeno, Kazuo Ohtake, Yasunori Morimoto
Oto-Rhino-Laryngology Tokyo 48 ( 1 ) 22 - 29 2005.08
Language：Japanese Publishing type：Research paper (scientific journal)
We investigated whether a poly-L-arginine (poly-L-Arg)-induced increase in the tight-junction permeability of FITC-labeled dextran (MW 4.4 kDa, FD-4) is associated with a transient increase in transepithelial electroresistance (PD) and short-circuit current (Isc) immediately after poly-L-Arg exposure and occurs following tight-junction disassembly-related cellular signaling in the rabbit nasal epithelium. A transient PD and Isc increase was associated with increased Cl - secretion induced by poly-L-Arg but not with the enhanced paracellular FD-4 permeability. Enhanced paracellular FD-4 permeability induced by poly-L-Arg was not changed by treatment with inhibitors of possible Ca 2+ mobilization pathways followed by poly-L-Arg exposure, suggesting that the promotional effect of poly-L-Arg is independent of Ca 2+-related signaling. Protein kinase C (PKC) and tyrosine phosphatase inhibitors suppress an increase in tight-junction permeability by poly-L-Arg, indicating that serine/threonine may have been phosphorylated via Ca 2+-independent PKC and tyrosine may have dephosphorylated the junction protein. ZO-1, a tight-junction-associated protein, and occludin, an integral membrane protein localized at the tight junction, were monitored immunofluorescently after preincubation with PKC and tyrosine phosphatase inhibitors followed by poly-L-Arg treatment, showing that ZO-1 and occludin were internalized via serin/threonine phosphorylation by PKC activation and via tyrosine dephosphorylation, providing tight-junction disassembly. We concluded that poly-L-Arg enhances the paracellular permeability of macromolecular drugs via serin/threonine phosphorylation of ZO-1 involving Ca 2+- independent PKC activation and tyrosine dephosphorylation of occludin in the rabbit nasal epithelium. Such phosphorylation and dephosphorylation disperses junction proteins, particularly ZO-1 and occludin, into cytoplasm, possibly followed by tight-junction disassembly. These findings should prove useful in developing transnasal delivery systems for macromolecular drugs with polycationic materials as solute transport enhancers.
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Tight junctionの調節を介した高分子医薬品の経鼻吸収性の改善 Reviewed
夏目 秀視, 前野 拓也, 大竹 一男, 森本 雍憲
耳鼻咽喉科展望 48 ( 補冊1 ) 22 - 29 2005.08
Language：Japanese Publisher：耳鼻咽喉科展望会
電気生理学的な手法を主に用い,家兎摘出鼻粘膜におけるポリ-L-アルギニン(poly-Arg)の細胞間隙開口作用について検討を行った.poly-Arg適用後のFITC標識したデキストラン(FD-4)の透過性および電気生理学的パラメータの変化,一過的な自発的膜電位(PD)および短絡電流(Isc)変化とその後の透過促進効果の関連性,poly-Arg適用後のみかけの透過係数(Pappa)と経上皮電気抵抗逆数(Gt)の増加に対する種々阻害剤の影響,poly-Arg適用後のtight junction関連タンパク質(ZO-1, occludin)に対する種々阻害剤の影響について調べた.その結果,poly-Argの促進効果はtight junctionの調節と深く関わり,水溶性高分子薬物の透過を促進することが明らかになった
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S-ニトロソアルブミンは循環血液中で最も豊富な高分子NOキャリアーとして働いている Invited
大竹 一男
ファルマシア 39 ( 12 ) 1200 - 1201 2003.12
Language：Japanese Publishing type：Research paper (other academic) Publisher：(公社)日本薬学会
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Effect of poly-L-arginine on the nasal absorption of FITC-dextran of different molecular weights and recombinant human granulocyte colony-stimulating factor (rhG-CSF) in rats. Reviewed
Miyamoto M, Natsume H, Satoh I, Ohtake K, Yamaguchi M, Kobayashi D, Sugibayashi K, Morimoto Y.
Int J Pharm. 226 127 - 138 2001.04
Language：English Publishing type：Research paper (scientific journal)
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Improved nasal absorption of drugs using poly-L-arginine: effects of concentration and molecular weight of poly-L-arginine on the nasal absorption of fluorescein isothiocyanate-dextran in rats. Reviewed
Miyamoto M, Natsume H, Iwata S, Ohtake K, Yamaguchi M, Kobayashi D, Sugibayashi K, Yamashina M, Morimoto Y.
Eur J Pharm Biopharm. 52 21 - 30 2001.04
Language：English Publishing type：Research paper (scientific journal)
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Screening of cationic compounds as an absorption enhancer for nasal drug delivery. Reviewed
Natsume H, Iwata S, Ohtake K, Miyamoto M, Yamaguchi M, Hosoya K, Kobayashi D, Sugibayashi K, Morimoto Y.
Int J Pharm. 185 1 - 12 1999.04
Language：English Publishing type：Research paper (scientific journal)
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Enhancing mechanism of macromolecule permeation by poly-L-arginine in rabbit nasal mucosa Reviewed
K. Ohtake, H. Natsume, Y. Morimoto
Proceedings of the Controlled Release Society ( 26 ) 206 - 207 1999
Language：English Publishing type：Research paper (international conference proceedings)
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Enhancing mechanism of poly-L-arginine in nasal absorption of fitc-dextran Reviewed
K. Ohtake, H. Natsume, M. Miyamoto, K. Sugibayashi, Y. Morimoto
Proceedings of the Controlled Release Society ( 25 ) 687 - 688 1998
Language：English Publishing type：Research paper (international conference proceedings) Publisher：Controlled Release Society