Springer Acta Medica Okayama 0236-5731 313 1 2017 In vitro analysis of radioprotective effect of monoterpenes 169 174 EN Ken-ichi Kudo Tadashi Hanafusa Department of Radiation Research, Advanced Science Research CenterOkayama University Toshiro Ono Department of Radiation Research, Advanced Science Research CenterOkayama University Monoterpenes are naturally occurring hydrocarbons composed of two units of isoprenes. They exhibit antioxidant activity to scavenge reactive oxygen species, such as hydroxyl radicals. We investigated the potential of monoterpenes such as thymol, linalool, and menthol to act as radioprotectants. The proliferation of EL4 cells, a mouse lymphoma cell line, treated with linalool at a concentration of 500 μM or more was not affected by X-ray irradiation. Plasmid-nicking assay performed using formamidopyrimidine-DNA glycosylase showed that linalool prevented single strand breaks and oxidized purines on pUC19 plasmid DNA. These findings indicate that linalool has the ability to scavenge reactive oxygen species and is a potential radioprotector. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 71 3 2017 The Immunological Impact of Chemotherapy on the Tumor Microenvironment of Oral Squamous Cell Carcinoma 219 226 EN Hiroaki Takakura Department of Oral and Maxillofacial Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Shohei Domae Department of Oral and Maxillofacial Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Toshiro Ono Department of Radiation Research, Advanced Science Research Center, Okayama University Akira Sasaki Department of Oral and Maxillofacial Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Original Article 10.18926/AMO/55204 　Anticancer drugs induce cell-cycle arrest and apoptosis not only in tumor cells, but also in immune cells. However, many preclinical and clinical findings show that some chemotherapeutic agents can improve the antitumor efficacy of immunotherapy. We immunohistochemically analyzed the degree of immune cell infiltration and the relevance of programmed cell death 1 ligand-1 (PD-L1) expression in surgically resected oral squamous cell carcinoma (OSCC) specimens from patients who had undergone pretreatment with certain chemotherapies and other patients without pretreatment. We divided the patients into the group of neoadjuvant chemotherapy (NAC) patients (n=8) and the nNAC (without NAC) patient group (n=10). We observed that NAC induced infiltrations of CD4, CD8 T cells and CD56 NK cells into the tumor microenvironment. Decreased numbers of Tregs and PD-1-positive cells were observed in the NAC group. No significant difference was observed in the degree of immune-cell infiltration between the patient groups except for CD56 NK cells in the stroma and PD-1 cells in cancer nests. Eighty percent of the nNAC specimens showed intermediate-to-strong PD-L1 protein expression, whereas 75% of the NAC specimens showed down-regulation of the PD-L1 protein, indicating the effectiveness of the chemotherapeutic treatment before surgery. No potential conflict of interest relevant to this article was reported.

Springer Netherlands Acta Medica Okayama 0236-5731 2015 Adsorption and removal of strontium in aqueous solution by synthetic hydroxyapatite EN Yuichi Nishiyama Tadashi Hanafusa Jun Yamashita Yoko Yamamoto Toshiro Ono Hydroxyapatite (HAP) is a main mineral constituent of bone and tooth and has an outstanding biocompatibility. HAP is a possible sorbent for heavy metals in wastewater due to its high adsorption capacity and low water solubility. We developed a removal system of 90Sr from aqueous solution by HAP column procedure. More than 90 % of 90Sr was adsorbed and removed from the 90Sr containing solution. Divalent cations, Ca2+, had little effect on the removal of 90Sr up to a concentration of 1 mmol L－1. This clearly indicates that the HAP column technique is advantageous with respect to the capacity to adsorb 90Sr from water present in the environment. No potential conflict of interest relevant to this article was reported.

Elsevier Ireland Ltd. Acta Medica Okayama 0304-3835 301 1 2011 Isolation and characterization of human lung cancer antigens by serological screening with autologous antibodies 57 62 EN Tadashi Hanafusa Ali Eldib Ali Mohamed Kenta Kitaoka Yoshihiro Ohue Eiichi Nakayama Toshiro Ono Serological analysis of a recombinant cDNA expression library (SEREX) derived from two lung adenocarcinoma cancer cell lines using autologous sera led to the isolation of 41 positive cDNA clones comprising 28 different antigens. They coded for a variety of nuclear and cytoplasmic proteins. Among the antigens, nucleoporin 107 (NUP107) was isolated most frequently (5 of 41 clones). The second most frequently isolated antigen was coded for by C21orf58 (4 of 41 clones). During serological analysis of selected antigens based on their reactivity to sera from normal individuals and lung cancer patients, none of the antigens showed a cancer-restricted recognition pattern. However, five genes including NUP107 showed higher expression when we examined the changes in gene expression in five different adenocarcinoma cell lines, including those used in SEREX, compared with their levels in normal lung tissues by cDNA microarray analysis. On the other hand, the expression levels of five genes including C21orf58 were down regulated in all adenocarcinoma cell lines. This SEREX study combining comprehensive gene expression assays has added to the growing list of lung cancer antigens, which may aid the development of diagnostic and immunotherapeutic reagents for patients with lung cancer. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 65 2 2011 An Assessment of Radioactivity Levels of 210Pb and 40K in Tobacco and Radiation Exposure from Smoking 91 95 EN Tomohiro Nagamatsu Akihiro Sakoda Takahiro Kataoka Toshiro Ono Kiyonori Yamaoka Original Article 10.18926/AMO/45267 No research has been conducted on the radiation influence of tobacco on the alimentary system, although there have been some previous works on the respiratory system. In this study, the radioactive concentrations of 210Pb and 40K in a cigarette sample were first measured. The transfer factors of the nuclides from tobacco into smoke and solution (saliva and/or alcohol) were then examined. Moreover, the radiation doses from smoke inhalation were also evaluated. The radioactive concentrations of 210Pb and 40K in the cigarette tobacco were 0.01 and 0.3 Bq/cigarette. Since this 210Pb activity and the 210Po activity previously reported for the same sample were comparable, it can be concluded that there was a radioactive equilibrium between the 2 nuclides. The observed transfer factor of 210Pb (12%) into smoke was almost the same as that of 40K (15%), whereas the reported value for 210Po (60%) was significantly higher. The radiation doses due to inhalation of cigarette smoke varied from organ to organ, depending on the organotropic properties of the nuclide. For example, the kidneys, respiratory tract, and spleen showed relatively high doses from 210Pb and 210Po. The leaching rates indicated an inconsistent tendency related to solution types. This result could suggest that alcohol drinking, which is common in smokers, does not especially enhance the leaching characteristics. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 48 5 1994 Allergens of the house dust mite, Dermatophagoides pteronyssinus, in patients with mite allergic rhinitis: a clinical investigation by intracutaneous skin tests and nasal provocation tests. 279 282 EN Keiko Nishioka Chisato Saito Toshiaki Nagano Mitsuhiro Okano Fumio Nakagawa Satoko Nishioka Yu Masuda Toshiro Ono Article 10.18926/AMO/31119 To determine the allergens of mite allergic rhinitis, we studied 31 patients with mite allergic rhinitis by skin tests and nasal provocation tests (15 for skin and 16 for nasal tests) using 6 fractions of Dermatophagoides pteronyssinus (Dp) extract differing in molecular weights (15, 25, 32, 53, 95 and 190 kDMW). In skin testing, patients showed intense positive reactions to the fractions of 15, 25, 32, 95 and 190 kDMW, among which the most patients showed positive reactions to the fractions of 15 and 25 kDMW. Significant differences were found in patients' positive reactivity among each fraction and between low (15 and 25 kD) and high (95 and 190 kD) molecular weight fractions as well. In nasal provocation tests, patients showed intense positive reactions to the fractions of 15, 32, 53 and 95 kDMW, especially to the fractions of 15 and 95 kDMW. Furthermore, the insidence of positive reactions to the 15 kDMW fraction was significantly higher than that to any other fraction in the skin tests (P < 0.05). From these results, the low molecular weight fraction, 15 kDMW, is considered to be the main allergen of this mite and the high molecular weight fractions, 95 and 190 kDMW, may also be considered to be allergens of this mite. No potential conflict of interest relevant to this article was reported.

Okayama University Medical School Acta Medica Okayama 0386-300X 48 5 1994 Allergens of the house dust mite, Dermatophagoides pteronyssinus, in patients with mite allergic rhinitis: a clinical investigation by intracutaneous skin tests and nasal provocation tests. 279 282 EN Keiko Nishioka Chisato Saito Toshiaki Nagano Mitsuhiro Okano Fumio Nakagawa Satoko Nishioka Yu Masuda Toshiro Ono Article 10.18926/AMO/31114 To determine the allergens of mite allergic rhinitis, we studied 31 patients with mite allergic rhinitis by skin tests and nasal provocation tests (15 for skin and 16 for nasal tests) using 6 fractions of Dermatophagoides pteronyssinus (Dp) extract differing in molecular weights (15, 25, 32, 53, 95 and 190 kDMW). In skin testing, patients showed intense positive reactions to the fractions of 15, 25, 32, 95 and 190 kDMW, among which the most patients showed positive reactions to the fractions of 15 and 25 kDMW. Significant differences were found in patients' positive reactivity among each fraction and between low (15 and 25 kD) and high (95 and 190 kD) molecular weight fractions as well. In nasal provocation tests, patients showed intense positive reactions to the fractions of 15, 32, 53 and 95 kDMW, especially to the fractions of 15 and 95 kDMW. Furthermore, the insidence of positive reactions to the 15 kDMW fraction was significantly higher than that to any other fraction in the skin tests (P < 0.05). From these results, the low molecular weight fraction, 15 kDMW, is considered to be the main allergen of this mite and the high molecular weight fractions, 95 and 190 kDMW, may also be considered to be allergens of this mite. No potential conflict of interest relevant to this article was reported.

BioMed Central Acta Medica Okayama 1471-2407 5 2005 Functional promoter upstream p53 regulatory sequence of IGFBP3 that is silenced by tumor specific methylation EN Tadashi Hanafusa Toshiyuki Shinji Hidenori Shiraha Kazuhiro Nouso Yoshiaki Iwasaki Eichiro Yumoto Toshiro Ono Norio Koide Background: Insulin-like growth factor binding protein (IGFBP)-3 functions as a carrier of insulinlike growth factors (IGFs) in circulation and a mediator of the growth suppression signal in cells. There are two reported p53 regulatory regions in the IGFBP3 gene; one upstream of the promoter and one intronic. We previously reported a hot spot of promoter hypermethylation of IGFBP-3 in human hepatocellular carcinomas and derivative cell lines. As the hot spot locates at the putative upstream p53 consensus sequences, these p53 consensus sequences are really functional is a question to be answered. Methods: In this study, we examined the p53 consensus sequences upstream of the IGFBP-3 promoter for the p53 induced expression of IGFBP-3. Deletion, mutagenesis, and methylation constructs of IGFBP-3 promoter were assessed in the human hepatoblastoma cell line HepG2 for promoter activity. Results: Deletions and mutations of these sequences completely abolished the expression of IGFBP-3 in the presence of p53 overexpression. In vitro methylation of these p53 consensus sequences also suppressed IGFBP-3 expression. In contrast, the expression of IGFBP-3 was not affected in the absence of p53 overexpression. Further, we observed by electrophoresis mobility shift assay that p53 binding to the promoter region was diminished when methylated. Conclusion: From these observations, we conclude that four out of eleven p53 consensus sequences upstream of the IGFBP-3 promoter are essential for the p53 induced expression of IGFBP-3, and hypermethylation of these sequences selectively suppresses p53 induced IGFBP-3 expression in HepG2 cells. No potential conflict of interest relevant to this article was reported.

岡山大学保健環境センター環境安全部門 Acta Medica Okayama 0917-1533 27 2005 埋蔵文化財発掘調査および建設工事が環境放射線に及ぼす影響に関する基礎的検討 11 16 EN Tomohiro Nagamatsu Katsumi Hanamoto Toshiro Ono Kiyonori Yamaoka In this paper, we investigated how the excavation of ruins and the construction work affected the environmental radiation in the Shikata campus of Okayama University (S-campus). The environmental radiation was steady in the S-campus until 1997, but began to change since 1998, while the ruin's survey and the construction work started frequently after 1998 in the S-campus. In general, the soil and the concrete include the natural radioisotope (uranium series and thorium series, etc). When ruins are surveyed, it is necessary to move a large amount of the soil. In addition, a large amount of the concrete is used for the construction work. To measure the environmental radiation, the monitor posts were set up on the east and west sides of the building of the Radioisotope Center (RIC). We sampled the soil in the container in each place. We used the high-purity germanium detector to analyze them. The monitor posts showed the difference in the in-air dose rate in each place in the S-campus. The in-air dose rate at the east side of the RIC was higher than that at the west side. The result of analysis showed that the soil includes the (40)K at the east side. While the specific activity of the 40K was 0.849Bq/g at the east side, the (40)K was hardly detected in the soil at the west side. The each soil included the 214-lead and 214-bismus ((214)Pb, (214)Bi), however, there was no significant difference in the specific activity of the each soil. The concrete included (40)K, whose specific activity was 0.492Bq/g. It was suggested that the change of the environmental radiation was attributed to radon and its daughter nuclides in the soil rather than the radiation from the concrete in the buildings. No potential conflict of interest relevant to this article was reported.

岡山大学環境管理センター Acta Medica Okayama 0917-1533 30 2008 岡山大学における核燃料物質の安全管理のための劣化ウランと天然ウランの鑑別について 33 37 EN Tomohiro Nagamatsu Tadashi Hanafusa Ikuo Kinno Akihiro Sakoda Katsumi Hanamoto Kiyonori Yamaoka Toshiro Ono In Japan, the Law for the Regulation of Nuclear Source Materials, Nuclear Fuel Materials and Reactors (Regulation Law) controls the nuclear fuel materials such as thorium (Th), uranium (U) and plutonium (Pu). Under the Regulation Law, all related materials and reactors are needed to register to the Government. In Okayama University, many nuclear fuel materials, mainly uranium compounds, are registered and stored in 11 departments, separately. Discrimination between depleted uranium and natural uranium is important for the observance of the Regulation Law and the safety management of the nuclear fuel materials in the Okayama University. However, the discrimination of the two kind of uranium has poorly analyzed. In this study, we analyzed several uranium compounds by using γ-ray spectrometry to determine whether the depleted uranium or not. No potential conflict of interest relevant to this article was reported.

岡山大学農学部 Acta Medica Okayama 0474-0254　 98 1 2009 積算温度によるブドウの芽の休眠完了予測 9 16 EN Naohiro Kubota Chaiwat Potjanapimon Fumio Fukuda Yuichiro Fujiia Toshiro Ono Yuki Kurafuji Atsuo Ogoro Yukihiro Kunugi Kazushi Kobayashi Izumi Shigehara Hiroyuki Yamashita Hiroyuki Fujishima 　This study was conducted to investigate the relationship between completion of dormancy of grapevine bud and temperature. Canes of ‘Kyoho’ and ‘Pione’ grapevines (Vitis labrusca×V. vinifera) grown in 7 vineyards with different temperature conditions, in Nagano, Northern Okayama, Yamanashi, Okayama, Okayama University, Fukuoka and Miyazaki, were collected at three different chilling exposures, December, January and February. These were then sent to Okayama University all at the same time. Cuttings with one bud were put into growth chambers kept at 25 or 30°C with 14 hours daylength, and budbreak in each cutting was surveyed at two day intervals for 60 days. Cumulative chilling hours (CCH) of exposure to below 7.2°C in each treatment time was largest in Nagano, followed in order by Northern Okayama, Yamanashi, Okayama, Okayama University, Fukuoka and Miyazaki. The CCH in Nagano was 2.5 to 4.8 times larger than in Miyazaki depending on the treatment time. The later the treatment time and the higher the temperature, the fewer were the number of days to first budbreak (NDFB) after treatment, irrespective of cultivar. A similar trend was observed in the number of days to 60% budbreak. In ‘Kyoho’ the NDFB was short in Nagano, Okayama University and Miyazaki, and longer in Okayama, Yamanashi and Fukuoka. In ‘Pione’ the NDFB was short in Fukuoka and Okayama University, and longer in Yamanashi and Okayama. The result was a weak negative correlation observed between CCH and NDFB in 4 of 7 vineyards. However, there was a strong positive correlation between NDFB and cumulative temperature (CT), a summation of temperature and hours of exposure to above 0°C from November 1 to treatment time and hours of exposure to 25 or 30°C from start of treatment to budbreak in each plot, in 6 vineyards excluding Miyazaki. The importance of estimating the completion of dormancy in grapevine bud based on CT is discussed. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 114 2 2002 ＳＥＲＥＸ法によるマウス移植腫瘍の抗原解析 125 130 EN No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 115 1 2005 プロアクロシン結合タンパク sp32 前駆体のヒトホモログ遺伝子の同定とがん・精巣 (CT) 抗原－岡山大学医学賞（林原賞）を受賞して－ 9 14 EN No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 00301558 119 2 2007 非小細胞肺癌における癌・精巣抗原XAGE-1の発現と免疫応答の解析 127 129 EN Kazuhiko Nakagawa Yuji Noguchi Hideo Okumura Shuichiro Sato Motoyuki Tanaka Michihide Shimono Ali Mohamed Ali Eldib Motoi Aoe Toshiro Ono Akiko Uenaka Nobuya Ohara Tadashi Yoshino Kazuki Yamashita Tsukasa Tsunoda Nobuyoshi Shimizu Eiichi Nakayama No potential conflict of interest relevant to this article was reported.