• Title/Summary/Keyword: In vitro irradiation

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Chromosomal Aberrations Induced in Human Lymphocytes by in vitro Irradiation with $^{60}Co\;{\gamma}-rays$ (체외 방사선조사시 인체 말초혈액 임파구의 염색체이상 빈도에 관한 연구)

  • Ahn, Yong-Chan;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.18 no.2
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    • pp.1-16
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    • 1993
  • As guides to decision-making in the management of the victims in case of acute whole body or partial body radiation exposure, we studied the relationship between radiation dose and the frequency of chromosomal aberrations observed in peripheral lymphocytes that were irradiated in vitro with $^{60}Co\;{\gamma}-rays$ at doses ranging from 2Gy to 12Gy. The yields of cells with unstable chromosomal aberrations (dicentric chromosomes, ring chromosomes, and acentric fragment pairs) were 32% at 2Gy, 47% at 4Gy, 80% at 6Gy, 94% at 8Gy, and 100% at 10Gy and over. Ydr, which reflect average dose to the whole body in case of acute whole body exposure, were 1.373 at 2Gy, 0.669 at 4Gy, 1.734 at 6Gy, 2.773 at 8Gy, 3.746 at 10Gy and 5.454 at 12Gy. The relationship between radiation dose (D) and the frequency of dicentric plus ring chromosomes per cell(Ydr) could be expressed as $Ydr=9.322{\times}10^{-2}/Gy {\times}D+2.975{\times}10^{-2}/Gy^2{\times}D^2$. Qdr, which are used in estimating dose of partial body exposure and dose of past exposure, were 1.166 at 2Gy, 1.436 at 4Gy, 2.173 at 6Gy, 2.945 at 8Gy, 3.746 at 10Gy and 5.454 at 12Gy. To see how confidently this dosimetry system may be used, we obtained Qdr values from those who received one fraction of homogenous partial body irradiation of 1.BGy, 2.5Gy, and 7.OGy therapeutically; in vivo Qdr values were 1.109, 1.222 and 2.222 respectively. The estimated doses calculated from these in vivo Qdr values using the equation $Qdr=Ydr/(1- e^{-Ydr})$ were 1.52Gy, 2.48Gy, and 6.54Gy respectively, which were very close to the doses actually given.

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Physiological Effects of Formulations Added with Black Garlic Extract on Skin Care: Oxidative Stress, Tyrosinase and Elastase Activities (흑마늘 추출물을 첨가한 화장품의 기능성 평가: Tyrosinase와 Elastase 저해 활성 및 산화 스트레스에 미치는 영향 중심으로)

  • Jung, Eun-Young;Hong, Yang-Hee;Kim, Seon-Hee;Suh, Hyung-Joo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.5
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    • pp.662-668
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    • 2010
  • We evaluated the physiological effects of formulations added with black garlic extract on skin care. Black garlic was made by spontaneous fermentation for 40 days at $60{\sim}70^{\circ}C$ and 85~95% RH without any additives. When black garlic extract was added to formulation, the tyrosinase activity was decreased significantly (p<0.05, p<0.01, p<0.001). Base formulation inhibited slightly elastase activity (<5%), while black garlic formulation had about 8-fold higher elastase inhibitory activity (p<0.01). It was observed that formulations, with or without black garlic extract, decreased thiobarbituric acid reactive substances (TBARS) and hydrogen peroxide which were increased by UVB irradiation, although there were no significant differences of these contents between black garlic formulation and base formulation. In conclusion, black garlic formulation had high inhibitory activities for tyrosinase and elastase, suggesting that black garlic may have beneficial properties as a material for cosmeceuticals.

Monoclonal Antibody-Based Indirect-ELISA for Early Detection, Diagnosis and Monitoring of Epiphytic Didymella bryoniae in Cucurbits.

  • Lee, Seon-Chul;Shim, Chang-Ki;Kim, Dong-Kil;Bae, Dong-Won;Kyo, Seo-Il;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.133.1-133
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    • 2003
  • Gummy stem blight, caused by Didymella bryoniae occurs exclusively on cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. Through this study, we optimized cultural conditions for mass-production of pycnidiospore by Metal Halide Lamp irradiation. In brief, the mycelial was cultured at $26^{\circ}C$ on PDA, for 2 days under the darkness, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$, a great number of pycnidia was simultaneously formed. Thus produced pycnidiospores were used as immunogen. From fusions of myeloma cell (v-653) with splenocytes from immunifed mice were car ried out. And, two hybridoma cell lines that recognized the immunogen Didymella bryoniae were obtained. One Monoclonal Antibody, Db1, recognized the supernatant and the other monoclonal antibody, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid two MAb Db1 and Db15, were immunotyped and identified as IgG1 and IgG2b, respectively. Titer of MAb Db1 and MAb Db15 was measured absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with Didymella bryoniae but none reacted with other of fungi and CMV, CGMMV Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{3}$ well by indirect ELISA characterization of the MAb Db1, Db15 antigen by heat and protease treatments show that the epitope recognized by the MAb Bb1, Db15 were a glycoprotein.

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Plant regeneration of Korean wild ginseng (Panax ginseng Meyer) mutant lines induced by ${\gamma}$-irradiation ($^{60}Co$) of adventitious roots

  • Zhang, Jun-Ying;Sun, Hyeon-Jin;Song, In-Ja;Bae, Tae-Woong;Kang, Hong-Gyu;Ko, Suk-Min;Kwon, Yong-Ik;Kim, Il-Woung;Lee, Jaechun;Park, Shin-Young;Lim, Pyung-Ok;Kim, Yong Hwan;Lee, Hyo-Yeon
    • Journal of Ginseng Research
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    • v.38 no.3
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    • pp.220-225
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    • 2014
  • An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

Induction of Apoptosis and Expression of Apoptosis-related Gene Products in Response to Radiation in Murine Tumors (방사선에 대한 종양의 반응에서 아포프토시스의 유도와 이에 관련되는 유전자 발현)

  • Seong, Jin-Sil;Hunter, Nancv;Milas, Luka
    • Radiation Oncology Journal
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    • v.15 no.3
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    • pp.187-195
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    • 1997
  • Purpose : To analyze the involvement of apoptosis regulatory genes p53, $p21^{wart/cip1}$ bax and bcl-2 in induction of apoptosis by radiation in murine tumors. Materials and methods : The radiation-sensitive ovarian carcinoma OCa-1, and the radiation-resistant hepatocarcinorna HCa-I were used. Tumors, 8 mm in diameter, were irradiated with 25 Gy and at various times after irradiation, ranging from 1 to 48 h, were analyzed histologically for apoptosis and by western blot for alterations in the expression of these genes. The p53 status of the tumors were determined by the polymerase chain reaction-single strand conformation polymorphism assay. Results : Both tumors were positive for wild-type p53. Radiation inducesd apoptosis in OCa-1 but not in HCa-1. Apoptosis developed rapidly, peaked at 2 h after irradiation and returned to almost the background level at 48 h In OCa-1 radiation upregulated the expression of p53, $p21^{wart/cip1}$. and the bcl-2/bax ratio was decreased. In HCa-1 radiation increased the expression of both p53 and $p21^{wart/cip1}$, although the increase of the latter was small The bcl-2/bax ratio was greatly increased. In general the observed changes occurred within a few hours after irradiation, and either preceded or coincided with development of apoptosis Conclusions : The development of apoptosis required upregulation of both p53 and $p21^{wart/cip1}$ as well as a decrease in bcl-2/bax ratio. In contrast, an increase in bcl-2/bax ratio Prevented apoptosis in the presence of upregulated p53 and $p21^{wart/cip1}$ . These findings indentified the involvement of multiple oncogenes in apoptosis regulation in vivo and demonstrate the complexity that may be associated with the use of a single oncogene assessment for Predicting the outcome of cancer therapy with cytotoxic agents.

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Photoprotection and Anti-inflammatory Effects of Chinese Medical Plants (약용식물추출물의 광보호 효과와 항염증 효과 연구)

  • Jin-Hwa, Kim;Sung-Min, Park;Gwan-Sub, Sim;Bum-Chun , Lee;Hyeong-Bae, Pyo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.227-233
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    • 2004
  • Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.

Evaluation of Cryptosporidiurn Disinfection by Ozone and Ultraviolet Irradiation Using Viability and Infectivity Assays (크립토스포리디움의 활성/감염성 판별법을 이용한 오존 및 자외선 소독능 평가)

  • Park Sang-Jung;Cho Min;Yoon Je-Yong;Jun Yong-Sung;Rim Yeon-Taek;Jin Ing-Nyol;Chung Hyen-Mi
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.534-539
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    • 2006
  • In the ozone disinfection unit process of a piston type batch reactor with continuous ozone analysis using a flow injection analysis (FIA) system, the CT values for 1 log inactivation of Cryptosporidium parvum by viability assays of DAPI/PI and excystation were $1.8{\sim}2.2\;mg/L{\cdot}min$ at $25^{\circ}C$ and $9.1mg/L{\cdot}min$ at $5^{\circ}C$, respectively. At the low temperature, ozone requirement rises $4{\sim}5$ times higher in order to achieve the same level of disinfection at room temperature. In a 40 L scale pilot plant with continuous flow and constant 5 minutes retention time, disinfection effects were evaluated using excystation, DAPI/PI, and cell infection method at the same time. About 0.2 log inactivation of Cryptosporidium by DAPI/PI and excystation assay, and 1.2 log inactivation by cell infectivity assay were estimated, respectively, at the CT value of about $8mg/L{\cdot}min$. The difference between DAPI/PI and excystation assay was not significant in evaluating CT values of Cryptosporidium by ozone in both experiment of the piston and the pilot reactors. However, there was significant difference between viability assay based on the intact cell wall structure and function and infectivity assay based on the developing oocysts to sporozoites and merozoites in the pilot study. The stage of development should be more sensitive to ozone oxidation than cell wall intactness of oocysts. The difference of CT values estimated by viability assay between two studies may partly come from underestimation of the residual ozone concentration due to the manual monitoring in the pilot study, or the difference of the reactor scale (50 mL vs 40 L) and types (batch vs continuous). Adequate If value to disinfect 1 and 2 log scale of Cryptosporidium in UV irradiation process was 25 $mWs/cm^2$ and 50 $mWs/cm^2$, respectively, at $25^{\circ}C$ by DAPI/PI. At $5^{\circ}C$, 40 $mWs/cm^2$ was required for disinfecting 1 log Cryptosporidium, and 80 $mWs/cm^2$ for disinfecting 2 log Cryptosporidium. It was thought that about 60% increase of If value requirement to compensate for the $20^{\circ}C$ decrease in temperature was due to the low voltage low output lamp letting weaker UV rays occur at lower temperatures.

The Effect of Plant Growth Regulators on Callus Induction and Shoot Regeneration from the Dwarf Type Variety, Hibiscus syriacus L.var. Ggoma (왜성 변이 품종 '꼬마' 무궁화의 캘러스 유도 및 지상부 형성에 식물생장조절물질이 미치는 영향)

  • Lee, Ji Yeon;Kang, Eun Jeong;Kim, Sang Hoon;Kim, Dong Sub;Kim, Jin-Baek;Ha, Bo-Keun;Kang, Si-Yong
    • Journal of Radiation Industry
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    • v.5 no.3
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    • pp.231-236
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    • 2011
  • 'Ggoma' is a new Hibiscus dwarf type variety developed by gamma irradiation at the Korea Atomic Energy Research Institute (KAERI). This study was conducted to determine the best optimum cultural callus formation and shoot regeneration condition. Sterilized leaf tissues were cultured on MS (Marashige and skoog's) medium containing 3% sucrose, 0.8% agar with different concentration and combination of TDZ, 2, 4-D, KT, BA, and 2iP for 4 weeks in vitro culture. MS medium containing 2,4-D $0.1mg\;l^{-1}$ and BA $0.5mg\;l^{-1}$ were most effective on callus formation and growth. After 4 weeks, callus was transferred on BA (0.5, 1, $1.5mg\;l^{-1}$) and TDZ (0.1, 0.2, $0.3mg\;l^{-1}$) for shoot formation. The best condition for inducing the shoot from callus was BA $1.5mg\;l^{-1}$ and TDZ $0.3mg\;l^{-1}$. This result will be useful for the rapid multiplication of Hibiscus syriacus L.var. Ggoma.

Differential Effects of Green Tea Polyphenol in the ${\gamma}-irradiation$ Induced Human Leukemic and Lymphoblastic Cell Damage (녹차 폴리페놀이 감마선조사에 의한 백혈병과 림프구모세포의 손상에 미치는 영향의 차이)

  • Jeong, Hwan-Jeong;Kim, Eun-Mi;Min, Jung-Jun;Bom, Hee-Seung;Kim, Young-Ho;Jeong, Young-Do;Kim, Chang-Guhn
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.5
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    • pp.308-316
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    • 2003
  • Purpose: The green tea polyphenol (GTPP) has been known to exert antioxidant activity as a radical scavenger as well as cancer preventive and cancer growth inhibition effect. The aim of this study was to identify whether GTPP not only potentiate the growth inhibition effect in ${\gamma}-irradiated$ human cancer cell but also exert protection action for irradiated human normal cell. Materials and Methods: GTPP (80% catechin including >45% EGCG) added in the HL60, human leukemia, and NC37, human lymphoblast, before irradiation. After establishing the amount of GTPP and the dose of radiation, the cells were treated with the GTPP for 6 hours and irradiated with the determined doses. Results: Viability when $10{\mu}g/ml$ GTPP added before ${\gamma}-irradiation$ with 1 Gy to NC37 cells was not different in comparison with control but it when was irradiated with 3 Gy significantly different (1 Gy;P=0.126, 3 Gy;P=0.010). $20{\mu}g/ml$ GTPP did not show significant difference in both NC37 cells irradiated with 1 Gy and 3 Gy (1 Gy;P=0.946, 3 Gy;P=0.096). Viabilities were significantly decreased with concentration of additional GTPP in HL60 with 1 or 3 Gy (1 Gy $69.0{\pm}1.7%\;vs\;42.4{\pm}1.3%,\;3\;Gy;\;66.9{\pm}3.9%\;vs\;44.2{\pm}1.6%$). Conclusion: In vitro study, we certified that when the cells were irradiated with dose below 3 Gy, GTPP provide not only anticancerous effect against cancer cells but also radioprotective effect in normal cells simultaneously. Theses results suggest the possibility that consumption of green tea could give the radioprotective effect and maximize the effect on internal radiation such as radioiodine therapy concomitantly.

Chromosomal Aberrations Induced by in Vitro Irradiation of $^{131}I$ ($^{131}I$의 체외 방사선조사에 의한 염색체이상)

  • Kim, Eun-Sil;Kim, Myung-Seon;Seo, Doo-Seon;Jeong, Nae-In;Rho, Seung-Ik;Lee, Jae-Yong;Kim, Chong-Soon;Han, Seung-Soo;Kim, Hee-Geun;Kang, Duck-Won;Song, Myung-Jae
    • The Korean Journal of Nuclear Medicine
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    • v.28 no.1
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    • pp.133-140
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    • 1994
  • Recently, there are many considerations and studies on biological effects of radiations in radiation workers, as well as in accidentally or therapeutically irradiated persons. The most practical and reliable method of dosimetry for radiation accidents is the scoring of gross chromosomal aberrations in human lymphocytes (Ydr) as a biological dosimetry. By the way, although usual doses of $^{131}I$ administered therapeutically for thyroid cancer are ranging from 100 mCi to 200 mCi, there are differences of absorbed doses and Ydr, ranging from 0.004 to 0.04, on equally administered $^{131}I$ due to variations in metabolic characteristics, stage of tumors and physical status of subjects. In this study, We exert to obtain the dose-response relationships of $^{131}I$, as a good guide to evaluating acute effects of accidental irradiations and radiation induced leukemia or solid tumor, by in vitro induction of chromosomal aberrations. we studied the relationship between radiation dose (D) and the frequency of chromosomal aberrations (Ydr) obserbed in peripheral lymphocytes that were irradiated in vitro with $^{131}I$ at doses ranging from 0.05 to 6.00 Gy. By scoring cells with unstable chromosomal aberrations (dicentric chromosomes and ring chromosomes) we obtained this linear-quadratic dose response equation Ydr=0.064351 $D^2$-0.13143 D+0.045684 This dose-response relationship may be useful for evaluating acute and chronic $^{131}I$ induced biological effects.

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