• Title/Summary/Keyword: 세포 배양

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Influence of Hypoxia on Catecholamine Secretion Evoked by DMPP, McN-A-343, Excess $K^+$ and ACh from The Perfused Rat Adrenal Gland (저산소증이 흰쥐 관류부신에서 DMPP, McN-A-343, Excess $K^+$ 및 Ach의 카테콜아민 분비작용에 미치는 영향)

  • Lim Dong-Yoon;Heo Jae-Bong;Park Yoo Han
    • The Korean Journal of Pharmacology
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    • v.31 no.1 s.57
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    • pp.63-74
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    • 1995
  • It has been known that, during hypoxia, the adrenal medulla is activated to release catecholamines (CA) while hypoxia also inhibits high $K^+$ -induced CA secretion in the cultured bovine adrenal chromaffin cells. The present study was attempted to examine the effect of hypoxia on CA secretion evoked by chlinergic stimulation and membrane-depolarization from the isolated perfused rat adrenal glands and also to clarify its mechanism of action. For this purpose, using the isolated rat adrenal glands, the effects of hypoxia on CA release evoked by nicotinic ($N_1$) and muscarinic ($M_1$) receptor agonists, membrane-depolarizing agent, $Ca^{++}$-channel activator, intracellular $Ca^{++}$-releaser and ACh were determined. Experiments were carried out, perfusing Krebs solution pre-equilibrated with a gas mixture of 95% N_2$ and 5% $CO_2$. Hypoxia was maintained for $3{\sim}4$ hours through the experiments. Hypoxia gradually caused a time-dependent seduction in CA secretion evoked by DMPP ($100{\mu}M$), McN-A-343 ($100{\mu}M$), ACh (5.32 mM), Bay-K-8644 ($10{\mu}M$) and high $K^+$ (56 mM) respectively. How-ever, it did not affect CA secretion evoked by cyclopiazonic acid ($10{\mu}M$). Hypoxia itself also did fail to produce any influence on spontaneous secretory response of CA. These experimental results suggest that hypoxia depresses CA release evoked by both cholinergic stimulation and membrane-depolarization from the isolated rat adrenal medulla, and that this inhibitory activity may be due to the result of the direct inhibition of $Ca^{++}$ influx into the chromaffin cells without any effect on the calcium mobilization from the intracellular store.

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Improvement of Productivity by Forest Tree Breeding Work in Korea (우리나라에서의 임목육종(林木育種)에 의(依)한 생산성(生産性) 증가(增加))

  • Ryu, Jang Bal;Shim, Sang Yung
    • Journal of Korean Society of Forest Science
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    • v.77 no.4
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    • pp.382-388
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    • 1988
  • Improvement of productivity by forest tree breeding work in Korea was estimated for a few important tree species. Progenies of 17 plus trees of red pine (Pines densiflora) outgrew by 57 percentage compared with progenies of unselected trees at age 15. If best three families are selected among the 17, more than double in volume grow-th is expected. The hybrid Pinus rigida ${\times}$ P. taeda showed more than double volume growth compare to P. rigida at a southern plantation at age 15. However, the superiority of the hybrid decreased at northern plantations, mainly because of low coldhardiness of the hybrid. At a northern plantation, the hybrid grew less than the P. rigida on upper hill, while the hybrid grew much better than the P. rigida on flat area. Another hybrid Populus alba ${\times}$ P. glandulosa grew faster than both parents by two to two and half times according to planting sites at age 10. Introduction of Pinus rigida also showed increased volume growth. Volume increase by selection of best five provenances among 45 at age 12 was estimated as 53 percent compare to progenies of plus trees in Korea, Additional 19 percent of volume increase was expected by selection of the best families within the best provenances. Annual production of chestnuts reached about 70,000 M/T by planting resistant clones to chestnut gall wasp (Dryocosmus kuriphilus), which killed almost all susceptible trees. Although polyploid trees and mutants have been produced by colchicine treatments in over 10 tree species, none of them is economically important Remarkable improvement of productivity is expected by biotechnology in future through selection, hybridization, introduction of foreign genes at cell, cell organelle and gene level, and gene transformation. At present, mass propagation of superior planting materials by tissue culture will increase the productivity.

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Pharmacological Activities of Coffee Roasted from Fermented Green Coffee Beans with Fungal Mycelia in Solid-state Culture (진균류 균사체의 고체발효 커피생두로부터 조제한 원두커피의 생리활성)

  • Shin, Ji-Young;Kim, Hoon;Kim, Dong-Gu;Baek, Gil-Hun;Jeong, Heon-Sang;Yu, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.487-496
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    • 2013
  • Green coffee beans (CB, Indonesian Mandheling) were fermented with three kinds of mushrooms (Phellinus linteus, PL; Hericium erinaceum, HE; Ganoderma lucidum, GL) or two kinds of mycelia from molds (Monascus purpureus, MP; Monascus ruber, MR) using solid-state culture to enhance physiological activity. After the roasting of fermented green coffee beans, roasted coffees were extracted with a hot-water decoction or 95% ethanol reflux. Yields from hot water extracts (HW, 17.7~25.3%) were higher than those from ethanolic extracts (EE, 9.5~12.2%). Hot-water extracts of roasted coffees from green coffee beans fermented with two molds (MP-CB-HW and MR-CB-HW) showed higher total polyphenols, flavonoids, and DPPH free radical scavenging activity than roasted coffees from non-fermented (CB-HW) or fermented green coffee beans with the three mycelia from mushrooms. MR-CB-HW also had the most potent macrophage stimulating and mitogenic activity (1.32 and 1.40-fold of CB-HW, respectively). In addition, MP-CB-EE and MR-CB-EE did not show any cytotoxicity to the RAW 264.7 cell at a concentration of $100{\mu}g/mL$, and these extracts significantly inhibited nitric oxide (NO) production from the LPS-stimulated RAW 264.7 cell line (38.6 and 37.0% of the LPS-treated group). Meanwhile, the chlorogenic acid concentrations of MP-CB-HW or MR-CB-HW highly increased (to 76.21 or $76.73{\mu}g/mL$, respectively), but caffeine concentrations were not affected by solid-state fermentation. In conclusion, the physiological activities of roasted coffees were enhanced by the solid-state culture of green coffee beans with M. purpureus or M. ruber, suggesting that these roasted coffees could possibly serve industrial applications as functional coffee beverages.

Chromosome Analysis from Papillary Carcinoma and Nodular Hyperplasia of the Thyroid Gland (결절성 갑상선종과 유두성 갑상선암의 염색체 분석)

  • Hwhang Dae-Won;Chung Ki-Yong;Kang Joong-Shin;Kim Hong-Tae;Chang Sung-Ik
    • Korean Journal of Head & Neck Oncology
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    • v.9 no.1
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    • pp.25-32
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    • 1993
  • The nodular hyperplasia of the thyroid is a common thyriod disease. Nodular hyperplasia does rarely progress to thyroid cancer. The differentiation of a nodular hyperplasia from a neoplasm may be simple or difficult, both clinically and anatomically. The papillary carcinoma of the thyroid is the most common type of thyroid malignancies. There were few studies about cytogenetic observation in thyroid cancer. But only one case of banding observation in nodular hyperplasia have been reported. In order to compare the chromosomal changes in the thyroid cancer and the noncancerous thyroid disease, we performed cytogenetic analysis in two papillary carcinoma and two nodular hyperplasia after cell culture. The chromosomal pattern of the nodular hyperplasia found was very heterogenous but no clonal abnormaly in both cases was observed. Case I : A modal chromosomal number was in 42-46 range. Chromosome 8, 19, 21. 22 were commonly lost. 9 structural anomalities among 51 analysed cells were observed but they were not clonal. Case II: A modal chromosomal number was 43. Chromosome 17 and 19 were commonly lossed. Common cytogenetic characters of this two nodular hyperplasia are hypodiploidity and very heterogenous chromosomal pattern. The result about the papillary carcinoma are as follow. In one case some numerical and structural chromosomal changes were observed. But they were not clonal abnormality. In another case the chromosomal pattern found was very heterogenous with a clonal abnormality of del(11)(q23). The modal number was 46. The del(11)(q23) a chromosomal change in papillary carcinoma of the thyroid have previously been reported(Eva Olah et al. 1989). We suggest that 11q deletion may be important role to pathogenesis of papillary carcinoma of the thyroid. According to this results, we could not find out specific differences about chromosomal changes and any relationship between the papillary carcinoma and the nodular hyperplasia.

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The Artificial Cultivation of Pleurotus eryngii (II) -Morphological Characteristics of Fruit Body and Cultural Conditions- (Pleurotus eryngii균의 인공재배 (II) -자실체의 형태적 특성 및 재배조건에 관하여-)

  • Kim, Han-Kyoung;Cheong, Jong-Chun;Seok, Soon-Ja;Kim, Gwang-Po;Cha, Dong-Yeul;Moon, Byung-Ju
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.311-319
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    • 1997
  • These studies were carried out to develop an artificial cultivation method. The diameter and thickness of pileus ranged $1.5{\sim}7.0\;cm$ and $0.8{\sim}3.0\;cm$, respectively. The diameters of stipe were $1.2{\sim}2.5\;cm$ and the lengthes were $4.5{\sim}9.0\;cm$. The spore fingerprint was white. The sizes were spore $10.8{\sim}12.2{\times}4.35{\sim}5.65\;{\mu}m$, basidia $50.0{\sim}59.2{\times}7.4{\sim}7.8\;{\mu}m$, nalsistidia $21.75{\sim}28.7{\times}4.8{\sim}6.1\;{\mu}m$, pileus hymenium cell $50.6{\sim}66.0{\times}4.4{\sim}6.7\;{\mu}m$, and stipe hymenium cell $28.6{\sim}33.0{\times}5.5{\sim}6.6\;{\mu}m$. The thirty percent mixture of rice and wheat bran into sawdust gives the high density of mycelia and the good development of fruiting structure. The optimum water contents of sawdust substrates were $60{\sim}65%$ in which condition the mycelium grows well and gives high density. In PP bottle cultivation, the first fruiting period was $6{\sim}8$ days earlier in nonscratching samples than scratching ones, but the quantity of fruiting body was higher in scratching samples than nonscratching ones. In the case of PP bag cultivation, the first fruiting was 10 days faster, and the quantity of fruiting bodies was 30% higher in samples with 30% wheat bran than those with rice bran. The fleshiness of stipe was $2{\sim}3$ times harder than that of pileus.

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In Vitro Phosphorylation of Nuclear Proteins in Isolated Liver Nuclei from Rats Maintained in a Starvation State, Following Refeeding, and from Diabetic Rats with Insulin Injection (단식(斷食), 재급식(再給食) 및 인슈린 투여(投與) 후(後)에 쥐의 간(肝)으로부터 분리된 세포핵의 핵단백질 인산화)

  • Lee, Hyo-Sa;Gibson, David M.
    • Applied Biological Chemistry
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    • v.23 no.1
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    • pp.23-30
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    • 1980
  • Labelling of chromatin proteins with 32P was observed after incubating isolated liver nuclei with $[{\gamma}-32P]$ ATP for 5 minutes at $37^{\circ}C$. The pattern of labelling with 32P was examined on SDS polyacrylamide gel electrophoresis with nuclei from rats maintained in a starvation state for 48 hours, following refeeding for 12 hours; and from fed streptozotocin-diabetic rats with insulin injection 6 hours before sacrifice. With 48h starved rat liver nuclei the level of phosphorylation for 0.14M NaCl soluble proteins was decreased in the molecular weights between 41,000 and 200,000 daltons relative to normal controls. Refeeding the starved rats reversed the change of phosphorylation pattern over 12 hour The level of phosphorylation for five phenol soluble non-histone proteins with molecular weights above 59,000 daltons was somewhat decreased with 48h starved rat liver nuclei as compared with that of normal controls. Starvation also decreased the phosphorylation level of major histones in relation to normal controls. The experiment with insulin injection into fed streptozotocin-diabetic rats showed the tendency to increase phosphorylation of 0.14M NaCl soluble proteins (130,000 dalton protein) and phenol soluble non-histone proteins (155,000 dalton protein). The phosphorylation level of histones appeared to be invariant under the experimental conditoins employed here. These results suggest the possibility that the phosphorylation and dephosphorylation of 0.14M NaCl soluble proteins and $H_1$ histone precede those of other chromatin associated nuclear proteins, It is of interest to find that insulin signal was correlated to phosphorylation of nuclear proteins while glucagon signet dephosphorylated nuclear proteins.

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EFFECT OF ENAMEL MATRIX DERIVATIVE (EMD, $EMDOGAIN^{(R)}$) ON THE DIFFERENTIATION OF CULTURED HUMAN PERIODONTAL LIGAMENT CELLS AND MESENCHYMAL STEM CELLS (배양된 사람 치주인대세포와 골수유래간엽줄기세포의 분화에 미치는 법랑기질유도체 (Enamel Matrix Derivative, EMD)의 영향)

  • Park, Sang-Gyu;Jue, Seong-Suk;Kwon, Yong-Dae;Choi, Byung-Joon;Kim, Young-Ran;Lee, Baek-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.4
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    • pp.281-286
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    • 2009
  • Introduction: Enamel matrix derivative (EMD) is a protein which is secreted by Hertwig root sheath and plays a major role in the formation of cementum and attachment of peridontium. Several studies have shown that EMD promoted the proliferation and differentiation of preosteoblasts, osteoblasts and periodontal ligament cells in vitro: however, reports showing the inhibition of osteogenic differentiation by EMD also existed. This study was designed to simultaneously evaluate the effect of EMD on the two cell lines (human mesenchymal stem cells: hMSC, human periodontal ligament derived fibroblasts: hPDLCs) by means of quantitative analysis of some bone related matrices (Alkaline phosphatase : ALP, osteopontin ; OPN, osteocalcin ; OC). Materials and Methods: hMSCs and hPDLCs were expanded and cells in the 4${\sim}$6 passages were adopted to use. hMSc and hPDLCs were cultured during 1,2,7, and 14 days with 0, 50 and 100 ${\mu}g/ml$ of EMD, respectively. ALP activity was assessed by SensoLyte ALP kit and expressed as values of the relative optical density. Among the matrix proteins of the bony tissue, OC and OPN were assessed and quantification of these proteins was evaluated by means of human OC immunoassay kit and human OPN assay kit, respectively. Results: ALP activity maintained without EMD at $1,2^{nd}$ day. The activity increased at $7^{th}$ day but decreased at $14^{th}$ day. EMD increased the activity at $14^{th}$ day in the hPDLCs culture. In the hMSCs, rapid decrease was noted in $7^{th}$ and $14^{th}$ days without regard to EMD concentrations. Regarding the OPN synthesis in hPDLCs, marked decrease of OPN was noted after EMD application. Gradual decrease tendency of OPN was shown over time. In hMSCs, marked decrease of OPN was also noted after EMD application. Overall concentration of OPN was relatively consistent over time than that in hPDLCs. Regarding the OC synthesis, in both of hPDLCs and hMSCs, inhibition of OC formation was noted after EMD application in the early stages but EMD exerted minimal effect at the later stages. Conclusion: In this experimental condition, EMD seemed to play an inhibitory role during the differentiation of hMSCs and hPDLCs in the context of OC and OPN formation. In the periodontium, there are many kinds of cells contributing to the regeneration of oral tissue. EMD enhanced ALP activity in hPDLCs rather than in hMSCs and this may imply that EMD has a positive effect on the differentiation of cementoblasts compared with the effect on hMSCs. The result of our research was consistent with recent studies in which the authors showed the inhibitory effect of EMD in terms of the differentiation of mineral colony forming cells in vitro. This in vitro study may not stand for all the charateristics of EMD; thus, further studies involving many other bone matrices and cellular attachment will be necessary.

Diagnostic Accuracy of 2-mm Minithoracoscopic Pleural Biopsy for Pleural Effusion (흉수 환자에서 Minithoracoscopy를 이용한 흉막 생검의 진단적 유용성)

  • Kim, Woo Jin;Lee, Hui Young;Lee, Sung Ho;Cho, Seong Joon;Park, Weon-Seo;Kim, Ja Kyoung;Lee, Seung-Joon
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.2
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    • pp.138-142
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    • 2004
  • Background : To evaluate exudative pleural fluid, thoracentesis for microbiological and cytological examination and pleural biopsy by using a Cope needle are traditionally performed. Even after these studies, about 20% of patients remain undiagnosed. We evaluated the diagnostic accuracy and complications of 2-mm minithoracoscopy instead of blind biopsy in patients with undiagnosed exudative pleural effusion. Method : Fifteen patients with exudative pleural effusion underwent thoracoscopy between April 2002 and August 2003. The indication was undiagnosed pleural effusions after having performed sputum and pleural fluid exami-nations both microbiologically and cytologically. Results : The median age of the patients was 56 years (range 21-77). Pleural effusions were lymphocyte-dominant in 11 patients (73.3%) and neutrophil-dominant in 3 (20.0%). The remaining patient (6.7%) had pleural-fluid eosinophilia. Minithoracoscopic biopsy revealed accurate diagnosis in 14 patients (93.3%), consisting of tuberculous pleurisy in 8 (66.7%), malignant effusions in 4 (33.3%), and parapneumonic effusions in 2 (13.3%). One was diagnosed as having paragonimiasis from thoracoscopic findings and clinical considerations. There was no procedure-associated mortality. There were six cases of new onset fever (40%) and one of pneumothorax (6.7 %). Conclusion : Two-millimeter minithoracoscopy, which is less invasive than conventional thoracoscopy, was an accurate and safe method for undiagnosed exudative pleural effusion.

Characterization of Salt Tolerant Rice Mutant Lines Derived from Azetidine-2-Carboxylic Acid Resistant Cell Lines Induced by Gamma Ray Irradiation (AZCA 저항성 돌연변이 세포주로부터 선발 육성만 내염성 벼 돌연변이 계통의 특성 검정)

  • Song, Jae-Young;Kim, Dong-Sub;Lee, Geung-Joo;Lee, In-Sok;Kang, Kwon-Kyoo;Yun, Song-Joong;Kang, Si-Yong
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.61-68
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    • 2007
  • To develop rice (Oryza sativa L.) cultivars to be planted on salt-affected sites, cell lines with enhanced proline content and resistance to growth inhibition by Azetidine-2-carboxylic acid (AZCA), a proline analogue, were screened out among calli irradiated with gamma ray of 50, 70, 90, and 120 Gy. The calli had been derived from embryo culture of the cultivar Donganbyeo. Selected AZCA resistant lines that had high proline accumulation were used as sources for selection of NaCl resistant lines. To determine an optimum concentration for selection of NaCl resistant lines, Donganbyeo seeds were initially cultured on the media containing various NaCl concentrations (0 to 2.5%) for 40 days, and 1.5% NaCl concentration was determined as the optimum concentration. One hundred sixteen salt-tolerant (ST) lines were selected from bulked 20,000 seeds of the AZCA resistant $M_{3}$ seeds in the medium containing 1.5% NaCl. The putative 33 lines ($M_{4}$ generation) considered with salt-tolerance were further analyzed for salt tolerance, amino acid and ion contents, and expression patterns of the salt tolerance-related genes. Out of the 33 lines, 7 lines were confirmed to have superior salt tolerance. Based on growth comparison of the entries, the selected mutant lines exhibited greater shoot length with average 1.5 times, root length with 1.3 times, root numbers with 1.1 times, and fresh weight with 1.5 times than control. Proline contents were increased maximum 20%, 100% and 20% in the leaf, seed and callus, respectively, of the selected lines. Compared to control, amino acid contents of the mutants were 24 to 29%, 49 to 143%, 32 to 60% higher in the leaf, seed and callus, respectively. The ratio of $Na^{+}/K^{+}$ for most of the ST-lines were lower than that of control, ranging from 1.0 to 3.8 for the leaf and 11.5 to 28.5 for the root, while the control had 3.5 and 32.9 in the leaf and root, respectively. The transcription patterns for the P5CS and NHXI genes observed by RT-PCR analysis indicated that these genes were actively expressed under salt stress. The selected mutants will be useful for the development of rice cultivar resistant to salt stress.

Frequency of Micronuclei in Lymphocytes Following Gamma and Fast-neutron Irradiations (방사선 조사량에 따른 인체 정상 림파구의 미세핵 발생빈도)

  • Kim Sung-Ho;Cho Chul-Koo;Kim Tae-Hwan;Chung In-Yong;Yoo Seong-Yul;Koh Kyoung-Hwan;Yun Hyong-Geun
    • Radiation Oncology Journal
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    • v.11 no.1
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    • pp.35-42
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    • 1993
  • The dose response of the number of micronuclei in cytokinesis-blocked (CB) lymphocytes after in vitro irradiation with $\gamma$-rays and neutrons in the 5 dose ranges was studied for a heterogeneous population of 4 donors. One thousand binucleated cells were systematically scored for micronuclei. Measurements performed after irradiation showed a dose-dependent increase in micronuclei (MN) frequency in each of the donors studied. The dose-response curves were analyzed by a linear-quadratic model, frequencies per 1000 CB cells were ($0.31{\pm}0.049$)D+($0.0022{\pm}0.0002)D^2+(13.19{\pm}1.854) (r^2=1.000,\;X^2=0.7074,\;p=0.95$) following $\gamma$ irradiation, and ($0.99{\pm}0.528$)\;D+(0.0093{\pm}0.0047)\;D^2+(13.31{\pm}7.309)\;(r^2=0.996,\;X^2=7.6834,\;p=0.11) following neutrons irradiation (D is irradiation dose in cGy). The relative biological effectiveness (RBE) of neutrons compared with $\gamma$-rays was estimated by best fitting linear-quadratic model. In the micronuclei frequency between 0.05 and 0.8 per cell, the RBE of neutrons was $2.37{\pm}0.17$. Since the MN assay is simple and rapid, it may be a good tool for evaluating the $\gamma$-ray and neutron response.

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