• Title/Summary/Keyword: genetic system

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Development of transgenic strawberry plants expressing monellin, a sweet protein (감미단백질 모넬린 발현 딸기 형질전환 식물체 개발)

  • Min, Sung Ran;Ko, Suk Min;Lyu, Jae Il;Park, Ji Hyun;Yi, So Young;Lee, In-Ha;Kim, Hyun Sook;Kim, Tae Il;Choi, Pil Son;Jeong, Won-Joong;Kim, Suk Weon;Kim, Jonghyun;Liu, Jang R.
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.180-185
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    • 2015
  • Leaf discs from 'Yeobong' and 'Maehyang' strawberry plants were used as explants for transformation. The Agrobacterium tumefaciens strain EHA105 harboring the monellin gene under the control of the CaMV 35S promoter was used in co-cultivation experiments. The frequencies of callus formation and plant regeneration from leaf explants after co-cultivation in 'Yeobong' were higher than those of 'Maehyang'. These transgenic plants showed normal growth patterns and flowering. PCR and Southern hybridization confirmed that 1 to 2 copies of the monellin gene were integrated into genome of the transgenic strawberry plants. Northern blot analysis confirm that the transcripts were expressed in transgenic strawberry plants. Although long-term subcultured transgenic strawberry plants showed a phenomenon to escape the transgene, the transformation system established in this study provides new opportunities for genetic improvement of strawberry plants.

Clinical Significance of the NQO1 C609T Polymorphism in Non Small Cell Lung Adenocarcinoma Patients

  • Masroor, Mirza;Jain, Amit;Javid, Jamsheed;Mir, Rashid;Prashant, Y;Imtiyaz, A;Mariyam, Z;Mohan, Anant;Ray, PC;Saxena, Alpana
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7653-7658
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    • 2015
  • Background: NAD(P)H:quinone oxidoreductase 1 (NQO1) is part of the antioxidant defence system involved in detoxification. This study aimed to analyze the influence of NQO1 (C609T) genetic polymorphism in non small cell lung cancer (NSCLC)as a putative risk factor. Materials and Methods: Present study included 100 cases of NSCLC (adenocarcinoma) patients and 100 age and sex matched healthy controls. NQO1 (C609T) genotyping was performed by allele specific PCR for assessment of putative associations with clinical outcome and genotypes of. The association of the polymorphism with the survival of NSCLC patients' was analyzed by Kaplan-Meier method. Results: In Indian NSCLC (adenocarcinoma) patients increased risk of developing NSCLC was found to be associated with NQO1 609TT genotype [OR 3.68(0.90-14.98), RR 2.04(0.78-5.31)] for CT [OR 2.91(1.58-5.34), RR 1.74(1.23-2.44) p=0.0005 for CT], for CT+TT [ OR 3.26(1.82-5.82), RR 1.87(1.34-2.61) p<0.0001 for CT+TT]. A significant difference (p=0.0009) was observed in genotype distribution among cases and healthy controls. Patients with CT+TT genotype exhibited a significant poor overall survival compared with patients displaying homozygous CC genotype (p=0.03) and when survival independently compared with CC, TT and CT genotype was also found to be significantly associated (p=0.02). Overall median survival times were CT 6.0 months, TT 8.2 months, and CT + TT (6.4 months)]. Conclusions: The present study revealed that NQO1 CT, TT and CT+TT genotypes may be associated with clinical outcome and risk of developing NSCLC in the Indian population.

Fatty Acid Synthesis Pathway Genetic Variants and Clinical Outcome of Non-Small Cell Lung Cancer Patients after Surgery

  • Jin, Xin;Zhang, Ke-Jin;Guo, Xu;Myers, Ronald;Ye, Zhong;Zhang, Zhi-Pei;Li, Xiao-Fei;Yang, Hu-Shan;Xing, Jin-Liang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7097-7103
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    • 2014
  • Over-expression of de novo lipogenesis (DNL) genes is associated with the prognosis of various types of cancers. However, the effects of single nucleotide polymorphisms (SNPs) in these genes on recurrence and survival of non-small cell lung cancer (NSCLC) patients after surgery are still unknown. In this study, a total of 500 NSCLC patients who underwent surgery treatment were included. Eight SNPs in 3 genes (ACACA, FASN and ACLY) of the DNL pathway were examined using the Sequenom iPLEX genotyping system. Multivariate Cox proportional hazards regression and Kaplan-Meier curves were used to analyze the association of SNPs with patient survival and tumour recurrence. We found that two SNPs in the FASN gene were significantly associated with the recurrence of NSCLC. SNP rs4246444 had a significant association with lung cancer recurrence under additive model (hazard ratio [HR], 0.82; 95% confidence interval [95%CI], 0.67-1.00; p=0.05). Under the dominant model, rs4485435 exhibited a significant association with recurrence (HR, 0.75; 95%CI, 0.56-1.01; p=0.05). Additionally, SNP rs9912300 in ACLY gene was significantly associated with overall survival in lung cancer patients (HR, 1.41; 95%CI, 1.02-1.94, p=0.04) under the dominant model. Further cumulative effect analysis showed moderate dose-dependent effects of unfavorable SNPs on both survival and recurrence. Our data suggest that the SNPs in DNL genes may serve as independent prognostic markers for NSCLC patients after surgery.

A molecular systematic study of Korean Iris (Iridaceae) based on RAPD analysis (RAPD에 의한 한국산 붓꽃속(Iris)의 계통분류학적 연구)

  • Park, Seon-Joo;Sim, Jeong-Ki;Park, Hong-Duok
    • Korean Journal of Plant Taxonomy
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    • v.32 no.4
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    • pp.383-396
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    • 2002
  • RAPD analyses were compared for 17 taxa of Korean Iris including the subgenus Sisyrinchium and Belamcanda. Eighty scorable RAPD markers were formed from the PCR reactions using 10 random oligoprimers. In this systematic analyses which used neighbor-joining methods including bootstrapping analyses with genetic coefficients, the Korean Iris were divided into three subgenera (Limniris, Crossiris, Pardanthopsis), or two genera (Limniris, Pardanthopsis). The molecular data agree with the previous classification system that recognized two sections and six series for the subgenus Limniris because the subgenus is comprised of four clades in the RAPD analyses. According to the molecula data, the series Chinensis should be divided into two groups. The minutoaurea group is composed of I. koreana, I. odaesanensis, and I. minitoaurea, while the rossi group is comprised of two varieties of I. rossi. The series Tripetalae is closely allied with the series Sibiricae, whereas the series Ensatae is recognized as a sister group to the series Ruthencae. The molecular phylogeny, which was based on RAPD analysis, for the most part agreed with the data proposed by previous authors. This is because the basis of morphological and ITS sequence data suggests that the RAPD markers should be very useful in addressing phylogenetic questions about the genus Iris.

Epigenetic control of LTR retrotransposons in plant germline and somatic cells

  • Lee, Seung Cho;Parent, Jean-Sebastien;Ernst, Evan;Berger, Frederic;Grimanelli, Daniel;Martienssen, Robert A.
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.20-20
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    • 2017
  • Plant genomes include heterochromatic loci that consist of repetitive sequences and transposable elements. LTR retrotransposon is the major class of transposons in advanced plants in terms of proportion in plant genome. The elements contribute not only to genome size but also to genome stability and gene expression. A number of cases have been reported transposon insertions near genic regions affect crop traits such as fruit pigments, stress tolerance, and yields. Functional LTR retrotransposons produce extrachromosomal DNA from genomic RNA by reverse transcription that takes place within virus-like-particles (VLPs). DECREASED DNA METHYLATION 1 (DDM1) plays important roles in maintaining DNA methylation of heterochromatin affecting all sequence contexts, CG, CHG, and CHH. Previous studies showed that ddm1 mutant exhibits massive transcription of retrotransposons in Arabidopsis, but only few of them were able to create new insertions into the genome. RNA-dependent RNA POLYMERASE 6 (RDR6) is known to function in restricting accumulation of transposon RNA by processing the transcripts into 21-22 nt epigenetically activated small interfering RNA (easiRNA). We purified VLPs and sequence cDNA to identify functional LTR retrotransposons in Arabidopsis ddm1 and ddm1rdr6 plants. Over 20 LTR copia and gypsy families were detected in ddm1 and ddm1rdr6 sequencing libraries and most of them were not reported for mobility. In ddm1rdr6, short fragments of ATHILA gypsy elements were detected. It suggests easiRNAs might regulate reverse transcription steps. The highest enriched element among transposon loci was previously characterized EVADE element. It has been reported that active EVADE element is more efficiently silenced through female germline than male germline. By genetic analyses, we found ddm1 and rdr6 mutation affect maternal silencing of active EVADE elements. DDM1-GFP protein accumulated in megaspore mother cell but was not found in mature egg cell. The fusion protein was also found in early embryo and maternal DDM1-GFP allele was more dominantly expressed in the embryo. We observed localization of DDM1-GFP in Arabidopsis and DDM1-YFP in maize and found the proteins accumulated in dividing zone of root tips. Currently we are looking at cell cycle dependency of DDM1 expression using maize system. Among 10 AGO proteins in Arabidopsis, AGO9 is specifically expressed in egg cell and shoot meristematic cells. In addition, mutation of AGO9 and RDR6 caused failure in maternal silencing, implying 21-22 nt easiRNA pathway is important for retrotransposon silencing in female gametophyte or/and early embryo. On the other hand, canonical 24 nt sRNA-directed DNA methylation (RdDM) pathways did not contribute to maternal silencing as confirmed by this study. Heat-activated LTR retrotransposon, ONSEN, was not silenced by DDM1 but the silencing mechanisms require RdDM pathways in somatic cells. We will propose distinct mechanisms of LTR retrotransposons in germline and somatic stages.

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Epigenetic control of LTR retrotransposons in plant germline and somatic cells

  • Lee, Seung Cho;Parent, Jean-Sebastien;Ernst, Evan;Berger, Frederic;Grimanelli, Daniel;Martienssen, Robert A.
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.97-97
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    • 2017
  • Plant genomes include heterochromatic loci that consist of repetitive sequences and transposable elements. LTR retrotransposon is the major class of transposons in advanced plants in terms of proportion in plant genome. The elements contribute not only to genome size but also to genome stability and gene expression. A number of cases have been reported transposon insertions near genic regions affect crop traits such as fruit pigments, stress tolerance, and yields. Functional LTR retrotransposons produce extrachromosomal DNA from genomic RNA by reverse transcription that takes place within virus-like-particles (VLPs). DECREASED DNA METHYLATION 1 (DDM1) plays important roles in maintaining DNA methylation of heterochromatin affecting all sequence contexts, CG, CHG, and CHH. Previous studies showed that ddm1 mutant exhibits massive transcription of retrotransposons in Arabidopsis, but only few of them were able to create new insertions into the genome. RNA-dependent RNA POLYMERASE 6 (RDR6) is known to function in restricting accumulation of transposon RNA by processing the transcripts into 21-22 nt epigenetically activated small interfering RNA (easiRNA). We purified VLPs and sequence cDNA to identify functional LTR retrotransposons in Arabidopsis ddm1 and ddm1rdr6 plants. Over 20 LTR copia and gypsy families were detected in ddm1 and ddm1rdr6 sequencing libraries and most of them were not reported for mobility. In ddm1rdr6, short fragments of ATHILA gypsy elements were detected. It suggests easiRNAs might regulate reverse transcription steps. The highest enriched element among transposon loci was previously characterized EVADE element. It has been reported that active EVADE element is more efficiently silenced through female germline than male germline. By genetic analyses, we found ddm1 and rdr6 mutation affect maternal silencing of active EVADE elements. DDM1-GFP protein accumulated in megaspore mother cell but was not found in mature egg cell. The fusion protein was also found in early embryo and maternal DDM1-GFP allele was more dominantly expressed in the embryo. We observed localization of DDM1-GFP in Arabidopsis and DDM1-YFP in maize and found the proteins accumulated in dividing zone of root tips. Currently we are looking at cell cycle dependency of DDM1 expression using maize system. Among 10 AGO proteins in Arabidopsis, AGO9 is specifically expressed in egg cell and shoot meristematic cells. In addition, mutation of AGO9 and RDR6 caused failure in maternal silencing, implying 21-22 nt easiRNA pathway is important for retrotransposon silencing in female gametophyte or/and early embryo. On the other hand, canonical 24 nt sRNA-directed DNA methylation (RdDM) pathways did not contribute to maternal silencing as confirmed by this study. Heat-activated LTR retrotransposon, ONSEN, was not silenced by DDM1 but the silencing mechanisms require RdDM pathways in somatic cells. We will propose distinct mechanisms of LTR retrotransposons in germline and somatic stages.

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Human Lung Insults due Air Pollutant -A Review for Priority Setting in the Research- (대기오염에 의한 폐장조직 손상 -연구방향의 설정을 위한 논의-)

  • 김건열;백도명
    • Journal of environmental and Sanitary engineering
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    • v.7 no.2
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    • pp.95-110
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    • 1992
  • Much progress has been made in understanding the subcellular events of the human lung injuries after acute exposure to environmental air pollutants. Host of those events represent oxidative damages mediated by reactive oxygen species such as superoxide, hydrogen peroxide, and the hydroxy, free radical. Recently, nitric oxide (NO) was found to be endogenously produced by endothelial cells and cells of the reticulo-endothelial system as endothelialderived relaxation factor (EDRF) which is a vasoactive and neurotransmitter substance. Together with superoxide, NO can form another strong oxidant, peroxonitrite. The relative importance of exogenous sources of $N0/N0_2$ and endogenous production of NO by the EDRF producing enzymes in the oxidative stresses to the heman lung has to be elucidated. The exact events leading to chronic irreversible damage are still yet to be known. From chronic exposure to oxidant gases, progressive epithelial and interstitial damages develop. Type I epithelial cells become thicker and cover a smaller average alveolar surface area while thee II cells proliferate instead. Under acute damages, the extent of loss of the alveolar epithelial cell lining, especially type II cells appears to be a good predictor of the ensuing irreversible damage to alveolar compartment. Interstitial matrix undergo remodeling during chronic exposure with increased collagen fibers and interstitial fibroblasts. However, Inany of these changes can be reversed after cessation of exposure. Among chronic lung injuries, genetic damages and repair responses received particular attention in view of the known increased lung cancer risks from exposure to several air pollutants. Heavy metals from foundry emission, automobile traffics, and total suspended particulate, especially polycystic aromatic hydrocarbons have been positively linked with the development of lung cancer. Asbestos in another air pollutant with known risk of lung cancer and mesothelioma, but asbestos fibers are nonauthentic in most bioassays. Studies using the electron spin resonance spin trapping method show that the presence of iron in asbestos accelerates the production of the hydroxy, radical in vitro. Interactions of these reactive oxygen species with particular cellular components and disruption of cell defense mechanisms still await further studies to elucidate the carcinogenic potential of asbestos fibers of different size and chemical composition. The distribution of inhaled pollutants and the magnitude of their eventual effects on the respiratory tract are determined by pollutant-independent physical factors such as anatomy of the respiratory tract and level and pattern of breathing, as well as by pollutant-specific phyco-chemical factors such as the reactivity, solubility, and diffusivity of the foreign gas in mucus, blood and tissue. Many of these individual factors determining dose can be quantified in vitro. However, mathematical models based on these factors should be validated for its integrity by using data from intact human lungs.

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The Lived Experience of Mothers of Children with Muscular Dystrophy (근디스트로피 자녀를 가진 어머니의 경험)

  • Oh Sang-Eun
    • Child Health Nursing Research
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    • v.7 no.4
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    • pp.421-433
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    • 2001
  • The purpose of this phenomenological study was to understand the lived experience of mothers of children with muscular dystrophy. The participants were 11 mothers recruited by snowball sampling, who agreed with the objectives of the research and could verbally communicate with the researcher and who lived with their sons who had muscular dystrophy. Data collection was done through in-depth interviews with participants in their own homes. Each interview took 50 to 120 minutes. All of the interviews were audiotaped and then transcribed. Data were analyzed using Colaizzi's (1978) method of phenomenology. From the transcripts 109 significant sentences and phrases were selected and 13 clusters of themes were categorized from 67 significant statements. These results were integrated into the essential structure of the lived experience of mothers of children with muscular dystrophy. 1. Anxious and relying on chance due to indefinite diagnosis. 2. Only able to cry with shock because of son's fatal disease. 3. Seeing the future as dismal and feeling resentment that this disease in transmitted through the mother. 4. Feeling caught between a husband who is distancing himself from his family and the family which is disintegrating. 5. Feeling like a sinner for transmission of genetic disease(Feeling guilt for conceiving a son with a genetically transmitted fatal disease). 6. Empathizing with other suffers of muscular dystropy and their parents in their grief and helping to dissipate their heavy feelings. 7. Deciding to sacrifice self as a way of taking responsibility for giving birth to a handi-capped son. 8. Losing hope (feeling despair) as son's condition deteriorates in spite of all attempts to help him. 9. Wishing to die with son who becomes progressively immobile. 10. Accumulating Han*(한, 恨), because of rising Hwa(화, anger), and becoming sick both mentally and physically. 11. Seeing events as destiny and finding self-control through faith. 12. Finally, giving up sacrificing self for son and becoming concerned(involved) with other children in the family. 13. Feeling fear at son's impending death. This is the first research on the experience of Korean mothers of children with muscular dystrophy. In applying the phenomenology research method, this study not only helps health professionals understand the experience of these mothers in the Korean patriarchal social system, but the researcher, as a nurse, can share their agony and grief through identification of their inner world through in-depth personal interviews. The results obtained in this study will not only help in the development of family nursing practice for families with genetically transmitted diseases but also prepare basic data for family nursing practice in the Korean sociocultural context.

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The Lived Experience of Mothers about Rearing of School Children With Cerebral palsy (뇌성마비 취학아동 어머니의 양육체험)

  • Baek Kyoung-Seon
    • Child Health Nursing Research
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    • v.7 no.4
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    • pp.434-450
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    • 2001
  • This study is designed to understand the meaning and nature of raising children with cerebral palsy. It researches the experience of mothers of schoolchildren with cerebral palsy by the research method of hermeneutic phenomenology. The study was conducted from November 10, 1999 to December 20, 2000. When children with cerebral palsy usually show symptoms in the early stage of cerebral palsy, mothers do not take children to a doctor for diagnosis. And, most of mothers have a difficult time to accept the reality; they usually respond to the initial diagnosis with shock, reproach, and deny. When mothers start recognizing the reality, they consider that their children have cerebral palsy due to the their mismanagement during pregnancy, delivery, nursing, and initial treatment. They shelter their children from view and feel guilty that they cannot afford to try folk remedies for their children. As time passes, mothers face conflicts between families in diverse ways. Families put the blame on genetic effects. Mothers-in-law give their daughters-in-law a hard time, husbands shift the responsibility of raising children onto their wives, and trouble arises between families-in-law and mothers native families. When children grow up, it is physically difficult for mothers to take care their children. In addition, they suffer from all the troubles in family due to childrens handicap. Mothers try the diverse methods of bringing up children. However, they start getting tired of raising children as they experience failures and financial difficulties. Mothers feel collapsed recalling the ways of raising children. They feel anxiety, miserable, lonely, and worrying when they think how children would attend school, make friends, and live in the future. In this stage, mothers do their best to raise their children with hope. They tend to compare their children with others without handicap and spend money and time in attempting all the treatments. When mothers and children join the society at school, they find that the society does not understand disabled people, teachers show inconsiderate attitude, friends avoid them, and children hardly follow classes. Such experiences make mothers feel angry and frustrated. However, when children adapt to school, mothers see the possibility that children could accomplish schoolwork. They appreciate teachers help and others consideration. Mothers place appropriate expectations on their children and help them to prepare for the future. I would make following suggestions based on the results. 1. As a primary basic course of rehabilitation nursing intervention, solution-centered nursing intervention system should be developed. The intervention needs to be based on the understanding of mothers, who raise children with cerebral palsy, through in-depth interview. 2. Advance researches on the development of individual nursing intervention should be conducted. Individual nursing intervention needs to prevent and release actual pain focusing on mothers raising children with cerebral palsy. 3. Integrated curriculum that help children with cerebral palsy lead a normal school life with ordinary children should be developed. 4. Basic research on using of facilities and effective application of service volunteer to help children with cerebral palsy in school needs to be conducted.

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Genotoxicity Studies on Corn Silk Extract Containing High Maysin (고메이신 함유 옥수수수염 추출물의 유전독성학적 안전성 연구)

  • Ha, Ae Wha;Kang, Hyeon Jung;Kim, Sun Lim;Kim, Myung Hwan;Kim, Woo Kyoung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.9
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    • pp.1045-1052
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    • 2017
  • In this study, a battery of genetic-toxicity studies on corn silk extract with high maysin content were performed according to internationally accepted protocols. In a mutation test using Salmonella Typhimurium TA1535, TA1537, TA98, and TA100, the number of mutant colonies did not significantly increase up to a maximum concentration of $5,000{\mu}g/plate$ in the presence or absence of the S9 metabolic activation system. In the chromosome aberration test using Chinese hamster lung fibroblasts, negative results were observed in the concentration up to $1,250{\mu}g/mL$ of corn silk extract. In the micronucleus test using ICR mice, incidence of polymorphonuclear erythrocytes with a maximum concentration of 2,000 mg/kg corn silk extract did not show any significant difference compared to the negative control group. Based on these results, the test substance, con silk extract, did not influence genotoxicity.