• 제목/요약/키워드: high-level expression

검색결과 1,204건 처리시간 0.028초

HIV-1 O형 항체 진단시료의 개발 (Development of Test System for Detection of Antibody to Human Immunodeficiency Virus Type 1 Subtype O)

  • 조영식;유승신;하건우;이상국;조명환;신형식;김선영
    • 대한바이러스학회지
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    • 제28권1호
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    • pp.31-38
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    • 1998
  • In Korea, all domestic made test systems for detecting antibodies in HIV-1 contain the antigens from human immunodeficiency type 1 (HIV-1) subtype B. However, because HIV-1 subtype O is significantly different in amino acid sequences from all other subtypes of HIV-1, there has been a need for developing a test for detecting antibodies in subtype O. For this purpose, the entire nucleotide sequence corresponding to the extracellular domain of the transmembrane glycoprotein of HIV-1 subtype O was synthesized with consideration of Escherichia coli condon usage. Various regions of the extracellular domain were cloned into E. coli expression vectors and tested for levels of protein production. The nucleotide sequence, named ECTM, that can encode a 129 amino acid-long peptide, was found to be expressed at a high level in E. coli. The protein of approximately 17 kDa specifically reacted with sera from individuals infected with HIV-1 subtype O. The ECTM protein was purified to near homogeneity by the CM-T gel chromatography, using concentrated, denatured inclusion bodies. In Western blot analysis, the purified viral antigen reacted with sera from individuals infected with subtype O more efficiently than subtype B. The enzyme linked immunoabsorbent assay (ELISA) system was developed using the subtype O viral protein and compared with the commercially available kit lacking the antigens from subtype O. The ELISA kit containing the subtype O antigen ECTM alone efficiently reacted with sera from individuals infected with subtype O. The subtype O antigen-containing kit produced a positive absorbence even when sera were diluted 512-fold, suggesting a high sensitivity. The commercially available kit also reacted with subtype O sera, but produced a negative result at a dilution of 8-fold. Our results suggest that the currently available kit may not be able to efficiently detect subtype O sera and that the viral protein developed in this study may be added to the current system to maximize the detection of sera from individuals infected with subtype O.

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Black ginseng extract ameliorates hypercholesterolemia in rats

  • Saba, Evelyn;Jeon, Bo Ra;Jeong, Da-Hye;Lee, Kija;Goo, Youn-Kyoung;Kim, Seung-Hyung;Sung, Chang-Keun;Roh, Seong-Soo;Kim, Sung Dae;Kim, Hyun-Kyoung;Rhee, Man-Hee
    • Journal of Ginseng Research
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    • 제40권2호
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    • pp.160-168
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    • 2016
  • Background: Ginseng (Panax ginseng Meyer) is a well-characterized medicinal herb listed in the classic oriental herbal dictionary as "Shin-nong-bon-cho-kyung." Ginseng has diverse pharmacologic and therapeutic properties. Black ginseng (BG, Ginseng Radix nigra) is produced by repeatedly steaming fresh ginseng nine times. Studies of BG have shown that prolonged heat treatment enhances the antioxidant activity with increased radical scavenging activity. Several recent studies have showed the effects of BG on increased lipid profiles in mice. In this study report the effects of water and ethanol extracts of BG on hypercholesterolemia in rats. To our knowledge, this is the first time such an effect has been reported. Methods: Experiments were conducted on male Sprague Dawley rats fed with a high-cholesterol diet supplemented with the water and ethanol extracts of BG (200 mg/kg). Their blood cholesterol levels, serum white blood cell levels, and cholesterol-metabolizing marker genes messenger RNA (mRNA) expression were determined. Liver and adipose tissues were histologically analyzed. Results: We found that BG extracts efficiently reduced the total serum cholesterol levels, low-density lipoprotein (LDL) levels with increased food efficiency ratio and increased number of neutrophil cells. It also attenuated the key genes responsible for lipogenesis, that is, acetyl-coenzyme A (CoA) acetyltransferase 2, 3-hydroxy-3-methyl-glutaryl-CoA reductase, and sterol regulatory element-binding protein 2, at the mRNA level inside liver cells. Furthermore, the BG extract also reduced the accumulation of fat in adipose tissues, and inhibited the neutral fat content in liver cells stained with hematoxylin and eosin and oil red O. Conclusion: Administration of BG extracts to Sprague Dawley rats fed with high-cholesterol diet ameliorated hypercholesterolemia, which was mediated via modulation of cholesterol-metabolizing marker genes. This data throw a light on BG's cardioprotective effects.

Physiological responses involved in reactive oxygen species (ROS) of rice plant under alone or multi artificial stress conditions

  • Kim, Yoonha;Waqas, Muhammad;Khan, Abdul Latif;Mun, Bong-Gyu;Yun, Byung-Wook;Lee, In-Jung
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.203-203
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    • 2017
  • The Earth's climate is rapidly changing because of increasing carbon dioxide content in atmosphere so, climate prediction models anticipate that earth surface temperature will rise by 3 to $5^{\circ}C$ in next 50 to 100 years. Therefore, frequency of un-expected weather events such as drought, salinity, low or high temperature and flooding etc. will be increasing worldwide. Furthermore, increased atmosphere temperature can influence pests and pathogens spread as well. Therefore, to protect enormous grain loss from unexpected weather conditions, studies related with combine stress conditions like abiotic plus biotic stress condition are really required. Thus, our research focused on physiological responses under combined abiotic and biotic stress condition in rice plant. To induce uniform stress condition, we used NaCl (100 mM) and salicylic acid (0.5 and 1.0 mM SA) as each stress a stimulator. Each artificial abiotic and biotic stress inducer was applied to hydroponically grown rice seedlings alone or together for four day. The data were collected in a time-dependent manner [1, 2, 3 and 4 day(s) after treatment (DAT)] and were matched with our anticipation that shoot length and shoot fresh weight was decreased in solo and combined abiotic and biotic stress condition. The lipid peroxidation content was significantly increased ($1.5{\pm}0.2$ to $2.7{\pm}0.1mg$ mg of $MDA\;g^{-1}FW$) in the first two days in both stress exposed plants, and showed the opposite trend ($0.5{\pm}0.01$ to $0.1{\pm}0.001mg$ of $MDA\;g^{-1}FW$) in last two days under multi stress condition. Superoxide dismutase (SOD) activity did not showed difference in only biotic stress condition (alone 0.5 and 1.0 mM SA) as compared to control however, it was significantly increased in multi stress condition or solo abiotic stress condition whereas, catalase (CAT), and ascorbate peroxidase (APX) activities were significantly decreased in solo biotic and combined abiotic and biotic condition. In particular, both enzymes activities were more decreased in multi stress condition as compared to solo biotic stress condition. The results for relative mRNA expression level of CAT and APX enzymes were in agreement with results of spectrophotometric values. Correlation value between each stress condition and phenotypic data showed that biotic stress condition showed high correlation with activity of CAT and APX whilst, abiotic stress condition revealed significant correlation with SOD activity.

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지역주민과 건강행태와 국민건강증진법에 대한 인식과 태도 (Health Behavior and Attitude of Residents toward the National Health Promotion Law in Kyungsan City)

  • 이관희;박재용;한창현;윤석옥
    • 보건교육건강증진학회지
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    • 제16권2호
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    • pp.19-40
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    • 1999
  • In order to ascertain the attitudes of residents to their health and the National Health Promotion Law, surveyors interviewed 1,220 subjects, 1% of men and women in Kyungsan city, who were twenty-year-old or more. The major findings are as follows: Men and women were 48.2% and 51.8%, respectively. The recognition rate of enacting and enforcing this law is 59.2% of men and 51.3% of women. With regard to the behavioral attitude to the health in the distinction of sex and age, current smokers are 31.2% of the interviewees, 61.6% of the men and 3.3% of the women. Current drinkers are 35.1%, 59.5% of the men and 12.3% of the women, but on the other hand there is little significance in the distinction of age. The acknowledgement proportion of enacting and enforcing this law is 59.2% of male and 51.3% of female. In terms of the recognition rate of the contents according to the general characteristics of interviewees, it appears that the indication of a warning expression on a packing paper of cigarette case and a liquor bottle is 92.4% and also the designation of a smoking free area in public facilities is 94.8%. Prohibition of cigarette-sale to the teenagers who are under 19, is 96.0%. Considering these facts, the recognition rate is high. On the contrary, 48.8% is accounted for encouraging a medical check-up before marriage which is in a low position. As a result of multiple behavior as a independent educational level, marital significant variables. In case of having undergone a periodic medical examination the recognition rate was high whereas frequent exercise led to the low recognition rate. Concerning the details of the undertaking in accordance with each factor of general characteristics, the greater part of them have been appraised successfully whether it is recognized or not. On the other side, no effect got answered about the result of the undertaking subjects to general and peculiar behavior attitude towards health was in effect or not. A great majority approved of more reinforcement of legal regulation about smoking and drinking regardless of whether they perceived the details of the law of promotion of National Health Promotion Law or not. Additionally there was significant difference in reinforcing legal regulation of smoking and drinking in compliance with the attitude of the substance of this law. With regard to education, public relations and evaluation about national health through public health centers by our government, the younger and the higher in education they are, the more deficient they feel. First of all, those who were aware of the enforcement of this law as well as plenty of scarcity answered that better service of disease prevention had to be expanded than ever. In consideration of the above-stated results, the education to public health and the business of public relations should be reinforced and a practical campaign for health life should also spread out for the purpose of encouraging to practise healthy life-style.

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Molecular Mechanisms of 5-Azacytidine-Induced Trifluorothymidine-Resistance In Chinese Hamster V79 Cells

  • Jin Kyong-Suk;Lee Yong-Woo
    • 대한의생명과학회지
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    • 제11권2호
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    • pp.165-173
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    • 2005
  • A potent demethylating agent, 5-Azacytidine (5-AzaC) has been widely used as in many studies on DNA methylation, regulation of gene expression, and cancer biology. The mechanisms of the demethylating activity were known to be formation of complex between DNA and DNA methyltransferase (MTase), which depletes cellular MTase activity. However, 5-AzaC can also induce hypermethylation of a transgene in a transgenic cell line, G12 cells and it was explained as a result of defense mechanisms to inactivate foreign gene(s) somehow. This finding evoked the question that whether the phenomenon of hypermethylation induced by 5-AzaC is limited to the transgene or it can be occurred in endogenous gene(s). In order to answer the question, mutagenicity test of 5-AzaC and molecular characterization of mutants obtained from the test were performed using an endogenous gene, thymidine kinase (tk) in Chinese hamster V79 cells. When V79 and V79-J3 subclone cells were treated with 1, 2.5 ,5, $10{\mu}M$ of 5-AzaC for 48 hours, their maximum mutant frequencies were revealed as $6\times10^{-3}\;at\;5{\mu}M$(350-fold induction over background) and $8\times10^{-3}\;at\;2.5{\mu}M$ (l,800-fold induction over background) respectively. Since the induction rates were too high to be induced by true mutations, many trifluorothymidine (TFT)-resistant $(TFT^R)$ cells were subjected to Northern blot analysis to check the presence of tk transcripts. Surprisingly, all clones tested possessed the transcripts in a similar level, that implicates the $TFT^R$ phenotype induced by 5-AzaC has not given rise to hypermethylation of the gene in spite of unusually high mutation frequency. In addition, it has shown that the TK activity in the pool of 5-AzaC-induced $TFT^R$ cells has about a half of that in spontaneously-induced $TFT^R$ cells or in non-selected parental V79-J3 cells. This result suggests that the mechanism(s) underlying the TFT-resistance between spontaneously occurred and 5-AzaC-induced cells may be different. These findings have shown that the $TFT^R$ phenotype induced by 5-AzaC has not given rise to hypermethylation of the tk gene, and 5-AzaC may be induced by one or combined pathways among many drug resistance mechanisms. The exact mechanisms for the 5-AzaC-induced $TFT^R$ phenotype remain to elucidate.

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Bacillus subtilis dgk (diacylglycerol kinase) 유전자의 생리적 자극에 의한 유도발현 (Expression Patterns of Bacillus subtilis Diacylglycerol Kinase Gene Induced by Physiological Stimuli)

  • Lee, Mi-Young;Suh, Seok-Jong;Lee, Jin-Hyung;Song, Bang-Ho;Kim, Jong-Cuk
    • 한국미생물·생명공학회지
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    • 제30권1호
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    • pp.15-20
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    • 2002
  • Diacylglycerol Kinase (DGK)는 E. coli 및 진핵세포에서의 신호전달에 관여하며, 또한 미생물에서 생리적 자극에 따라 다른 발현 양상을 보이는 것으로 밝혀져 있다. Bacillus subtilis에서 이 유전자에 대한 환경에서의 자극 신호들, 즉 pH 변화, 삼투압의 변화 및 온도의 변화에 따른 발현양상을 연구하였다. 이미 동정된 dgk locus의 KpnI-HindIII의 0.45 kb의 DNA fragment를 probe로 하여 Dot blot, Northern blot analysis를 통해 발현량을 조사해 본 결과 dgk 유전자는 pH변화, 삼투압의 변화 및 온도의 변화에 대응하여 발현되는 유전자임을 알 수 있었다. 특히 낮은 pH, 고 삼투압, 및 저온에서 dgk 유전자의 발현량이 많아짐을 확인 할 수 있었다. Northern hybridization에서 약 2.5kb의 mRNA가 관찰되었다. dgk gene의 ORF size는 약 0.4 kb로 관찰된 transcript size와는 일치하지 않았다. 따라서 Streptococcus mutans의 dgk gene과 마찬가지로 B. subtilis의 dgk 유전자도 polycistronic mRNA로 발현되는 것을 추정할 수 있었으며, 염색체상의 dgk gene에서 상류의 ORF2까지의 크기가 약 2.5 kb로 관찰된 mRNA size와 동일하였다. dgk gene 상류의 ORF2영역의 0.6 kb의 DNA fragment를 probe로 하여 northern blot hybridization을 수행한 결과, 2.5 kb의 mRNA가 관찰되었으며 발현되는 형태도 dgk probe를 이용한 결과와 동일하였다. 따라서 dgk gene은 상류부위의 ORF2 gene과 operon을 형성하여 polycistronic mRNA로 전사되는 것으로 판단된다.

Aspergillus nidulans에서 MsnA 하위 유전자로 선별된 단당류 수송자 mstB의 기능 분석 (Characterization of a Monosaccharide Transporter mstB Isolated as a Downstream Gene of MsnA in Aspergillus nidulans)

  • 전미향;채순기
    • 미생물학회지
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    • 제47권4호
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    • pp.281-288
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    • 2011
  • 스트레스 반응에 관여하는 Saccharomyces cerevisiae 전사인자인 Msn2/4의 $C_2H_2$ zinc finger 부위와 아미노산 서열 유사성을 보이는 Aspergillus nidulans MsnA의 하위 유전자 획득을 위하여 msnA 결손 돌연변이체 또는 과발현 균주에서 야생주와 비교하여 차별적으로 발현되는 유전자(Differentially Expressed Gene, DEG)들을 분리하였다. 선별된 DEG들은 염기서열 결정을 통해 해당 유전자들을 동정하였고 이들 중 DEG6는 단당류 수송자(monosaccharide transporter)로 예측된 mstB 유전자로 밝혀졌다. mstB의 발현은 MsnA 과발현에 의하여 증가되었으며 MsnA는 in vitro에서 mstB 프로모터 부위에 직접적으로 결합하였다. MstB는 12개의 막결합 부위를 가지며 A. niger의 고친화성 단당류 수송자(high-affinity monosaccharide transporter)인 MstA와 80%의 높은 아미노산 서열 동일성을 보였다. mstB 결손 돌연변이체의 표현형은 야생주와 유사하였으나 MstB가 과발현된 균주는 낮은 당 농도인 0.1% glucose 배지에서 유성생식 기관인 cleistothecia의 형성이 증가하였다. 이러한 결과는 단당류 수송자인 MstB가 유성분화 과정에서 요구되는 당의 수송에 관여하고 있음을 시사한다.

한국인 일반인구에서의 사회 불안 증상의 특성 (The Characteristics of Social Anxiety Symptoms in the Korean Nonclinical Population)

  • 강여진;하주원;임세원;오강섭
    • 대한불안의학회지
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    • 제7권1호
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    • pp.22-28
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    • 2011
  • Objective : Social anxiety disorder (SAD) has been characterized by a marked sense of anxiety provoked by exposure to one or more social situations. It has been well-known that the cultural characteristics of a society strongly affect the expression of SAD symptoms. The current study attempted to explore clinical and cultural characteristics of social anxiety symptoms in the Korean nonclinical population. Methods : 801 volunteers who participated in the national social anxiety screening day, which was held by Korean academy of anxiety disorders, were invited to complete the self-report version of Liebowitz social anxiety scale (LSAS-SR). A total of 691 participants completed the LSAS-SR and other information related to this study. Results : Social anxiety symptoms were reported by a great percentage of subjects, as displayed by the relatively high mean LSAS scores (44.58 ; SD=25.60). There were no significant differences in total LSAS score according to sex, education, or marital status. The three most fearful or avoided situations for participants, as measured by the percentage of subjects who had a score of more than 3 on each LSAS item were as follows: item 6 (acting, performing, or giving a talk in front of an audience ; 29.67% for fear and 22.72% for avoidance), item 16 (speaking up in a meeting ; 25.62% for fear and 21.56% for avoidance), and item 15 (being the center of attention; 13.89% for fear and 9.70% for avoidance). Conclusion : The Korean participants in this study demonstrated a high level of SAD symptoms. Our subjects reported that they were more likely to be fearful or avoid passive situations such as being the center of attention as compared with results from a study of western society which indicated that western people were more likely to be fearful or avoid active situations. These findings emphasize the importance of considering a patient's cultural background when evaluating SAD symptoms.

공동배양과정의 배지조성과 배양조건이 벼 형질전환효율에 미치는 영향 (The Effects of Co-cultivation Medium and Culture Conditions on Rice Transformation Efficiency)

  • 김율호;박향미;최만수;윤홍태;최임수;신동범;김정곤;이장용
    • 한국육종학회지
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    • 제41권3호
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    • pp.252-260
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    • 2009
  • 1. 본 연구에서는 공동배양 배지에 Agrobacterium 성장 억제물질인 silver nitrate를 첨가하고 변온과 여과지처리를 추가하여 공동배양 기간을 7일로 늘였으며, 또한 항산화 물질 3종을 공동배양 배지에 첨가하여 세포의 oxidative burst를 최소화함으로써 벼 형질전환효율을 높일 수 있었다. 또한 이 방법을 적용하여 형질전환이 어려운 품종을 대상으로도 형질전환 식물체를 작성할 수 있었다. 2. 벼 형질전환체의 70%에서 도입유전자 수가 1copy인 것으로 나타나, 적은 수의 유전자가 안정적으로 도입됨을 확인 하였다. 3. 이러한 결과를 바탕으로, 새로운 공동배양 방법을 사용하여 우수한 농업적 형질을 가진 벼 육종 소재 및 품종을 신속하게 개발할 수 있을 것으로 기대된다.

쥐 모델에서 고지방사료로 악화된 대장 건강에 대한 콩의 개선 효과 (Improvement effect of cooked soybeans on HFD-deteriorated large intestinal health in rat model)

  • 최재호;신태균;류명선;양희종;정도연;운노타쯔야
    • Journal of Applied Biological Chemistry
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    • 제64권4호
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    • pp.383-389
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    • 2021
  • 비만은 숙주의 전신 염증 및 대사 기능 장애에 기여하는 장 상피 장벽 기능 저하와 관련이 있다. 한국의 전통 식품으로 식이섬유가 풍부한 콩 제품은 항염증 반응을 비롯한 다양한 생물학적 활성을 나타내어 왔으나 대장 건강에 대해서는 보고된 바가 없다. 본 연구에서는 고지방 식이(HFD)를 섭취한 비만 모델에서 콩(CSB)에 대한 장 건강 증진 효과를 조사하였다. SD 쥐에게 동물 실험 기간 동안 HFD 또는 10.6% CSB가 함유된 HFD (HFD + CSB)를 제공하였다. CSB의 섭취는 HFD로 유발된 체중과 지방 축적 증가를 현저하게 감소시켰다. 또한, CSB의 섭취는 대장 조직에서 HFD에 의해 감소된 밀착 결합 지표(ZO-1, Claudin-1 및 Occludin-1)의 mRNA 발현을 개선시켰다. 또한, 조직병리학적 평가에서도 CSB 섭취는 대장 조직에서 HFD에 의해 증가된 염증 세포 침윤과 대장 상피 조직 붕괴를 개선하는 것으로 나타났다. Genus 수준에서 HFD 섭취에 의해 Lactobacillus, Duncaniella, Alloprevotella 등 미생물 종의 abundance 차이는 확인되었으나, CSB 섭취로 인한 영향은 명확하게 나타나지 않았다. NMDS 분석에서 HFD 섭취에 의해 유의적인 장내미생물 생태 이동을 보여주었지만 CSB 섭취는 차이가 없었다. HFD와 CSB 간 유의적으로 차이가 나타나는 genera를 조사하기 위해 LEfSe를 수행한 결과, CSB는 Anaerotignum, Enterococcus, Clostridium sensu stricto 및 Escherichia/Shigella 속의 풍부함을 증가시킨 반면 Longicatena 및 Ligilactobacillus의 풍부함을 감소시켰다. 이러한 결과는 CSB 섭취는 긴밀한 접합 성분을 개선하여 HFD로 악화된 대장 건강을 개선하는 반면 장내미생물생태에 긍정적인 효과를 미치는지에 대해서는 명확하지 않았다.