• Title/Summary/Keyword: 생물검정법

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Construction of a Transgenic Tobacco Expressing a Polydnaviral Cystatin (폴리드나바이러스 유래 시스타틴 유전자 발현 형질전환 담배 제작)

  • Kim, Yeongtae;Kim, Eunsung;Park, Youngjin;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.54 no.1
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    • pp.7-15
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    • 2015
  • CpBV (Cotesia plutellae bracovirus) is a polydnavirus and encodes a cystatin (CpBV-CST1) gene. Its overexpression suppresses insect immunity and alters insect developmental processes. This study aimed to construct a genetically modified (GM) tobacco to further explore the physiological function of the viral cystatin and to apply to control insect pests. To this end, the transgenic tobacco lines were screened in expression of the target gene and assessed in insecticidal activity. A recombinant vector (pBI121-CST) was prepared and used to transform a bacterium, Agrobacterium tumefasciens. The transformed bacteria were used to generate transgenic tobacco lines, which were induced to grow callus and resulted in about 92% of shoot regeneration. The regenerated plants were screened by PCR analysis to confirm the insertion of the target gene in the plant genome. In addition, the expression of the target gene was assessed in the regenerated plants by quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis showed that the transgenic line plant expressed the target gene about 17 times more than the control tobacco, indicating a stable insertion and expression of the target gene in the transgenic tobacco line. The insecticidal activity was then analyzed using the screened transgenic tobacco lines against the teneral 1st instar larvae of the oriental tobacco budworm, Helicoverpa assulta. Though there was a variation in the insecticidal efficacy among transgenic lines, T9 and T12 lines exhibited more than 95% mortality at 7 days after feeding treatment. These results suggest that CpBV-CST1 is a useful genetic resource to be used to generate GM crop against insect pests.

Action properties and insecticidal effects of thiamethoxam to the melon aphid, Aphis gossypii, and diamondback moth, Plutella xylostella (목화진딧물과 배추좀나방에 대한 thiamethoxam의 살충효과 및 작용특성)

  • Jang, Cheol;Hwang, In-Cheon;Yu, Yong-Man;Choe, Kwang-Ryul
    • The Korean Journal of Pesticide Science
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    • v.2 no.3
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    • pp.126-136
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    • 1998
  • For the purpose of effective control strategy of the melon aphid, Aphis gossypii and the diamondback moth, Plutella xylostella, thiamethoxam and 3 other insecticides in different classes were used with bioassay test methods in laboratory and greenhouse. They were examined to evaluated and compared with contact toxicity, stomach toxicity, rapid action, systemic action, and residual effect of imidacloprid, thiamethoxam (nicotinoids), acephate (organophosphorates), and carbosulfan (carbamates). As results of contact toxicity responses of A. gassypii against 4 insecticides using a spray application method, $LC_{50}$ values of acephate, carbosulfan, imidacloprid and thiamethoxam were 41.9, 5.2, 1.1, and 0.7 ppm. respectively. In the evaluation of stomach toxicity response of P. xylostella using a leaf-dipping method, with the 2nd instar larva $LC_{50}$ values of imidacloprid, thiamethoxam and acetamiprid were 64.9, 24.6 and 15.2 ppm, with the 3rd instar larva were 125.2, 42.7 and 27.8 ppm. and with the 4th instar larva were 241.1, 44.5 and 23.9 ppm, respectively. In the case of rapid action to A. gossypii using a spray application method after inoculation, $LT_{50}$ values of imidacloprid, thiamethoxam, carbosulfan, and acephate were 26.6, 28.0, 30.3, and 41.7 min. respectively. Otherwise, in the inoculation after applying compounds, $LT_{50}$ values of thiamethoxam, imidacloprid, and carbosulfan were 95.5, 118.0, and 122.9 min. respectively. Evaluating to systemic action from the abaxial surface to the adaxial surface of red pepper leaf with spray method, $LT_{50}$ values of thiamethoxam, imidacloprid, and carbosulfan were 162.2, 168.9, and 564.1 min. respectively. For the systemic action from the lower leaves to the upper leaves on red pepper, $LT_{50}$ values of carbosulfan, thiamethoxam, imidacloprid, and acephate were 2.3, 2.9, 3.0, and 8.8 days, respectively. In red pepper plant, $LT_{50}$ values of carbosulfan, imidacloprid, thiamethoxam, and acephate on the systemic action from the roots to the upper leaf were 0.6, 1.0, 1.0, and 13.8 days, respectively. As these results, it might be that thiamethoxam was excellent on systemic effect in red pepper. For the evaluation of residual effect on red pepper with A. gossypii, thiamethoxam and imidacloprid maintained high control effects as over 80% upto 10 days after treating compounds.

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Management of the Development of Insecticide Resistance by Sensible Use of Insecticide, Operational Methods (실행방식 측면에서 살충제의 신중한 사용에 의한 저항성 발달의 관리)

  • Chung, Bu-Keun;Park, Chung-Gyoo
    • Korean journal of applied entomology
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    • v.48 no.2
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    • pp.123-158
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    • 2009
  • An attempt was made to stimulate future research by providing exemplary information, which would integrate published knowledge to solve specific pest problem caused by resistance. This review was directed to find a way for delaying resistance development with consideration of chemical(s) nature, of mixture, rotation, or mosaics, and of insecticide(s) compatible with the biological agents in integrated pest management (IPM). The application frequency, related to the resistance development, was influenced by insecticide activity from potentiation, residual period, and the vulnerability to resistance development of chemical, with secondary pest. Chemical affected feeding, locomotion, flight, mating, and predator avoidance. Insecticides with negative cross-resistance by the difference of target sites and mode of action would be adapted to mixture, rotation and mosaic. Mixtures for delaying resistance depend on each component killing very high percentage of the insects, considering allele dominance, cross-resistance, and immigration and fitness disadvantage. Potential disadvantages associated with mixtures include disruption of biological control, resistance in secondary pests, selecting very resistant population, and extending cross-resistance range. The rotation would use insecticides in high and low doses, or with different metabolic mechanisms. Mosaic apply insecticides to the different sectors of a grid for highly mobile insects, spray unrelated insecticides to sedentary aphids in different areas, or mix plots of insecticide-treated and untreated rows. On the evolution of pest resistance, selectivity and resistance of parasitoids and predator decreased the number of generations in which pesticide treatment is required and they could be complementary to refuges from pesticides To enhance the viability of parasitoids, the terms on the insecticides selectivity and factors affecting to the selectivity in field were examined. For establishment of resistant parasitoid, migration, survivorship, refuge, alternative pesticides were considered. To use parasitoids under the pressure of pesticides, resistant or tolerant parasitoids were tested, collected, and/or selected. A parasitoid parasitized more successfully in the susceptible host than the resistant. Factors affecting to selective toxicity of predator are mixing mineral oil, application method, insecticide contaminated prey, trait of individual insecticide, sub-lethal doses, and the developmental stage of predators. To improve the predator/prey ratio in field, application time, method, and formulation of pesticide, reducing dose rate, using mulches and weeds, multicropping and managing of surroundings are suggested. Plant resistance, predator activity, selective insect growth regulator, and alternative prey positively contributed to the increase of the ratio. Using selective insecticides or insecticide resistant predator controlled its phytophagous prey mites, kept them below an economic level, increased yield, and reduced the spray number and fruits damaged.

Environmental Pollutants in Streams of Andong District and Insect Immune Biomarker (안동지역 하천의 환경오염물질과 곤충면역 생체지표 분석)

  • Ryoo Keon Sang;Ko Seong-Oon;Cho Sunghwan;Lee Hwasung;Kim Yonggyun
    • Korean journal of applied entomology
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    • v.44 no.2
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    • pp.97-108
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    • 2005
  • Samples of water, soil, and sediment were taken from 10 streams near Andong, Korea in May 2004. To assess the degree of environmental pollution of each stream, chemical pollutants such as total notrogen (T-N), total phosphorus (T-P), chemical oxygen demand (COD), heavy metals, organophosphorus pesticides, organochlorine pesticides, and dioxin-like PCB congeners were analyzed by standard process tests or U.S. EPA methods. In addition, biomarkers originated from insect immune systems of beet armyworm, Spodoptera exigua, were used to analysis of the environmental samples. Except Waya-chun stream showing T-N content of 9.12 mg/L, most streams were contaminated with relatively low levels of overall pollutants in terms of T-N, T-P, and COD, compared to their acceptable environmental levels designated by the Ministry of Environment. Contents of Pb and Cd in samples of each stream were much lower than environmentally permissible levels. However, several times higherconcentrations of Pb and Cd were found in locations at Mi-chun, Kilan-chun, and Hyunha-chun streams, in comparison with other streams. Diazinon, parathion, and phenthoate compounds among organophosphorus pesticides were detected as concentrations of 0.19, 0.40, and $1.13\;{\mu}g/g$, respectively, from soil sample collected in the vicinity of Mi-chun stream. On the other hand, 16 organochlorine pesticides and 12 dioxin-like PCB congeners, known as endocrine disrupting chemicals, selected in this study were not found above the limit of detection. Biomarker analyses using insect immune responses indicated that Waya-chun stream was suspected as exposure to environmental pollutants. Limitation and compensation of both environmental analysis techniques are discussed.

The effect of bracket width on frictional force between bracket and arch wire during sliding tooth movement (치아의 활주 이동시 브라켓 폭이 브라켓과 호선 사이의 마찰력에 미치는 효과)

  • Choi, Won-Cheul;Kim, Tae-Woo;Park, Joo-Young;Kwak, Jae-Hyuk;Na, Hyo-Jeong;Park, Du-Nam
    • The korean journal of orthodontics
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    • v.34 no.3 s.104
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    • pp.253-260
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    • 2004
  • Frictional force between the orthodontic bracket and arch wire during sliding tooth movement is related to many factors, such as the size, shape and material of both the bracket and wire, ligation method and the angle formed between the bracket and wire. There have been clear conclusions drawn in regard to most of these factors, but as to the effect of bracket width on frictional force there are only conflicting studies. This study was designed to investigate the effect of bracket width on the amount of frictional forces generated during clinically simulated tooth movement. Three different widths of brackets $(0.018{\times}0.025'\;standard)$ narrow (2.40mm), medium (3.00mm) and wide (4.25mm) were used in tandem with $0.016{\times}0.022'$ stainless steel wire. Three bracket-arch wire combinations were drawn on for 4 minutes on a testing apparatus with a head speed of 0.5mm/min and tested 7 times each. To reproduce biological conditions, dentoalveolar models were designed with indirect technique using a material with similar elastic properties as periodontal ligament (PDL). In addition, to minimize the effect of ligation force, elastomer was used with added resin, which was attached to the bracket to make up for the discrepancies of bracket width. The results were as follows: 1. Maximum frictional force for each bracket-arch wire combination was: Narrow (2.40mm): $68.09\pm4.69gmf$ Medium (3.00mm): $72.75\pm4.98 gmf$ Wide (4.25mm): $72.59\pm4.54gmf$ 2. Frictional force was increased with more displacement of wire through the bracket slot. 3. The ANOVA psot-hoc test showed that the bracker width had no significant effect on frictional force when tested under clinically simulated conditions(p>0.05).

Biological Activity of Ixeris dentata Nakai juice Extracts (씀바귀(Ixeris dentata Nakai) 생즙 추출물의 생리활성)

  • 김명조;김주성;강원희;조미애;함승시;정동명
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.924-930
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    • 2002
  • Ixeris dentata extracts exllibited antimicrobial activity against some bacteria and fungi. Also EtOH extracts showed strong antioxidant activity and RC$_{50}$ value was 28 $\mu\textrm{g}$/mL. The inhibitory effect of Ixeris dentata on the mutagenicity in Salmonella and cytotoxicity on cancer cell were studied. Ixeris dentata extracts showed anti-mutagenic effects of 78.83 and 75.96% on B(a)P in S. typhimurium TA98 and Th100, respectively. These extracts showed 78.72% antimutagenicity on TA100 against MNNG. The Ixeris dentata extract with strong antimutagenic activities was further fractionated by hexane, ethyl acetate, butanol and water. Butanol fraction was found to be highest in antimutagenic activity against MNNG than the other fractions. Butanol fraction of Ixreis dentate revealed the highest cytotoxicity against AS49 human lung carcinoma cells in which cell growth was inhibited by 93.75% at 375 $\mu\textrm{g}$/mL. Hexane fraction of ixeris dentate exhibited 68.56% inhibition against MCF-7 human breast adenocarcinoma cells at 500 $\mu\textrm{g}$/mL. Hexane fraction of Ixeris dentata exhibited 84.91% inhibition against Hep 3B human hepatocellular carcinoma cells at 500 $\mu\textrm{g}$/mL. From these results, it is considered that Ixeris dentata has strong antimutagenic and anticancer effects in vitro. However, these extracts and fractions did not show any cytotoxic effect against 293 cells.

Assessment of the Specificity of A Hybridization of Surfactant Protein A by Addition of Non-specific Rat Spleen RNA (Surfactant Protein A mRNA을 이용한 유전자 재결합 반응에서 비특이성 RNA의 첨가에 의한 특이성 검정)

  • Kim, Byeong Cheol;Kim, Mi Ok;Kim, Tae-Hyung;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.4
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    • pp.393-404
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    • 2004
  • Background : Nucleic acid hybridization has become an essential technique in the development of our understanding of gene structure and function. The quantitative analysis of hybridization has been used in the measurement of genome complexity and gene copy number. The filter hybridization assay is rapid, sensitive and can be used to measure RNAs complementary to any cloned DNA sequence. Methods : The authors assessed the accuracy, linearity, correlation coefficient and specificity of the hybridization depending on the added dose(0, 1, 5, and $10{\mu}g$) of non-specific rat spleen RNA to hybridization of surfactant protein A mRNA. Filter hybridization assays were used to obtain the equation of standard curve and thereby to quantitate the mRNA quantitation. Results : 1. Standard curve equation of filter hybridization assay between counts per minute (X) and spleen RNA input (Y) was Y=0.13X-19.35. Correlation coefficient was 0.98. 2. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) was Y=0.00066X-0.046. Correlation coefficient was 0.99. 3. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $1{\mu}g$ spleen RNA was Y=0.00056X-0.051. Correlation coefficient was 0.99. 4. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $5{\mu}g$ spleen RNA was Y=0.00065X-0.088. Correlation coefficient was 0.99. 5. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $10{\mu}g$ spleen RNA was Y=0.00051X-0.10. Correlation coefficient was 0.99. Conclusions : Comparison of cpm/filter in a linear range allowed accurate and reproducible estimation of surfactant protein A mRNA copy number irrespective of the addition dosage of non-specific rat spleen RNA over the range $0-10{\mu}g$.

Isolation and Evaluation of an Antiviral Producing Serratia spp. Strain Gsm01 against Cucumber mosaic virus in Korea (한국에서 CMV에 항바이러스 효과를 나타내는 Serratia spp. Gsm01 균주의 분리 동정 및 효과 검정)

  • Ipper, Nagesh S.;Lee, Seon-Hwa;Suk, Jung-Ki;Shrestha, Anupama;Seo, Dong-Uk;Park, Duck-Hwan;Cho, Jun-Mo;Park, Dong-Sik;Hur, Jang-Hyun;Lim, Chun-Keun
    • The Korean Journal of Pesticide Science
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    • v.10 no.4
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    • pp.344-350
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    • 2006
  • An Antiviral producing bacterial strain was isolated from ginseng root environment in Hongcheon, Kangwon province of Republic of Korea. Identification of this bacterial strain was performed by physiological and biochemical tests along with 16S rRNA analyses. The results revealed that the bacterium was closer to genus Serratia, which was named as Gsm01. The strain was grown in Mannitol-Glutamate-Yeast (MGY) broth for 48 h. The culture was centrifuged and the filtrate obtained was tested for its ability to control Cucumber mosaic virus strain Y (CMV-Y) in greenhouse and field experiments. In the green house experiments, CF was evaluated for its ability to protect local host, Chenopodium amaranticolor and systemic host of CMV, Nicotiana tabacum cv. Xanthi-nc. It was found that, CF treatment reduced viral infection by 98% in local host; C. amaranticolor. The N. tabacum cv. Xanthi-nc plants treated with CF did not show visible viral symptoms 15 days post inoculation (dpi) and remained symptomless throughout the periods of the study. To evaluate effectiveness of CF under field conditions, experiment was carried out in a polyvinyl house. It was observed that, 52% plants were protected from viral diseases compared to non-treated plants, increasing the crop yield. This is the first report showing antiviral activity of a Serratia spp. against CMV.

Yearly Occurrence of Thrips Infesting Hot Pepper in Greenhouses and Differential Damages of Dominant Thrips (시설 고추재배지 총채벌레 연중 발생 및 주요 총채벌레의 차등 해충성)

  • Kim, Chulyoung;Choi, Duyeol;Lee, Donghyun;Khan, Falguni;Kwon, Gimyon;Ham, Eunhye;Park, Jungjoon;Kil, Eui-Joon;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.61 no.2
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    • pp.319-330
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    • 2022
  • Andong is a place to culture the great amount of hot pepper (Capsicum annuum L.) in Korea. This study reports a yearly occurrence (March 31~October 25, 2021) of thrips infesting the hot pepper in Andong. Thrips caught to yellow sticky traps were diagnosed by morphological characters and showed two dominant species: Frankliniella occidentalis and F. intonsa. During this period, a total of 107,874 thrips were caught and included F. occidentalis at about 82%, F. intonsa at about 17%, and the other thrips at about 0.3%. There were two main peaks at May~June and at September~October, respectively, in which the total number of thrips was higher in the second peak and most were F. occidentalis. Interestinly, a low level of thrips occurred during July~August was observed and explained by their susceptibility to high temperatures. A laboratory experiment by exposing thrips to high temperatures showed that thrips were susceptible to temperatures higher than 35℃ and not tolerant to 45℃ for 1 h. Indeed, high temperatures higher than 45℃ were recorded in the greenhouses in Andong during July~August. F. occidentalis was more tolerant to the high temperatures than F. intonsa. On the other hand, the thrips showed the highest occurrence peak at July~August in hot pepper-culturing greenhouse in Kangwon, where the average temperatures were mostly lower than those of Andong and no high temperatures higher than 45℃ were recorded during July~August. A viral disease caused by tomato spotted wilt virus (TSWV) was observed in the hot peppers cultured in Andong greenhouses. Multiplex PCR was used to detect the virus along with identification of thrips. With a high record of about 30%, the virus-infected thrips were detected during all the monitoring period. The virulent thrips were identified to be only F. intonsa. These results suggest that F. occidentalis gives a direct damage especially during harvesting period with their high populations while F. intonsa gives indirect damage by transmitting TSWV.

Association of osteoarthritis and bone mineral density in women -The health and nutritional examination survey in Kuri- (여성의 골관절염과 골밀도간의 관련성 분석 -구리시민 건강.영양진단 조사결과를 바탕으로-)

  • Sheen, Seung-Soo;Lee, Soon-Young;Min, Byung-Hyun;Suh, Il
    • Journal of Preventive Medicine and Public Health
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    • v.30 no.4 s.59
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    • pp.669-685
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    • 1997
  • Previous studies, reporting the inverse relationship between osteoarthritis and osteoporosis suggest the existence of possible pathophysiologic mechanisms between them. To examinine the hypothesis that 'bone mineral densities of women with osteoarthritis are significantly higher than that of women without osteoarthritis in Korea', subjects from the health and nutritional examination survey in Kuri city were sampled. Samples were selected through multi-stage sampling frame using established clusters in Kuri city. From August 18 to September 10,1997, the survey was conducted. Among the. total number of selected sample population (1,656 people), response .ate was 52.4 percent (348 men and 519 women). 420 women who took BMD measurement, radiologic exam, and anthropometric exam were selected for the analysis. The analytic results are as follows. 1. General characteristics: Mean BMD was $0.493g/cm^2$, mean age was 43.0, mean BMI was $23.9kg/m^2$. The number of women who experienced menopause was 106, hysterectomy was 19. There were 0 case of osteoarthritis of hip, 64 cases of osteoarthritis of knee, and 2 cases of osteoarthritis of hand. 2. Univariate analysis results: Mean BMD of women with the osteoarthritis of knee was significantly lower than that of women without the osteoarthritis of knee(0.4269 vs. $0.5057g/cm^2$). But, there were too few cases of osteoarthritis of hip and hand, so comparative studies of BMD in osteoarthritis of hip and hand could not be conducted. There were significant differences of BMD among pre-menopause group(0.5204), post-menopause group(0.4206), and hysterectomy group(0.4881). Additionally, there were significant differences of BMD among diabetes group(0.4297), impaired glucose tolerance group(0.4874), and normal group(0.5057). Furthermore, age, parity, BMI, bioimpedance were significantly related with BMD. 3. Multivariate analysis results: To examinine the relationship between osteoarthritis and BMD while controlling the other variables' effects which were significant in the univariate analyses, multiple linear regression analysis was done. But, it was found that osteoarthritis of knee was not a significant variable to BMD anymore. While age and menopause had significant negative relationship with BMD. Diabetes, parity, BMI, and bioimpedance did not have significant relationships with BMD. After stratification of subjects according to menopause, multiple linear regression analyses were done to each strata. Consequently, age in post-menopause group, age and osteoarthritis of knee in hysterectomy group showed significant negative relationship with BMD. The results did not support the many results of other previous studies done with white men and women. further studies of biological plausibility to Korean women are recommended. Also it is suggested that longitudinal study to verify the relationship between osteoarthritis and BMD will be valuable.

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