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Purification and Enzymatic Characteristics of the Bacillus pasteurii Urease Expressed in Escherichia coli (Escherichia coli에서 발현된 Recombinant Bacillus pasteurii Urease의 정제 및 효소학적 특성)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.519-526
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    • 1992
  • The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

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Evaluation of Water Quality after Rehabilitation of Cheonggye Stream using AGP Test (조류성장잠재력 조사를 이용한 청계천 복원 이후 수질 평가)

  • Park, Myung-Hwan;Hwang, Soon-Jin;Suh, Mi-Yeon;Kim, Yong-Jae;Kim, Baik-Ho
    • Korean Journal of Ecology and Environment
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    • v.40 no.2
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    • pp.234-243
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    • 2007
  • Algal growth potential (AGP) test was performed to evaluate the water quality and changes in phytoplankton communities before and after a heavy rain event at six sampling sites in Cheonggye Stream (St. 1 and 2), Jungnang Stream (St. 3 and 4), and Lower part of Han River System (St. 5 and 6) after rehabilitation of Cheonggye Stream, October 2005. To test AGP on each sampling site, cyanobacterium Microcystis aeruginosa was applied as a standard alga. Total nitrogen (TN) showed high values at Jungnang Stream, while St. 4 recorded highest values in this study. However, TN values of Cheonggye Stream and Lower Part of Han River showed similar levels. Total phosphate (TP) also showed high values at Jungnang Stream, while St. 4 recorded highest. However, TP in Cheonggye Stream were extremely low levels. Although chlorophyll-${\alpha}$ (chi-${\alpha}$) contents before the rain event were similar through the sampling sites, chl-${\alpha}$ after the rain increased dramatically at Jungnang Stream and Lower part of Ban River. In particular, after the rain, TP was the highest at St. 4, where Cyclotella sp. dominated the phytoplankton community. When compared with control, AGP values before the rain were comparatively low in all sites, while those after the rain highly increased with the dose-dependently of field water added, due perhaps to the increased nutrients by rainfall. Similar results were observed in Cheonggye Stream. Therefore, for the aesthetic fostering for the citizens, although Cheonggye Stream was Presently being sustained by treated water supply, they have a potential of outbreak of phytoplankton by the increased nutrients supply when a heavy rain comes.

Effect of Dietary Fish Oil on Lipid Peroxidation in Rats Liver and Brain During Postnatal Development (어유섭취가 출생후 발달과정의 흰 쥐의 간과 뇌조직의 지질과산화와 그 관련기능에 미치는 영향)

  • 박명희
    • Journal of Nutrition and Health
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    • v.20 no.2
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    • pp.111-121
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    • 1987
  • Lipie peroxide formation, antiperoxidative s system and body adaptability for handling lipid p peroxide were examined in the first and second g generations of rats fed fish oil. Mackerel oil(MO) was used and four other dietary oils and fat, i.e. soybean oil(SO), perilla oil(PO), rapeseed oil(RO) and beef tallow(BT) were also employed to compare the effect of fish oil. Synthetic diets containing these five dietary fats at the level of 1O%(w/w), were given to the correspond­m ing groups of male and female rats weighing about 70 grams. After 34 days of feeding, male a and female rats were mated and their offsprings were raised throughout suckling (17, 26 days) and weanling (39 days) periods. Liver lipid pero­x xide level was highest in MO group of both first (mother rats after lactation) and second genera­t tions of 17 and 26 days old, but not of 39 days old. During suckling period, liver lipid peroxide level was well matched to total unsaturation of dietary fat. Brain lipid peroxide levels were not different among five groups. Liver $alpha$-tocopherol a and reduced glutathione (GSH) levels were lowest in MO fed first generation. In second generation, $alpha$-tocopherol level was also low in MO group, although the effect was less pronoun­c ced, but GSH level was not different from other groups. Oxidized glutathione (GSSG) level did not consistently vary by change in dietary fat. Glutathione peroxidase activity increased as young rats grew up to 39 days. Superoxide d dismutase activity change was insignificant by a age, but was shown as lowest in MO group. At the age of 26 and 39 days, liver glutatione peroxidase activity was increased as was level of lipid peroxide, suggesting that this is the one of the mechanisms responsible for body adapta­b bility for protection against the accumulation of lipid peroxide.

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A Study on the Effect of Luffae Fructus Retinervus Herbal-Acupuncture at ST36 on Collagen-induced Arthritis in Mice (족삼리(足三里) 사과락약침(絲瓜絡藥鍼)이 생쥐의 Collagen-induccd Arthritis에 미치는 영향)

  • Chae, Choong-heon;Choi, Sun-mi;Yim, Yun-kyoung
    • Journal of Acupuncture Research
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    • v.22 no.1
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    • pp.131-144
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    • 2005
  • Objective : The aim of this study is to observe the effect of Herbal-acupuncture(HA) with Luffae Fructus Retinervus Herbal-Acupuncture Solution(LFR-HAS) at ST36(Joksamni) on Collagen Ⅱ -induced arthritis(CIA) in mice. Methods : DBA1/J mice were immunized with bovine type Ⅱ collagen(CⅡ) on days 0 and 21 to induce an arthritis. The mice were divided into 5 groups. They were Normal group(wild type), Control group(CIA), Saline group(CIA +saline injection), Needle Prick group(CIA +single Prick with an injection needle) and LFR-HA group(CIA +LFR-HA treatment). The saline injection, needle prick and LFR-HA were made on the right ST36(Joksamni) of mice for 5 weeks, 3 times a week beginning 4 weeks after the booster immunization. Results : 1. The highest synovial rate of lung fibroblasts was measured in the 1% LFR-HAS. 2. TNF-${\alpha}$ expression of survival cells from CIA mouse joint was significantly reduced in the 1% LFR-HAS. 3. The incidence of arthritis and the spleen weight of CIA mouse were significantly reduced by the Luffae Fructus Retinervus Herbal-Acupuncture (LFR-HA) at ST36. 4. The concentrations of IL-6, INF-${\alpha}$, INF-${\alpha}$, IgG, IgM, and anti-collagen Ⅱ in the CIA mouse serum were significantly reduced by the LFR-HA at ST36. 5. The histological examination showed that, in the LFR-HA group, the cartilage destruction and the synoviocyte proliferation in the CIA mouse joint were not significant compared to the control group, and the collagen fiber was similarly expressed as the normal group. 6. In the LFR-HA group, the ratio of CD3e+ to CD19+ cell, and the ratio of CD4+ to CD8+ cell in the lymph node were similarly maintained as those of the normal group. 7. CD69+/CD3e+ and CD11a+/CD19+ cells in the CIA mouse lymph node were significantly reduced by the LFR-HA at ST36. 8. CD11b+/Gr-1+ cells in the CIA mouse joint were significantly reduced by the LFR-HA at ST36. Conclusions : These results indicate that Luffae Fructus Retinervus Herbal-Acupuncture (LFR-HA) at ST36 may regulate the immune system and have a therapeutic effect on Collagen-induced arthritis(CIA) in mice.

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Characterization of Entomopathogenic Fungus from Trialeurodes vaporariorum and Evaluation as Insecticide (온실가루이 병원성 곰팡이의 특성 및 살충제 개발을 위한 평가)

  • Yoon, Hwi Gun;Shin, Tae Young;Yu, Mi Ra;Lee, Won Woo;Ko, Seung Hyun;Bae, Sung Min;Choi, Jae Bang;Woo, Soo Dong
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.64-70
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    • 2013
  • The greenhouse whitefly, Trialeurodes vaporariorum, is an economically important pest for greenhouse crops because they cause direct damage by feeding on plant nutrients and indirect damage as transmits many virus vectors. It has recently become a serious problem because of the continuous use of insecticide resulting in resistance among greenhouse whitefly population. To overcome these problems, in this study, the biological characteristics and virulence of an entomopathogenic fungus isolated from the cadaver of nymph greenhouse whitefly were investigated. Isolated fungus was identified as Isaria fumosorosea by morphological examinations and genetic identification using sequences of the ITS, ${\beta}$-tubulin, and EF1-${\alpha}$ regions. This fungus was named as I. fumosorosea SDTv and tested for the virulence against nymphs T. vaporariorum and the cold activity, the thermotolerance and the stability of UV-B irradiation on conidia. Mortality rate of greenhouse whitefly showed from 84 to 100% and the virulence increased with increasing conidial concentrations, $1{\times}10^5$ to $10^8$ conidia/ml. Conidia were stable at $35^{\circ}C$, 0.1 $J/cm^2$ of UV irradiation and germinated after 8 days at $4^{\circ}C$. Additionally, the activities of chitinases and proteases produced by I. fumosorosea SDTv were varied according to the medium. In conclusion, I. fumosorosea SDTv which showed high mortality rate against greenhouse whitefly will be used effectively in the integrated pest management programs against the greenhouse whitefly.

Application of Geographic Information System for Risk Analysis on Gas Industrial facilities (가스산업시설에서의 위험성 평가분야에서 지리정보시스템의 적용)

  • Lee Jeung-Woo;Kim Yun-Hwa;Kim Ky-Soo;Ko Jae-Wook
    • 한국가스학회:학술대회논문집
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    • 1997.09a
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    • pp.121-126
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    • 1997
  • 우리나라는 그동안 가스산업에서 고속성장을 이룩해 왔으나, 근래에는 중대가스산업사고가 빈번하게 일어나고 있다. 그런 이유로 1996년도에는 중대산업사고 예방제도가 전면적으로 실시되고 공정안전보고서를 자성하여 제출하도록 함으로서 위험성 평가의 중요성이 점차 높아가고 있으며, 위험성 평가 기법들에 대하여 여러 연구 단체에서 연구${\cdot}$ 시도되고 있다. 또한 이러한 연구${\cdot}$보고된 위험성 평가 기법들을 프로그램화하고 적용하려는 시도가 많이 있어 왔다. 본 연구에서는 기존에 연구${\cdot}$개발된 위험성 평가 기법들에 최근 컴퓨터 산업의 발달에 힘입어 각광을 받고 있는 지리정보시스템을 적용하고자 한다. 이러한 가스산업시설의 위험성 평가 시스템은 위험성을 평가하기 위해서 필요한 여러 가지 정보들을 지리정보시스템이 속성 데이터로서 저장하고 있으며, 가스산업시설에 관련된 주변의 도민들을 공간 데이터로서 저장하고 있다. 그리고 위험성 평가 시스템의 세부적인 기능을 모듈화하였다. 우선 위에 언급한 속성 데이터와 공간 데이터를 관리하는 모듈과 이러한 데이터를 가지고 사고영향 범위를 산출해내기 위한 모듈, 그리고 산출된 사고 영향 범위를 도면에 나타내는 모듈로 나뉘어져 있다. 이렇게 지리정보시스템에 구축되어 있는 도면에 위험성을 평가한 결과치를 나타냄으로서 위험성 평가 숙련자가 아니더라도 위험성 평가를 할 수 있고 결과를 분석하도록 도와 줄 수 있도록 할 수 있다. 또한 향후 재난관리시스템에서는 도면상의 도로에 교통량 가중치와 인근 소방서와 경찰서등의 위치를 관리하도록 지리정보시스템을 적용할 수 있으며, 가스시설물 관리시스템에서는 최근 대형가스사고의 대부분이 타공사에 의한 것임을 고려하여 가스배관망을 포함하여 기타 다른 지하배관망을 관리하도록 지리정보시스템을 적용할 수 있다.기질로서 이용하지 못하므로 invertase 및 ${\alpha}-glucosidase$ 활성이 없는 것으로 나타났다.)의 모양은 배양에 의해 형성된 균핵(菌核)의 모양과 다소 차이가 있었다. R. cerealis의 균사생장(菌絲生長) 적온(適溫)은 $23^{\circ}C$였으며, 다른 균핵진균(菌核眞菌)들의 균사생장(菌絲生長) 적온(適溫)은 $27-33^{\circ}C$였다. 일부 균핵진균(菌核眞菌)들의 균사생장(菌絲生長) 최고온도(最高溫度)는 $41^{\circ}C$ 로서 높은 반면, R. cerealis 의 균사생장(菌絲生長) 최고온도(最高溫度)는 $31^{\circ}C$로서 낮았다. R. cerealis 의 균사생장(菌絲生長) 최저온도(最低溫度)는 $2^{\circ}C$ 였으며, 다른 균핵진균(菌核眞菌)들의 균사생장(菌絲生長) 최저온도(最低溫度)는 $6-10^{\circ}C$였다. 시술(施術)하였다. 10. 한양방향시치료시(韓洋方同時治療時) 양호이상(良好以上)의 효과(效果)를 보인 경우(境遇)가 30건중(件中) 21건(件)으로 70%의 치료효과(治療效果)를 보여 한방단일진료시(韓方單一診療時)보다 다소 높은 치료효과(治療效果)를 나타냈다. 11. 1일평균(日平均) 한방진료건수(韓方診療件數)는 26.95명(名)이었고, 1일평균(日平均) 한방초진환자수(韓方初診患者數)는 3.9명(名)으로 나타났으며, 한방진료(韓方診療)가 양한방(洋韓方) 총진료(總診療)의 23.20%를 차지했고, 1일(日) 평균(平均) 한양방(韓洋方) 총진료건수(總診療件數)에 대(對)해 1일(日) 평균(平均) 한방초진건수(韓方初診件數)가 21.51%를 차지하는 것으로 나타났다.험이 완료된 31명에서는 부작용이 없거나 경도의 부작용이 있었으나 임상적

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Characterization of Bacteriocin, lacticin YH-10, Produced by Lactococcus lactis subsp. lactis YH-10 Isolated from Kimchi (김치유산균인 Lactococcus lactis subsp. lactis YH-10가 생산하는 박테리오신의 특성)

  • Park, Eun-Min;Kim, Young-Hwa;Park, So-Jin;Kim, Yun-Im;Ha, Yu-Mi;Kim, Sung-Koo
    • Journal of Life Science
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    • v.14 no.4
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    • pp.683-688
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    • 2004
  • A bacteriocin-producing lactic acid bacteria was isolated from Kimchi on MRS selective media with the use of Lactobacillus delbrueckii subsp. delbrueckii as an indicator strain. The strain YH-10 was identified as Lactococcus lactis subsp. lactis through the API test. The crude bacteriocin (freeze-dried 50% ammonium sulfate precipitate of culture supernatant) produced by the strain was named as lacticin YH-10. Lacticin YH-10 showed the growth inhibitory activity against Gram positive pathogenic bacteria and other lactic acid bacteria. The bacteriocin was inactivated by proteases such as protamex and aroase AP-10 and partially inactivated by amylase, proteinase K, trypsin, and papain. The lacticin YH-10 remained its activity with the treatment of heat at 10$0^{\circ}C$ for 60 min or the changes of pH 2 to 11. However, the activity was lost at high pH combined with the exposure to 10$0^{\circ}C$. The bacteriocin production of the strain was started in the exponential phase and stopped in the stationary phase. The approximate molecular mass of the bacteriocin produced by the strain was approximate 14 kDa in the analysis on SDS-PAGE.

Association of Cytochrome-17 (MspA1) Gene Polymorphism with Risk of Gall Bladder Stones and Cancer in North India

  • Dwivedi, Shipra;Agrawal, Sarita;Singh, Shraddha;Madeshiya, Amit Kumar;Singh, Devendra;Mahdi, Abbas Ali;Chandra, Abhjeet
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5557-5563
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    • 2015
  • Background: Cholelithiasis is associated in 54%-98% of patients with carcinoma of the gallbladder, and a high incidence among females suggests a role of female hormones in the etiology of the disease. Cytochrome $P450C17{\alpha}$ (CYP-17) is a key enzyme involved in estrogen metabolism and polymorphisms in CYP-17 are associated with altered serum levels of estrogens. Thus, we investigated whether the CYP-17 MspA1 gene polymorphism might impact on risk of gall bladder cancers or gallstones, as well as to determine if this gene polymorphism might be linked with estrogen serum levels and lipid profile among the North Indian gall bladder cancer or gallstone patients. Materials and Methods: CYP-17 gene polymorphisms (MspA1) were genotyped with PCR-RFLP in cancer patients (n=96), stone patients (n=102), cancer + stone patients (n=52) and age/sex matched control subjects (n= 256). Lipid profile was estimated using a commercial kit and serum estrogen was measured using ELISA. Results: The majority of the patients in all groups were females. The lipid profile and estrogen level were significantly higher among the study as compared to control groups. The frequency of mutant allele A2 of CYP17 MspA1 gene polymorphism was higher among cancer (OR=5.13, 95% CI+3.10-8.51, p=0.0001), stone (OR=5.69, 95%CI=3.46-9.37, p=0.0001) and cancer + stone (OR=3.54, 95%CI=1.90-6.60, p=0.0001) when compared with the control group. However there was no significant association between genotypes of CYP17 MspA1 gene polymorphism and circulating serum level of estrogen and lipid profile. Conclusions: A higher frequency of mutant genotype A1A2 as well as mutant allele A2 of CYP-17 gene polymorphism is significantly associated with risk of gallbladder cancer and stones. Elevated levels of estrogen and an altered lipid profile can be used as predictors ofgall bladder stones and cancer in post menopausal females in India.

Effect of Etomidoline on the Isolated smooth Muscle of Rabbit (Etomidoline이 각종 평활근에 미치는 영향)

  • Kim, W.J.;Kim, J.H.;Sheen, Y.Y.
    • The Korean Journal of Pharmacology
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    • v.16 no.2 s.27
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    • pp.25-29
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    • 1980
  • Etomidoline ($Nonspa^{\circledR}$), which is chemically related to tertiary amine, is new synthetic antispasmodic agent with analgesic action. Antispasmodic effect of this agent is stronger than hyoscine butylbromide ($Buscopan^{\circledR}$), quaternary amine, and the absorption from intestine is also much higher. This study was undertaken to determine the effect of etomidoline on duodenal motility and other smooth muscles of rabbit. Strips of various isolated smooth muscle, 2 cm long from adult rabbits weighting about 2 kg, were suspended in a muscle chamber containing Tyrode's solution, which was bubbled with oxygen gas, and the temperature of the solution was kept constant at $38^{\circ}C$. After being washed with fresh solution several times the strips of smooth muscle attained constant motility and tonus. Etomidoline and other drugs were added in various concentrations to the chamber. Contractility of the strips was measured by using polygraph (Grass, model 7). The results are as follows: 1) In isolated rabbit atrium etomidoline produces a slight depression of contractility and the rate is also decreased. 2) On the other hand, etomidoline relaxed isolated strips of stomach, duodenal, and detrusor of rabbit. This relaxing effect of etomidoline on isolated duodenal strip of rabbit was not blocked by ${\alpha}$-adrenergic blocking agent, phenoxybenzamine, but by ${\beta}$-adrenergic blocking agent, propranolol. 3) Etomidoline did not exert any effect on isolated aorta, gall bladder, and trigone of rabbit. From the above results, it may be concluded that the relaxing effect of etomidoline on duodenal strip is related ${\beta}$-adrenergic receptor.

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A Similitude Study of Soil-Wheel System for Identifying the Dimension of Pertinent Soil Parameter(I) -Pull Prediction Analysis- (구동륜(驅動輪)의 성능예측(性能豫測)에 적합한 토양변수(土壤變數)의 차원해석(次元解析)을 위한 차륜(車輪)-토양(土壤) 시스템의 상사성(相似性) 연구(硏究)(I) -견인력(牽引力) 예측(豫測) 분석(分析)-)

  • Lee, K.S.;Chung, C.J.
    • Journal of Biosystems Engineering
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    • v.14 no.2
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    • pp.67-79
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    • 1989
  • This study was conducted to investigate the applicability of true model theory for pull prediction in a powered lugged wheel-soil system and to examine the possibility of using principles of similitude in investigating the dimensions of soil parameters pertinent to a powered lugged wheel-soil system concerning the pull prediction. The following conclusions were derived from the study; 1) The pull of prototype wheels proved to be predicted by those of the model wheels for the range of the dynamic weight tested. The pull curves of models and prototype were respectively very similar in the shape. From this basic knowledge, it was enabled to apply the similitude theory to the performance prediction of the true model. 2) A conditional equation which can be used for the prediction of pull of prototype by model test was derived as follows. $n_f=n_1^{-b}$ where $n_f$ : force scale = $w/w_m$ $n_1$ : length scale = ${\ell}/{\ell}_m$ b : exponent on the length dimension of the soil property ${\alpha}$ The range of the numerical value of b, which was determined by the least square method, was found to be -2.0~-2.6. 3) Considering a relatively wide variation of b values in the pull prediction, b is considered to be a function of many variales. Thus it was concluded that there are several soil properties which are pertinent to the powered lugged-wheel-soil system concerning the pull prediction, and these soil properties may have the different effects on the pull of model and protytype wheels, to give the different dimension on the soil parameters.

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