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Establishment of an Analytical Method for Novobiocin in Livestock Products Using HPLC-UVD (HPLC-UVD를 이용한 축산식품 중 Novobiocin의 시험법 확립)

  • Park, Hee-Ra;Kwon, Chan-Hyeok;Lee, Jong-Goo;Kim, Hyung-Soo;Chae, Young-Sik;Oh, Jae-Ho;Kwon, Ki-Sung
    • Korean Journal of Food Science and Technology
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    • v.44 no.3
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    • pp.263-268
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    • 2012
  • Novobiocin is a coumarin-containing antibiotic, and has a longer half-life in various animals than other veterinary medicines. A simple and rapid high-performance liquid chromatography assay for the determination of residual novobiocin levels in chicken, beef and milk has been developed and validated. The separation condition for HPLC/UVD was optimized by a MG II $C_{18}$ (4.6 mm $ID{\times}250$ mm, 5 ${\mu}m$) column with 0.1% formic acid in $H_2O$/0.1% formic acid in Acetonitrile (40/60, v/v) as the mobile phase at a flow rate of 1.0 mL/min and the detection wavelength was set at 340 nm. Residues were extracted from tissue by blending with methanol. After liquid-liquid partitioning, lipid materials were removed with n-hexane and purification as Silica (1 g, 6 mL) cartridge with 10 mL acetone/dichloromethane (10/90, v/v). Limit of quantification and linearity performed by the analytical method were 0.02 mg/kg and 0.999 ($r^2$), and the recovery range was $88.8{\pm}5.6-100.3{\pm}4.4$, $88.8{\pm}7.2-97.0{\pm}3.2$ and $88.1{\pm}4.3-92.8{\pm}3.6%$. It is expected that this analytical method with regards to novobiocin in chicken, beef and milk could be applied as an official method to administer food safety on veterinary medicines.

Isolation and Identification of a Photosensitizer from Pueraria thunbergiana Leaves that Induces Apoptosis in SK-HEP-1 Cells (P. thunbergiana 잎으로부터 SK-HEP-1세포에 대한 apoptosis를 유도하는 광과민성물질의 분리 및 구조동정)

  • Lee, Jun Young;Kim, Mi Kyeong;Ha, Jun Young;Kim, Yong Gyun;Hong, Chang Oh;Kim, So Young;Kim, Chung-Hwan;Kim, Keun Ki
    • Journal of Life Science
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    • v.24 no.3
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    • pp.242-251
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    • 2014
  • The objective of this study was to isolate a photosensitizer from Pueraria thunbergiana leaves that induces apoptosis in SK-HEP-1 cells. Column chromatography and thin layer chromatography were used to isolate active compounds from extracts of P. thunbergiana leaves. The structures of the isolated compounds were determined by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. A substance, named M4-3, was purified from the leaves of P. thunbergiana using various chromatography methods, and the absorbance of the substance was measured. The absorbance was highest at 410 nm, suggesting that the M4-3 substance was a different compound from chlorophyll a and b, which absorb at 410, 502, 533, and 607 nm. Further analyses revealed that the M4-3 compound was a $13^2$-hydoxy pheophorbide, a methyl ester with a molecular weight of 662. M4-3 was identified as a derivative compound of pheophorbide, with a structure that magnesium comes away from the porphyrin ring. The results of the analysis of the cytotoxicity of the M4-3 substance against the SK-HEP-1 cells revealed that it inhibited rates of cell growth by 40% and 80% at a concentration of 0.04 ${\mu}M$ and 0.08 ${\mu}M$, respectively. The M4-3 compound was found to be a photosensitizer for cytotoxicity because it was appeared only in light condition as examining activity in different irradiation conditions (light condition and nonlight condition) under the same concentration. Analysis of morphological changes in the cells following cell death induced by exposure to the M4-3 substance reveled representative phenomena of apoptosis (nuclear condensation, vesicle formation, and fragmentation of DNA). The induction of apoptosis was attributed to the compound's photodynamic activity.

Establishment of a Murine Model for Radiation-induced Bone Loss in Growing C3H/HeN Mice (성장기 마우스에서 방사선 유도 골소실 동물모델 확립)

  • Jang, Jong-Sik;Moon, Changjong;Kim, Jong-Choon;Bae, Chun-Sik;Kang, Seong-Soo;Jung, Uhee;Jo, Sung-Kee;Kim, Sung-Ho
    • Journal of Radiation Protection and Research
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    • v.40 no.1
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    • pp.10-16
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    • 2015
  • Bone changes are common sequela of irradiation in growing animal. The purpose of this study was to establish an experimental model of radiation-induced bone loss in growing mice using micro-computed tomography (${\mu}CT$). The extent of changes following 2 Gy gamma irradiation ($2Gy{\cdot}min^{-1}$) was studied at 4, 8 or 12 weeks after exposure. Mice that received 0.5, 1.0, 2.0 or 4.0 Gy of gamma-rays were examined 8 weeks after irradiation. Tibiae were analyzed using ${\mu}CT$. Serum alkaline phosphatase (ALP) and biomechanical properties were measured and the osteoclast surface was examined. A significant loss of trabecular bone in tibiae was evident 8 weeks after exposure. Measurements performed after irradiation showed a dose-related decrease in trabecular bone volume fraction (BV/TV) and bone mineral density (BMD), respectively. The best-fitting dose-response curves were linear-quadratic. Taking the controls into accounts, the lines of best fit were as follows: BV/TV (%) = $0.9584D^2-6.0168D+20.377$ ($r^2$ = 0.946, D = dose in Gy) and BMD ($mg{\cdot}cm^{-3}$) = $8.8115D^2-56.197D+194.41$ ($r^2$ = 0.999, D = dose in Gy). Body weight did not differ among the groups. No dose-dependent differences were apparent among the groups with regard to mechanical and anatomical properties of tibia, serum ALP and osteoclast activity. The findings provide the basis required for better understanding of the results that will be obtained in any further studies of radiation-induced bone responses.

Antioxidant Activity of Different Parts of Lespedeza bicolor and Isolation of Antioxidant Compound (싸리나무(Lespedeza bicolor) 부위별 추출물의 항산화 활성 및 항산화물질 분리)

  • Lee, Jae-Hak;Jhoo, Jin-Woo
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.763-771
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    • 2012
  • In this study, total antioxidant properties of extracts from different parts of Lespedeza bicolor were determined using techniques of measuring 1,1-diphenyl-2-picryl hydrazyl/2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)-radical scavenging activity and total phenolic contents. The total antioxidant activities of leaf, stem and root extracts from various solvents (water, 50, 70, 100% ethanol, and hot-water) indicated that 50 and 70% ethanol extracts have high radical scavenging activities and phenolic contents. A systematic approach was used to determine the total antioxidant activity of different solvent fractions of the Lespedeza bicolor extracts, partitioning with chloroform, ethyl acetate, n-butanol, and water, and the ethyl acetate fraction was found to have the strongest antioxidant activity. Antioxidant assay-guided isolation was carried out to isolate potential antioxidant compounds. The ethyl acetate fraction of the leaf extract was subjected to silica gel, LH-20 and RP-18 column chromatography successively, and afforded compound 1, which was identified as eriodictyol by NMR and MS analysis, after which its antioxidant activity was determined.

Apoptotic Cell Death by Pectenotoxin-2 in p53-Deficient Human Hepatocellular Carcinoma Cells (종양억제유전자 p53 결손 인체간암세포에서 Pectenotoxin-2에 의한 Apoptosis 유도)

  • Shin, Dong-Yeok;Kim, Gi-Young;Choi, Byung-Tae;Kang, Ho-Sung;Jung, Jee-H.;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1447-1451
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    • 2007
  • Through the screening of marine natural compounds that inhibit cancer cell proliferation, we previously reported that pectenotoxin-2 (PTX-2) isolated from marine sponges exhibits selective cytotoxicity against several cell lines in p53-deficient tumor cells compared to those with functional p53. However, the molecular mechanisms of its anti-proliferative action on malignant cell growth are not completely known. To further explore the mechanisms of its anti-cancer activity and to test whether the status of p53 in liver cancer cells correlates with their chemo-sensitivities to PTX-2, we used two well-known hepatocarcinoma cell lines, p53-deficient Hep3B and p53-wild type HepG2. We have demonstrated that PTX-2 markedly inhibits Hep3B cell growth and induces apoptosis whereas HepG2 cells are much more resistant to PTX-2 suggesting that PTX-2 seems to act by p53-independent cytotoxic mechanism. The apoptosis induced by PTX-2 in Hep3B cells was associated with the modulation of DNA fragmentation factor (DFF) family proteins, up-regulation of pro-apoptotic Bcl-2 family members such as Bax and Bcl-xS and activation of caspases (caspase-3, -8 and -9). Blockade of the caspase-3 activity by caspase-3 inhibitor, z-DEVD-fmk, prevented the PTX-2-induced growth inhibition in Hep3B cells. Moreover, treatment with PTX-2 also induced phosphorylation of AKT and extracellular-signal regulating kinase (ERK), but not c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MARK). Specific inhibitors of PI3K inhibitor (LY294002) and ERK1/2 inhibitor (PD98059) significantly blocks PTX-2-induced-anti-proliferative effects, whereas a JNK inhibitor (SP600125) and a p38 MAPK inhibitor (SB203580) have no significant effects demonstrating that the pro-apoptotic effect of PTX-2 mediated through activation of AKT and ERK signal pathway in Hep3B cells.

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.

Growth Characteristics and Ingredient Analysis According to the Transplanting Date and Distance of Dendranthema (L.). (감국 정식시기와 거리에 따른 생육특성과 성분 분석)

  • Jung, J.H.;Park, N.B.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.22 no.1
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    • pp.65-77
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    • 2020
  • In order to industrialize of Dendranthema indicum (L.) DesMoul., which is a lot of commercially available and is synonymous with chrysanthemum tea, in the autumn of 2018, Dendranthema indicum (L.) DesMoul. seeds were collected from its own native region, and the seeds were germinated after refrigerated storage. Young seedlings were subjected to experiments in February, March, and April in the open field to examine the effects on the harvesting of leaves by distance and the growth of leaves and stems. The results of analyzing the components by collecting the leaves+stem after collecting the flower of Dendranthema indicum (L.) DesMoul. are as follows. 1. When D. indicum (L.) DesMoul. seedlings were planted according to the transplanting date, the number of flowers was 17.1 in the transplanting date in April. The diameter of the flower was 2.9cm, 16ea, 6.5~6.6g in the fresh weight, and the dry weight of the case was 1.1~1.2g. The leaves were 46~47ea in March and April in the planted area, 5.2~5.3cm in leaf length and 3.5~3.6cm in leaf width. 2. When planted D. indicum (L.) DesMoul. seedlings according to transplanting distance, the number of flowers was 16.2 when planted at 20×20cm intervals and, 16.8~17.1 at 30×30~50×50cm intervals. The diameter of the flower was 2.7~2.8cm, the number of petals was 8, the length of the petal was 0.8 cm, and fresh weight was 6.5~6.6g per flower. Leaves had the largest number of 47 of 30×30cm and 40×40cm, and leaf length appeared at the longest 6.2cm in the 50×50cm treatment area, but 5.2cm in the other treatment areas. 3. The extraction yield of D. indicum (L.) DesMoul. leaves+stems was 7.93%, and the extraction solvent colors were light green at 50, 60% and green at 70, 80, 90, 100%. The extraction yield of D. indicum (L.) DesMoul. flowers was 7.58%, the color of the extraction solvent was light yellow at 50, 60 and 70%, yellow at 80 and 90%, and dark yellow at 100%. 4. We confirmed 11 kinds of ingredients such as in D. indicum (L.) DesMoul. flowers are gallic acid, 4-hydroxy benzoic acid, methyl gallate, 4-hydroxy-3-methoxy benzoic, caffeic acid, salicylic acid, p-coumaric acid, sinapic acid, naringin, 4-melthoxyben, flavone. The content was 29.200-36.900ppm. 5. The components contained in the D. indicum (L.) DesMoul. leaf+stem, salicylic acid appeared at 6,129.526ppm, and the next 4-methoxyben was 1,966.714ppm. It was methyl gallate 8.197ppm, 4-hydroxy-3-methoxy benzoic 6.994ppm, caffeic acid 5.566ppm, flavone 4.522ppm, p-coumaric acid 3.787ppm, gallic acid 1.893ppm that appeared in the content below 10ppm.

Effects of Gibberellic Acid and Abscisic Acid on Proteolysis of Senescing Leaves from Rice Seedlings (노화 수도유묘엽의 단백질분해에 미치는 GA$_3$과 ABA의 영향)

  • Kang, S. M;Kang, N. J;Cho, J. L;Kim, Z. H;Kwon, Y. W
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.38 no.4
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    • pp.350-359
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    • 1993
  • The effect of gibberellic acid ($GA_3) and abscisic acid (ABA) on KCl-enhanced proteolysis of senescing leaves of rice(Oryza sativa L. cv. Chilsung) was studied. Emphasis was given to their effects on KCI-enhanced efflux of amino acids and proteinase activity. When treated singly, $GA_3 affected leaf proteolysis little, while ABA increased proteolysis, the rate of amino acid efflux, and ribulose -1,5 -bisphosphate carboxylase / oxygenase (Rubisco)-degrading endoproteinase activity. An additive increase in all three parameters mentioned above was observed when leaves were treated with ABA and KCl. No such an additive effect was found when $GA_3 was treated with KCl. Both $GA_3 and ABA helped to alleviate the KCI-suppressed activity of Rubisco-degrading exoproteinases. The additive increase in proteolysis of rice leaves in the presence of both ABA and KCl could thus be ascribed to a further increase in the efflux of protein hydrolyzates and Rubisco-degrading endoproteinase activity. An increase in proteolysis was accompanied by a decrease in water absorption, and the combined treatment of ABA with KCl resulted in a further reduction of water absorption.

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Field Survey on Pig Slurry Utilization for Crop Cultivation in the Agricultural Farm (양돈분뇨 액비를 이용한 경종농가의 작물재배 실태조사)

  • Choi, D.Y.;Noh, J.S.;Lee, S.C.;Kim, H.N.;Ahn, K.J.;Cho, I.K.
    • Journal of Animal Environmental Science
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    • v.12 no.3
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    • pp.141-150
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    • 2006
  • To optimise the efficient use of nutrients in pig slurry is to cultivate friendly environmental crops. This field survey is to investigate the actual conditions of pig slurry utilization for cultivation of crops in the agricultural farm, based on the survey for 407 selected farms in 9 provinces included 78 counties in Korea. The results obtained in this survey were summarized as follow ; The motive which came to use pig slurry in the agricultural farm were production of friendly environmental crops (29.7%), economy of chemical fertilizer (25.1%), spontaneously (19.2%), inducement of neighboring farmhouse (16.0%), increase of soil fertility (9.3%), and the others (0.7%), respectively. The proportions of pig slurry application land were 56.5% for.ice paddy, 22.6% for dry field, 13.3% for orchard, 4.4% for controlled agriculture and 3.2% for other, respectively. The number of times of pig slurry utilization per year were once (48.9%), twice (31.9%), thrice (14.0%), and the others (5.2%), respectively. The controversial points of pig slurry utilization were malodor (54.1%), insufficiency of spread equipment (22.1%), inconvenience (14.5%), over application (3.4%), over cost (2.9%), heavy metal (1.7%), sanitation (1.0%) and the other (0.2%), respectively. The results indicated that pig slurry could be used as fertilizer source of friendly environmental crops, but further studies are needed to determine the application method and value of the pig slurry for crop cultivation.

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Studies on the Mechanical Properties of Weathered Granitic Soil -On the Elements of Shear Strength and Hardness- (화강암질풍화토(花崗岩質風化土)의 역학적(力學的) 성질(性質)에 관(關)한 연구(硏究) -전단강도(剪斷强度)의 영향요소(影響要素)와 견밀도(堅密度)에 대(對)하여-)

  • Cho, Hi Doo
    • Journal of Korean Society of Forest Science
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    • v.66 no.1
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    • pp.16-36
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    • 1984
  • It is very important in forestry to study the shear strength of weathered granitic soil, because the soil covers 66% of our country, and because the majority of land slides have been occured in the soil. In general, the causes of land slide can be classified both the external and internal factors. The external factors are known as vegetations, geography and climate, but internal factors are known as engineering properties originated from parent rocks and weathering. Soil engineering properties are controlled by the skeleton structure, texture, consistency, cohesion, permeability, water content, mineral components, porosity and density etc. of soils. And the effects of these internal factors on sliding down summarize as resistance, shear strength, against silding of soil mass. Shear strength basically depends upon effective stress, kinds of soils, density (void ratio), water content, the structure and arrangement of soil particles, among the properties. But these elements of shear strength work not all alone, but together. The purpose of this thesis is to clarify the characteristics of shear strength and the related elements, such as water content ($w_o$), void ratio($e_o$), dry density (${\gamma}_d$) and specific gravity ($G_s$), and the interrelationship among related elements in order to decide the dominant element chiefly influencing on shear strength in natural/undisturbed state of weathered granitic soil, in addition to the characteristics of soil hardness of weathered granitic soil and root distribution of Pinus rigida Mill and Pinus rigida ${\times}$ taeda planted in erosion-controlled lands. For the characteristics of shear strength of weathered granitic soil and the related elements of shear strength, three sites were selected from Kwangju district. The outlines of sampling sites in the district were: average specific gravity, 2.63 ~ 2.79; average natural water content, 24.3 ~ 28.3%; average dry density, $1.31{\sim}1.43g/cm^3$, average void ratio, 0.93 ~ 1.001 ; cohesion, $ 0.2{\sim}0.75kg/cm^2$ ; angle of internal friction, $29^{\circ}{\sim}45^{\circ}$ ; soil texture, SL. The shear strength of the soil in different sites was measured by a direct shear apparatus (type B; shear box size, $62.5{\times}20mm$; ${\sigma}$, $1.434kg/cm^2$; speed, 1/100mm/min.). For the related element analyses, water content was moderated through a series of drainage experiments with 4 levels of drainage period, specific gravity was measured by KS F 308, analysis of particle size distribution, by KS F 2302 and soil samples were dried at $110{\pm}5^{\circ}C$ for more than 12 hours in dry oven. Soil hardness represents physical properties, such as particle size distribution, porosity, bulk density and water content of soil, and test of the hardness by soil hardness tester is the simplest approach and totally indicative method to grasp the mechanical properties of soil. It is important to understand the mechanical properties of soil as well as the chemical in order to realize the fundamental phenomena in the growth and the distribution of tree roots. The writer intended to study the correlation between the soil hardness and the distribution of tree roots of Pinus rigida Mill. planted in 1966 and Pinus rigida ${\times}$ taeda in 199 to 1960 in the denuded forest lands with and after several erosion control works. The soil texture of the sites investigated was SL originated from weathered granitic soil. The former is situated at Py$\ddot{o}$ngchangri, Ky$\ddot{o}$m-my$\ddot{o}$n, Kogs$\ddot{o}$ng-gun, Ch$\ddot{o}$llanam-do (3.63 ha; slope, $17^{\circ}{\sim}41^{\circ}$ soil depth, thin or medium; humidity, dry or optimum; height, 5.66/3.73 ~ 7.63 m; D.B.H., 9.7/8.00 ~ 12.00 cm) and the Latter at changun-long Kwangju-shi (3.50 ha; slope, $12^{\circ}{\sim}23^{\circ}$; soil depth, thin; humidity, dry; height, 10.47/7.3 ~ 12.79 m; D.B.H., 16.94/14.3 ~ 19.4 cm).The sampling areas were 24quadrats ($10m{\times}10m$) in the former area and 12 in the latter expanding from summit to foot. Each sampling trees for hardness test and investigation of root distribution were selected by purposive selection and soil profiles of these trees were made at the downward distance of 50 cm from the trees, at each quadrat. Soil layers of the profile were separated by the distance of 10 cm from the surface (layer I, II, ... ...). Soil hardness was measured with Yamanaka soil hardness tester and indicated as indicated soil hardness at the different soil layers. The distribution of tree root number per unit area in different soil depth was investigated, and the relationship between the soil hardness and the number of tree roots was discussed. The results obtained from the experiments are summarized as follows. 1. Analyses of simple relationship between shear strength and elements of shear strength, water content ($w_o$), void ratio ($e_o$), dry density (${\gamma}_d$) and specific gravity ($G_s$). 1) Negative correlation coefficients were recognized between shear strength and water content. and shear strength and void ratio. 2) Positive correlation coefficients were recognized between shear strength and dry density. 3) The correlation coefficients between shear strength and specific gravity were not significant. 2. Analyses of partial and multiple correlation coefficients between shear strength and the related elements: 1) From the analyses of the partial correlation coefficients among water content ($x_1$), void ratio ($x_2$), and dry density ($x_3$), the direct effect of the water content on shear strength was the highest, and effect on shear strength was in order of void ratio and dry density. Similar trend was recognized from the results of multiple correlation coefficient analyses. 2) Multiple linear regression equations derived from two independent variables, water content ($x_1$ and dry density ($x_2$) were found to be ineffective in estimating shear strength ($\hat{Y}$). However, the simple linear regression equations with an independent variable, water content (x) were highly efficient to estimate shear strength ($\hat{Y}$) with relatively high fitness. 3. A relationship between soil hardness and the distribution of root number: 1) The soil hardness increased proportionally to the soil depth. Negative correlation coefficients were recognized between indicated soil hardness and the number of tree roots in both plantations. 2) The majority of tree roots of Pinus rigida Mill and Pinus rigida ${\times}$ taeda planted in erosion-controlled lands distributed at 20 cm deep from the surface. 3) Simple linear regression equations were derived from indicated hardness (x) and the number of tree roots (Y) to estimate root numbers in both plantations.

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