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Isolation of Isoflavones and Soyasaponins from the Germ of Soybean (콩 배아로 부터 Isoflavone과 Soyasaponin의 동시 분리)

  • Kim, Sun-Lim;Lee, Jae-Eun;Kim, Yul-Ho;Jung, Gun-Ho;Kim, Dea-Wook;Lee, Choon-Ki;Kim, Mi-Jung;Kim, Jung-Tae;Lee, Yu-Young;Hwang, Tae-Young;Lee, Kwang-Sik;Kim, Wook-Han;Kwon, Young-Up;Kim, Hong-Sig;Chung, Ill-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.2
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    • pp.149-160
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    • 2013
  • The objective of present study was to simultaneously isolate of isoflavone and soyasaponin compounds from the germ of soybean seeds. Soy germ flours were defatted with hexane for 48h at room temperature, and methanolic extracts were prepared using reflux apparatus at $90^{\circ}C$ for 6h, two times. After extraction, extracts were separated with preparative RP-$C_{18}$ packing column ($125{\AA}$, $55-105{\mu}m$, $40{\times}150mm$), and collected 52 fractions were identified with TLC plate (Kieselgel 60 F-254) and HPLC, respectively. Among the identified isoflavone and soyasaponin fractions, isoflavone fractions were re-separated using a recycling HPLC with gel permeation column (Jaigel-W252, $20{\times}500mm$). Final fractions were air-dried, and the purified compounds of two isoflavones (ISF-1-1, ISF-1-2) and four soyasaponins (SAP-1, SAP-2, SAP-3, SAP-4) were obtained. Two isoflavone compounds (ISF-1-1, ISF-1-2) were acid-hydrolyzed for the identification of their aglycones, and confirmed by comparing with 12 types of isoflavone isomers. While the four kinds of soyasaponins were identified by using a micro Q-TOF mass spectrometer in the ESI positive mode with capillary voltage of 4.5kV, and dry temperature of $200^{\circ}C$. Base on the obtained results, it was conclude that ISF-1-1 is the mixture isomers of daidzin (43.4%), glycitin (47.0%), and genistin (9.6%), but ISF-1-2 is the single compound of genistin (99.8% <). On the other hand, soyasaponin SAP-1 is the mixture compounds of soyasaponin A-group (Aa, Ab, Ac, Ae, Af); SAP-2 is soyasaponin B-group (Ba, Bb, Bc) and E-group (Bd, Be); SAP-3 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$); SAP-4 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$, ${\beta}a$), respectively.

Effect of Feeding Rare Earth on Egg Production and Hatchability Broiler Growth (희토(稀土)의 급여가 종계의 산란율 및 부화율에 미치는 영향)

  • Ham, S.K.;Song, T.H.;Zhang, G.Q.;Hur, S.N.;Park, H.S.
    • Korean Journal of Poultry Science
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    • v.33 no.3
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    • pp.225-231
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    • 2006
  • To investigate the effect of feeding rare earth (RE) on the performance of breeder hens a feeding trial with sixteen thousands of 158 day old Ross broiler breeder hens was conducted for thirty weeks. A mixture of RE- chlorides containing mainly La, Ce, and Pr was mixed into corn-soy based diet at two levels, 300 and 600 mg/kg, All the hens were housed in flat layer houses and the ratio of male to female was maintained at one to ten. Dietary supplementation of the of RE at a level of 300 mg/kg made the hens reach egg production peak higher by about 6% and earlier by about two weeks. As a result, it made higher hatchable egg production by 3.5%. It also reduced dramatically the mortality of both male and female breeders(P<0.05). Egg weight was slightly increased but egg qualify was not much influenced by dietary supplementation of RE. Egg albumin hight and Haugh unit were significantly improved while egg shell thickness, egg breaking strength and yolk color were similar to those of the control. Experimental results appeared to show a good possibility that egg fertility and hatchability were improved by feeding RE. Dietary supplementation of RE at a level of 300 mg/kg should be acceptable but not the 600 mg/kg level for breeder hens. Further studies on the effect of RE on egg fertility and hatchability appears to be necessary.

Antioxidative and Cytoprotective Effects of Annona muricata (Graviola) Extract for HDF Cell Damage Induced by Hydrogen Peroxide (H2O2에 의해 유도된 HDF 세포 손상에 대한 그라비올라 추출물의 항산화 및 세포 보호 효과)

  • Shin, Yun-Mi;Kim, You-Jeong;You, Seon-Hee
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.3
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    • pp.568-576
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    • 2017
  • As interest in functionality and environmentally friendly cosmetics is growing in recent years, materials that use safe and effective plant extracts have been developed. Therefore, this study also attempted to check the possibility of the graviola extract, which is known to have various efficacy mainly as a health functional material as a functional cosmetic material. In order to find out the antioxidant activity of graviola, we measured total polyphenol, total flavonoid content and DPPH radical scavenging activity and measured the ROS activity inhibition effect and cytoprotective effect on oxidative stress by treating HDF with hydrogen peroxide cells at an appropriate concentration after checking cytotoxicity in HDF cells. Based on the results of this experiment, the graviola extract was found to contain as high as 26.6 mg(CA)/100g, 14.3 mg(Q)/100g of total polyphenol and flavonoid, which are the antioxidant indexes and to have the high radical scavenging activity. The cell survival rate of the HDF cells was measured, and as a result, no significant cytotoxicity was observed at all concentrations and the experiment was carried out at a concentration of $100{\mu}g/mL$ afterwards. Inhibition of ROS activity in HDF cells induced by hydrogen peroxide was measured and the concentration-dependent inhibition of ROS activity was found and the cell protection effect of graviola was measured after hydrogen peroxide was treated for 4, 24 and 48 hours. As a result, the cell protection effect as high as 89.92% was confirmed at a $25{\mu}g/mL$ concentration up to 24 hours. As these results show that the graviola extract has excellent antioxidant activity, almost no toxicity to HDF cells, an effective activity inhibitory effect on active oxygen generated by hydrogen peroxide and excellent cytoprotective effect, the possibility as various functional materials with antioxidant and cytoprotective effects was confirmed.

Biochemical Characterizations of Phenylalanine Ammonia-Lyase and its Mutants to Develop an Enzymatic Therapy for Phenylketonuria (페닐케톤뇨증의 효소치료 개발을 위한 phenylalanine ammonia-lyase 및 유전자 변이형의 생화학적 특성)

  • Kim, Woo-Mi
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1226-1231
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    • 2009
  • Enzyme substitution with recombinant phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is currently being explored for treatment of phenylketonuria (PKU), an autosomal recessive genetic disorder with mutations of the gene encoding phenylalanine-4-hydroxylase (EC 1.14.16.1). However, oral administration of PAL is limited because of proteolytic digestion in the gastrointestinal tract. The aim of this study was to determine the biochemical properties of PAL and delinate the susceptibility of wild-type PAL to pancreatic proteolysis by exploring several mutants, and to develop therapeutic drugs with PAL for PKU. The specific activity of PAL was assayed and its optimal pH, temperature stability, and intestinal protease susceptibility were investigated. Its $V_{max}$ values for phenylalanine and tyrosine were 1.77 and $0.47{\mu}mol$/ min/mg protein, respectively, and its $K_m$ values were $4.77{\times}10^{-4}$ and $4.37{\times}10^{-4}\;M$, respectively. PAL showed an optimal pH at 8.5, corresponding to the average pH range of the small intestine. It showed no loss of activity at $-80^{\circ}C$ for 5 months and possessed 93.4% of its activity under $4^{\circ}C$ for 4 wks. PAL was susceptible to chymotrypsin digestion and, to a lesser extent, to trypsin, elastase, carboxypeptidase A, and B. The trypsin and chymotrypsin cleaving sites were mutated to investigate protection from pancreatic digestion and the specific activities of these mutants were evaluated. The six mutants displayed low specific activities compared to the wild-type, suggesting that the primary trypsin and chymotrypsin cleaving sites may be essential for catalytic reaction. The PAL mutants could therefore be applied as a pretreatment modality without susceptibility to proteolytic attack, however, additional modification for enhancing enzymatic activity is needed to reduce the Phe levels effectively.

Effects of Physical Parameters on Water Quality in Agricultural Reservoirs (농업용 저수지의 물리적 인자가 수질에 미치는 영향)

  • Jeon, Ji-Hong;Ham, Jong-Hwa;Kim, Ho-Il;Hwang, Soon-Jin;Yoon, Chun-Gyeong
    • Korean Journal of Ecology and Environment
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    • v.35 no.1 s.97
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    • pp.28-35
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    • 2002
  • The effect of physical parameters on water quality was analyzed using monitoring data of 193 agricultural reservoirs. The retention time of reservoirs ($t_d$) ranged between 10 and 140 days, and the ratio of drainage area (DA) to reservoir surface area (SA) was between 10 and 120. Both ratios of DA/SA and total area (TA)/ reservoir storage (ST) in Korean agricultural reservoirs were relatively greater than those in natural lakes in other countries. As retention time was plotted against DA/SA ratio, it was shorter in Korean reservoirs than natural lakes. The semi-logarithmic relationship between TA/SA and t>$t_d$ was $t_d\;=\;42.21(TA/ST)^{-1}$ (n = 50, $R^2\;=\;0.89$). While areal loading of total phosphorus (TP) was below $4\;gTP{\cdot}m^{-2}{\cdot}yr^{-1}$ in general, it exceeded $10\;gTP{\cdot}m^{-2}{\cdot}yr^{-1}$ in reservoirs where DA/SA ratio was greater than 100, which implies that areal loading of TP increases as DA/SA ratio increases. Chl-a concentration was positively related with the mean depth of reservoir, implying the higher Chl-a concentration with deeper the mean depth. Therefore, the deeper reservoir might be advantageous in water quality management perspective if other morphological conditions are similar. The empirical regression equation using physical parameters was also suggested in the estimation of TP concentration in the reservoirs. Combined information presented in this paper might be applicable to the water quality management in agricultural reservoirs.

RALY RNA Binding Protein-like Reduced Expression is Associated with Poor Prognosis in Clear Cell Renal Cell Carcinoma

  • Cui, Zhi-Wen;Xia, Ye;Ye, Yi-Wang;Jiang, Zhi-Mao;Wang, Ya-Dong;Wu, Jian-Ting;Sun, Liang;Zhao, Jun;Fa, Ping-Ping;Sun, Xiao-Juan;Gui, Yao-Ting;Cai, Zhi-Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3403-3408
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    • 2012
  • The molecular mechanisms involved in the progression of clear cell renal cell carcinomas (ccRCCs) are still unclear. The aim of this study was to analyse the relationships between expression of RALYL and clinical characteristics. In 41 paired samples of ccRCCs and adjacent normal tissues, we used real-time qPCR to evaluate the expression of RALYL mRNA. RALYL protein levels were determined in 146 samples of ccRCC and 37 adjacent normal tissues by immunohistochemistry. Statistical analysis was used to explore the relationships between expression of RALYL and the clinical characteristics (gender, age, tumor size, T stage, N stage, M stage, survival times and survival outcome) in ccRCC. In addition, these patients were follow-up period 64 months (range: 4~116months) to investigate the influence on prognosis. We found significantly differences between ccRCC tissues and normal tissues (p<0.001, paired-sample t test) in mRNA levels of RALYL. Immunohistochemistry analyses in 146 ccRCC samples and 37 adjacent normal tissues showed significantly lower RALYL protein levels in ccRCC samples (${\chi}^2$-test, p<0.001), inversely correlating with tumour size (p=0.024), T stage (0.005), N stage (p<0.001) as well as M stage (p=0.019), but not age (p=0.357) and gender (p=0.348). Kaplan-Meier survival analysis demonstrated that people with lower level of RALYL expression had a poorer survival rate than those with a higher level of RALYL expression, significantly different by the log-rank test (p=0.011). Cox regression analysis indicated that RALYL expression (p=0.039), N stage (p=0.008) and distant metastasis (p<0.001) were independent prognosis factors for the overall survival of ccRCC patients. We demonstrated that the expression of RALYL was significantly low in ccRCC and correlated with a poor prognosis in a large number of clinical samples. Our findings showed that RALYL may be a potential therapeutic target as well as a poor prognostic factor.

A Study on the Neumann-Kelvin Problem of the Wave Resistance (조파저항에서의 Neumann-Kelvin 문제에 대한 연구)

  • 김인철
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.21 no.2
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    • pp.131-136
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    • 1985
  • The calculation of the resulting fluid motion is an important problem of ship hydrodynamics. For a partially immersed body the condition of constant pressure at the free surface can be linearized. The resulting linear boundary-value problem for the velocity potential is the Neumann-Kelvin problem. The two-dimensional Neumann-Kelvin problem is studied for the half-immersed circular cylinder by Ursell. Maruo introduced a slender body approach to simplify the Neumann-Kelvin problem in such a way that the integral equation which determines the singularity distribution over the hull surface can be solved by a marching procedure of step by step integration starting at bow. In the present pater for the two-dimensional Neumann-Kelvin problem, it has been suggested that any solution of the problem must have singularities in the corners between the body surface and free surface. There can be infinitely many solutions depending on the singularities in the coroners.

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The Effects of Repeated Restraint Stress on the Rat Parotid Glands, Ultramicroscopical and Histochemical Study (구속 스트레스에 대한 백서 타액선 조직의 미세구조적, 조직화학적 변화)

  • Yoon, In-Jong;Kang, Soo-Kyung;Auh, Q-Schick;Chun, Yang-Hyun;Hong, Jung-Pyo
    • Journal of Oral Medicine and Pain
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    • v.38 no.2
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    • pp.121-136
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    • 2013
  • It has been known that saliva may affect the most of oral diseases. On the contrary, several systemic conditions may affect salivary flow and cause oral dryness and psychosocial stress especially may a crucial role in the etiology of hyposalivation and oral dryness. Many studies have focused on macroscopic effects of the stress on the salivary glands by autonomic response, but on the other hand it has hardly been reported on cellular microscopic effects of the stress on the salivary glands. Therefore, this study was performed to examine clusterin, a antiapoptotic and cytoprotective protein, in the parotid glands under restraint stress condition. For this study, 18 rats were divided into 3 groups; 1) 2 rats of group I were selected as a normal control. 2) 2 rats of group II, as a experimental control were placed in the restraint cone for 2 hours 3) 14 rats of group III were placed in the restraint cone for 2 hours once a day. The rats were sacrificed immediately(group II, as a experimental control), 1, 2, 3, 4, 5, 6 and 7 days after application of the stress and the both parotid glands were excised. Immunohistochemistry and electron microscopy were performed. The finding were as follows: 1. In parotid glands, repeated stress denaturalize the acinar cells, interacinous tissues and interacinous connective tissues were separated to individual acinar cells. After 4 days of experiment, there were lots of vacuoles and intercalated ducts. 2. In parotid glands, repeated stress make the rER which is in acinar cells swollen after 3 days of experiment and it was intensified to 4 days. After 5 days of experiment the edema got worse and degenerated. 3. In parotid glands, clusterin was reduced in ductal cell cytoplasm but in intercalated duct clusterin was slightly stained until 3 days prominently increased until 4 days and then decreased again after 5 days of experiment.

Measurement of thermal expansion characteristic of root canal filling materials : Gutta-percha and Resilon (수 종의 근관충전재의 열팽창 특성 측정 : Gutta-percha와 Resilon)

  • Jeon, Kyung-A;Lee, In-Bog;Bae, Kwang-Shik;Lee, Woo-Cheol;Baek, Seung-Ho
    • Restorative Dentistry and Endodontics
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    • v.31 no.5
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    • pp.344-351
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    • 2006
  • The purpose of this study was to evaluate the thermal expansion characteristics of injectable ther-moplasticized gutta-perchas and a Resilon. The materials investigated are Obtura gutta-percha, Diadent gutta-percha, E&Q Gutta-percha Bar and Epiphany (Resilon). The temperature at the heating chamber orifice of an Obtura II syringe and the extruded gutta-percha from the tip of both 23- and 20-gauge needle was determined using a Digital thermometer. A cylindrical ceramic mold was fabricated for thermal expansion test, which was 27 mm long, with an internal bore diameter of 3 mm and an outer diameter of 10 mm. The mold was filled with each experimental material and barrel ends were closed with two ceramic plunger. The samples in ceramic molds were heated in a dilatometer over the temperature range from $25^{\circ}C$ to $75^{\circ}C$. From the change of specimen length as a function of temperature, the coefficients of thermal expansion were deter-mined. There was no statistical difference between four materials in the thermal expansion in the range from $35^{\circ}C$ to $55^{\circ}C$ (p > 0.05). However, Obtura Gutta-percha showed smaller thermal expansion than Diadent and Metadent ones from $35^{\circ}C$ to $75^{\circ}C$ (p < 0.05). The thermal expansion of Epiphany was similar to those of the other gutta-percha groups.

Plant Community Structure of Abies holophylla Community from Sinseongam to Jungdaesa in Odaesan National Park (오대산국립공원 신성암~중대사 전나무림 식물군집구조 특성)

  • Kim, Dong-Wook;Han, Bong-Ho;Kim, Jong-Yup;Yeum, Jung-Hun
    • Korean Journal of Environment and Ecology
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    • v.29 no.6
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    • pp.895-906
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    • 2015
  • This study was carried out to the structure of plant community from Sinseongam to Jungdaesa in Odaesan National Park, furthermore, it seeks to curate the basic data for planning of the Abies holophylla's forest management in Odaesan National Park. In order to identify the current ecological environment, this study explored the actual vegetation as primary research and set to twenty plots(i.e. $400m^2$) for analysing detailed structure of plant communities. The research methodology was qualitative analysis, therefore it used TWINSPAN and DCA analysis tools. Especially, TWINSPAN performed well in several comparisons of classification techniques, DCA is one of the ordination technique showed that the plant communities. The plant community was analysed classification and ordination by TWINSPAN and DCA, moreover it was analysed the structure of plant community such as importance percentage of woody species, DBH class distribution, the index of diversity and rate of sample tree growth. The main vegetation was A. holophylla-Quercus mongolica forest and Deciduous broad-leaved forest in the communities where located in low altitude and valley, whereas main vegetation where located in high altitude and slope was Q. mongolica forest. The research site's plant communities were classified four groups. In all of communities, A. holophylla was dominant species in main canopy layer, furthermore, the three communities (community I, II, III) are growing up next generation of A. holophylla excluding community IV. The communities (community I, II, III) can be sustained current status which dominates the A. holophylla communities, simultaneously, there might be expanded the Deciduous broad-leaved communities by Carpinus cordata, Betula schmidtii and so on. While, it showed that the community IV tended to be weaken the forces of A. holophylla, therefore the community IV can be transferred to C. cordata-Deciduous broad-leaved communities in the future. The age of sample trees was 79~128(i.e. A. holophylla), 75~87(i.e. Pinus koraiensis) and 190 years(i.e. Ulmus davidiana var. japonica). The index of Shannon's Species diversity (H') were ranged from 0.3889 to 1.3332 in the communities.