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Development of a Simple and Reproducible Method for Removal of Contaminants from Ginseng Protein Samples Prior to Proteomics Analysis (활성탄을 이용한 불순물제거에 의한 효과적인 인삼 조직 단백질체 분석 방법 개선 연구)

  • Gupta, Ravi;Kim, So Wun;Min, Chul Woo;Sung, Gi-Ho;Agrawal, Ganesh Kumar;Rakwal, Randeep;Jo, Ick Hyun;Bang, Kyong Hwan;Kim, Young-Chang;Kim, Kee-Hong;Kim, Sun Tae
    • Journal of Life Science
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    • v.25 no.7
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    • pp.826-832
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    • 2015
  • This study describes the effects of activated charcoal on the removal of salts, detergents, and pigments from protein extracts of ginseng leaves and roots. Incubation of protein extracts with 5% (w/v) activated charcoal (100-400 mesh) for 30 min at 4℃ almost removed the salts and detergents including NP-40 as can be observed on SDS-PAGE. In addition, analysis of chlorophyll content showed significant depletion of chlorophyll (~33%) after activated charcoal treatment, suggesting potential effect of activated charcoal on removal of pigments too along with the salts and detergents. 2-DE analysis of activated charcoal treated protein samples showed better resolution of proteins, further indicating the efficacy of activated charcoal in clearing of protein samples. In case of root proteins, although not major differences were observed on SDS-PAGE, 2-DE gels showed better resolution of spots after charcoal treatment. In addition, both Hierarchical clustering (HCL) and Principle component analysis (PCA) clearly separated acetone sample from rest of the samples. Phenol and AC-phenol samples almost overlapped each other suggesting no major differences between these samples. Overall, these results showed that activated charcoal can be used in a simple manner to remove the salts, detergents and pigments from the protein extracts of various plant tissues.

Breaking Physical Dormancy with Sulfuric Acid in Seeds of Lespedeza tomentosa (Thunb.) Siebold ex Maxim (황산처리를 이용한 개싸리 종자의 물리적 휴면 타파)

  • Rhie, Yong Ha;Choi, Han;Lee, Su Gwang;Lee, Jeong Ho;Lee, Ki Cheol
    • Korean Journal of Plant Resources
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    • v.29 no.1
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    • pp.136-142
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    • 2016
  • Lespedeza species are mainly used for wildlife food and cover and for erosion control. The germination of these species can be enhanced after a fire occurrence in forest, which is known as fire-activated seeds to germinate. While the heat treatment could break seed dormancy of Lespedeza, its germination rate was quite low. We investigated that chemical scarification could promote germination of L. tomentosa. Seeds were soaked in 100% sulfuric acid (H2SO4) for 0, 1, 3, 6, 12, 24, 48, 96, 192, and 384 min, and then washed in distilled water for 24 h. Very few seeds were germinated in control (H2SO4 for 0 min). More than 90% of seeds were germinated in H2SO4 for 24, 48, and 92 min. However, some damage was observed in roots and cotyledons of seedling dipped in H2SO4 for a long time. To search the optimal soaking time in H2SO4 without defects, seeds scarified in H2SO4 for 30, 60, 90, 120, 150, 180, and 300 min were sown the commercial soil medium. Seeds treated with H2SO4 for 90 min and 150 min emerged by about 92% and 84%, respectively. Therefore, H2SO4 treatment could break the seed dormancy of Lespedeza species, and especially in case of L. tomentosa the optimal treatment time in sulfuric acid was one to two hours. Germination of L. tomentosa began promptly following the scarification and was completed within about one month, indicating that seeds has no physiological dormancy, just has physical dormancy.

Inhibitory Effect of the Hexane Extract of Saussurea lappa on the Growth of LNCaP Human Prostate Cancer Cells (목향 헥산추출물의 LNCaP 전립선암세포 증식 억제 효과)

  • Park, So-Young;Kim, Eun-Ji;Lim, Do-Young;Kim, Jong-Sang;Lim, Soon-Sung;Shin, Hyun-Kyung;Yoon Park, Jung-Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.1
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    • pp.8-15
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    • 2008
  • Saussurea lappa (SL) has been used to reduce abdominal pain and tenesmus in traditional oriental medicine. SL and major compounds of SL, sesquiterpene lactones, have been suggested to possess various biological effects, including anti-tumor, anti-ulcer, anti-inflammatory, anti-viral and cardiotonic activities. Recently, it has been reported that ethanol extracts from roots of SL have antiproliferative effects on gastric cancer cells. To explore the possibility that SL has chemopreventive effects on prostate cancer, we examined whether the hexane extract of SL (HESL) inhibits the growth of LNCaP human prostate cancer cells. Cells were incubated with various concentrations ($0{\sim}4$ mg/L) of HESL in DMEM/F12 containing 5% charcoal stripped fetal bovine serum. HESL substantially decreased viable cell numbers and induced apoptosis of LNCaP cells in dose-dependent manners. HESL increased the levels of cleaved caspase-8, -9, -7 and -3, and poly (ADP-ribose) polymerase. HESL increased the levels of the pro-apoptotic Bak and truncated-Bid proteins whereas it had no effect on the anti-apoptotic Bcl-2, Bcl-xL, or Mcl-1. The present results indicate that HESL inhibits the growth of human prostate cancer cells by inducing apoptosis, which involves the activation of the caspase cascades.

Effects of Inoculation with Mycorrhizal Fungi, Pisolithus tinctorius and Glomus sp. on the Rooting of Quercus acutissima Carr. Cuttings at Various Ortet Ages (모래밭 버섯균(菌)과 Glomus 균근균(菌根菌)의 인공접종(人工接種)이 연령(年齡)이 다른 상수리나무에서 채취(採取)한 삽수(揷穗)의 삽목발근(揷木發根)에 미치는 영향(影響))

  • Kim, Jong Ju;Lee, Kyung Joon
    • Journal of Korean Society of Forest Science
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    • v.79 no.3
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    • pp.302-308
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    • 1990
  • This study was conducted to determine the effects of mycorrhizal inoculation and ortet ages on the rooting of Quercus acutissima cuttings. The cuttings taken from 12-week-, 2-year-, 4-year-, and 20-year-old plants were rooted in mid summer in the rooting medium (vermiculite 2 : peatmoss 1 by volume) with or without Pisolithus tinctorius (Pt) and Glomus sp. inocula and IBA under intermittent misting system in an open shed. The average percentages of rooting were about 82%, 49%, 29%, and 13% for cuttings taken from 12-week-, 2-year-. 4-year-, and 20-year-old seedlings, respectively. Pt inoculation, enhanced rooting of cuttings at all age classes, except 12-week-old seedlings, with the highest enhancement(22%) observed in cuttings taken from 20-year-old trees. The highest percentage of rooting in each age group eras 88.9% in 12-week-old seedlings treated with Pt plus 3.000ppm IBA, 75% in 2-Year-old plants with 1,000ppm IBA, 58.3% in 4-year-old plants with 3.000ppm IBA and 22% in 20-year-old plants. The addition of Glomus sp. fungus inoculum failed to enhance rooting. Pt mycorrhizal inoculation enhanced root dry weight, length, and diamter of adventitious roots at cuttings taken from 12-week- and 20-year-old trees, except the cuttings taken from 4-year-old seedlings. Rooted cuttings had more total nitrogen content in the leaves than unrooted cuttings, and the greater rooting response was associated with the higher phosphorus content in the leaves.

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Initial Root Development of Larix leptolepis Gordon Cuttings as Related to Organic Substances and Cutting Date (낙엽송(落葉松)의 삽목발근(揷木發根)에 관련된 물질(物質) 및 발근과정(發根過程))

  • Koo, Yeong Bon;Hyun, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.85 no.2
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    • pp.300-308
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    • 1996
  • This study was conducted to identify factors involved in rooting of Japanese larch cuttings. Cuttings were taken every week from June 24th to July 25th before and after the 10th of July which supposed to be best time for cutting. The content of IAA, carbohydrate, nitrogen and inorganic elements such as $Mg^{{+}{+}}$, $K^+$ and $Ca^{{+}{+}}$ within cutting materials were analyzed. After the cuttings were planted, the root development was observed every week. Cuttings of easy-to-root clones were higher in IAA contents than the cuttings of hard-to-root clones at the time of cuttings were taken. There were significant differences in moisture content of cuttings taken on June 24th and July 25th between easy-to-root and hard-to-root clones, however no significant differences in moisture contents were observed between them taken on the 8th and 15th of July which supposed to be the best time for cutting. Inorganic matters and carbohydrate contents of cuttings taken from hedged trees were higher than those of non-hedged trees. The ratios K/C and C/N of cuttings taken from hedged trees were higher than those of non-hedged trees. Cuttings taken from easy-to-root clones showed higher content of carbohydrate and nitrogen than those of hard-to-root clones. The contents of inorganic elements, such as $Mg^{{+}{+}}$, $K^+$, and $Ca^{{+}{+}}$ in cutting taken from hard-to-root clones were slightly higher than that of easy-to-root clones. The ratio C/N and K/C of cuttings which were taken from easy-to-root clones have shown a similar trend of carbohydrate and nitrogen. These data indicate that there was no one dominant factor that affect rooting of cutting, but many factors such as contents of IAA, carbohydrate, nitrogen, phosphate, and ratios of C/N and K/C appeared to affect on rooting of cuttings variously. Most of root initials were formed from phloem tissue within one week after cutting, and few were formed from callus tissue. Root primordia were developed from root initials within two weeks after cutting, and most of new roots were developed from root primordia within about five weeks after cutting.

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Comparison of Above-ground Growth Characteristics Between Naturally Regenerated and Planted Stands of Pinus densiflora for. erecta Uyeki in Gangwon Province (천연갱신과 인공조림된 강원지역 금강소나무의 지상부 생장특성 비교)

  • Na, Sung-Joon;Woo, Kwan-Soo;Kim, Chang-Soo;Yoon, Jun-Hyuck;Lee, Heon-Ho;Lee, Do-Hyung
    • Journal of Korean Society of Forest Science
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    • v.99 no.3
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    • pp.323-330
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    • 2010
  • This study was undertaken to compare early growth characteristics, such as height, diameter at root collar (DRC), and the dry weights of stem, branches, needles and roots between naturally regenerated and planted stands of Pinus densiflora for. erecta in Dae-gi and Bo-gwang site, Gangwon province, Republic of Korea. Average DRC growth was similar between the naturally regenerated and the planted stands in Dae-gi area, however, difference was statistically significant (P<0.01) between the naturally regenerated and the planted stands in Bo-gwang area, 4.66 cm and 2.61 cm, respectively. Average height of the naturally regenerated stands was significantly greater (P<0.01) than that of planted stands in both Dae-gi and Bo-gwang areas. The composition ratio of the dry weights for each part of measured trees was the highest in the stem in the naturally regenerated stands, but in the planted stands, the needle was the highest in both sites. The root/shoot ratio in the Dae-gi and Bo-gwang areas in the planted stands were much greater than those in the naturally regenerated stands, 0.25-0.30 and 0.17-0.18, respectively. The ratio of needle to stem biomass (N/S), branch to stem biomass (B/S), and needle to branch biomass (N/B) was significantly greater in the planted stands than those in the naturally regenerated stands in both sites. The results of this study represent that the regeneration methods, natural and artificial regenerations, can have an influence on the early growth of P densiflora for. erecta. This results not only provide the need for a correct planting method, but also will be useful in studies on the environmental adaptability of trees.

The Effects of Estrogen on Experimental Tooth Movement in Ovariectomized Rats (난소적출 백서에서 estrogen투여가 실험적 치아이동에 미치는 영향)

  • Jin, Keun-Ho;Kim, Jong-Ghee;Park, Byung-Keon;Kim, Oh-Hwan
    • The korean journal of orthodontics
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    • v.27 no.4 s.63
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    • pp.585-597
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    • 1997
  • The purpose of this study was to evaluate the effect of estrogen on the periodontium and alveolar bone tissue response during experimental tooth movement in ovariectomized rats. Eighty female rats, 250gm in body weight, were classified into four groups ; sham operated group(NN), ovariectomized group(ON), ovariectomized & estrogen injected group(OE), sham operated & estrogen injected group(NE). flats were ovariectomized before 3 weeks to begin the experiment, which resulted in estrogen-deficient osteoporosis. In OE group & NE group, estrogen was injected $50{\mu}g/kg\;B.W.$ every other days. The left maxillary 1st molar was moved mesially with 60g force. Each foot rats were sacrificed after 1, 3, 7, 15 days from application of orthodontic appliance and alter additional 7 days from removal of orthodontic appliance. Histological findings on mesial roots of upper 1st molar in pressure and tension side are observed. The results were summarized as follows ; 1. In pressure side of alveolar bone, the number of osteoclasts and Howship's lacuna of ON group was significantly more than that of NN group from 1 day to 15 days(P<0.05). Especially the number of Howship's lacuna of ON group was significantly more than that of OE group during all experimental period(P<0.05). 2. In tension side of alveolar tune, the number of osteoclasts of ON group was significantly increased from 1 day to 3 days and decreased after 7 days. But the number of osteoclast of ON group was significantly mote than that of NN group during all experimental period(P<0.05). Also the number of Howship's lacuna of all groups was abruptly increased at 1 day, but slowly decreased till experimental 15 days. And the number of Howship's lacuna of of group was significantly more than that of NN group from 0 hr to 7 days(P<0.05). 3. The speed of tooth movement of OE group & NE group was similar to that of NN group(P>0.05). The amount of tooth movement of ON group between 7 days and 15 days was significantly greater than those of other groups(P<0.05). 4. The degree of relapse of ON group after 7 days from removal of orthodontic appliance was similar to those of other groups(P>0.05).

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Changes in the Titer of Tooth Root Antibodies Accompanying Root Resorption Associated with Orthodontic Tooth Movement (치아이동시 치근 흡수에 따른 치근항체의 역가 변화)

  • Park, Soo-Byung;Son, Woo-Sung
    • The korean journal of orthodontics
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    • v.24 no.2
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    • pp.303-317
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    • 1994
  • This study was designed to measure the changes in the titer of tooth root antibodies accompanying root resorption associated with orthodontic tooth movement in dogs to explore a role of the specific immune response in root resorption during orthodontic tooth movement. Five adult mongrel dogs, 2 years of age, were used in the study. Six lower incisors were extracted as sources of homologous antigen in the dogs. Tooth root antigen preparations were made from a 6M Guanidine-HCl-10% EDTA(pH5.0) extract of these root dentins. Root resorption was elicited by intrusion of six maxillary incisors with 200-250gm intrusive force. In 9th week, resorbing six maxillary anterior teeth were extracted. Serum samples were taken from each dog prior to intrusion and weekly for 11 consecutive weeks. Serum autoantibody titers were determined with an enzyme-linked immunosorbent assay. As controls for antibody specificity, sera which were previously incubated with tooth root antigen as well as sera to an unrelated bacterial antigen (Porphyromonas gingivalis 33277) for 3 hours at 25 were measured in all runs. Root resorption was monitored monthly using occlusal radiographs. And then root resorption patterns were observed with a zoom stereo microscope (Model SZH-121, Olympus optical Co. Ltd.). Incisors did not show clear radiographic evidence of significant and progressive root resorption, but periodontal ligament space had widened. But root resorption was observed on the apical regions of the maxillary incisors with a zoom stereo microscope. Teeth showed the shallow depression generally accompanying deep resorption. These demonstrate a slight tendency for an immediate decrease followed by rebound to levels above the pre-treatment baseline. A peak titer of autoantibody to dentin antigen occurred on day 28, then steadily decreased during the 9th week period as the roots resorbed and then rapidly spiked in animals when the resorbing teeth were extracted. When sera is incubated with tooth root antigen, serum activity in the ELISA was almost absent. This is because serum activity in the ELISA could be removed by absorption of the serum with dog dentin antigen. Serum ELISA activity to the unrelated bacterial antigen remained essentially unchanged in all animals throughout the experimental period. When the time course of changes in autoantibody to homologous tooth root antigen prepatration and unrelated bacterial antigen was compared, no significant differences were found(${\alpha}=0.05$). In general, the overall pattern of changes in autoantibody was similar to the two antigens. These findings suggest the possibility that these immunologic changes precede a significant development of root resorption lesions rather than merely reflecting their presence. Therefore, this suggests that the changes of antibody levels may have some predictive value for root resorption.

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Effects of Arbuscular Mycorrhizal Fungus, Glomus intraradices, on the Growth, Photosynthesis and Phosphorus Content of Robinia pseudoacacia Seedlings Treated with Simulated Acid Rain (Glomus 내생균근균 접종이 인공산성우를 처리한 아까시나무 묘목의 생장, 광합성, 인 함량에 미치는 영향)

  • Kim, Eun Ho;Lee, Kyung Joon;Lee, Kyu Hwa
    • Journal of Korean Society of Forest Science
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    • v.95 no.6
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    • pp.735-742
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    • 2006
  • The objective of this study was to elucidate the tolerance of woody plants to simulated acid rain in relation to mycorrhizal inoculation. Germinating seedlings of Robinia pseudoacacia were planted in 1I pots with autoclaved soil mixture of vermiculite, sand and nursery soil at 1:1:1 ratio. Each pot was inoculated with both crushed root nodules from a wild tree of the same species and commercial arbuscular mycorrhizal inoculum of Glomus intraradices at the time of planting the seedlings. Simulated acid rains at pH 2.6, 3.6, 4.6, and 5.6 were made by mixing sulfuric acid and nitric acid at 3: 1 ratio. Each pot received nutrient solution without N and P, and was also supplied with 180 ml of the one pH level of the acid rains once a week for 50 days. The plants were grown in the green house. At the end of experimental period, plants were harvested to determine contents of chlorophyll, mineral nutrients and net photosynthesis in the tissues, dry weight of the plants, and mycorrhizal infection in the roots. Mycorrhizal infection rate was significantly reduced only at pH 2.6, which meant vitality of G intraradices was inhibited at extremely low pH. Height growth, dry weight production, nodule production and chlorophyll content were increased by mycorrhizal infection in all the pH levels except pH 3.6. Particularly, mycorrhizal inoculation increased root nodule production by 85% in pH 5.6 and 45% in 4.6 treatments. But the stimulatory effect of mycorrhizal inoculation on nodule production was reduced at pH 3.6 and 2.6. Net photosynthesis was increased by mycorrhizal infection in all the pH levels. The phosphorus(P) content in the tissues was increased by 43% in average by mycorrhizal inoculation, which was statistically significant except in pH 2.6. It was concluded that mycorrhizal inoculation of Robinia pseudoacacia would enhance growth and resistance of the plants to acid rain by improving the photosynthesis, phosphorus nutrition, and more nodule production.

Effects of Controlling the Pine Needie Gall Midaes by Salicylic Acid Content in Needles of Some Pinus spp. (소나무류(類) 침엽내(針葉內)의 salicylic acid에 의한 솔잎혹파리의 방제효과(防除效果))

  • Son, Doo-Sik;Eom, Tae-Jin;Choi, Chang-Ok;Zhang, Ruo Ming
    • Journal of Korean Society of Forest Science
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    • v.88 no.1
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    • pp.31-37
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    • 1999
  • The frequency of gall formation by pine needle gall midges ranged from 35% to 40% in both P. densiflora and P. thunbergii species. However, there was no indication of gall development from larva on both P. virgiana and P. rigida species suggesting that some specific compounds might play role in preventing gall formation of larva. The susceptible species to pine needle gall midges including P. densiflora and P. thunbergii contained a little salicylic acid under the free phenolic compound conditions while the resistant species against such insects including P. virginiana and P. rigida species contained about 37ppm to 50ppm of salicylic acid. Thus, this compound might have important roles in insect resistance. The contents of internal salicylic acid in the needles of susceptible pines increased from 9.5ppm to 20.6ppm after direct external irrigation of salicylic acid solution and flour treatment on roots. As a result, the frequency of gall formation decreased dramatically 17~19 times lower when compared with that of control. According to our results, the application of salicylic acid for effective prevention against insect damages should be performed before hatching eggs. The frequency of gall formation in the hybrid pines of P. thunbergii(susceptible) and P. virginiana(resistant) showed approximately 24% as average value of those in both species. In the case of specific individuals, it ranged from 2.8% to 11.5% in hybrids of both species. Therefore, effective production of insect resistant pines can be obtained through hybrids by crossing between susceptible and resistant species including P. thunbergii and P. virginiana by increasing internal salicylic acid contents of the needles.

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