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Mass Culture and Ginsenoside Production of Ginseng Hairy Root by Two-Step Culture Process (2계단 배양방법을 이용한 인삼 모상근의 대량배양과 Ginsenoside 생산)

  • Ko, Kyeong-Min;Yang, Deok-Chun;Park, Ji-Chang;Choi, Kang-Ju;Choi, Kwang-Tae;Hwang, Baik
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.63-69
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    • 1996
  • A hairy root clone of Panax ginseng C.A. Meyer, HRB-15 was cultured iu various conditions with 3 L bubble type bioreactor to enhance both growth and ginsenoside production. The hairy roots were more rapidly grown under the dark condition than under the light condition. However, total amount of ginsenoside of hairy roots cultured under the light for 30 days increased 2 folds as compared with the dark condition and was 1.10% based on 6 ginsenosides. Especially, ginsenoside-Re was significantly increased and some ginsenosides except for ginsenoside-Re was slightly reduced. Also, the growth of hairy roots decreased about 30% as compared with the dark condition. In contrast, addition of sodium acetate led to decreased production of ginsenoside and growth of hairy roots under light condition. The influence of potassium dihydrogenphosphate concentration was examined in MS medium and a 1.25 mM concentration was found to be the most appropriate for growth and ginsenoside production under light condition. Two-step process of hairy roots culture with yeast elicitation or without ammonia in culture medium was developed to enhance growth and giusenoside synthesis. $50\;\mu\textrm{g}$ of yeast elicitor per g of fresh weight showed a synergistic effect on the ginsenoside synthesis of hairy roots on 20 days after culture. At that time, the content of total ginsenoside was 1.15%, while the growth of hairy roots decreased 21 % as compared with the dark condition. In addition, when elimination of ammonia on 20 days after culture, the content of total ginsenoside was 1.26% with significant increment of ginsenoside-Rd (0.27%) in addition to ginsenoside-Re and the growth of hairy roots decreased 10% as compared with the dark condition. In this system, we have demonstrated a unique two-step process of hairy root cultures to maximize biomass and secondary metabolites. It has found possibility to enhance ginsenosides production by growing hairy roots in this method.

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Potential of River Bottom and Bank Erosion for River Restoration after Dam Slit in the Mountain Stream

  • Kang, Ji-Hyun;So, Kazama
    • Proceedings of the Korea Water Resources Association Conference
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    • pp.46-46
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    • 2011
  • Severe sediment erosion during floods occur disaster and economic losses, but general sediment erosion is basic mechanism to move sediment from upstream to downstream river. In addition, it is important process to change river form. Check dam, which is constructed in mountain stream, play a vital role such as control of sudden debris flow, but it has negative aspects to river ecosystem. Now a day, check dam of open type is an alternative plan to recover river biological diversity and ecosystem through sediment transport while maintaining the function of disaster control. The purpose of this paper is to verify sediment erosion progress of river bottom and bank as first step for river restoration after dam slit by cross-sectional shear stress and critical shear stress. Study area is upstream reach of slit check dam in mountain stream, named Wasada, in Japan. The check dam was slit with two passages in August, 2010. The transects were surveyed for four upstream cross-sections, 7.4 m, 34 m, 86 m, and 150 m distance from dam in October 2010. Sediment size was surveyed at river bottom and bank. Sediment of cobble size was found at the wetted bottom, and small size particles of sand to medium gravel composed river bank. Discharge was $2.5\;m^3/s$ and bottom slope was 0.027 m/m. Excess shear stress (${\tau}_{ex}$) was calculated for hydraulic erosion by subtracting the values of critical shear stress (${\tau}_{c}$) from the value of shear stress (${\tau}$) at river bottom and bank (${\tau}_{ex}=\tau-{\tau}_c$). Shear stress of river bottom (${\tau}_{bottom}$) was calculated using the cross-sectional shear stress, and bank shear stress (${\tau}_{bank}$) was calculated from the method of Flintham and Carling (1988). $${\tau}_{bank}={\tau}^*SF_{bank}((B+P_{bed})/(2^*P_{bank}))$$ where $SF_{bank}=1.77(P_{bed}/p_{bank}+1.5)^{-1.4}$, B is the water surface width, $P_{bed}$ and $P_{bank}$ are wetted parameter of the bed and bank. Estimated values for ${\tau}_{bottom}$ for a flow of $2.5\;m^3/s$ were lower as 25.0 (7.5 m cross-section), 25.7 (34 m), 21.3 (86 m) and 19.8 (150 m), in N/$m^2$, than critical shear stress (${\tau}_c=62.1\;N/m^2$) with cobble of 64 mm. The values were insufficient to erode cobble sediment. In contrast, even if the values of ${\tau}_{bank}$ were lower than the values for ${\tau}_{bottom}$ as 18.7 (7.5 m), 19.3 (34 m), 16.1 (86 m) and 14.7 (150 m), in N/$m^2$, excess shear stresses were calculated at the three cross-sections of 7.5 m, 34 m, and 86 m distances compare with ${\tau}_c$ is 15.5 N/$m^2$ of 16mm gravel. Bank shear stresses were sufficient for erosion of the medium gravel to sand. Therefore there is potential to erode lateral bank than downward erosion in a flow of $2.5\;m^3/s$. Undercutting of the wetted bank can causes bank scour or collapse, therefore this channel has potential to become wider at the same time. This research is about a potential of sediment erosion, and the result could not verify with real data. Therefore it need next step for verification. In addition an erosion mechanism for river restoration is not simple because discharge distribution is variable by snow-melting or rainy season, and a function for disaster control will recover by big precipitation event. Therefore it needs to consider the relationship between continuous discharge change and sediment erosion.

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High Glucose Induces Apoptosis through Caspase-3 Dependent Pathway in Human Retinal Endothelial Cell Line (인간망막 내피세포주에서 고농도 포도당이 caspase-3 경로를 통해 세포자연사 유도)

  • Seo, Eun-Sun;Chae, Soo-Chul;Kho, Eun-Gyeong;Lee, Jong-Bin
    • Korean Journal of Environmental Biology
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    • v.27 no.1
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    • pp.66-72
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    • 2009
  • Diabetic Retinopathy (DR) is a leading cause of blindness among adults in the western countries. Hyperglycemia is a condition, that induces apoptotic cell death in a variety of cell types in diabetes, but the mechanism remains unclear. The aim of the study is to understand the effects of high Glucose on Human Retinal Endothelial Cells. Retinal endothelial cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 5, 25 and 50 mM Glucose, incubated for 24, 36 and 48 hours in humidified 5 % CO$_2$ incubator at 37$^{\circ}C$. Human Retinal Endothelial Cell Line (HREC) were characterized for morphology with different treatment by phase contrast microscopic analysis. Number of dead and viable cells was counted by trypan blue exclusion and supported by MTT assay. The intracellular Hydrogen peroxide (H$_2$O$_2$), a Reactive Oxygen Species (ROS) generation in high glucose conditions was assessed by FOX II assay and apoptosis by caspase-3 assay. The high glucose treated cells undergoing DNA fragmentation was witnessed by Agarose gel electrophoresis. We found that the cells incubated with 25 and 50 mM glucose containing medium for 48 hours altered the morphology of the cell, induced apoptosis and DNA fragmentation. The dead cell number were high in 25 and 50 mM when compared to the cells incubated with 5 mM glucose for 24, 36, and 48 hours. Also, the H$_2$O$_2$ levels and the activity of caspase-3 were increased in high glucose treated cells. Conclusions/interpretation: Our results demonstrated that elevated glucose induces apoptosis in cultured HREC. The hyperglycemia-induced increase in apoptosis may be dependent on caspase activation. The association between ROS generation and caspase-3 activation on high glucose treated cells is yet to be investigated.

Impacts of Climate Change and Follow-up Cropping Season Shift on Growing Period and Temperature in Different Rice Maturity Types (미래 기후변화 및 그에 따른 재배시기 조정이 벼 생태형별 생육기간과 생육온도에 미치는 영향)

  • Lee, Chung-Kuen;Kwak, Kang-Su;Kim, Jun-Hwan;Son, Ji-Young;Yang, Won-Ha
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.3
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    • pp.233-243
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    • 2011
  • This experiment was conducted to investigate the effect of future climate change on growing period and temperature in different rice maturity types as global warming progressed, where Odaebyeo, Hwaseongbyeo, Ilpumbyeo were used as a representative cultivar of early, medium, and medium-late rice maturity type, respectively, and A1B scenario was applied to weather data for future climate change at 57 sites in Korea. When cropping season was not adjusted to climate change, entire growing period and growing temperature were shorten and risen, respectively, as global warming progressed. On the other side, when cropping season was adjusted to climate change, growing period and temperature after heading date were not changed in contrast to growing period and growing temperature before heading which were more seriously shortened and risen as global warming progressed than in not adjusted cropping season. It is supposed that adjusting cropping season to climate change can alleviate rice yield reduction and quality deterioration to some degree by improving growing temperature condition during grain-filling period, but also still have a limit such as seriously shortened growing period indicating that there need to develope actively new rice cultivation methods and varieties for future climate change.

Co-Culture Model Using THP-1 Cell and HUVEC on AGEs-Induced Expression of Cytokines and RAGE (THP-1 Cell과 HUVEC을 이용한 Co-Culture Model System에서 최종당화산물에 의한 Cytokines와 RAGE 발현)

  • Lee, Kwang-Won;Lee, Hyun-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.3
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    • pp.385-392
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    • 2011
  • Although monoculture methods have been remarkably useful due to their simplicity, they have serious limitation because of the different types of cells communication with each other in many physiological situations. We demonstrated levels of markers of endothelial dysfunction such as tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$) as well as stimulation of receptor of advanced glycation endproducts (AGEs) on monoand co-culture system such as only monocyte (THP-1) cultivation system, only endothelial cell (HUVEC) cultivation system, and co-cultivation system of THP-1 and HUVEC. The mRNA levels of TNF-$\alpha$ and IL-1$\beta$ on HUVEC increased by the co-culture with monocyte after 4 hr at 100 ${\mu}g/mL$ glyceraldehyde-AGE. The secreted protein contents into medium of TNF-$\alpha$ and IL-1$\beta$ increased after 8 hr approximately 2~2.5 times compared to mono-cultivation. In contrast, the mRNA level of receptor of AGE (RAGE) was relatively insensitive on the co-culture system. The mediators by which monocytes activate endothelial cell have not been fully elucidated. In this study we confirmed production of soluble cytokines such as TNF-$\alpha$ and IL-1$\beta$ by monocytes. Use of monocyte conditioned medium, which contains both cytokines, can activate endothelial cell.

Acclimatization of in vitro Plantlets of Wasabia japonica(Miq.) Matsum. Derived from the Apical Meristem Culture (고추냉이(Wasabia japonica (Miq.) Matsum.)의 정단분열조직유래 기내묘의 순화)

  • 은종선
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.257-261
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    • 1998
  • The repeated subcultures of in vitro plant materials in wasabi became highly vitrified and the capacity for multiple shoot formation from the vitrified plant materials was very low. In order to improve the quality of in vitro propagated planting materials, the experiments were carried out using culture vessels capped with membrane filter(MF). When vitrified shoots were cultured on MS medium with 0.2mg/L BA in the vessels with MF or without MF for 60 days, the shoots in the vessels with MF did not vitrified. In contrast, the shoots grown in the vessels without MF vitrified at 65%. The stomates of vitrified leaves were circular and inflated, whereas those of normal leaves acclimatizated in the vessels with MF were ovate in shape. The hardened shoots were also cultured on MS media without sucrose containing 0.01mg/L IBA in vessels with(photoautotrophic culture) or without(control) MF. Sucrose was necessary for survival of the in vitro plantlets in the vessels without MF. After 20 days of culture, the shoots in the vessels without MF on the sucrose-free media turned yellow and died. But the shoots in the vessels with MF in the sucrose-free media produced a lot of roots. When shoots were cultured on MS medium with 2% sucrose containing 0.01mg/L IBA in the vessels with(photomixotrophic culture) or without(heterotrophic culture) MF, best growth occured in photomixotrophic culture.

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Efficiency of microspore embryogenesis in Brassica rapa using different genotypes and culture conditions (배추 유전자원의 소포자 유래 배 발생 효율에 미치는 배양 조건 구명)

  • Seo, Mi-Suk;Sohn, Seong-Han;Park, Beom-Seok;Ko, Ho-Cheol;Jin, Mina
    • Journal of Plant Biotechnology
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    • v.41 no.3
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    • pp.116-122
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    • 2014
  • Total of fifty accessions of Brassica rapa with various morphological characteristics were used for production of double haploid plants though microspore culture in Brassica rapa. Among them, only 30 accessions induced embryos from microspores. The highest efficiency of embryo induction of 1.194 per bud was obtained from IT135449 of turnip type, while 3 accessions of sarson (winter oil) type did not generate embryo. The effect of heat shock periods for embryogenesis was also investigated with 4 accessions (IT135449; Turnip type, IT199710; Chinese cabbage type, IT212886; Pak choi type, IT218043; Summer oil type). The high productions of embryos were observed in IT135449, IT199710 and IT212886 when microspores were pre-cultured to $32^{\circ}C$ for 2 days. In IT218043, high embryogenesis was observed at the 3 days of heat shock treatment. The optimal condition of shoot regeneration for IT199710 was observed in MS medium supplemented with NAA $0.5mg{\cdot}L^{-1}$ and BAP $1mg{\cdot}L^{-1}$. In contrast, the IT135449 and IT212886 were observed high regeneration frequency in MS medium without plant growth regulators. All the plantlets regenerated from microspore-derived embryos have been successfully transplanted to soil, and bud self-pollinated seeds were produced from doubled haploid plants. This indicated that double-haploid genotype was likely generated naturally during embryogenesis process.

Effects of Activation Treatments and Culture Condition on In Vitro Development of Caprine In Vivo and In Vitro Oocytes (재래산양의 체내 및 체외유래 난자의 활성화 처리방법 및 배양조건이 단위발생란의 체외발달에 미치는 영향)

  • Park H. S.;Kim T. S.;Lee Y. H.;Jung S. Y.;Lee M. Y.;Jin J. I.;Park J. K.;Lee J. S.;Kim C. H.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.181-185
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    • 2004
  • This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.

A STUDY OF NI-RESISTANT BACTERIA ON THE RESTORED STAINLESS STEEL CROWN (기성금관 수복어린이에서의 니켈내성균에 관한 연구)

  • Chung, Sat-Byul;Lee, Keung-Ho
    • JOURNAL OF THE KOREAN ACADEMY OF PEDTATRIC DENTISTRY
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    • v.27 no.2
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    • pp.274-282
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    • 2000
  • Stainless steel crowns are widely used for restoration for primary molars. The material used for the crowns is an alloy of $70\sim80%$ nickel and $5\sim15%$ chromium. Nickel has been known to cause allergic reaction, cancer and cell toxicity. Little is known about nickel with respect to the relationship between Ni-contained SS crown and graining of Ni-resistance in oral microorganisms. The purpose of this study is to examine whether use of Ni-contained SS crown leads to occurrence of Ni-resistant microorganism, especially enterococci. The gingival crevicular fluid of two different groups was taken. Experimental group included patients wearing SS crown, and control group comprised individuals without SS crown. The samples were plated in BHI agar, BHI agar supplemented with nickel chloride at the concentration of 3mM and bile esculin azide (BEA) agar. The cultured enterococci on BEA agar medium were tested their Ni-resistance in nickel-containing media increasing concentrations from 3mM to 50mM. The results were as follows: 1. In experimental group, a total of 507,350 strains were isolated on BHI agar, of which 53,864(10.62%) strains were found to be resistant to 3mM nickel. In control group, of 414,590 isolates on BHI agar, 37,523 isolates were resistant to 3mM nickel. 2. A total of 95 enterococci were isolated on BEA agar in experimental group, while 20 were isolated in control group. of the enterococci, 68 and 12 isolates were found to be nickel-resistant in experimental and control group, respectively. 3. Of 68 nickel-resistant isolates in experimental group, one survived 50mM nickel. In contrast, none of the isolates in control group was observed to grow at the concentrations over 30mM nickel.

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Changes In Protein and Its Components of High and Low Protein Varieties during Seed Development and Maturation of Soybeans (대두(大豆) 고저단백질(高低蛋白質) 품종(品種)의 성숙중(成熟中) 종실(種實)의 발육(發育)과 단백질(蛋白質) 및 그 구성분(構成分)의 변화(變化))

  • Lee, Jong Suk
    • Korean Journal of Agricultural Science
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    • v.5 no.2
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    • pp.56-67
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    • 1978
  • This study was conducted to characterize comparatively the accumulative patterns of protein and oil, temporal changes in electrophoretic components of proteins during seed development and maturation for the soybean varieties with high, medium and low protein contents. 1. The dry matter of the developing seed increases slowly for the first 22 days after flowering, followed by rapid linear increase for 20 to 30 days and further slow increase for 5 to 15 days attaining its maximum. During the period 12 to 27 days after flowering the protein content of seed increases rapidly while oil content increases rapidly. Following this period of rapid changes, there was period of slow increase until 40 to 47 days after flowering and no seizable further change in the content of both protein and oil. 2. The high protein variety, Saikai # 20, was characterized by shorter period and lower rate of decrease in protein content during the early period, followed by longer period and higher rate of increase in protein content, with earlier stop of oil accumlation during the seed development. 3. The low protein and high oil variety, Shelby, was characterized by longer period of decrease in protein content and shorter period of increase in protein content in contrast to the longer period of slow oil increase during seed development. 4. The temporal pattern of protein component accumulation during seed development was distinctly different among varieties differing in protein content. The time of distinct appearance of all the protein components identifiable in the matured seeds was in accordance with the end of d crease in the protein content of seed. A component having Rm of 0.03 which was absent in the matured seeds was identifiable during the first 17 days after flowering. 5. The high protein variety, Saikai # 20, had much higher compositioral ratio of the component a from the early days of seed development and it continued to increase until 47 days after flowering, while the increase in the composition of the component a stopped as early as 27 days after flowering in the other lower protein varieties. 6. The composition of the component b increased during the period from 17 to 42 days after flowering in all the varieties tested, but the rate of increase during the period was lowest in the high protein variety, Saikai # 20.

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