• Title/Summary/Keyword: Carbon-Dioxide

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Effects of $TGF-{\beta}1$ on Cellular Activity of Minocycline-Pretreated Human Periodontal Ligament Cells (($TGF-{\beta}$)이 Minocycline을 전처리한 사람 치주인대세포의 활성에 미치는 영향)

  • Yang, Seung-Oh;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.469-490
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    • 1996
  • The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

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Soil CO2 Efflux Dynamics in Response to Fertilization in Pinus densiflora and Quercus variabilis Stands (소나무와 굴참나무 임분의 시비에 따른 토양 CO2 방출 동태)

  • Baek, Gyeongwon;Kim, Choonsig
    • Journal of Korean Society of Forest Science
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    • v.109 no.3
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    • pp.271-280
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    • 2020
  • This study compared soil CO2 efflux rates after fertilization, in Pinus densiflora and Quercus variabilis stands. Compound fertilizers were applied to the forest floor in March 2016, following a one-year calibration period (from March 2015 to February 2016). In situ soil CO2 efflux rates were measured every month during the two-year study periods, using an infrared gas analyzer with a closed chamber system. Mean annual soil CO2 efflux rates were higher following fertilizer application in the P. densiflora and Q. variabilis stands (P. densiflora: 2.180 μmol m-2 s-1; Q. variabilis: 1.977 μmol m-2 s-1) as compared with the rates measured during the calibration period (P. densiflora: 1.620 μmol m-2 s-1; Q. variabilis: 1.557 μmol m-2 s-1). The mean annual soil CO2 efflux rates in the unfertilized treatments of both stands were not significantly different between the two-year study periods. The Q10 values of fertilized treatments in Q. variabilis stands were higher in the fertilization period (3.41) than in the calibration period (3.14), whereas the Q10 values in P. densiflora stands did not change between the fertilization and calibration periods. The Q10 values of unfertilized treatments in the Q. variabilis stands were lower during the 2016-2017 period (3.69), than in the 2015-2016 period (3.85), whereas the Q10 values in P. densiflora stands were higher during the 2016-2017 period (3.65), than in the 2015-2016 period (3.15). These results indicate that the increase in soil CO2 efflux rates in P. densiflora stands could be more sensitive to fertilization compared with the rates in Q. variabilis stands.

Legal Review on the Regulatory Measures of the European Union on Aircraft Emission (구주연합의 항공기 배출 규제 조치의 국제법적 고찰)

  • Park, Won-Hwa
    • The Korean Journal of Air & Space Law and Policy
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    • v.25 no.1
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    • pp.3-26
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    • 2010
  • The European Union(EU) has recently introduced its Directive 2008/101/EC to include aviation in the EU ETS(emissions trading system). As an amendment to Directive 2003/87/EC that regulates reduction of the green house gas(GHG) emissions in Europe in preparation for the Kyoto Protocol, 1997, it obliges both EU and non-EU airline operators to reduce the emission of the carbon dioxide(CO2) significantly in the year 2012 and thereafter from the level they made in 2004 to 2006. Emission allowances allowed free of charge for each airline operator is 97% in the first year 2012 and 95% from 2013 and thereafter from the average annual emissions during historical years 2004 to 2006. Taking into account the rapid growth of air traffic, i.e. 5% in recent years, airlines operating to EU have to reduce their emissions by about 30% in order to meet the requirements of the EU Directive, if not buy the emissions right in the emissions trading market. However, buying quantity is limited to 15% in the year 2012 subject to possible increase from the year 2013. Apart from the hard burden of the airline operators, in particular of those from non-European countries, which is not concern of this paper, the EU Directive has certain legal problems. First, while the Kyoto Protocol of universal application is binding on the Annex I countries of the Climate Change Convention, i.e. developed countries including all Member States of the European Union to reduce GHG at least by 5% in the implementation period from 2008 to 2012 over the 1990 level, non-Annex I countries which are not bound by the Kyoto Protocol see their airlines subjected to aircraft emissions reductions scheme of EU when operating to EU. This is against the provisions of the Kyoto Protocol dealing with the emissions of GHG including CO2, target of the EU Directive. While the Kyoto Protocol mandates ICAO to set up a worldwide scheme for aircraft emissions to contribute to stabilizing GHG concentrations in the atmosphere at a level that would prevent dangerous anthropogenic interference with the climate system, the EU ETS was drawn up outside the framework of the international Civil Aviation Organization(ICAO). Second, EU Directive 2008/101 defines 'aviation activities' as covering 'flights which depart from or arrive in the territory of a Member State to which the [EU] Treaty applies'. While the EU airlines are certainly subject to the EU regulations, obliging non-EU airlines to reduce their emissions even if the emissions are produced during the flight over the high seas and the airspace of the third countries is problematic. The point is whether the EU Directive can be legally applied to extra-territorial behavior of non-EU entities. Third, the EU Directive prescribes 2012 as the first year for implementation. However, the year 2012 is the last year of implementation of the Kyoto Protocol for Annex I countries including members of EU to reduce GHG including the emissions of CO2 coming out from domestic airlines operation. Consequently, EU airlines were already on the reduction scheme of CO2 emissions as long as their domestic operations are concerned from 2008 until the year 2012. But with the implementation of Directive 2008/101 from 2012 for all the airlines, regardless of the status of the country Annex I or not where they are registered, the EU airlines are no longer at the disadvantage compared with the airlines of non-Annex I countries. This unexpected premium for the EU airlines may result in a derogation of the Kyoto Protocol at least for the year 2012. Lastly, as a conclusion, the author shed light briefly on how the Korean aviation authorities are dealing with the EU restrictive measures.

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Operating Characteristics of 1 $Nm^3/h$ Scale Synthetic Natural Gas(SNG) Synthetic Systems (1 $Nm^3/h$ 규모 합성천연가스(SNG) 합성 시스템의 운전 특성)

  • Kim, Jin-Ho;Kang, Suk-Hwan;Ryu, Jae-Hong;Lee, Sun-Ki;Kim, Su-Hyun;Kim, Mun-Hyun;Lee, Do-Yeon;Yoo, Yong-Don;Byun, Chang-Dae;Lim, Hyo-Jun
    • Korean Chemical Engineering Research
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    • v.49 no.4
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    • pp.491-497
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    • 2011
  • In this work, we proposed the three different reactor systems for evaluating of synthetic natural gas(SNG) processes using the synthesis gas consisting of CO and $H_2$ and reactor systems to be considered are series adiabatic reaction system, series adiabatic reaction system with the recirculation and cooling wall type reaction system. The maximum temperature of the first adiabatic reactor in series adiabatic reaction system raised to 800. From the these results, carbon dioxide in product gas as compared to other systems was increased more than that expected due to water gas shift reaction(WGSR) and the maximum $CH_4$ concentration in SNG was 90.1%. In series adiabatic reaction system with the recirculation as a way to decrease the temperature in catalyst bed, the maximum $CH_4$ concentration in SNG was 96.3%. In cooling wall type reaction system, the reaction heat is absorbed by boiling water in the shell and the reaction temperature is controlled by controlling the amount of flow rate and pressure of feed water. The maximum $CH_4$ concentration in SNG for cooling wall type reaction system was 97.9%. The main advantage of the cooling wall type reaction system over adiabatic systems is that potentially it can be achieve almost complete methanation in one reactor.

Photosynthetic Characteristics and Cellular Tissue of Chinese Cabbage are Affected by Temperature and $CO_{2}C$ Concentration (온도와 $CO_{2}C$ 농도에 따른 배추의 광합성특성 및 세포조직의 변화)

  • Lee, Sang-Gyu;Moon, Ji-Hye;Jang, Yoon-Ah;Lee, Woo-Moon;Cho, Ill-Hwan;Kim, Seung-Yu;Ko, Kwan-Dal
    • Journal of Bio-Environment Control
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    • v.18 no.2
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    • pp.148-152
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    • 2009
  • Numerous studies have presented evidence that global atmospheric carbon dioxide ($CO_{2}$ ) concentration and temperature is increasing every year. Both of the $CO_{2}$ and temperature are important components for photosynthesis activity of plants and thusgrowth and yield. However, little information is available in terms of the reaction of vegetable plants to increased $CO_{2}$ concentration and temperature, and also the reaction to a complex condition of both increased $CO_{2}$ concentration and temperature. The aim of this research was therefore to investigate changes in growth, photosynthetic activity and ultra-cellular structure of leaf tissue of Chinese cabbage. Plants were grown under either of elevated $CO_{2}$ concentration (elevated $CO_{2}$, 2-fold higher than atmospheric $CO_{2}$ ) or elevated temperature (elevated temp, 4$^{\circ}C$ higher than atmospheric temperature), under both of elevated $CO_{2}$ concentration and elevated temperature (elevated temp+$CO_{2}$), and under atmospheric $CO_{2}$ concentration and temperature (control). The treatment of 'elevated temp' negatively affected leaf area, fresh weight, chlorophyll and starch content. However, when the treatment of 'elevated temp' was applied coincidently with the treatment of 'elevated $CO_{2}$', growth and photosynthetic performance of plants were as good as those in the treatment of 'elevated $CO_{2}$', Microscopic study resulted that the highest starch content and density of cells were observed in the leaf tissue grown at the treatment of 'elevated $CO_{2}$', whereas the lowest ones were observed in the leaf tissue grown at the treatment of 'elevated temp'. These results suggest that when Chinese cabbage grows under a high-temperature condition, supplement of $CO_{2}$ would improve the growth and yield. In our knowledge, it is the first time to determine the effect of a complex relationship between the increased $CO_{2}$ concentration and temperature on the growth of Chinese cabbage.

The effect of using laser for ceramic bracket bonding of porcelain surfaces (세라믹 브라켓 부착 시 레이저를 이용한 포세린 표면처리 효과)

  • An, Kyung-Mi;Sohn, Dong-Seok
    • The korean journal of orthodontics
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    • v.38 no.4
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    • pp.275-282
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    • 2008
  • Objective: The purpose of this study was to investigate the effect of using laser for ceramic bracket bonding of porcelain surfaces and to compare it with conventional treatment of porcelain surfaces. Methods: Ninety feldspathic porcelain specimens were divided into 9 groups of 10, with each group having different surface treatments performed. Surface treatment groups were orthophosphoric acid, orthophosphoric acid with silane, hydrofluoric acid, hydrofluoric acid with silane, sandblasted, sandblasted with silane, laser etched, laser etched with silane, and glazed surface served as a control group. In the laser etched groups, the specimens were irradiated with 2-watt superpulse carbon dioxide ($CO_2$) laser for 20 seconds. Ceramic brackets were bonded with light-cure composite resin and all specimens were stored in water at $37^{\circ}C$ for 24 hours. Shear bond strength was determined in megapascals (MPa) by shear test at 1 mm/minute crosshead speed and the failure pattern was assessed. For statistical analysis, one-way ANOVA and tukey test were used. Results: Statistical analysis showed significant differences between the groups. The HFA + S group showed the highest mean shear bond strength ($13.92{\pm}1.94\;MPa$). This was followed by SB + S ($10.16\;{\pm}\;1.27\;MPa$), HFA ($10.09\;{\pm}\;1.07\;MPa$), L + S ($8.25\;{\pm}\;1.24\;MPa$), L ($7.86\;{\pm}\;0.96\;MPa$), OFA + S ($7.22\;{\pm}\;1.09\;MPa$), SB ($3.41\;{\pm}\;0.37\;MPa$), OFA ($2.81\;{\pm}\;0.37\;MPa$), G ($2.46\;{\pm}\;1.36\;MPa$), Bond failure patterns of HFA and silane groups, except L + S, were cohesive modes in porcelain while adhesive failure was observed in the control group and the rest of the groups. Conclusions : A 2-watt superpulse $CO_2$ laser etching of porcelain surfaces can provide a satisfactory result for porcelain surface treatment for ceramic bracket bonding. Laser irradiation may be an alternative conditioning method for the treatment of porcelain surfaces.

Enhancement of the Working Capacity and Selectivity Factor of Calcium-Exchanged Y Zeolites for Carbon Dioxide Pressure Swing Adsorption (이산화탄소 압력순환흡착을 위한 칼슘 이온교환 Y 제올라이트의 작업용량과 선택계수 향상)

  • Kim, Moon Hyeon
    • Clean Technology
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    • v.24 no.1
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    • pp.41-49
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    • 2018
  • Y zeolites with different extra-framework cations, such as $Na^+$, $N^+$, $Ca^{2+}$, and $Cu^{2+}$, with different charge and ionic radius have been investigated to greatly enhance a working capacity (W) of $CO_2$ adsorption at $25^{\circ}C$ and a $CO_2/CO$ selectivity factor (S). A sample of NaY with a very small amount of 0.012% $Ca^{2+}$ was fully reversible for seven times repeated $CO_2$ adsorption/desorption cycles, thereby forming no surface carbonates unlikely earlier reports. Although at pressures above 4 bar, 2.00% CaY, 1.60% CuY and 1.87% LiY all showed a $CO_2$ adsorption very similar to that measured for NaY, they gave a significant decrease in the adsorption at lower pressures, depending on the metal ion. At 0.5 ~ 2.5 bar, the extent of $CO_2$ adsorption was in the order NaY > 1.60% CuY > 2.00% CaY > 1.87% LiY. All the $Na^+-based$ metals-exchanged zeolites have a FAU (faujasite) framework and a Si/Al value near 2.6; thus, there is no discernible difference in the framework topology, framework chemical compositions, effective aperture size, and channel structure between the zeolite samples. Therefore, the distinctive behavior in the adsorption of $CO_2$ with a character as a weak Lewis acid is associated with the site basicity of the zeolites, and the interaction potentials of the cations. Different trend was shown for a CO adsorption due to weaker quadrupole interactions. Adsorption of $CO_2$ and CO on samples of CaY with 0.012 to 5.23% Ca disclosed a significant dependence on the Ca loading. The $CO_2$ adsorption increased when the cation exists up to ca. 0.05%, while it decreased at higher Ca amounts. However, values for both W and S could greatly increase as the bare zeolite is enriched by $Ca^{2+}$ ions. The 5.23% CaY had $W=2.37mmol\;g^{-1}$ and S = 4.37, and the former value was comparable to a benchmark reported in the literature.

THE COMPARATIVE STUDY ON THE SEDATIVE EFFECT OF DIFFERENT INTRAVENOUS MIDAZOLAM DOSAGES FOR PEDIATRIC DENTAL PATIENTS (소아환자의 Midazolam의 정맥투여 용량에 따른 진정 효과에 관한 비교연구)

  • Kim, Eun-Young;Kim, Jong-Soo;You, Seung-Hoon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.3
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    • pp.416-426
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    • 2005
  • Intravenous sedation have many advantages of rapid onset and recovery, ability of control sedation levels and duration through titration. Midazolam is most commonly used intravenous medication for sedation in pediatrics, endoscopy, oncologic procedures and so on. But in dentistry, midazolam intravenous sedation is usually for adult, and there are few reports for children. Todays, children who need sedation become more and older, intravenous sedation technique is going a matter of concern in pediatric dentistry. The purpose of this paper is to evaluate the efficacy of sedation and clinical success for different initial dosage of midazolam in intravenous sedation for pediatric dental patients. 16 healthy children (male 10, female 6), mean age $54.7{\pm}10.7$ months, who needed at least two separate treatment visits requiring local anesthesia were chosen for this study. Every children were taken 0.3mg/kg, maximum 5mg of midazolam by intramuscular route, and then 30~50% $N_2O-O_2$ for 10 minutes was given. On every visits, one of the following 2 different initial dosage was given by intravenous route : (1) Group I : 0.1mg/kg Midazolam (2) Group II : 0.2mg/kg Midazolam. Additional dosage was half of the first dose. Physiologic parameters (oxygen saturation, heart rate, respiratory rate, end-tidal carbon dioxide pressure) was recorded by ten procedure steps. Behavior was videotaped and rated using Ohio State University Behavioral Rating Scale and Automated Counting System by one investigator, blind to administered dosage. After the treatment, operator evaluated the clinical success. Physiologic parameters were stable and within normal range during treatment in both groups. The analyzed sedative effect, in behavioral evaluation, ratio of favorable Quiet was higher in group II, and clinical success rate of group II was better than group I. Induction time was rapid in group II, and recovery time was rapid in group I. And there was no statistically difference between two groups in every results.

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Recent Progress in Air Conditioning and Refrigeration Research - A Review of Papers Published in the Korean Journal of Air-Conditioning and Refrigeration Engineering in 2004 and 2005 - (공기조화, 냉동 분야의 최근 연구 동향 -2004년 및 2005년 학회지 논문에 대한 종합적 고찰-)

  • Choi, Yong-Don;Kang, Yong-Tae;Kim, Nae-Hyun;Kim, Man-Hoe;Park, Kyoung-Kuhn;Park, Byung-Yoon;Park, Jin-Chul;Hong, Hi-Ki
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.19 no.1
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    • pp.94-131
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    • 2007
  • A review on the papers published in the Korean Journal of Air-Conditioning and Refrigerating Engineering in 2004 and 2005 has been done. Focus has been put on current status of research in the aspect of heating, cooling, air-conditioning, ventilation, sanitation and building environment. The conclusions are as follows. (1) Most of fundamental studies on fluid flow were related with heat transportation of facilities. Drop formation and rivulet flow on solid surfaces were interesting topics related with condensation augmentation. Research on micro environment considering flow, heat, humidity was also interesting for comfortable living environment. It can be extended considering biological aspects. Development of fans and blowers of high performance and low noise were continuing topics. Well developed CFD and flow visualization(PIV, PTV and LDV methods) technologies were widely applied for developing facilities and their systems. (2) The research trends of the previous two yews are surveyed as groups of natural convection, forced convection, electronic cooling, heat transfer enhancement, frosting and defrosting, thermal properties, etc. New research topics introduced include natural convection heat transfer enhancement using nanofluid, supercritical cooling performance or oil miscibility of $CO_2$, enthalpy heat exchanger for heat recovery, heat transfer enhancement in a plate heat exchanger using fluid resonance. (3) The literature for the last two years($2004{\sim}2005$) is reviewed in the areas of heat pump, ice and water storage, cycle analysis and reused energy including geothermal, solar and unused energy). The research on cycle analysis and experiments for $CO_2$ was extensively carried out to replace the Ozone depleting and global warming refrigerants such as HFC and HCFC refrigerants. From the year of 2005, the Gas Engine Heat Pump(GHP) has been paid attention from the viewpoint of the gas cooling application. The heat pipe was focused on the performance improvement by the parametric analysis and the heat recovery applications. The storage systems were studied on the performance enhancement of the storage tank and cost analysis for heating and cooling applications. In the area of unused energy, the hybrid systems were extensively introduced and the life cycle cost analysis(LCCA) for the unused energy systems was also intensively carried out. (4) Recent studies of various refrigeration and air-conditioning systems have focused on the system performance and efficiency enhancement. Heat transfer characteristics during evaporation and condensation are investigated for several tube shapes and of alternative refrigerants including carbon dioxide. Efficiency of various compressors and expansion devices are also dealt with for better modeling and, in particular, performance improvement. Thermoelectric module and cooling systems are analyzed theoretically and experimentally. (5) According to the review of recent studies on ventilation systems, an appropriate ventilation systems including machenical and natural are required to satisfied the level of IAQ. Also, an recent studies on air-conditioning and absorption refrigeration systems, it has mainly focused on distribution and dehumidification of indoor air to improve the performance were carried out. (6) Based on a review of recent studies on indoor environment and building service systems, it is noticed that research issues have mainly focused on optimal thermal comfort, improvement of indoor air Quality and many innovative systems such as air-barrier type perimeter-less system with UFAC, radiant floor heating and cooling system and etc. New approaches are highlighted for improving indoor environmental condition as well as minimizing energy consumption, various activities of building control and operation strategy and energy performance analysis for economic evaluation.

Extraction Characteristics of Flavonoids from Lonicera flos by Supercritical Fluid Carbon Dioxide ($SF-CO_2$) with Co-solvent (초임계유체 $CO_2$ 및 Co-solvent 첨가에 따른 금은화(Lonicera fles)의 Flavonoid류 추출특성)

  • Suh, Sang-Chul;Cho, Sung-Gill;Hong, Joo-Heon;Choi, Yong-Hee
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.183-188
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    • 2005
  • Effects of co-solvent polarity, citric acid, pressure, temperature, run time, and co-solvent ratio on extraction of major flavonoids from Lonicera Flos were investigated using supercritical fluid $CO_{2}(SF-CO_{2})$. HPLC analysis revealed addition of pure methanol resulted in low extraction yield of major flavonoids, luteoloin (Lu), Quercetin (Qu), Apigenin (Ap). Under same condition, as co-solvent polarity increased, yields of major flavonoids increased gradually, At optimum co-solvent extraction condirion of 60% aqueous methanol (10%, v/v), yields of Lu, Qu, and Ap were 42.09, 28.18, and 3.49 mg/100 g, respectively. Addition of citric acid to 60% aqueous methanol gave higher, with addition of 1% citrie acid resulting in highest yields of 63.2 (Lu), 39.35 (Qu), and 5.79 (Ap) mg/100 g. Optimum extraction conditions of major flavonoids were 200 bar, $50^{\circ}C$, 60 min, and $CO_{2}$-methanol-water(20: 1.8: 1.2).