• Title/Summary/Keyword: 최적해

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Preparation of Pure CO2 Standard Gas from Calcium Carbonate for Stable Isotope Analysis (탄산칼슘을 이용한 이산화탄소 안정동위원소 표준시료 제작에 대한 연구)

  • Park, Mi-Kyung;Park, Sunyoung;Kang, Dong-Jin;Li, Shanlan;Kim, Jae-Yeon;Jo, Chun Ok;Kim, Jooil;Kim, Kyung-Ryul
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.18 no.1
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    • pp.40-46
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    • 2013
  • The isotope ratios of $^{13}C/^{12}C$ and $^{18}O/^{16}O$ for a sample in a mass spectrometer are measured relative to those of a pure $CO_2$ reference gas (i.e., laboratory working standard). Thus, the calibration of a laboratory working standard gas to the international isotope scales (Pee Dee Belemnite (PDB) for ${\delta}^{13}C$ and Vienna Standard Mean Ocean Water (V-SMOW) for ${\delta}^{18}O$) is essential for comparisons between data sets obtained by other groups on other mass spectrometers. However, one often finds difficulties in getting well-calibrated standard gases, because of their production time and high price. Additional difficulty is that fractionation processes can occur inside the gas cylinder most likely due to pressure drop in long-term use. Therefore, studies on laboratory production of pure $CO_2$ isotope standard gas from stable solid calcium carbonate standard materials, have been performed. For this study, we propose a method to extract pure $CO_2$ gas without isotope fractionation from a solid calcium carbonate material. The method is similar to that suggested by Coplen et al., (1983), but is better optimized particularly to make a large amount of pure $CO_2$ gas from calcium carbonate material. The $CaCO_3$ releases $CO_2$ in reaction with 100% pure phosphoric acid at $25^{\circ}C$ in a custom designed, evacuated reaction vessel. Here we introduce optimal procedure, reaction conditions, and samples/reactants size for calcium carbonate-phosphoric acid reaction and also provide the details for extracting, purifying and collecting $CO_2$ gas out of the reaction vessel. The measurements for ${\delta}^{18}O$ and ${\delta}^{13}C$ of $CO_2$ were performed at Seoul National University using a stable isotope ratio mass spectrometer (VG Isotech, SIRA Series II) operated in dual-inlet mode. The entire analysis precisions for ${\delta}^{18}O$ and ${\delta}^{13}C$ were evaluated based on the standard deviations of multiple measurements on 15 separate samples of purified $CO_2$. The pure $CO_2$ samples were taken from 100-mg aliquots of a solid calcium carbonate (Solenhofen-ori $CaCO_3$) during 8-day experimental period. The multiple measurements yielded the $1{\sigma}$ precisions of ${\pm}0.01$‰ for ${\delta}^{13}C$ and ${\pm}0.05$‰ for ${\delta}^{18}O$, comparable to the internal instrumental precisions of SIRA. Therefore, we conclude the method proposed in this study can serve as a way to produce an accurate secondary and/or laboratory $CO_2$ standard gas. We hope this study helps resolve difficulties in placing a laboratory working standard onto the international isotope scales and does make accurate comparisons with other data sets from other groups.

Incidence of Chronic Pathologic Nephrotoxicity of Cyclosporine A in Pediatric Nephrotic Syndrome (소아 신증후군에서 Cyclosporine A에 의한 만성 조직학적 신독성의 발현빈도에 대한 연구)

  • Kim Ji-Hong;Jeong Hyun-Ju;Choi In-Jun;Kim Pyung-Kil
    • Childhood Kidney Diseases
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    • v.3 no.2
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    • pp.130-144
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    • 1999
  • Purpose : Long-term use of Cyclosporine(CsA) reduce renal blood flow by afferent arteriolar vasoconstriction and lead to chronic pathologic changes of CsA nephrotoxicity - 1) interstitial nephritis(IN); tubular atrophy (TA) and/or interstitial fibrosis(IF),2) arteriolopathy(AP). The Object of this study is to estimate the incidence of chronic pathologic CsA nephrotoxicity by duration of treatment and type of renal disease, relationship between histologic and clinical nephrotoxicity, and optimal duration of CsA therapy. Methods : 102 children with steroid resistant or dependent nephrotic syndrome confirmed by renal biopsy and treated with CsA from 1986 to 1997 were enrolled in this study(58 MCNS, 10 FSGS, 10 MGN, 15 $Henoch-Sch\"{o}nlein$ purpura nephritis with nephrotic syndrome (HSPN) and 9 IgA nephropathy with nephrotic syndrome(IgAN)). CsA was administered for 1yr, 1.5yr, 2yr in 24, 12, 22 MCNS patients and 2, 2, 6 FSGS patients respectively, 1yr, 2yr in MGN and 1yr in HSPN and IgAN. Sequential biopsies were done in all 102 patients after CsA treatment for evaluation of pathologic nephrotoxicity. Results : Complete remission rate was 92.2% (100% in MCNS and MGN, 80% in FSGS, 86.6% in HSPN and 55.5% in IgAN). Incidence of relapse during 6months after CsA treatment was significantly decreased compaed with relapsing spisodes during 6months before CsA treatment in MCNS(P<0.0001) and FSGS(P<0.0001). According to pathologic changes, 71 patients(69.6%) showed no pathological change, 24 patients(23.5%) showed IN and 7 patients(6.8%) showed AP. IN was 16.6%, 33.3%, 27.2% in 1, 1.5, 2 year of CsA treatment group in MCNS. AP was 0%, 16.6%, 9% in 1, 1.5, 2 year of CsA treatment group in MCNS. 14 out of 58 MCNS(24.1%) showed IN and 4 out of 58 MCNS(6.8%) showed AP. Incidence of pathologic change was significantly lower in CsA therapy of <1yr than >1yr(P=0.03). There were no significant difference of incidence of pathologic change in original renal disease, age and sex. Conclusion : Duration of CsA treatment was significant risk factor for nephrotoxicity and optimal duration seemed to be 1 year. Pathologic change due to nephrotoxicity did not correlate with deterioration of renal function and only detectable by renal biopsy.

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Development of Conformal Radiotherapy with Respiratory Gate Device (호흡주기에 따른 방사선입체조형치료법의 개발)

  • Chu Sung Sil;Cho Kwang Hwan;Lee Chang Geol;Suh Chang Ok
    • Radiation Oncology Journal
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    • v.20 no.1
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    • pp.41-52
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    • 2002
  • Purpose : 3D conformal radiotherapy, the optimum dose delivered to the tumor and provided the risk of normal tissue unless marginal miss, was restricted by organ motion. For tumors in the thorax and abdomen, the planning target volume (PTV) is decided including the margin for movement of tumor volumes during treatment due to patients breathing. We designed the respiratory gating radiotherapy device (RGRD) for using during CT simulation, dose planning and beam delivery at identical breathing period conditions. Using RGRD, reducing the treatment margin for organ (thorax or abdomen) motion due to breathing and improve dose distribution for 3D conformal radiotherapy. Materials and Methods : The internal organ motion data for lung cancer patients were obtained by examining the diaphragm in the supine position to find the position dependency. We made a respiratory gating radiotherapy device (RGRD) that is composed of a strip band, drug sensor, micro switch, and a connected on-off switch in a LINAC control box. During same breathing period by RGRD, spiral CT scan, virtual simulation, and 3D dose planing for lung cancer patients were peformed, without an extended PTV margin for free breathing, and then the dose was delivered at the same positions. We calculated effective volumes and normal tissue complication probabilities (NTCP) using dose volume histograms for normal lung, and analyzed changes in doses associated with selected NTCP levels and tumor control probabilities (TCP) at these new dose levels. The effects of 3D conformal radiotherapy by RGRD were evaluated with DVH (Dose Volume Histogram), TCP, NTCP and dose statistics. Results : The average movement of a diaphragm was 1.5 cm in the supine position when patients breathed freely. Depending on the location of the tumor, the magnitude of the PTV margin needs to be extended from 1 cm to 3 cm, which can greatly increase normal tissue irradiation, and hence, results in increase of the normal tissue complications probabiliy. Simple and precise RGRD is very easy to setup on patients and is sensitive to length variation (+2 mm), it also delivers on-off information to patients and the LINAC machine. We evaluated the treatment plans of patients who had received conformal partial organ lung irradiation for the treatment of thorax malignancies. Using RGRD, the PTV margin by free breathing can be reduced about 2 cm for moving organs by breathing. TCP values are almost the same values $(4\~5\%\;increased)$ for lung cancer regardless of increasing the PTV margin to 2.0 cm but NTCP values are rapidly increased $(50\~70\%\;increased)$ for upon extending PTV margins by 2.0 cm. Conclusion : Internal organ motion due to breathing can be reduced effectively using our simple RGRD. This method can be used in clinical treatments to reduce organ motion induced margin, thereby reducing normal tissue irradiation. Using treatment planning software, the dose to normal tissues was analyzed by comparing dose statistics with and without RGRD. Potential benefits of radiotherapy derived from reduction or elimination of planning target volume (PTV) margins associated with patient breathing through the evaluation of the lung cancer patients treated with 3D conformal radiotherapy.

Studies on the Determination Method of Natural Sweeteners in Foods - Licorice Extract and Erythritol (식품 중 감초추출물 및 에리스리톨 분석법에 관한 연구)

  • Hong Ki-Hyoung;Lee Tal-Soo;Jang Yaung-Mi;Park Sung-Kwan;Park Sung-Kug;Kwon Yong-Kwan;Jang Sun-Yaung;Han Ynun-Jeong;Won Hye-Jin;Hwang Hye-Shin;Kim Byung-Sub;Kim Eun-Jung;Kim Myung-Chul
    • Journal of Food Hygiene and Safety
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    • v.20 no.4
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    • pp.258-266
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    • 2005
  • Licorice Extract and Erythritol, food additives used in korea, are widely used in foods as sweetener. Its application for use in food is regulated by the standard and specification for food additives but official analytical method far determination of these sweetener in food has not been established. Accordingly, we has been carried out to set up analytical method of the glycyrrhizic acid in several foods by the way of thin layer chromatography and high performance liquid chromatography glycyrrhizic acid is qualitative anaylsis technique consists of clean-up with a sep-pak $C_{18}$ cartridge, separation of the sweeteners by Silica gel 60 F254 TLC plate using 1-butanol:4Nammonia solution:ethanol (50:20:10) as mobile solvent. Also, the quantitative analysis for glycyrrhizic acid, was performed using Capcell prk $C_{18}$ column at wavelength 254nm and DW:Acetonitrile (62:38 (pH2.5)) as mobile phase. and we has been carried out to set up analytical method of the erythritol in several foods by the way of high performance liquid chromatography. erythritol is qualitative anaylsis technique consists of clean-up with a DW and hexane. The quantitative analysis for erythritol, was performed using Asahipak NH2P-50 column, Rl and DW:Acetonitrile (25:75) as mobile phase. The glycyrrhizic acid results determined as glycyrrhizic acid in 105 items were as follows; N.D$\∼$48.7ppm for 18 items in soy sauce, N.D$\∼$5.3ppm for 12 items in sauce, N.D$\∼$988.93ppm for 15 items in health food, N.D$\∼$180.7ppm for 26 items in beverages, N.D$\∼$2.6ppm for 8 items in alcoholic beverages repectively and ND for 63 items in the ethers. The erythritol results determined as erythritol in 52 items were as follows; N.D$\∼$155.6ppm for 13 items in gm, N.D$\∼$398.1ppm for 12 items in health foods repectively and ND for 45 items in the others.

Abundance of Harmful Algae, Cochlodinium polykrikoides, Gyrodinium impudicum and Gymnodinium catenatum in the Coastal Area of South Sea of Korea and Their Effects of Temperature, Salinity, Irradiance and Nutrient on the Growth in Culture (남해안 연안에서 적조생물, Cochlodinium polykikoides, Gyrodinium impudicum, Gymnodinium catenatum의 출현상황과 온도, 염분, 조도 및 영양염류에 따른 성장특성)

  • LEE Chang Kyu;KIM Hyung Chul;LEE Sam-Geun;JUNG Chang Su;KIM Hak Gyoon;LIM Wol Ae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.5
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    • pp.536-544
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    • 2001
  • Three harmful algal bloom species with similar morphology, Cochlodinium polykrikoides, Gyodinium impudicum and Gymodinium catenatum have damaged to aquatic animals or human health by either making massive blooms or intoxication of shellfishes in a food chain. Eco-physiological and hydrodynamic studies on the harmful algae offer useful informations in the understanding their bloom mechanism by giving promising data for the prediction and modelling of harmful algal blooms event. Thus, we studied the abundance of these species in the coastal area of South Sea of Korea and their effects of temperature, salinity, irradiance and nutrient on the growth for the isolates. The timing for initial appearance of the three species around the coastal area of Namhaedo, Narodo and Wando was between Bate July and late August in 1999 when water temperature ranged from $22.8^{\circ}C\;to\;26.5^{\circ}C$ Vegetative cells of C. polykrikoides and G. impudicum were abundant until late September when water temperature had been dropped to less than $23^{\circ}C$. By contrast, vegetative cell of G. catenatum disappeared before early September, showing shorter period of abundance than the other two species in the South Sea. Both G. impudicum and G. catenatum revealed comparatively low density with a maximal cell density of 3,460 cells/L and 440 cells/L, respectively without making any bloom, while C. polykrikoides made massive blooms with a maximal cell density more than $40\times10^6$cells/L, The three species showed a better growth at the relatively higher water temperature ranging from 22 to $28^{\circ}C$ with their maximal growth rate at $25^{\circ}C$ in culture, which almost corresponded with the water temperature during the outbreak of C. polykrikoides in the coastal area of South Sea. Also, they all showed a relatively higher growth at the salinity from 30 to $35\%$. Specially, G. impudicum showed the euryhalic characteristics among the species, On the other hand, growth rate of G. catenatum decreased sharply with the increase of water temperature at the experimental ranges more than $35\%$. The higher of light intensities showed the better growth rates for the three species, Moreover, C. polykrikoides and G. impudirum continued their exponential growth even at 7,500 lux, the highest level of light intensity in the experiment, Therefore, It is assumed that C. polykrikoides has a physiological capability to adapt and utilize higher irradiance resulting in the higher growth rate without any photo inhibition response at the sea surface where there is usually strong irradiance during its blooming season. Although C. poiykikoides and G. impudicum continued their linear growth with the increase of nitrate ($NO_3^-$) and ammonium ($NH_4^-$) concentrations at less than the $40{\mu}M$, they didn't show any significant differences in growth rates with the increase of nitrate and ammonium concentrations at more than $40{\mu}M$, signifying that the nitrogen critical point for the growth of the two species stands between 13.5 and $40{\mu}M$. Also, even though both of the two species continued their linear growth with the increase of phosphate ($PO_4^{2-}$) concentrations at less than the $4.05{\mu}M$, there were no any significant differences in growth rates with the increase of phosphate concentrations at more than $4.05{\mu}M$, signifying that the phosphate critical point for the growth of the two species stands between 1.35 and $4.05{\mu}M$. On the other hand, C. polykrikoides has made blooms at the oligotrophic environment near Narodo and Namhaedo where the concentration of DIN and DIP are less than 1.2 and $0.3{\mu}M$, respectively. We attributed this phenomenon to its own ecological characteristics of diel vertical migration through which C. polykrikoides could uptake enough nutrients from the deep sea water near bottom during the night time irrespective of the lower nutrient pools in the surface water.

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Study on Spring Cocoon Crops with the Leaf Produced in the Mulberry Field close to the Totacco Field (개량 Mulching 담배밭 부근뽕잎이 춘잠작에 미치는 영향에 관한 연구)

  • 이상풍;김정배;김계명;박광준
    • Journal of Sericultural and Entomological Science
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    • v.16 no.1
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    • pp.67-75
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    • 1974
  • The studies are to know how much cocoon crops is damaged by the stained leaf with nicotine produced from the tobacco field cultivated in mulching system in spring season and by residual nicotine in autumn season. Furthermore, the new knowledges are to make both industries keep up with their development. In spring season mulberry Held is located higher on the West-North of tobacco held below 20 degrees of slope and with 36 per cent of East-South wind and 18 per cent of South wind blowing from tobacco fold to the mulberry fold. In addition, silkworm larvae are fed with the mulberry leaf produced in the different plots placing by the different distances, l0m, 25m, 50m, 80m, and loom far from the tobacco Held as a control and it is also considered that narcotic larvae including the dead larvae are not observed. On the other hand, it is noted that better leaf quality and abundant growth of mulberry tree is produced from the mulberry fold closer to the tobacco field and with a low slope. 1) Maximum weight of larval body at the 5th stage is damaged by the stained leaf with the nicotine up to 25m far from the tobacco held. 2) The larvae fed with the mulberry leaf in mulberry Held up to 25m far from the tobacco fold produce small number of the fresh cocoons per 1 liter. 3) Low single cocoon weight and low cocoon shell weight are produced by the poison damaged larvae fed with the mulberry. leaf up to 25m far from the tobacco field and weight of cocoon shell is damaged higher than the single cocoon weight. It is resulted in low percentage of cocoon shell. 4) Cocoon yield including the double cocoon from 10,000 larvae is decreased by the larvae fed with the stained leaf in the mulberry fold up to 25m far from the tobacco fold and 19 per cent of cocoon yield is decreased with 2.4kg of cocoon yield in l0m plot and with 2.5kg of cocoon yield in 25m plot at the first season and at the 2nd season with 1.8kg o( cocoon yield in l0m plot and with 11.5kg of cocoon yield in 25m plot, 11 per cent and 9 per cent of cocoon yield including double cocoon from 10,000 larvae is decreased, as compared with the control, respectively. With these results, it is observed that nicotine damage is occurred to the silkworm larvae if the larvae are fed with the leaf in the mulberry Held within 25m-50m far from the tobacco field.

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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.

Decomposition Characteristics of Fungicides(Benomyl) using a Design of Experiment(DOE) in an E-beam Process and Acute Toxicity Assessment (전자빔 공정에서 실험계획법을 이용한 살균제 Benomyl의 제거특성 및 독성평가)

  • Yu, Seung-Ho;Cho, Il-Hyoung;Chang, Soon-Woong;Lee, Si-Jin;Chun, Suk-Young;Kim, Han-Lae
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.9
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    • pp.955-960
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    • 2008
  • We investigated and estimated at the characteristics of decomposition and mineralization of benomyl using a design of experiment(DOE) based on the general factorial design in an E-beam process, and also the main factors(variables) with benomyl concentration(X$_1$) and E-beam irradiation(X$_2$) which consisted of 5 levels in each factor was set up to estimate the prediction model and the optimization conditions. At frist, the benomyl in all treatment combinations except 17 and 18 trials was almost degraded and the difference in the decomposition of benomyl in the 3 blocks was not significant(p > 0.05, one-way ANOVA). However, the % of benomyl mineralization was 46%(block 1), 36.7%(block 2) and 22%(block 3) and showed the significant difference of the % that between each block(p < 0.05). The linear regression equations of benomyl mineralization in each block were also estimated as followed; block 1(Y$_1$ = 0.024X$_1$ + 34.1(R$^2$ = 0.929)), block 2(Y$_2$ = 0.026X$_2$ + 23.1(R$^2$ = 0.976)) and block 3(Y$_3$ = 0.034X$_3$ + 6.2(R$^2$ = 0.98)). The normality of benomyl mineralization obtained from Anderson-Darling test in all treatment conditions was satisfied(p > 0.05). The results of prediction model and optimization point using the canonical analysis in order to obtain the optimal operation conditions were Y = 39.96 - 9.36X$_1$ + 0.03X$_2$ - 10.67X$_1{^2}$ - 0.001X$_2{^2}$ + 0.011X$_1$X$_2$(R$^2$ = 96.3%, Adjusted R$^2$ = 94.8%) and 57.3% at 0.55 mg/L and 950 Gy, respectively. A Microtox test using V. fischeri showed that the toxicity, expressed as the inhibition(%), was reduced almost completely after an E-beam irradiation, whereas the inhibition(%) for 0.5 mg/L, 1 mg/L and 1.5 mg/L was 10.25%, 20.14% and 26.2% in the initial reactions in the absence of an E-beam illumination.

Immuno-regulatory Activities of Various Fractions from Ehpedrae Sinica STAPF, Rubus Coreanus Miq. and Angelica gigas Nakai Extracts with Ultrasonification (초음파 병행 추출을 이용한 마황과 복분자, 당귀 분획물의 면역활성 조절 효과)

  • Kim, Jung-Hwa;Kim, Dae-Ho;You, Jin-Hyun;Kim, Cheol-Hee;Kwon, Min-Chul;Seong, Nak-Sul;Lee, Seung-Eun;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.4
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    • pp.161-170
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    • 2005
  • This study was performed to examine immuno-regulatory activities of Ehpedrae Sinica STAPF, Rubus Coreanus Miq. and Angelica gigas Nakai extracts in conbination with ultrasonification. The extract yields of plants were the highest in the extraction system of $60^{\circ}C$ and 40 kHz of ultrasonification. The immune cell growth ratio of human immune B and T cells was increased compared to other fractions by the water fraction of the plants at $60^{\circ}C$ and 40 kHz. The water fractions of the plants at $60^{\circ}C$ and 40 kHz increased the specific secretion of IL-6 and $TNF-{\alpha}$ of human immune B and T cells compared to other fractions of the plants. The water fraction of Ehpedrae Sinica STAPF among the plants was observed to show the highest specific secretion of IL-6 and $TNF-{\alpha}$. Also, NK-92 MI cells growth was increased in adding the water fractions of the plants at $60^{\circ}C$ and 40 kHz. The water fraction of Ehpedrae Sinica STAPF among the plants showed the highest in NK-92 MI cell growth ratio. The differentiation activity of the HL-60 cells significantly increased in adding the water fraction of Ehpedrae Sinica STAPF compared to other fractions of the plants. These results suggest that the water fractions of the plants in extraction system of temperature $60^{\circ}C$ and ultrasonification 40 kHz have marked useful immuno-stimulatory activities.

Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus (가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성)

  • Park, Eun-Mi;Yum, Jung-Joo
    • Journal of Life Science
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    • v.20 no.2
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    • pp.260-268
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    • 2010
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Channa argus were purified and characterized by biochemical, immunochemical and kinetic methods. The activity of LDH in skeletal muscle was the highest at 380.4 units and those in heart, eye and brain tissues were 13.4, 3,5 and 5.4 units, respectively. Citrate synthase (EC 4.1.3.7, CS) activity in heart tissue was the highest at 20.7 units. LDH/CS in skeletal muscle, heart, eye and brain tissues were 172.9, 0.6, 0.32 and 0.47. Protein concentration in skeletal muscle tissue was 14.7 mg/g and specific activities of LDH in skeletal muscle, heart, eye and brain tissues were 25.88, 0.79, 0.31 and 1.38 units/mg, respectively. Therefore, skeletal muscle tissue was anaerobic and heart tissue was aerobic. The LDH isozymes in tissues were identified by polyacrylamide gel electrophoresis, immunoprecipitation and Western blot with antiserum against $A_4$, $B_4$, and eye-specific $C_4$. LDH $A_4$, $A_3B$, $A_2B_2$. $AB_3$ and $B_4$ isozymes were detected in every tissue, $C_4$, $AC_3$, $A_2C_2$ and $A_3C$ were detected in eye tissue, and $A_3C$ was found in brain tissue. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ isozymes were purified by affinity chromatography and Preparative PAGE Cells. The LDH $A_4$ isozyme was purified in the fraction from elution with $NAD^+$ containing buffer of affinity chromatography. Eye-specific $C_4$ isozyme was eluted right after $A_4$, after which $B_4$ isozyme was eluted with plain buffer. As a result, one part of molecular structures in $A_4$, $B_4$ and eye-specific $C_4$ were similar, but were different from each other in $B_4$ and $C_4$. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The activity of LDH $A_4$, $A_2B_2$, $B_4$, and eye-specific $C_4$ isozymes remained at 39.98, 21.28, 19.67 and 16.87% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The $Km^{PYR}$ values were 0.17, 0.27 and 0.133 mM in $A_4$, $B_4$ and eye-specific $C_4$ isozymes, respectively. The optimum pH of LDH $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$, and $AB_3$ were pH 6.5, pH 8.5, pH 5.5, pH 6.0-6.5, pH 5.0 and pH 7.5. The $A_4$ and heterotetramer isozymes stabilized a broad range of pH. Especially, LDH activities in skeletal muscle tissue were high, resulting in a high degree of muscle activity.LDH metabolism in eye tissue seems to be converted faster from pyruvate to lactate by eye-specific $C_4$ isozyme as eye-specific $C_4$ have the highest affinity for pyruvate, and right after the conversion, oxidation of lactate was induced by $A_4$ isozyme. It was found that expression of Ldh-C, affinity to substrate and reaction time of $C_4$ isozyme were different according to the ecological environmental and feeding capturing patterns.