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Smoking-Induced Dopamine Release Studied with $[^{11}C]Raclopride$ PET ($[^{11}C]Raclopride$ PET을 이용한 흡연에 의한 도파민 유리 영상 연구)

  • Kim, Yu-Kyeong;Cho, Sang-Soo;Lee, Do-Hoon;Ryu, Hye-Jung;Lee, Eun-Ju;Ryu, Chang-Hung;Jeong, In-Soon;Hong, Soo-Kyung;Lee, Jae-Sung;Seo, Hong-Gwan;Jeong, Jae-Min;Lee, Won-Woo;Kim, Sang-Eun
    • The Korean Journal of Nuclear Medicine
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    • v.39 no.6
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    • pp.421-429
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    • 2005
  • Purpose: It has been postulated that dopamine release in the striatum underlies the reinforcing properties of nicotine. Substantial evidence in the animal studies demonstrates that nicotine interacts with dopaminergic neuron and regulates the activation of the dopaminergic system. The aim of this study was to visualize the dopamine release by smoking in human brain using PET scan with $[^{11}C]raclopride$. Materials and Methods: Five male non-smokers or ex-smokers with an abstinence period longer than 1 year (mean age of $24.4{\pm}1.7$ years) were enrolled in this study $[^{11}C]raclopride$, a dopamine D2 receptor radioligand, was administrated with bolus-plus-constant infusion. Dynamic PET was performed during 120 minutes ($3{\times}20s,\;2{\times}60s,\;2{\times}120s,\;1{\times}180s\;and\;22{\times}300s$). following the 50 minute-scanning, subjects smoked a cigarette containing 1 mg of nicotine while in the scanner. Blood samples for the measurement of plasma nicotine level were collected at 0, 5, 10, 15, 20, 25, 30, 45, 60, and 90 minute after smoking. Regions for striatal structures were drawn on the coronal summed PET images guided with co-registered MRI. Binding potential, calculated as (striatal-cerebellar)/cerebellar activity, was measured under equilibrium condition at baseline and smoking session. Results: The mean decrease in binding potential of $[^{11}C]raclopride$ between the baseline and smoking in caudate head, anterior putamen and ventral striatum was 4.7%, 4.0% and 7.8%, respectively. This indicated the striatal dopamine release by smoking. Of these, the reduction in binding potential in the ventral striatum was significantly correlated with the cumulated plasma level of the nicotine (Spearman's rho=0.9, p=0.04). Conclusion: These data demonstrate that in vivo imaging with $[^{11}C]raclopride$ PET could measure nicotine-induced dopamine release in the human brain, which has a significant positive correlation with the amount or nicotine administered bt smoking.

Effects of Motion Correction for Dynamic $[^{11}C]Raclopride$ Brain PET Data on the Evaluation of Endogenous Dopamine Release in Striatum (동적 $[^{11}C]Raclopride$ 뇌 PET의 움직임 보정이 선조체 내인성 도파민 유리 정량화에 미치는 영향)

  • Lee, Jae-Sung;Kim, Yu-Kyeong;Cho, Sang-Soo;Choe, Yearn-Seong;Kang, Eun-Joo;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul;Kim, Sang-Eun
    • The Korean Journal of Nuclear Medicine
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    • v.39 no.6
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    • pp.413-420
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    • 2005
  • Purpose: Neuroreceptor PET studies require 60-120 minutes to complete and head motion of the subject during the PET scan increases the uncertainty in measured activity. In this study, we investigated the effects of the data-driven head mutton correction on the evaluation of endogenous dopamine release (DAR) in the striatum during the motor task which might have caused significant head motion artifact. Materials and Methods: $[^{11}C]raclopride$ PET scans on 4 normal volunteers acquired with bolus plus constant infusion protocol were retrospectively analyzed. Following the 50 min resting period, the participants played a video game with a monetary reward for 40 min. Dynamic frames acquired during the equilibrium condition (pre-task: 30-50 min, task: 70-90 min, post-task: 110-120 min) were realigned to the first frame in pre-task condition. Intra-condition registrations between the frames were performed, and average image for each condition was created and registered to the pre-task image (inter-condition registration). Pre-task PET image was then co-registered to own MRI of each participant and transformation parameters were reapplied to the others. Volumes of interest (VOI) for dorsal putamen (PU) and caudate (CA), ventral striatum (VS), and cerebellum were defined on the MRI. Binding potential (BP) was measured and DAR was calculated as the percent change of BP during and after the task. SPM analyses on the BP parametric images were also performed to explore the regional difference in the effects of head motion on BP and DAR estimation. Results: Changes in position and orientation of the striatum during the PET scans were observed before the head motion correction. BP values at pre-task condition were not changed significantly after the intra-condition registration. However, the BP values during and after the task and DAR were significantly changed after the correction. SPM analysis also showed that the extent and significance of the BP differences were significantly changed by the head motion correction and such changes were prominent in periphery of the striatum. Conclusion: The results suggest that misalignment of MRI-based VOI and the striatum in PET images and incorrect DAR estimation due to the head motion during the PET activation study were significant, but could be remedied by the data-driven head motion correction.

Assessment of Cerebral Hemodynamic Changes in Pediatric Patients with Moyamoya Disease Using Probabilistic Maps on Analysis of Basal/Acetazolamide Stress Brain Perfusion SPECT (소아 모야모야병에서 뇌확률지도를 이용한 수술전후 혈역학적 변화 분석)

  • Lee, Ho-Young;Lee, Jae-Sung;Kim, Seung-Ki;Wang, Kyu-Chang;Cho, Byung-Kyu;Chung, June-Key;Lee, Myung-Chul;Lee, Dong-Soo
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.3
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    • pp.192-200
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    • 2008
  • To evaluate the hemodynamic changes and the predictive factors of the clinical outcome in pediatric patients with moyamoya disease, we analyzed pre/post basal/acetazolamide stress brain perfusion SPECT with automated volume of interest (VOIs) method. Methods: Total fifty six (M:F = 33:24, age $6.7{\pm}3.2$ years) pediatric patients with moyamoya disease, who underwent basal/acetazolamide stress brain perfusion SPECT within 6 before and after revascularization surgery (encephalo-duro-arterio-synangiosis (EDAS) with frontal encephalo-galeo-synangiosis (EGS) and EDAS only followed on contralateral hemisphere), and followed-up more than 6 months after post-operative SPECT, were included. A mean follow-up period after post-operative SPECT was $33{\pm}21$ months. Each patient's SPECT image was spatially normalized to Korean template with the SPM2. For the regional count normalization, the count of pons was used as a reference region. The basal/acetazolamide-stressed cerebral blood flow (CBF), the cerebral vascular reserve index (CVRI), and the extent of area with significantly decreased basal/acetazolamide- stressed rCBF than age-matched normal control were evaluated on both medial frontal, frontal, parietal, occipital lobes, and whole brain in each patient's images. The post-operative clinical outcome was assigned as good, poor according to the presence of transient ischemic attacks and/or fixed neurological deficits by pediatric neurosurgeon. Results: In a paired t-test, basal/acetazolamide-stressed rCBF and the CVRI were significantly improved after revascularization (p<0.05). The significant difference in the pre-operative basal/acetazolamide-stressed rCBF and the CVRI between the hemispheres where EDAS with frontal EGS was performed and their contralateral counterparts where EDAS only was done disappeared after operation (p<0.05). In an independent student t-test, the pre-operative basal rCBF in the medial frontal gyrus, the post-operative CVRI in the frontal lobe and the parietal lobe of the hemispheres with EDAS and frontal EGS, the post-operative CVRI, and ${\Delta}CVRI$ showed a significant difference between patients with a good and poor clinical outcome (p<0.05). In a multivariate logistic regression analysis, the ${\Delta}CVRI$ and the post-operative CVRI of medial frontal gyrus on the hemispheres where EDAS with frontal EGS was performed were the significant predictive factors for the clinical outcome (p =0.002, p =0.015), Conclusion: With probabilistic map, we could objectively evaluate pre/post-operative hemodynamic changes of pediatric patients with moyamoya disease. Specifically the post-operative CVRI and the post-operative CVRI of medial frontal gyrus where EDAS with frontal EGS was done were the significant predictive factors for further clinical outcomes.

Establishment of Analytical Method for Dichlorprop Residues, a Plant Growth Regulator in Agricultural Commodities Using GC/ECD (GC/ECD를 이용한 농산물 중 생장조정제 dichlorprop 잔류 분석법 확립)

  • Lee, Sang-Mok;Kim, Jae-Young;Kim, Tae-Hoon;Lee, Han-Jin;Chang, Moon-Ik;Kim, Hee-Jeong;Cho, Yoon-Jae;Choi, Si-Won;Kim, Myung-Ae;Kim, MeeKyung;Rhee, Gyu-Seek;Lee, Sang-Jae
    • Korean Journal of Environmental Agriculture
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    • v.32 no.3
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    • pp.214-223
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    • 2013
  • BACKGROUND: This study focused on the development of an analytical method about dichlorprop (DCPP; 2-(2,4-dichlorophenoxy)propionic acid) which is a plant growth regulator, a synthetic auxin for agricultural commodities. DCPP prevents falling of fruits during their growth periods. However, the overdose of DCPP caused the unwanted maturing time and reduce the safe storage period. If we take fruits with exceeding maximum residue limits, it could be harmful. Therefore, this study presented the analytical method of DCPP in agricultural commodities for the nation-wide pesticide residues monitoring program of the Ministry of Food and Drug Safety. METHODS AND RESULTS: We adopted the analytical method for DCPP in agricultural commodities by gas chromatograph in cooperated with Electron Capture Detector(ECD). Sample extraction and purification by ion-associated partition method were applied, then quantitation was done by GC/ECD with DB-17, a moderate polarity column under the temperature-rising condition with nitrogen as a carrier gas and split-less mode. Standard calibration curve presented linearity with the correlation coefficient ($r^2$) > 0.9998, analysed from 0.1 to 2.0 mg/L concentration. Limit of quantitation in agricultural commodities represents 0.05 mg/kg, and average recoveries ranged from 78.8 to 102.2%. The repeatability of measurements expressed as coefficient of variation (CV %) was less than 9.5% in 0.05, 0.10, and 0.50 mg/kg. CONCLUSION(S): Our newly improved analytical method for DCPP residues in agricultural commodities was applicable to the nation-wide pesticide residues monitoring program with the acceptable level of sensitivity, repeatability and reproducibility.

Studies on Grouping of the Varieties by Plant Type and their Ecological Variation for Peanut(Arachis hypogaea L.) (땅콩의 초형을 주로한 품종군분류 및 그들의 생태적 변이에 관한 연구)

  • Eun-Sup Lee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.18
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    • pp.124-155
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    • 1975
  • To obtain the fundamental informations on the varietal improvement of peanut and to study the ecological variations of the important agronomic traits and to the relationship between the traits studied, an investigation was made on varietal classification of 489 introduced on the basis of their morphological and ecological differences at Crop Experiment Station, Suweon in 1968, and the other study conducted at some location as above in 1969 was to investigate the ecological variations of the materials in accordance with changes of seeding date using classified varietal group under 5 different seeding times from April 16 to July 7 with twenty days interval. The results obtained were summarized as follows: 1. Peanut varieties tested were classified into Spanish, Virginia Erect, Virginia Runner, Southeast Runner. Valencia and Semirunner, on the basis of plant type, flowering time, number of grains per pod and grain size. 2. Characteristics of varietal group classified are as followings. (1) Spanish; erect, small grained and early maturing type. (2) Virginia Erect; erect, large grained and late flowering type. (3) Virginia Runner; runner, large grained and late maturing type. (4) Southeast Runner; runner, small grained and early maturing type. (5) Valencia; erect, small grained and early flowering type with 3-4 grains per pod. (6) Semi runner; semirunner, large grained and late flowering type. 3. Flowering period in respective varietal group was consistently shorted by delayed seeding date and the degree of shortening was more serious in late flowering varietal group. 4. Number of branches per plant was generally decreased in late seeding date in respective group. However, Spanish and Virginia Runner exhibited lower number of branches in the first seeding rather than the second seeding and the lowest number of branches was found in Spanish while the highest were Virginia Erect in all seeding date. 5. Shelling ratio was high in Spanish and Southeast Runner in any seeding date and decreased remarkedly by seeding after May. 6. Number of pod per plant in all varietal groups was remarkedly decreased by delayed seeding date and the degree of decreasing was more serious in large grain varietal group. 7. The higher pod weight per plant was found in second seeding date rather than first seeding and pod weight per plant was decreased obviously in all late seeding after the second. Therefore, among the cultivars tested, Southeast Runner noted the highest pod weight per plant while Virginia Runner showed the lowest. 8. Grain number per plant expressed the similar tendency as the pod weight per plant but was low in large grain group and high in small grain group in all seeding date employed. 9. 100 grain weight was heaviest in second seeding and was decreased remarkedly after the second and even the first seeding date. 10. Yield per 10a noted considerable variations in accordance with seeding date in all groups classified. However, the yield was increased in second seeding date (May 7) and decreased in the others. 11. Length of main stem and branches were exceptionally decreased in the first seeding date compare to the second in Spanish while other varieties were tend to be same between the indicated seeding date, but. these two traits were strikingly decreased in all seeding after the second. This tendency, however, strongly suggested the importance of environmental effects on peanut growth in terms of their changes due to the different seeding date. 12. Highly significant positive correlations were showed between yield and yield componets such as pod weight per plant, 100 grain weight and the number of grains per plant in all varietal groups except, Virginia Runner. However, the other characters were almost not correlated with yield and differences in correlation coefficients among the seeding dates were found. 13. Path coefficients estimated for yield components to yield was higher in number of grains per plant pod weight per plant and 100 grain weight in terms of direct effect and the other components were negligible in all varietal groups. 14. Heritabilities estimated were generally high in pod number per plant, shelling ratio, 100 grain weights and number of grains per pod and the other traits were relatively low.

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Protective Effect of Plantago asiatica L. Leaf Ethanolic Extract Against Ferric Nitrilotriacetate-Induced Prostate Oxidative Damage in Rats (랫드에서의 Fe-NTA 유발 산화스트레스에 대한 차전초 에탄올 추출물의 전립선보호 효과)

  • Hong, Seung-Taek;Hong, Chung-Oui;Nam, Mi-Hyun;Ma, Yuan-Yuan;Hong, Yun-Jin;Son, Da-Hee;Chun, Su-Hyun;Lee, Kwang-Won
    • Journal of Food Hygiene and Safety
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    • v.26 no.3
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    • pp.260-265
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    • 2011
  • Plantago asiatica L. (P. asiatica) has been used as one of the popular folk medicines in Asia for human health care practices. Various activities of P. asiatica have been reported, such as anti-oxidant, anti-glycation, anti-inflammatory and hepatoprotective activity. Therefore, the potential of P. asiatica to reduce oxidative stress has been studied in several ways for over 20 years, especially at liver and kidney. However no investigation has been reported revealing its protective effect on prostate. Method: Treatment of P. asiatica leaf ethanolic extract (PLE) (1 g/kg body weight (b.w.), 2 g/kg b.w., or 4 g/kg b.w.) were given separately to animals for pretreatment once per day for 7 days, and on the seventh day ferric nitrilotriacetate (Fe-NTA; 0.24 mmol Fe/kg b.w.), which is known as an oxidative stress-inducer at prostate, was administrated by i.p to negative control group. At the end of the study period, dissection was carried out for detecting the prostate protective effect of PLE. Result: Fe-NTA-treated animals produced reactive oxygen species (ROS) resulting in depletion of antioxidant biomaker, such as glutathione (GSH), glutathione reductase (GR), and glutathione s-transferase (GST) and increase of lipid peroxidation in prostate. However, PLE pretreatment resulted in an increase in the GSH, GST and GR levels concentration dependent manner and in an significant decrease in the levels of lipid peroxidation. Conclusion: Our data suggest that PLE may be effective in protecting oxidative stress-induced damage of prostate, and PLE may be an chemopreventive agent against Fe-NTA-mediated prostate oxidative damage.

A Report on the Shigella Cultures Isolated in Korea(1971) (1971년(年) 한국(韓國)에서 분리(分離)한 이질균(痢疾菌)에 관(關)한 보고(報告))

  • Ryu, Young-Hat;Kim, Soon-Hee
    • The Journal of the Korean Society for Microbiology
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    • v.7 no.1
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    • pp.9-15
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    • 1972
  • The authors identified 45 Shigella cultures among 63 suspectable cultures obtained from the cases showing dysentery or the like symptoms in various parts of the country during the period from February to September, 1971. Of 45 cultures, 36 cultures belonged to Subgroup B, 7 cultures to Subgroup C and 2 cultures to Subgroup D. There was none of cultures belonging to Subgroup A in 1971, although the authors detected one culture of that among 41 identified Shigella in 1967. Of 36 cultures belonging to Subgroup B, 27 cultures were $B_{2a}$ and 3 cultures of each $B_{3c},\;B_{4a}$ and $B_y$ were serotyped, respectively. Of 7 cultures beloning to Subgroup C, one was $C_{11}$ and other six cultures were $C_{15}$ and all S. boydii were isolated in Seoul area. It might be possible to suggest that there was a tendency of decreasing prevalence of S. dysenteriae and of increasing prevalence of S. boydii in Korea, although the number of cultures. tested were very much limited to conclude, if it was compared with the results obtained by the authors in 1967 that the fourty-one identified Shigella were composed of one culture of S. dysenteriae, 36 cultures of S. flexneri, 4 cultures of S. sonnei and none of S. boydii. According to the results obtained from the biochemical tests, the positive ratio of Indol tests in Subgroup B was 31/36, which could be higher than 9/36 observed in 1961, but which was nearly correlated with the result published by Ewing and his colleagues in U.S.A. The positive rates of both sorbitol and raffinose were lower than that observed by Ewing and his colleagues in S. flexneri. Regarding with the sensitivity of Shigella cultures to the antibiotics being widely used in the country, 46.2% were sensitive to chloramphenicol, which was very much higher than that observed by Park, and 85.8% were sensitive to ampicillin, which would be the drug of choice according to the results from the In Vitro tests.

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Studies on the Changes of N-Compounds during the Fermentation Process of the Korean Daenjang (한국된장의 발효과정(醱酵過程)에 따른 N-Compounds의 소장(消長)에 관(關)한 연구(硏究))

  • Yoon, Il-Sup;Kim, Hyun-Oh;Youn, Se-Eok;Lee, Kap-Sang
    • Korean Journal of Food Science and Technology
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    • v.9 no.2
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    • pp.131-137
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    • 1977
  • This study was conducted to examine the changes of pH, proteolytic enzymic activity, and every kind of nitrogen compounds during their fermentation of the three groups of meju for 90days. Among the three groups, the first group was conventional Korean meju which was proved to be good quality (sample J), the second group was prepared with soybean paste using B. subtilis (sample K), and the third group was an improved meju which was fermented with the soybean and wheat (7 : 3) mixtured paste with Asp. sojae (sample L). These groups were analyzed at an interval of 10 day and the results are summarized as follows: 1) The pH of the all three groups was lowered from $6.45{\sim}6.75\;to\;4.85{\sim}5.20$ in just the 30 days and maintained the weak acidity during this fermentation. 2) The proteolytic enzymic activity was increased as soon as the three groups of meju were fermented and marked the maximum value in 30 days. The maximum value of the three groups of meju J.K. and L was 147, 112, and 52 respectively. The proteolytic enzymic activity of sample J and K was decreased to 23.5 and 20.5 in 20 days, while that of sample L was decreased to 18.0 in 40 days, and maintained the volues to the end of fermentation for 90 days. The conventional meju J and the improved meju K showed sparkling activity at the pH 7, while the activity of improved meju L was strong at the pH 10. 3) The PAA-N content of sample J and K was increased and reached to the peak point with 1.55% and 1.49% respectively in 60 days. But the sample L marked the maximum value with 1.28% after 80 days. 4) The amino-N content of sample J was increased and reached to 0.36% after 60 days, and that of sample K and L was increased and reached to 0.29% and 0.21% respectively after 40 days. After reaching to the peak point, the contents were decreased. 5) The content of ammonia-N was most abundant in sample K which was fermented with soybean paste using B. subtilis. 6) The peptide-N content of sample K and L was increased after decreasing in the middle of fermentation period, while that of sample J was increased gradually during fermentation. 7) The changes of nitrogen compounds were seemed to complete in 60 days of fermentation.

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Effects of Application of Fermented Swine Manure with Additional Nitrogen Fertilizer on Productivity of Corn and Leaching of Nitrogen and Phosphorous in Corn Cultivation Soil (돈분발효 퇴액비 시용과 첨가적인 질소비료 시용이 옥수수의 생산성과 질소 및 인의 용탈에 미치는 영향)

  • Choi, Ki-Choon;Jo, Nam-Chul;Jung, Min-Woong;Yook, Wan-Bang
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.30 no.1
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    • pp.15-24
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    • 2010
  • This study was conducted to investigate the effects of swine manure (SM) application with additional nitrogen (N) fertilizer on productivity of corn and environmental pollution in com cultivation soil. The experiment was conformed in lysimeter which was constructed with 30 cm diameter, and 100 cm height. Swine manures used in this study were the compost of swine manure fermented with sawdust (SMFWS) and soft rice hulls treated with high temperature and high pressure (SRH), and swine slurry (SS). The application rates of the additional N, as urea, with swine manure were 50 and 100 kgN/ha on each plot. This study was arranged in completely randomized design with three replication. DM yields in SM treatments with mineral N were increased significantly compared to those in SM treatment without mineral N (zero-mineral N) (P<0.05) and increased as the rate of mineral N application increased (P<0.05). DM yields in SMFWS and SS treatments with mineral 100 kgN/ha showed trends similar to those of chemical fertilizer (control) but higher than those of 50 kg N/ha. DM yields tended to be higher in SMFWS and SS treatments than in SRH treatment. Total N contents in SMFWS, SRH and SS treatments with mineral N were increased, compared with SM treatment without mineral N. N contents in SM treatments with mineral N were lower than those of chemical fertilizer treatment, but N content of chemical fertilizer treatment showed trends similar to that of SS treatments with mineral 100 kgN/ha. $NO_3$-N concentration in SM treatments with mineral N were increased significantly, compared to those in SM treatment without mineral N and in chemical fertilizer (P<0.05). $NH_4$-N concentrations in SMFWS and SS treatments with mineral 100 kgN/ha showed trends similar to those of chemical fertilizer, but higher than those of 50 kg N/ha. $PO_4$-P concentration in SM treatments with mineral N were increased significantly, compared to those in SM treatment without mineral N (P<0.05). $PO_4$-P concentration in chemical fertilizer treatment showed trends similar to that of SS treatments with mineral 100 kgN/ha. The concentrations of $NO_3$-N $NH_4$-N and $PO_4$-P increased as the rate of mineral N application increased (P<0.05). The concentrations of $NO_3$-N $NH_4$-N and $PO_4$-P were highly elevated in the concentrated rainy season in the early stage among experimental period. The maximum $NO_3$-N $NH_4$-N and $PO_4$-P concentrations in the leaching water were 3.46 mg/L, 1.11 mg/L and 0.14 mg/L, respectively.

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.