• Title/Summary/Keyword: isotope dilution

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Dynamic MRM Measurements of Multi-Biomarker Proteins by Triple-Quadrupole Mass Spectrometry with Nanoflow HPLC-Microfluidics Chip

  • Ji, Eun-Sun;Cheon, Mi-Hee;Lee, Ju-Yeon;Yoo, Jong-Shin;Jung, Hyun-Jin;Kim, Jin-Young
    • Mass Spectrometry Letters
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    • v.1 no.1
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    • pp.21-24
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    • 2010
  • The development of clinical biomarkers involves discovery, verification, and validation. Recently, multiple reaction monitoring (MRM) coupled with stable isotope dilution mass spectrometry (IDMS) has shown considerable promise for the direct quantification of proteins in clinical samples. In particular, multiple biomarkers have been tracked in a single experiment using MRM-based MS approaches combined with liquid chromatography. We report here a highly reproducible, quantitative, and dynamic MRM system for validating multi-biomarker proteins using Nanoflow HPLC-Microfluidics Chip/Triple-Quadrupole MS. In this system, transitions were acquired only during the retention window of each eluting peptide. Transitions with the highest MRM-MS intensities for the five target peptides from colon cancer biomarker candidates were automatically selected using Optimizer software. Relative to the corresponding non-dynamic system, the dynamic MRM provided significantly improved coefficients of variation in experiments with large numbers of transitions. Linear responses were obtained with concentrations ranging from fmol to pmol for five target peptides.

Effect of N Application Rate on Fixation and Transfer from Vetch to Barley in Mixed Stands. (질소시용수준이 베치-보리 혼파 사초의 질소고정 및 베치에서 보리로 질소이동에 미치는 영향)

  • Lee Hyo Won;Kim Won Ho;Park Hyung Soo;Ko Han Jong;Kim Su Gon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.1
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    • pp.1-6
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    • 2005
  • With recent interest organic farming the use of legumes including vetch and clover to provide N to adjacent crops is increasing in Korea. In the present studies, we conducted a trial to investigate the effects of the application of N rate on nitrogen fixation and transfer from vetch to barley in mixed stands. The experiment was arranged in a randomized complete block design with three replications. Four different N rates(0, 75, 113 and 150/ha) was used and vetch+barley was broadcasted manually on 1.5 $\times$2 m plot in Oct. 2001. Half of urea and K$_{2}O, 200 Phosphate and 75 kg potash per ha were applied as basal dressing md half of N md 75 potash were used for topdressing to soil surface on MarctL 2002. The equivalent of 1kg ha$^{-1}$ at($^{15}$NH$_{4}$)$_{2}$SO$_{4}$ solution at 99.8 atom $\%$$^{15}$N excess was applied to the microplot in mid April. Forage was harvested from each plot at ground level and separated into barley and vetch. Total N content and It values of samples were determined using a continuous flow stable isotope ratio mass spectrometry(IsoPrime-EA. Micromass, UK.). The percentage of legume H fixed from atmospheric N2 were 95.0, 93.8, 94.4 and $84.8\%$ with increment of N levels. The percentage of N transfer from vetch to barley by N-difference method with increment of N fertilizer were from 58 to$49\%$ while 39 to $23\%$ in $^{15}$N-dilution method. The amount of transfer from vetch to barley were 87 to 68 kg/ ha with N level by N-difference moth여 and 58 to -56/ha with N application levels by $^{15}$N dilution method. The amount of nitrogen fixation per ha were from 150 kg / ha to 219 kg / ha by different method, but on the other side 49 to 105kg/ha by N$^{15}$-dilution.

Geochemical and Isotopic Study of the Kumho River (금호강 하천수의 지구화학 및 동위원소 연구)

  • Kim, Yeong-Kyoo;Nam, Eun-Kyung
    • Economic and Environmental Geology
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    • v.42 no.6
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    • pp.527-539
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    • 2009
  • The Kumho River flows through volcanic and sedimentary rocks at upstream and downstream regions and also through industrial district including dyeing complex before it meets the Nakdong River, and as a result, many factors can influence the geochemistry of river water. The concentrations of dissolved ions generally increased as it flows downstream. The concentrations of cations are in the order of Ca>Na>Mg>K, and those of anions are $HCO_3$>$SO_4$>Cl>$NO_3$. These results show that the weathering of sandstone and shale containing carbonate including calcite caused the enrichment of Ca and $HCO_3$. At first 4 sampling sites, Si contents are relatively high mainly due to the weathering of silicate minerals of volcanic rocks. However, Na and $SO_4$ contents are higher at downstream sites due to the industrial and municipal sewage. Piper diagram also shows that the geochemical patterns changed from Ca-$HCO_3$ to Ca-Cl/Ca-$SO_4$ and Na-Cl/Na-$SO_4$ type. When comparing the samples collected in May and July, the concentrations of dissolved ions in July are generally lower than those in May, which indicates that dilution by precipitation played an important role. In July the relative concentration of Ca increased, indicating that Ca in soils probably from fertilizer were mixed into the river water by precipitation. The river waters are mainly from precipitation. The dissolved ions are mainly from weathering of carbonate minerals and pollutants from municipal sewage and discharged water from industrial complex. The composition of oxygen and deutrium isotope in July showed higher values, which is contrary to the amount effect, maybe due to Youngchon Dam. The nitrogen isotope showed lower values in July than those in May, which can be interpreted to indicate mixing of nitrate from soils and fertilizer in the cultivated land by the heavy rain. The isotope composition of nitrate increased downstream, indicating that the influence of sewage and animal manure also increased downstream.

The Sr and Pb Isotopic and Geochemical Properties of the Atmospheric Bulk Deposition of Jeonju, Gunsan, and Namweon Areas (전주, 군산, 남원지역 강수의 Sr, Pb동위원소 지화학)

  • Jeon Seo-Ryeong;Chung Jae-il
    • Economic and Environmental Geology
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    • v.38 no.4 s.173
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    • pp.463-479
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    • 2005
  • The Sr and Pb isotopic ratios and chemical composition were measured for atmospheric bulk deposition samples collected in the Jeonju, Gunsan and Namweon areas over a period of one year. Acidity of deposition ranged pH $4\~7$ with little higher in dry season, and around pH 5.0 in rainy season. The EC and TDS of rainy season was low showing dilution effect, and increased during dry season. Sulfate $(SO_4)\;and\;NO_3$ are atmospheric aerosols largely of anthropogenic origin in winter. Sodium was concentrated in winter deposition, Ca was concentrated in spring to summer deposition. Namweon has lower EC and TDS than those of other, and Jeonju has higher. Namweon was concentrated in $HCO_3$ and Cunsan was concentrated in Cl. Aluminium, Cu, and Zn show good correlation index with TDS, indicating of their origin atmospheric. $^{87}Sr/^{86}Sr$ ratios of bulk deposition ranged from 0.7109 to 0.7128. The isotopic variations are correlated with mixing of isotopic compositions of local soils, road deposit and biogenic aerosol. In order to constrain further the origin of aerosols in rainwater, it will be necessary to collect additional Sr isotopic data for aerosols. Lead isotope ratios for all areas were similar and belonged to Pb isotope ratios of Seoul's aerosols, but little different with Beijing's aerosols. It showing that Pb in the Korea mainly derived from the gasoline combustion, not exclusively from the Beijing.

Effects of Non-protein Energy Intake on Whole Body Protein Synthesis, Nitrogen Retention and Glucose Turnover in Goats

  • Fujita, Tadahisa;Kajita, Masahiro;Sano, Hiroaki
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.536-542
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    • 2007
  • The responses of whole body protein and glucose kinetics and of nitrogen (N) metabolism to non-protein energy intake (NPEI) were determined using an isotope dilution approach and measurement of N balance in three adult male goats. The diets containing 1.0, 1.5 and 2.0 times ME maintenance requirement, with fixed intake of CP (1.5 times maintenance) and percentage of hay (33%), were fed twice daily for each 21 d experimental period. After an adaptation period of 11 d, N balance was determined over 3 d. On day 17, whole body protein synthesis (WBPS) and glucose irreversible loss rate (ILR) were determined during the absorptive state by a primed-continuous infusion of [$^2H_5$]phenylalanine, [$^2H_2$]tyrosine, [$^2H_4$]tyrosine and [$^{13}C_6$]glucose, with simultaneous measurements of plasma concentrations of metabolites and insulin. Ruminal characteristics were also measured at 6 h after feeding over 3 d. Nitrogen retention tended to increase (p<0.10) with increasing NPEI, although digestible N decreased linearly (p<0.05). Increasing NPEI decreased (p<0.01) ammonia N concentration, but increased acetate (p<0.05) and propionate (p<0.05) concentrations in the rumen. Despite decreased plasma urea N concentration (p<0.01), increased plasma tyrosine concentration (p<0.05), and trends toward increased plasma total amino N (p<0.10) and phenylalanine concentrations (p<0.10) were found in response to increasing NPEI. Increasing NPEI increased ILR of both glucose (p<0.01) and phenylalanine (p<0.05), but did not affect ($p{\geq}0.10$) that of tyrosine. Whole body protein synthesis increased (p<0.05) in response to increasing NPEI, resulting from increased utilization rate for protein synthesis (p<0.05) and unchanged hydroxylation rate of phenylalanine ($p{\geq}0.10$). These results suggest that increasing NPEI may enhance WBPS and glucose turnover at the absorptive state and improve the efficiency of digestible N retention in goats, with possibly decreased ammonia and increased amino acid absorption. In addition, simultaneous increases in WBPS and glucose ILR suggest stimulatory effect of glucose availability on WBPS, especially when sufficient amino acid is supplied.

Lead-Zinc-Tin-Silver Mineralization of Tangguanpu Mine, Hunan Province, China: Fluid Inclusion and Sulfur Isotope Studies (중국 호남성 당관포 광산의 연-아연-주석-은 광화작용: 유체포유물 및 황동위원소 연구)

  • 허철호;윤성택;소칠섭
    • Economic and Environmental Geology
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    • v.34 no.2
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    • pp.157-166
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    • 2001
  • Numerous base-metal bearing hydrothermal quartz vein deposits occur in the Hunan province of southern China. The Tangguanpu lead-zinc-tin-silver mine is the major producer among these deposits. Lead-zinc-tin-silver mineralization occurs in a single stage of massive quartz veins which filled fractures in fault zones within Paleozoic metasedimentary rocks. Sphalerite, chalcopyrite, galena, pyrite, arsenopyrite and pyrrhotite are the principal sulphide minerals in the Tangguanpu lead-zinc ores with minor amounts of tin- and antimony-bearing sulphides (stannite, teallite, boulangerite and tetrahedrite). Based on the iron and zinc partitioning between coexisting stannite and sphalerite, the formation temperature for this mineral assemblage range from 300$^{\circ}$ to 330$^{\circ}$C, which relatively agree with the upper part of homogenization temperature of fluid inclusion in quartz (20T-358$^{\circ}$C). Fluid inclusion data show that main lead-zine-tin-silver mineralization occurred from $H_{2}O$-NaCl fluids with relatively low salinities (11.2-7.3 wl.% eg. NaCI) at temperatures between 207$^{\circ}$ and 358$^{\circ}$C. The relationship between homogenization temperature and salinity suggests a history of cooling and dilution followed by initial boiling. Evidence of initial fluid boiling may indicate the fluid trapping pressures of 180 bars. The ${\delta}^{34}S{{\Sigma}S}$ values of -5.0 to 1.1 %, indicate an igneous source of sulfur in the Tangguanpu lead-zinc-tin-silver hydrothermal fluids.

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Evolution and Origin of the Geothermal Waters in the Busan Area, Korea: 1. Cooling and Dilution by Groundwater Mixing after Heated Seawater-Rock Interaction (부산지역 지열수의 기원과 진화: 1.가열된 해수-암석 반응과 지하수의 혼입에 따른 희석과 냉각)

  • 성규열;박맹언;고용권;김천수
    • Economic and Environmental Geology
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    • v.34 no.5
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    • pp.447-460
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    • 2001
  • The geothermal waters from the Busan area belong to Na-CI type and are characterized by much higher EC (921 ~6,520${\mu}$S/cm) and TDS (608-3,390 mg/L) than other geothermal waters in Korea. The concentration of majorions shows a weakly positive relationship with temperature except for Mg ion. The concentrations of the major cat ions have the order of Na>Ca>K>Mg. Ca ion is enriched and Mg ion is depleted compared with seawater. All Br concentrations of geothermal water are lower than those of seawater, showing a positive relationship with temperature. Generally geochemical characteristics of geothermal waters of the Busan area indicate that these waters have relatively increased Ca and Sr contents and depleted Mg, Na and K contents caused by seawater interaction with wall rock at depth during deep circulation of seawater. Base on the relationship between major ions and temperature, saline geothermal waters are diluted and are cooled by mixing of groundwaters during ascent. Isotope study and reaction path modeling of the overall geochemical system are required in order to better quantify the evolution and origin of geothermal waters in the Busan area.

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Preliminary study on the use of near infrared spectroscopy for determination of plasma deuterium oxide in dairy cattle

  • Purnomoadi, Agung;Nonaka, Itoko;Higuchi, Kouji;Enishi, Osamu;Amari, Masahiro;Terada, Fuminori
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.4101-4101
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    • 2001
  • Information of body composition (fat and protein) in living animal is important to determine the nutrients requirement. Deuterium oxide (D2O) dilution techniques, as one of isotope dilution techniques have been useful for the prediction of body composition. However, the determination of D2O concentration is time consuming and complicated. Therefore this study was conducted to develop a new method to predict D2O concentration in plasma using near infrared spectroscopy technique (NIRS). Four dairy cows in early lactation were used. They were fed total mixed ration containing conr silage, timothy hay, and concentrates to make 17.0%CP and 14.0 MJDE/kgDM. Dosing D2O was at week 1,3 and 5 after parturition. After dosing D2O, the blood was collected from hour 0 to 72. Blood samples were then centrifuge at 3,000 rpm for 10 minutes to obtain plasma. D2O concentration was analyzed by gas chromatograph (deuterium oxide analyzable system, HK102, Shokotsusyou) after extracted from plasma by liophilization. Plasma sample was scanned by NIRS using Pacific Scientific (Neotec) model 6500 (Perstorp Analytical, Silver Spring, MD) in the range of wavelength from 1100 to 2500 nm. Calibration equation was developed using multiple linear regression. Sample from one animal (cow #550; n: 74) was used for developing the calibration while the rest three animals were used for validating the equation. The range, R and SEC of the calibration set samples were 135-925 ppm, 0.93 and 48.1 ppm, respectively. Validation of the calibration equation for three individual cows was done and the average of NIR predicted value of D2O at each collection time from three weeks injection showed a high correlation. The range, r and 53 of plasma from cow #474 were 322-840 ppm,0.93 and 53.1; cow #478 were 146-951 ppm,0.95 and 39.8; cow #942 were 313-885 ppm,0.95 and 37.2, respectively. Judgement of accuracy based on ratio of standard deviation and standard error in validation set samples (RPD) for cow #474, #478 and #942 were 2.2,4.3 and 3.4, respectively. The error in application due to the variation between individual was considered smaller than the bias from collection period, however, this prediction can be overcome with correction of standard zero-minute concentration of blood. The results of this preliminary study on the use of NIRS for determination of D2O in plasma showed very promising as shown by a convenient and satisfy accuracy. Further study on various physiological stage of animal should be done.

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Effect of feeding garlic leaves on rumen fermentation, methane emission, plasma glucose kinetics, and nitrogen utilization in sheep

  • Panthee, Arvinda;Matsuno, Ayana;Al-Mamun, Mohammad;Sano, Hiroaki
    • Journal of Animal Science and Technology
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    • v.59 no.6
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    • pp.14.1-14.9
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    • 2017
  • Background: Garlic and its constituents are reported to have been effective in reducing methane emission and also influence glucose metabolism in body; however, studies in ruminants using garlic leaves are scarce. Garlic leaves contain similar compounds as garlic bulbs, but are discarded in field after garlic bulb harvest. We speculate that feeding garlic leaves might show similar effect as garlic constituents in sheep and could be potential animal feed supplement. Thus, we examined the effect of freeze dried garlic leaves (FDGL) on rumen fermentation, methane emission, plasma glucose kinetics and nitrogen utilization in sheep. Methods: Six sheep were fed Control diet (mixed hay and concentrate (60:40)) or FDGL diet (Control diet supplemented with FDGL at 2.5 g/kg $BW^{0.75}$ of sheep) using a crossover design. Methane gas emission was measured using open-circuit respiratory chamber. Plasma glucose turnover rate was measured using isotope dilution technique of [$U-^{13}C$]glucose. Rumen fluid, feces and urine were collected to measure rumen fermentation characteristics and nitrogen utilization. Result: No significant difference in rumen fermentation parameters was noticed except for rumen ammonia tended to be higher (0.05 < P < 0.1) in FDGL diet. Methane emission per kg dry matter ingested and methane emission per kg dry matter digested were lower (P < 0.05) in FDGL diet. Plasma glucose concentration was similar between diets and plasma glucose turnover rate tended to be higher in FDGL diet (0.05 < P < 0.1). Nitrogen retention was higher (P < 0.05) and microbial nitrogen supply tended to be higher (0.05 < P < 0.1) in FDGL diet. Conclusion: FDGL diet did not impair rumen fermentation, improved nitrogen retention; while absence of significant results in reduction of methane emission, glucose turnover rate and microbial nitrogen supply, further studies at higher dose would be necessary to conclude the merit of FDGL as supplement in ruminant feedstuff.

Analysis of Polybrominated Diphenyl Ethers (PBDEs) in Sediments from the Southeastern Coastal Areas of Korea

  • Choi, Hee-Gu;Kim, Sang-Soo;Jeong, Seung-Ryul;Lee, Pil-Yong;Moon, Hyo-Bang
    • Journal of the korean society of oceanography
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    • v.36 no.4
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    • pp.101-108
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    • 2001
  • Polybrominated diphenylether (PBDEs) congeners, 2,4,4'-TrBDE, 2,2',4,4'-TeBDE, 2,2',4,4',5-PeBDE, 2,2',4,4',5,5'-HXBDE, and 2,2',4,4',5,6-HxBDE, were measured in sediments from 52 stations in the southeastern coastal areas of Korea. Sediment samples were analyzed using gas chromatography coupled to mass spectrometer detector (GC/MSD) with positive electron impact (PEI) mode. New analytical methodology for PBDEs by the isotope dilution method was established using a multi-layer silica gel column chromatography. Total PBDEs levels in sediments for Pohang coast ranged from 1,1 to 33.8 ng/g dry weight (mean 5.3 ng/g dry), from 1.6 to 36.4 ng/g dry weight (mean 5.7 ng/g dry) for Ulsan coast, from 0.8 to 20.3 ng/g dry weight (mean 4.9 ng/g dry) for Busan coast, and from 0.8 to 10.3 ng/g dry weight (mean 4.4 ng/g dry) for Jinhae Bay. PBDEs contamination in surface sediments from Korean southeastern coasts was relatively moderate in comparison to that of other marine environment in the world. The predominant PBDE congeners were 2,2',4,4'-TeBDE and 2,2',4,4',5-PeBDE.

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