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Quantification of Odorants from Animal Husbandry using Solid-phase Microextraction (고상(固相) 미세 추출법에 의한 축산 관리시설에서 발생하는 악취성 가스 화합물의 정량적 평가)

  • Kim, Jae-Hyuck;Choi, Hong-Lim;Kown, So-Young;Lim, Hong-Lae;McConnell, Laura L.;Arispe, Susana;Park, Chul-Hwi;Kim, Hyun-Ook
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.2
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    • pp.158-164
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    • 2006
  • Offensive odor from CAFO(concentrated animal feeding operation) and its control have become a significant issue in Korea. Control of odors from the CAFO requires to identify major odorant and their generation mechanisms. In this study, an easy method to collect gas sample and to quantify its odorants is proposed. The method involves on-site odorant extraction with solid-phase microextraction and quantitation with GC/MSD or GC/FID. Analytes of the current study include: trimethylamine(TMA), carbon disulfide($CS_2$), dimethyl sulfide(DMS), dimethyl disulfide(DMDS), acetic acid(AA), propionic acid(PA) and n-butyric acid(BA). The resulting linearity($R^2$) of calibration curve for each analyte was good over the range from several ppbv to ppmv; 0.984 for TMA(0.056-1.437), 0.996 for $CS_2$(0.039-0.999), 0.994 for DMS(0.029-0.756), 0.995 for DMDS(0.024-0.623), 0.992 for AA(0.068-1.314), 0.955 for PA(0.047-0.940), and 0.976 for BA(0.036-0.712). Method detection limits were 5.67, 6.39, 5.78, 25.2, 0.098, 0.363 and 0.099 ppbv for AA, PA, BA, TMA, DMS, $CS_2$, and DMDS, respectively. With the developed method, odorants from poultry, swine, and cattle barns were analysed. All the compounds but DMDS were detected from the sample collected in the poultry barn, and their levels exceeded the representative published human olfactory threshold.

Studies on the Petroleum hydrocarbon-utilizing Microorganisms(Part 1) -On the Production of Protein from the Yeast-cell- (석유(탄화수소) 이용미생물에 관한 연구(제 1보) -효모세포에 의한 석유로부터 단백질 생성에 관하여-)

  • Lee, Ke-Ho;Shin, Hyun-Kyung
    • Applied Biological Chemistry
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    • v.13 no.1
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    • pp.43-50
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    • 1970
  • To study the productivity of single cell protein from the petroleum hydrocarbon utilizing yeasts, 242 soil samples, such as oil soaked soil of gas stations and garage, coal, farm soil, and sewage, from 135 places in Korea were collected. From these samples 468 yeast strains which utilize petroleum hydrocarbon as a sole organic carbon source were isolated and identified by observing the growth rates. For the identified strains optimum culture conditions were determined and analysis of cell components were performed. 1. 90.8% of petroleum hydrocarbon utilizing yeast strains were found from oil soaked soil and about 10% from coal, farm soil and sewage etc. 2. The yeast strain of the highest cell productivity was isolated from oil soaked soil and was identified as Candida curvata HY-69-19. 3. The optimum culture conditions for the selected yeast strain were found to be pH 5.0, $28^{\circ}C$ and affluent aerated state. 4. Candida curvata HY-69-19 was found to utilize favorably the heavy gas oil fractionated at above $268.9^{\circ}C$ as carbon source and urea as inorganic nitrogen source. 5. The growth curve of this strain on heavy gas oil medium showed that the yeast has a lag phase up to 18 hours and logarithmic growth phase between 24 to 42 hours. Generation time was found to be between 3.8 and 4.5 hours during the logarithmic growth phase. 6. About 300 mg dried cells per heavy gas oil was harvested under the culture conditions of adjusted pH to 5.0 at time intervals of 6 hours for 54 hours and heavy gas oil urea for shaking culture medium. 7. Chemical composition of the yeast cell was found to be 40.25%, 14.81%, 24.32% and 10.63% for crude protein, crude lipid, carbohydrate and ashes, respectively.

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Simultaneous analysis for 2-thiothiazolidine-4-carboxilic acid and thiocarbamide using butanol extraction method (부타놀 추출법을 이용한 2-thiothiazolidine-4-carboxilic acid와 thiocarbamide의 동시정량에 관한 연구)

  • Lee, Sanghoi;Song, Jaesok;Yoon, youngshik;Kim, Chinyon;Won, Jonguk;Roh, Jaehoon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.10 no.1
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    • pp.208-222
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    • 2000
  • This study was conducted to supplement limit of previous study, The objectives of this study were to select optimal conditions of high performance liquid chromatography(HPLC) operation for detecting urinary 2-thiothiazolicline-4-carboxylic acid(TTCA) and thiocarbamide simultaneously, and to evaluate recovery rates for various liquid-liquid extration method of these metabolites, The results are as follows : 1. The urinary TTCA and thiocarbamide were separate sharply when flow rate is $0.7m{\ell}/min$, using a series $C_8$ and $C_{18}$ column, 50 mM $KH_2PO_4$ : acetonitrile (93.5 : 6.5) and pH 3.5 as a mobile phase. The retention time was TTCA, $12.07{\pm}0.11$(mean${\pm}$SD, n=06), thiocarbamide, $7.85{\pm}0.01$ (mean${\pm}$SD, n=6), respectively. The calibration curve for TTCA and thiocarbamide was linear within the range 0.05 to $30{\mu}g/m{\ell}$. 2. By the liquid-liquid extration, butanol extration with $(NH_4)_2$ as a salting-out reagent was used as a simultaneous extration method for these metabolites in acid state, and recovery rates of this method are urinary TTCA, $49.6{\pm}17.7$ (mean${\pm}$SD, n=16), thiocarbamide, $43,9{\pm}5.50$ (mean${\pm}$SD, n=16), respectively 3. The precision(pooled coefficients of variation for 4 concentration) of the urinary thiocarbamide analysis was 0.03754 by butanol liquid-liquid extraction with $(NH_4)_2$ as a salting-out reagent, and TTCA was 0.04082 by ethyl acetate liquid-liquid extration with $(NH_4)_2$ as a salting out reagent The above results show that the butanol liquid-liquid extraction with $(NH_4)_2$ as a salting-out reagent in acid state, and using a series $C_8$ and $C_{18}$ column, 50 mM $KH_2PO_4$ : acetonitrile (93.5 : 6.5) and pH 3.5 as a mobile phase are suitable for the analysis of urinary TTCA and thiocarbamide simultaneously. The detection limit of TTCA and thiocarbamide was about $0.17{\mu}g/m{\ell}$, $0.07{\mu}g/m{\ell}$.

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Analytical Determination of Vitamin B12 Content in Infant and Toddler Milk Formulas by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS)

  • Lee, Jung-Hoon;Shin, Jin-Ho;Park, Jung-Min;Kim, Ha-Jung;Ahn, Jang-Hyuk;Kwak, Byung-Man;Kim, Jin-Man
    • Food Science of Animal Resources
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    • v.35 no.6
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    • pp.765-771
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    • 2015
  • The development of a sample preparation method and optimization of the analytical instrumentation conditions were performed for the determination of the vitamin B12 content in emulsified baby foods sold on the Korea market. After removal of the milk protein and fats by chloroform extraction and centrifugation, the vitamin B12 was water extracted from the sample. Following filtration of the solution through a nylon filter, the water-soluble extract was purified by solid-phase extraction using a Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). The solution eluted from the cartridge was dried under a stream of nitrogen gas and reconstituted with 1 mL of water. The sample solution was injected into an LC-MS/MS system after optimizing the mobile phase for vitamin B12 detection. The calibration curve showed good linearity with the coefficient of correlation (r2) value of 0.9999. The limit of detection was 0.03 µg/L and the limit of quantitation was 0.1 µg/L. The method of detection limit was 0.02 µg/kg. The vitamin B12 recovery from a spiking test was 99.62% for infant formula and 99.46% for cereal-based baby food. The sample preparation method developed in this study would be appropriate for the rapid determination of the vitamin B12 content in infant formula and baby foods with emulsified milk characteristics. The ability to obtain stable results more quickly and efficiently would also allow governments to exercise a more extensive quality control inspection and monitoring of products expected to contain vitamin B12. This method could be implemented in laboratories that require time and labor saving.

Quantification of Ceruloplasmin in Wale Rats Exposed to ${\gamma}$-radiation by Enzyme Linked Immunosorbent Assay (ELISA방법에 의한 방사선 피폭 후 흰쥐 혈액 내에서의 ceruloplasmin 정량)

  • Kim, In-Gyu;Park, Seon-Young;Kim, Kug-Chan;Lee, Kang-Suk
    • Journal of Radiation Protection and Research
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    • v.22 no.2
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    • pp.103-109
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    • 1997
  • Adult male rats were exposed to a whole body with a single dose of 1.0, 2.0, 3.0, 5.0, and 7.0 Gy. The animals were sacrificed 48, 72, 96 and 216 hours following exposure. A competitive enzyme linked immunosorbent assay(ELISA) with antigen immobilized on the solid phase has been developed to measure ceruloplasmin in rat serum and complete dose response curves. Ceruloplasmin was purified from the plasma of turpentine treated male rats. Coating of ceruloplasmin had more effectiveness in 10 mM Tris-HCI, 150 mM sodium chloride, pH 7.4 than in 50 mM carbonate/bicarbonate buffer, pH 9.6. The coating range for ceruloplasmin was $70{\sim}140ng$/well. Levels of ceruloplasmin increased to maximum on the $72{\sim}96$ hours after irradiation. Slope of between response and dose was greatest value 96 hours following irradiation. Normal ceruloplasmin levels were not recorded 216 hours following exposure. In 0.1 Gy irradiated group, levels of ceruloplasmin also increased to maximum on the $72{\sim}96$ hours following irradiation. The concentration of this protein remained significantly different from control value, 196 hours after exposure. Ceruloplasmin was identified as one of the major acute phase protein following irradiation and further studies about gene expression and regulation would be necessary for radiation protection.

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Environmentally friendly determination of urinary trans, trans-muconic acid for biological monitoring of benzene exposure by green high-performance liquid chromatography (소변 중 뮤콘산의 친환경 HPLC 분석방법을 이용한 벤젠 노출 생물학적 노출평가)

  • Yang, Dong-Hyug;Lee, Mi-Young;Won, Yong Lim
    • Analytical Science and Technology
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    • v.25 no.6
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    • pp.460-466
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    • 2012
  • An environmentally friendly, so-called green, high performance liquid chromatography method was developed and validated for the determination of trans, trans-muconic acid (t,t-MA) in human urine as a biomarker of benzene exposure. After urinary t,t-MA was extracted and enriched using solid-phase extraction, a MF-Ph1 SG80 ($150mm{\times}2.0mm$ I.D., 5 ${\mu}m$) column with a mobile phase of 10 mM $KH_2PO_4$ containing 0.1% $H_3PO_4$ was used for isocratic separation of t,t-MA with UV detection at 259 nm. The calibration curve was constructed in the range of 0.1-5.0 mg/L with good linearity ($r^2$=0.9992). The intra-day and inter-day precision (as RSD) were 0.9-8.5% and 3.1-4.5%, respectively. The average recovery ranged from 97.5% to 101.7%. The green sample preparation and separation with no organic solvents were successfully achieved. The validated method would be suitable for the routine biological monitoring of benzene exposure in the occupational settings.

Studies on the chlorella in Korea (한국 Chlorella에 대한 연구)

  • 이주식;심종호
    • Korean Journal of Microbiology
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    • v.1 no.1
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    • pp.38-44
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    • 1963
  • According to the experiment on pure-isolation and the related contaminants of Chlorella, the phenomena of the ecological distributions of Chlorella in Korea have been manifested in several areas and also the aim that in going to do culture, biological and physiological study of Chlorella is carried out. Contaminants very oftenly occupied on the colony of the strains taken in order to fulfil pure-isolation of Chlorella, but in accordance with being piled up the minute research on this subject, I can obtain the desirable results as follows: 1. For the pure-isolation, the duration chose the time from May to September 1957 so that may easily isolate from contaminant water with utilizing the antibiotic substances. 2. To take long time, 36-48 hours until growth of nascent through the non-sporulated, it originates from the difference of the cultured media. In addition to the above mention, the mechanism of growth until nascent through the sporulated must not always require the ligh. However the supply of metabolic energy depend upon its nutritional conditions per phase. 3. The culture of Chlorella should be based on the lower culturing except adding especial conditions such as reagent concentration of media, artifical shake of media and other facts due to the natural conditions. And also these strains grew not only in distilled water but 2% NaCl solution without any abnormality in cell it self. I, therefore, guess it is possible to culture in sea-water under phasic environment. 4. In the experiment of ammonia detection, it is caused by the sampling surroundings to contain the minute quantity of ammonia in strain No. M 918; that is the place to be plenty of Carbohydrate on behalf of protein. 5. To compare the absorption curve of chlorophyll of higher plant with that of Chlorella, the absorption zone made mostly the Same ones each other but a little absorption grade dose not clearly appear. The colony which formed giant type grows with intensive colour and green band on surrounding of the colony and after that it was changed into all the green colour and developed up to end. 6. At first phase for a week, the development of Chlorella suspends the normal condition as in vivo but after a few days, the colour of chlorophyll gradually changed into blue-yellow which secrete the mucous substances on the agar media. The cell was flew out the contained substances itself on leaving the cell wall only, or the various micro-organism diffused on the outer-region of the cell.

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A Screening Method for Src Homology 3 Domain Binding Blockers Based on Ras Signaling Pathway

  • Ko, Woo-Suk;Yoon, Sun-Young;Kim, Jae-Won;Lee, Choong-Eun;Han, Mi-Young
    • BMB Reports
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    • v.30 no.5
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    • pp.303-307
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    • 1997
  • Grb2, which is composed of a Src homology 2 (SH2) domain and two Src homology 3 (SH3) domains, is known to serve as an adaptor protein in signaling for Ras activation. Thus, a blocker of the Grb2 interactions with other proteins can be a potential candidate for an anticancer drug. In this study, we have developed a high throughput screening method for SH3 domain binding ligands and blockers. Firstly, we made and purified the glutathione S-transferase (GST)-fusion proteins with the Grb2 SH2 and SH3 domains, and the entire Grb2. This method measures the binding of a biotin-labeled oligopeptide, derived from a Grb2/SH3 binding motif in the hSos, to the GST-fusion proteins, which are precoated as glutathione S-transferase fusion protein on a solid phase. When $1\;{\mu}g$ of each fusion protein was used to coat the wells, both N- and C- terminal SH3 the domains as well as the whole of Grb2 were able to interact with the biotin-conjugated ligand peptide, while the SH2 domain and GST alone showed no binding affinity. Although N- and C- terminal SH3 domains showed an increase of binding to the ligand peptide in proportion to the amount of peptide, the GST fusion protein with Grb2 demonstrated much higher binding affinity. GST-Grb2 coating on the solid phase showed a saturation curve; 66 and 84% of the maximal binding was observed at 100 and 300 ng/$100\;{\mu}l$, respectively. This binding assay system was peptide sequence-specific, showing a dose-dependent inhibition with the unlabeled peptide of SH3 binding motif. Several other peptides, such as SH2 domain binding motifs and PTB domain binding motif, were ineffective to inhibit the binding to the biotin-conjugated ligand peptide. These results suggest that our method may be useful to screen for new anticancer drug candidates which can block the signaling pathways mediated by SH3 domain binding.

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A Search for New Variable Stars in the Open Cluster NGC 129 using a Small Telescope (소형망원경을 이용한 산개성단 NGC 129 영역의 변광성 탐사)

  • Lee, Eun-Jung;Jeon, Young-Beom;Lee, Ho;Park, Hong-Suh
    • Journal of the Korean earth science society
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    • v.28 no.1
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    • pp.87-104
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    • 2007
  • As part of the SPVS (Short-Period Variability Survey) which is a wide-field $(90'{\times}60')$ photometric monitering program at Bohyunsan Optical Astronomy (BOAO), we performed V band time-series CCD photometric observations ofthe young open cluster NGC 129 for 11 nights between October 12, 2004 and November 3, 2005 using the 155mm refractor equipped with $3K{\times}2K$ CCD camera. From the observation we obtained 2400 V band CCD frames and color-magnitude diagram of the cluster. To transform instrumental magnitude to standard magnitude, we applied ensemble normalization technique to all observed time-series data. After the photometric reduction process, we examined variations of 9537 stars. As a result, sixty six of the new variable stars were discovered. To determine the periods of the sevariables, we used DFT(Discrete Fourier Transform) and phase-matching technique. According to light curve shape, period, amplitude and the position on a C-M diagram, we classified these variables as 9 SPB type, 9 ${\delta}$ Scuti type, 29 eclipsing, 17 long term variables. However, two of them were not classified. From this study, we learned that small telescopes could be a very useful tool to observe variable stars in the open cluster in survey program.

Isolation and Characterization of Bacillus subtilis CA105 from Spent Mushroom (Pleurotus ostreatus) Substrates (느타리버섯 수확후배지로부터 분리한 Bacillus subtilis CA105의 특성)

  • Kim, Hye Soo;Kim, Chul Hwan;Kwon, Hyun Sook;Lee, Chan-Jung;Kong, Won-Sik;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.4
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    • pp.305-309
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    • 2015
  • In order to isolate compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated CA105 was selected by agar diffusion method. The strain CA105 was identified as members of the Bacillus subtilis by biochemical characteristics using VITEK 2 system. Comparative 16S rRNA gene sequence analysis showed that isolate CA105 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rRNA gene sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, isolate CA105 was classified within the genus Bacillus subtilis, for which the name Bacillus subtilis CA105 is proposed. The cellulase and xylanase activity of B. subtilis CA105 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for Bacillus sp. CA105.