• Title/Summary/Keyword: $L_3$ phase

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Functional Analysis of an Antibiotic Regulatory Gene, afsR2 in S. lividans through DNA microarray System (DNA 마이크로어레이 시스템 분석을 통한 S. lividans 유래 항생제 조절유전자 afsR2 기능 분석)

  • Kim, Chang-Young;Noh, Jun-Hee;Lee, Han-Na;Kim, Eung-Soo
    • KSBB Journal
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    • v.24 no.3
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    • pp.259-266
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    • 2009
  • AfsR2 in Streptomyces lividans, a 63-amino acid protein with limited sequence homology to Streptomyces sigma factors, has been known for a global regulatory protein stimulating multiple antibiotic biosynthetic pathways. Although the detailed regulatory mechanism of AfsK-AfsR-AfsR2 system has been well characterized, very little information about the AfsR2-dependent down-stream regulatory genes were characterized. Recently, the null mutant of afsS in S. coelicolor (the identical ortholog of afsR2) has been characterized through DNA microarray system, revealing that afsS deletion regulated several genes involved in antibiotic biosynthesis as well as phosphate-starvation. Through comparative DNA microarray analysis of afsR2-overexpressed S. lividans, here we also identify several afsR2-dependent genes involved in phosphate starvation, morphological differentiation, and antibiotic regulation in S. lividans, confirming that the AfsR2 plays an important pleiotrophic regulatory role in Streptomyces species.

High-Level Expression of T4 Endonuclease V in Insect Cells as Biologically Active Form

  • Kang, Chang-Soo;Son, Seung-Yeol;Bang, In-Seok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1583-1590
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    • 2006
  • T4 endonuclease V (T4 endo V) [EC 3. 1. 25. 1], found in bacteriophage T4, is responsible for excision repair of damaged DNA. The enzyme possesses two activities: a cyclobutane pyrimidine dimer DNA glycosylase (CPD glycosylase) and an apyrimidic/apurinic endonuclease (AP lyase). T4 denV (414 bp cDNA) encoding T4 en do V (138 amino acid) was synthesized and expressed using either an expression vector, pTriEx-4, in E. coli or a baculovirus AcNPV vector, pBacPAK8, in insect cells. The recombinant His-Tag/T4 endo V (rHis-Tag/T4 endo V) protein expressed from bacteria was purified using one-step affinity chromatography with a HiTrap Chelating HP column and used to make rabbit anti-His-Tag/T4 endo V polyclonal antibody for detection of recombinant T4 endo V (rT4 endo V) expressed in insect cells. In the meantime, the recombinant baculovirus was obtained by cotransfection of BacPAK6 viral DNA and pBP/T4 endo V in Spodoptera frugiperda (Sf21) insect cells, and used to infect Sf21 cells to overexpress T4 endo V protein. The level of rT4 endo V protein expressed in Sf21 cells was optimized by varying the virus titers and time course of infection. The optimal expression condition was set as follows; infection of the cells at a MOI of 10 and harvest at 96 h post-infection. Under these conditions, we estimated the amount of rT4 endo V produced in the baculovirus expression vector system to be 125 mg/l. The rT4 endo V was purified to homogeneity by a rapid procedure, consisting of ion-exchange, affinity, and reversed phase chromatographies, based on FPLC. The rT4 endo V positively reacted to an antiserum made against rHis-Tag/T4 endo V and showed a residual nicking activity against CPD-containing DNA caused by UV. This is the first report to have T4 endo V expressed in an insect system to exclude the toxic effect of a bacterial expression system, retaining enzymatic activity.

In vitro rumen fermentation kinetics, metabolite production, methane and substrate degradability of polyphenol rich plant leaves and their component complete feed blocks

  • Aderao, Ganesh N.;Sahoo, A.;Bhatt, R.S.;Kumawat, P.K.;Soni, Lalit
    • Journal of Animal Science and Technology
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    • v.60 no.11
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    • pp.26.1-26.9
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    • 2018
  • Background: This experiment aimed at assessing polyphenol-rich plant biomass to use in complete feed making for the feeding of ruminants. Methods: An in vitro ruminal evaluation of complete blocks (CFB) with (Acacia nilotica, Ziziphus nummularia leaves) and without (Vigna sinensis hay) polyphenol rich plant leaves was conducted by applying Menke's in vitro gas production (IVGP) technique. A total of six substrates, viz. three forages and three CFBs were subjected to in vitro ruminal fermentation in glass syringes to assess gas and methane production, substrate degradability, and rumen fermentation metabolites. Results: Total polyphenol content (g/Kg) was 163 in A. nilotica compared to 52.5 in Z. nummularia with a contrasting difference in tannin fractions, higher hydrolysable tannins (HT) in the former (140.1 vs 2.8) and higher condensed (CT) tannins in the later (28.3 vs 7.9). The potential gas production was lower with a higher lag phase (L) in CT containing Z. nummularia and the component feed block. A. nilotica alone and as a constituent of CFB produced higher total gas but with lower methane while the partitioning factor (PF) was higher in Z. nummularia and its CFB. Substrate digestibility (both DM and OM) was lower (P < 0.001) in Z. nummularia compared to other forages and CFBs. The fermentation metabolites showed a different pattern for forages and their CFBs. The forages showed higher TCA precipitable N and lower acetate: propionate ratio in Z. nummularia while the related trend was found in CFB with V. sinensis. Total volatile fatty acid concentration was higher (P < 0.001) in A. nilotica leaves than V. sinensis hay and Z. nummularia leaves. It has implication on widening the forage resources and providing opportunity to use forage biomass rich in polyphenolic constituents in judicious proportion for reducing methane and enhancing green livestock production. Conclusion: Above all, higher substrate degradability, propionate production, lower methanogenesis in CFB with A. nilotica leaves may be considered useful. Nevertheless, CFB with Z. nummularia also proved its usefulness with higher TCA precipitable N and PF. It has implication on widening the forage resources and providing opportunity to use polyphenol-rich forage biomass for reducing methane and enhancing green livestock production.

Behavior of Implanted Dopants and Formation of Molybdenum Siliclde by Composite Sputtering (Composite target으로 증착된 Mo-silicide의 형성 및 불순물의 거동)

  • Cho, Hyun-Choon;Paek, Su-Hyon;Choi, Jin-Seog;Hwang, Yu-Sang;Kim, Ho-Suk;Kim, Dong-Won;Shim, Tae-Earn;Jung, Jae-Kyoung;Lee, Jong-Gil
    • Korean Journal of Materials Research
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    • v.2 no.5
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    • pp.375-382
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    • 1992
  • Molybdenum silicide films have been prepared by sputtering from a single composite MoS$i_2$ source on both P, B$F_2$respectively implanted (5${\times}10^{15}ions/cm^2$ single crystal and P implanted (5${\times}10^{15}ions/cm^2$) polycrystalline silicon substrates followed by rapid thermal annealing in the ambient of argon. The heat treatment temperatures have been varied in the range of 600-l20$0^{\circ}C$ for 20 seconds. The properties of Mo-silicide and the diffusion behaviors of dopant after the heat treatment are investigated using X-ray diffraction, scanning electron microscopy(SEM) , secondary ions mass spectrometry(SIMS), four-point probe and $\alpha-step.$ Annealing at 80$0^{\circ}C$ or higher resulted in conversion of the amorphous phase into predominantly MoS$i_2$and a lower sheet resistance. There was no significant out-diffusion of dopants from both single crystal and polycrystalline silicon substrate into molybdenum silicide layers during annealing.

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Prognostic Value of Sarcopenia and Myosteatosis in Patients with Resectable Pancreatic Ductal Adenocarcinoma

  • Dong Wook Kim;Hyemin Ahn;Kyung Won Kim;Seung Soo Lee;Hwa Jung Kim;Yousun Ko;Taeyong Park;Jeongjin Lee
    • Korean Journal of Radiology
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    • v.23 no.11
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    • pp.1055-1066
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    • 2022
  • Objective: The clinical relevance of myosteatosis has not been well evaluated in patients with pancreatic ductal adenocarcinoma (PDAC), although sarcopenia has been extensively researched. Therefore, we evaluated the prognostic value of muscle quality, including myosteatosis, in patients with resectable PDAC treated surgically. Materials and Methods: We retrospectively evaluated 347 patients with resectable PDAC who underwent curative surgery (mean age ± standard deviation, 63.6 ± 9.6 years; 202 male). Automatic muscle segmentation was performed on preoperative computed tomography (CT) images using an artificial intelligence program. A single axial image of the portal phase at the inferior endplate level of the L3 vertebra was used for analysis in each patient. Sarcopenia was evaluated using the skeletal muscle index, calculated as the skeletal muscle area (SMA) divided by the height squared. The mean SMA attenuation was used to evaluate myosteatosis. Diagnostic cutoff values for sarcopenia and myosteatosis were devised using the Contal and O'Quigley methods, and patients were classified according to normal (nMT), sarcopenic (sMT), myosteatotic (mMT), or combined (cMT) muscle quality types. Multivariable Cox regression analyses were conducted to assess the effects of muscle type on the overall survival (OS) and recurrence-free survival (RFS) after surgery. Results: Eighty-four (24.2%), 73 (21.0%), 75 (21.6%), and 115 (33.1%) patients were classified as having nMT, sMT, mMT, and cMT, respectively. Compared to nMT, mMT and cMT were significantly associated with poorer OS, with hazard ratios (HRs) of 1.49 (95% confidence interval, 1.00-2.22) and 1.68 (1.16-2.43), respectively, while sMT was not (HR of 1.40 [0.94-2.10]). Only mMT was significantly associated with poorer RFS, with an HR of 1.59 (1.07-2.35), while sMT and cMT were not. Conclusion: Myosteatosis was associated with poor OS and RFS in patients with resectable PDAC who underwent curative surgery.

NUWARD SMR safety approach and licensing objectives for international deployment

  • D. Francis;S. Beils
    • Nuclear Engineering and Technology
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    • v.56 no.3
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    • pp.1029-1036
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    • 2024
  • Drawing on the deep experience and understanding of the principles of nuclear safety, as well as many years of nuclear power plant design and operation, the EDF led NUWARD SMR Project is developing a design for a Small Modular Reactor (SMR) of 340 MWe composed of two 170 MWe independent units, that will supplement the offering of high-output nuclear reactors, especially in response to specific needs such as replacement of fossil-fuelled power plants. NUWARD SMR is a mix of proven and innovative design features that will make it more commercially competitive, while integrating safety features that comply with the highest international standards. Following the principles of redundancy and diversity and rigorous application of Defence in Depth (DID), with an international view on nuclear safety licensing, the Project also incorporates new safety approaches into its design development. The NUWARD SMR Project has been in development for a number of years, it entered conceptual design formally in mid-2019 and entered Basic Design in 2023. The objective of the concept design phase was to confirm the project technological choices and to define the first design configuration of the NUWARD SMR product, to document it, in order to launch pre-licensing with the French Safety Authority (ASN) and to define its estimated cost and its subsequent development and construction schedules. As a delivery milestone the Safety Options file (called the Dossier d'Options de Sûreté (DOS)) has been submitted to ASN in July 2023 for their opinion. An integral part of the NUWARD SMR Project, is not only to deliver a design suitable for France and to satisfy French regulation, but to develop a product suitable and indeed desirable, for the international market, with a first focus in Europe. In order to achieve its objectives and realise its market potential, the NUWARD SMR Project needs to define and realise its safety approach within an international environment and that is the key subject of this paper. The following paper: • Summarises the foundation principles and technological background which underpin the design; • Contextualises the key design features with regard to the international safety regulatory framework with particular emphasis on innovative passive safety aspects; • Illustrates the Project activities in preparation for first licensing in France, and also a wider international view via the ASN led Joint Early Review of the NUWARD SMR design, including Finnish and Czech Republic regulators, recently joined by the Swedish, Polish and Dutch regulators; • Articulates the collaborative approach to design development from involvement with the Project partners (the Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Naval Group, TechnicAtome, Framatome and Tractebel) to the establishment of the International NUWARD Advisory Board (INAB), to gain greater international insight and advice; • Concludes with the focus on next steps into detailed design development, standardisation of the design and its simplification to enhance its commercial competitiveness in a context of further harmonisation of the nuclear safety and licensing requirements and aspirations.

Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling

  • Shi, Qing-Qiang;Zuo, Guo-Wei;Feng, Zi-Qiang;Zhao, Lv-Cui;Luo, Lian;You, Zhi-Mei;Li, Dang-Yang;Xia, Jing;Li, Jing;Chen, Di-Long
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7849-7855
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    • 2014
  • Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.

The Effects of Hypercapnia and High Flow on Cerebral Metabolism During Cardiopulmonary Bypass (심폐바이패스 시 고탄산분압과 고관류법이 뇌대사에 미치는 영향)

  • 강도균;최석철;윤영철;최국렬;정신현;황윤호;조광현
    • Journal of Chest Surgery
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    • v.36 no.7
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    • pp.472-482
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    • 2003
  • Recent studies have demonstrated that cerebral desaturation during rewarming period of CPB was associated with postoperative neurologic dysfunction. The prevention of cerebral desaturation during CPB may reduce the incidences of neurologic and neuropsychological complications. The present study was prospectively undertaken to compare the clinical effects between two strategies (hypercapnic CPB and high flow CPB) to prevent cerebral desaturation for establishing a proper CPB technique. Material and Method: Thirty-six adult patients scheduled for elective cardiac surgery were randomized into either hypercapnic (Pa$CO_2$ 45~50mmHg, n=18) or high flow group (flow rate 2.75 L/ $m^2$/min and Pa$CO_2$ 35~40mmHg, n=18) during rewarming period of CPB. In each patient, middle cerebral artery blood flow velocity ( $V_{MCA}$), cerebral arteriovenous oxygen content difference (C(a-v) $O_2$), modified cerebral metabolic rate for oxygen (MCMR $O_2$), cerebral oxygen transport rate ( $T_{E}$ $O_2$), incidence of cerebral desaturation (internal jugular bulb blood oxygen saturation $\leq$ 50%), increased rate of S-100 $\beta$ concentration, and arterial and internal jugular bulb blood gas were measured during the five phases of the operation; Pre-CPB, CPB-10 min (steady-state CPB, nasopharyngeal temperature 29~3$0^{\circ}C$), Rewarm-1 (rewarming phase, nasopharyngeal temperature 33$^{\circ}C$), Rewarm-2 (nasopharyngeal temperature 37$^{\circ}C$), and CPB-off. Incidence of postoperative delirium and duration were assessed in all patients. All variables were compared between the two groups. Result: $V_{MCA}$ (157.88$\pm$10.87 vs 120.00$\pm$6.18%, p=0.006), internal jugular bulb $O_2$ saturation (68.01$\pm$2.75 vs 61.28$\pm$2.87%, p=0.03) and $O_2$ tension (41.01$\pm$2.25 vs 32.02$\pm$ 1,67 mmHg, p=0.03), and $T_{E}$ $O_2$(110.84$\pm$7.41 vs 81.15$\pm$8.11%, p=0.003) at rewarming periods were higher in the hypercapnic group than in the high flow group. C(a-v) $O_2$ (4.0$\pm$0.30 vs 4.84$\pm$0.38 mg/dL, p=0.04), COE (0.36$\pm$0.03 vs 0.42$\pm$0.03, p=0.04), increased rate of S- 100$\beta$ (391.67$\pm$23.40 vs 940.0$\pm$17.02%, p=0.003), and incidence of cerebral desaturation (2 vs 4 patients, p=0.04) at rewarming periods, and duration of postoperative delirium (18 vs 34 hr, p=0.02) were low in the hypercapnic group compared to the high flow group. Conclusion: These results indicate that hypercapnic CPB may provide relatively diminished cerebral injury and beneficial effects for cerebral metabolism relatively compared to high flow CPB.low CPB.

Synthesis of ${\alpha}$-Alumina Nanoparticles Through Partial Hydrolysis of Aluminum Chloride Vapor (염화알미늄 증기의 부분가수분해를 통한 알파 알루미나 나노입자 제조)

  • Park, Hoey Kyung;Yoo, Youn Sug;Park, Kyun Young;Jung, Kyeong Youl
    • Korean Chemical Engineering Research
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    • v.49 no.5
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    • pp.664-668
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    • 2011
  • Spherical alumina precursors represented by $AlO_xCl_y(OH)_z$, 30~200 nm in particle diameter, were prepared by partial hydrolysis of $AlCl_3$ vapor in a 500 ml reactor. Investigated on the particle morphology and size were the effects of the reaction time, the stirring speed and the reaction temperature. The particle morphology and size was insensitive to the reaction time in the range 20 to 300 s. The variation of the stirring speed from 0 to 300 and 800 rpm showed that the particle size was the largest at 0 rpm. As the temperature was varied from 180 to 190, 200, $140^{\circ}C$, the particle size showed a maximum at $190^{\circ}C$. By calcination of the as-produced particles at $1,200^{\circ}C$ for 6h with a heating rate of $10^{\circ}C$/min, ${\alpha}$-alumina particles 45 nm in surface area equivalent diameter were obtained. The particle shape after calcination turned wormlike due to sintering between neighboring particles. A rapid calcination at $1400^{\circ}C$ for 0.5 h with a higher heating rate of $50^{\circ}C$/min reduced the sintering considerably. An addition of $SiCl_4$ or TMCTS(2,4,6,8-tetramethylcyclosiloxane) to the $AlCl_3$ reduced the sintering effectively in the calcination step; however, peaks of ${\gamma}$ or mullite phase appeared. An addition of $AlF_3$ to the particles obtained from the hydrolysis resulted in a hexagonal disc shaped alumina particles.

Optimization of HPLC Method and Clean-up Process for Simultaneous and Systematic Analysis of Synthetic Color Additives in Foods (식품 중 타르색소의 동시분석 및 계통분석을 위한 HPLC 분석조건 및 정제과정 확립)

  • Park, Sung-Kwan;Hong, Yeun;Jung, Yong-Hyun;Lee, Chang-Hee;Yoon, Hae-Jung;Kim, So-Hee;Lee, Jong-Ok
    • Korean Journal of Food Science and Technology
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    • v.33 no.1
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    • pp.33-39
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    • 2001
  • To develop a method for separation process using Sep-pak $C_18$, simultaneous and systematic analysis of 8 permitted and 11 non-permitted synthetic food colors in Korea, optimization of analysis conditions for reverse phase ion-pair high performance liquid chromatography was carried out. For the best result of Sep-pak $C_18$ separation the pH of color standard mixture solution was $5{\sim}6$ and 0.1% HCl-methanol solution were set as eluent. The colors eluated from Sep-pak $C_18$ cartridge were determined and confirmed by high performance liquid chromatography with a photodiode array detector at 420 nm for yellow colors type, at 520 nm for red colors type, at 600 nm for blue and green colors type and at 254 nm for mixed colors. Conditions for HPLC analysis were as follows: column, Symmetry $C_18$ (5 m, 3.9 mm $i.d.{\times}150\;mm$); mobile phase, 0.025 M ammonium acetate (containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (65 : 25 : 10) and 0.025 M ammonium acetate(containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (40 : 50 : 10); flow rate, 1 mL/min. It takes 35 minutes for simultaneaus analysis and 18 minutes for systematic analysis. The detection limits range of each colors were $0.01{\sim}0.05\;{\mu}g/g$.

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