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Development of HPLC Determination Method for Trace Levels of 1-, 2-Nitropyrenes and 2-Nitrofluoranthene in Airborne Particulates and Its Application to Samples Collected at Noto Peninsula

  • Hayakawa, Kazuichi;Tang, Ning;Sato, Kosuke;Izaki, Akihiko;Tatematsu, Michiya;Hama, Hirotaka;Li, Ying;Kameda, Takayuki;Toriba, Akira
    • Asian Journal of Atmospheric Environment
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    • v.5 no.3
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    • pp.146-151
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    • 2011
  • 1-Nitropyrene (1-NP), 2-NP and 2-nitrofluoranthene (2-NFR) are useful markers for studying the atmospheric behaviors of polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs). However, present methods for measuring trace levels of these compounds are lesssensitive and laborious. Here we describe several improvements to a previously reported high-performance liquid chromatography-chemiluminescence detection system that allows it to determine trace levels of 1-, 2-NPs and 2-NFR. The proposed system was equipped with a reducer column packed with Pt/Rh instead of zinc whose life-time was limited. The combination of Cosmosil MS-II (monomeric ODS) and AR-II (polymeric ODS) columns was used instead of polymeric ODS columns as the separator column to improve the separation. An ethanol mixture with acetate buffer (pH 5.5) was used in place of an acetonitrile mixture with the same buffer to activate the reducer column. The same ethanol mixture was used as the mobile phase for the clean-up column. The switching time of the column switching valve was optimized to concentrate the amino-derivatives of above NPAHs quantitatively on the concentrator column. The concentrations of bis(2,4,6-trichlorophenly) oxalate and hydrogen peroxide in the chemiluminescence reagent solution were optimized to 0.4 mM and 30 mM, respectively, to increase the sensitivity. Under the above conditions, the detection limits (S/N=3) of 1-, 2-NPs and 2-NFR were 1 fmol (0.25 pg), 10 fmol (2.5 pg) and 4 fmol (1 pg), respectively. The proposed system was effectively used to determine trace levels of 1-, 2-NPs and 2-NFR in airborne particulates collected at Noto Peninsula. The atmospheric concentrations of 1-, 2-NPs and 2-NFR were not more than sub pg $m^{-3}$ levels. They were higher in winter (January) than in summer (July). In both seasons, the concentrations were in decreasing order, [2-NFR]>[1-NP]>[2-NP].

A Comparative Study of Physiological Activity of Glycyrrhiza uralensis Fischer Stems and Leaves by Processing Methods (감초 경엽(莖葉)의 포제방법에 따른 생리활성 비교)

  • Park, Jeong Seob;Park, Sun Hee;Oh, Il Soo;Chang, Young Nam;Bang, Keuk Soo;Byeon, Eun Ju;Lee, Jeong Ho
    • Korean Journal of Plant Resources
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    • v.26 no.5
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    • pp.539-547
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    • 2013
  • This study investigates the change of chemical components, antioxidant and antimutagenic activity in Glycyrrhiza uralensis Fischer stems and leaves (GU) by the various processing methods, as follows: fresh (GU-1), dried under the shade (GU-2), blanched (GU-3), roasted 3 times (GU-4), roasted 4 times (GU-5). The components have been identified and quantified through the use of gas chromatograph-mass spectrometry (GC-MS). At results, cis-1,3-dimethyl-2-methylenecyclohexaneat at 19.7 min and n-hexadecanoic acid at 21.5 min were detected in five kinds of extract. 3-O-Methyl-D-fructose at 17.8 min was observed in four extracts except in GU-1 and aminopyrazine at 19.8min was observed in four extracts except in GU-4. The total phenolics contents was high in GU-4(3.38 g/100 g). ABTS radical scavenging was high in GU-5($EC_{50}$, 0.57 ug) and DPPH radical scavenging was high in GU-4($EC_{50}$, 2.66 ug). The extracts of GU-3, GU-4, and GU-2 were most potent in anti-mutagenicity activity against 1-NP, Trip-P-1, and Trip-P-2, respectively. GU-3 and GU-4 also showed most potent effect of anti-mutagenicity on 2-AA and AFB1, respectively.

Adventitious Shoot Formation and Plant Regeneration from Explants of Solanum nigrum L. (까마중(Solanum nigrum L.)의 유식물 절편체에서 부정아 형성 및 식물체 재분화)

  • Oh, Soon-Ja;Koh, Seok-Chan
    • Korean Journal of Plant Resources
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    • v.25 no.2
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    • pp.277-284
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    • 2012
  • In the present study, the effects of plant growth regulators on adventitious shoot and root formation of various explants of $in$ $vitro$ seedlings of $Solanum$ $nigrum$ L. were investigated to determine the optimum conditions for the high-efficiency plant regeneration of this species. The formation of adventitious shoots was higher in leaf explants than in cotyledon, hypocotyl, or epicotyl explants at low concentrations (0.5~2.0 mg $L^{-1}$ ) of 6-benzylaminopurine (BAP). The number of adventitious shoots and the shoot length were also higher in both leaf and cotyledon explants. In particular, 2.0 mg $L^{-1}$ BAP was most effective for stimulating the induction and multiplication of adventitious shoots. In terms of root formation and root development from shoots that were separated from multiple shoots, indole butyric acid (IBA) and indole acetic acid (IAA) were more effective than ${\alpha}$-naphthalene acetic acid (NAA). The percentage of rooting as well as the number of roots per shoot (4.0), root length (7.82 cm), and shoot length (8.76 cm) was highest on MS media supplemented with 0.05 mg $L^{-1}$ IAA. Furthermore, 100% of the regenerated plantlets survived when transplanted to compost soil. These results suggest that leaf explants are the best source for the high-efficiency regeneration of $S.$ $nigrum$ L., and that 2.0 mg $L^{-1}$ BAP and 0.05 mg $L^{-1}$ IAA are the best conditions for shoot and root induction, respectively.

Effect of the amino acid mixture on freeze-drying and preservation of Lactobacillns casei YIT 9018 (아미노산 혼합용액이 Lactobacillus casei YIT 9018의 동결건조 및 저장성에 미치는 영향)

  • 윤성식;이해옥;유주현
    • Microbiology and Biotechnology Letters
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    • v.14 no.5
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    • pp.421-426
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    • 1986
  • This study was attempted to find out effective storage methods of Lactobacillus casei YIT 9018, industrial strain for fermented mil k production, without severe bacterial death and activity deteriorations. The cryoprotective effect of the ammo acid mixture consisting of glycine and DL-g1utamic acid on the test strain were examined and also compared with those other protectants already reported. The apparent protective effect by the amino acid mixture was observed to controls. Both glycine and DL-glutamic acid prevented the freezing death of test strain and his effect of 1. casei YIT 9018 had reached stationary stage in MRS-broth 18h after inoculation. Cells harvested from stationary stage were most resistant to freezing damage. The viability of the test strain was affected by rehydration media and the recovery of viable cells was increased about threefold when amino acid mixture was used for rehydration. The presence of non-fat milk solid (NFMS), sucrose and lactose in amino acid mixture increased viability of the test strain up to 85%. In this case, optimal concentrations of NFMS, sucrose and lactose were 10%, 7.5-10%, 7.5-10%, respectively.

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Transient neonatal diabetes mellitus with macroglossia diagnosed by methylation specific PCR (MS-PCR) (메틸화 특이 PCR로 진단된 거설증을 동반한 일과성 신생아 당뇨병)

  • Jin, Hye Young;Choi, Jin-Ho;Kim, Gu-Hwan;Yoo, Han-Wook
    • Clinical and Experimental Pediatrics
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    • v.53 no.3
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    • pp.432-436
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    • 2010
  • Transient neonatal diabetes mellitus (TNDM) has been associated with paternal uniparental isodisomy of chromosome 6, paternally inherited duplication of 6q24, or a methylation defect at a CpG island of the ZAC or HYMAI gene. We experienced a case of TNDM in which the patient presented with hyperglycemia, macroglossia, and intrauterine growth retardation, caused by a paternally derived HYMAI. An 18-day-old female infant was admitted to the hospital because of macroglossia and recurrent hyperglycemia. In addition to the macroglossia, she also presented with large fontanelles, micrognathia, and prominent eyes. Serum glucose levels were 200-00 mg/dL and they improved spontaneously 2 days after admission. To identify the presence of a maternal methylated allele, bisulfite-treated genomic DNA from peripheral blood was prepared and digested with BssHII after polymerase chain reaction (PCR) amplification with methylation-specific HYMAI primers. PCR and restriction fragment length polymorphism analysis showed that the patient had only the paternal origin of the HYMA1 gene. TNDM is associated with a methylation defect in chromosome 6, suggesting that an imprinted gene on chromosome 6 is responsible for this phenotype.

Effects of Supplemental Synbiotics Composed of Anaerobic Bacteria, Yeast and Mold on the Change of Chemical Composition and Fermentation Characteristics of Total Mixed Ration for Cattle (혐기성 박테리아, 효모 및 곰팡이로 제조된 synbiotics 첨가 축우용 완전혼합사료의 성분 변화 및 발효 특성에 미치는 영향)

  • Lee, Shin-Ja;Shin, Nyeon-Hak;Jung, Ho-Sik;Moon, Yea-Hwang;Lee, Sang-Suk;Lee, Sung-Sill
    • Journal of Life Science
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    • v.19 no.2
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    • pp.241-248
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    • 2009
  • In order to investigate the effects of synbiotics on change of chemical composition and fermentation characteristics of total mixed ration (TMR), eight TMRs fermented by synbiotics composing the anaerobic microbes (bacteria, yeast, mold) were alloted to the experimental treatments. Treatments were composed of untreated synbiotics(US), bacterial synbiotics (BS), yeast synbiotics (YS), mold synbiotics (MS), bacterial and mold synbiotics (BMS), yeast and mold synbiotics (YMS), bacterial and yeast synbiotics (BYS), and bacterial, yeast and mold synbiotics (BYMS). After 7 days of anaerobic fermentation, fermented-TMRs were exposed to air during 1, 3, 5, 7, 14 and 21 days. One hundred forty four (8 treatments ${\times}$ 6 exposing days ${\times}$ 3 replications) fermented- TMRs were manufactured by vinyl bag sized of 43 cm by 58 cm. The results obtained were as follows. Moisture contents of the fermented TMRs anaerobically ranged from 41% to 45%, and was similar to those of basal TMRs. As results of anaerobic fermentation, the concentration of crude protein was decreased by 11.7% to 14.8% in the untreated sample, while was rather increased by 11% when the TMR was fermented with BMYS. And also BMYS treatment showed decreases by 32% for crude fiber, 15.5% for NDF and 26.1% for ADF. Internal temperature of fermented-TMRs was highest at 7 day of exposing in the air. The pH of fermented-TMR juice was significant difference betweentreatments after 7 day of exposing in air, and that of BMS was highest at 14 day after exposing in air (P<0.05). Acid buffering capacity was increased in proportion to the exposing day of TMR, and peaked at 7 or 14 days after exposing. Ammonia concentration of fermented-TMRs was highest at 5 day after exposing in the air. Individual volatile fatty acid of fermented-TMR juice was very low level in all treatments. Although BMYS treatment to TMR inclined to increase in crude protein and decrease in fibers, but there were no positive effects on the fermentation characteristics after exposing in the air by supplementation of anaerobic synbiotics to TMR.

In vitro Callus and Somatic Embryo Induction of Six Hosta Species Native to Korea

  • Choi, Han;Lee, Seung Youn;Ryu, Sun Hee;Yoon, Sae Mi;Kim, Sang Yong;Lee, Jong Suk;Yang, Jong Cheol
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.80-80
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    • 2018
  • Hosta is a genus of the family Asparagaceae and distributed in East Asia. There are six Hosta species (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) native to Korea and among them, four species (H. minor, H. jonesii, H. venusta and H. yingeri) are endemic to the Korea peninsula. Hosta is generally propagated by seed, crown division or tissue culture. However, tissue culture is a more efficient method to mass proliferation, a new cultivar development and disease-free plantlet production in a limit time. Hence, we conducted this study to evaluate the influence of various plant growth regulators (PGRs) treatments on the induction of callus and somatic embryo of the six Hosta species. Leaf, petiole and root were used to select optimum tissue culture explants. Petiole explants thus only were used for callus induction and somatic embryogenesis with TDZ (0.1, 0.5 or 1.0mg/L) and NAA (0.1 or 0.5 mg/L) combinations. After 12 weeks of culture, the highest rate of somatic embryogenesis was achieved on modificated MS medium containing 1.0 mg/L TDZ and 0.1 mg/L NAA in H. capitata and H. minor (15.5%, respectively), 0.1 or 0.5 mg/L TDZ and 0.1 mg/L NAA in H. jonesii (22.2%), 1.0 mg/L TDZ and 0.5 mg/L NAA in H. yingeri (26.7%), and 0.1 mg/L TDZ and 0.5 mg/L NAA in H. venusta (53.3%). H. clausa showed very low effect on somatic embryogenesis by PGRs; 2.2%. There was interspecies difference to PGRs respond for callus and somatic embryo induction. Regenerated multiple shoots and plantlet of H. minor, H. jonesii, H. venusta and H. yingeri were obtained via somatic embryogenesis.

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Analysis of Chemical Composition of Bulro Kugi (Lycium chinense Mill) Fruit, Leaf, and Root (불로 구기의 부위별 화학적 성분분석)

  • Kim, Eun-Hae;Lee, Joo-Chan;Kim, Hyeon-Wee;Lee, Cherl-Ho;Koh, Kyung-Hee
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.154-163
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    • 2005
  • Chemical and flavor components of Bulro Kugi (Lycium chinense Mill) fruit leaf, and root were compared. Fructose and glucose were detected in fruit and leaf, and sucrose in root, respectively. Citrate was the highest among organic acids in fruits, and malate in leaf and root Capsaicin was detected in leaf and root. Volatile flavor compounds were extracted by simultaneous distillation and extraction method using Likens and Nickerson's extraction apparatus. Concentrated flavor extract was analysed, and 128 compounds, including 22 acids 15 alcohols, 12 aldehydes, 8 esters, 10 furans, 26 hydrocarbons, 4 phenols, 2 pyrroles, 1 pyrazine, and 28 miscellaneous components. were identified by GC and GC-MS. Main volatile compounds were hexadecanoic acid and 2-furancarboxaldehyde in hot-air dried fruit, hexadecanoic acid and 1-hexadecene in fresh fruit, 3, 7, 11, 15-tetranethyl-2-hexadecan-1-ol and hexadecanoir acid in leaf, and hexadecanoic acid in root.

Isolation and Characterization of Bacillus Strains for Biological Control

  • Kim, Han-Soo;Park, Jiyong;Cho, Sung-Won;Park, Kee-Hyun;Lee, Gung-Pyo;Ban, Soo-Jung;Lee, Chang-Roo;Kim, Chung-Sun
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.196-201
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    • 2003
  • The object of this study was to characterize Bacillus strains GB-017 and GB-0356, which produce antifungal substances, especially for plant pathogens. In addition, this study was undertaken to characterize the culture conditions required for the production of antifungal substances and to document some of the properties of the antifungal substance produced by these soil-isolated strains. Strains GB-0365 and GB-017 were found to be bacillus-shaped, gram-positive and motile, and to inhibit Botrytis cineria, Fusarium sp., Pythium sp., and Rhizoctonia solani. Antagonistic activity was maintained up to pH 9.0, and the antifungal activity was stable to heat at 80$^{\circ}C$ for 1 h. Antifungal substances were separated and purified using ion exchange and adsorption columns including WK-I0(H$\^$+) (pH 7.0), HP20 column (pH 3.0) and IPA (pH 3.0). and IPA. Its UV absorption spectrum showed major peaks at 231 and 259 nm, corresponding to polyene and lactone. A fast atom bombardment mass spectrum (FAB MS) showed a highest peak at 441 m/z and major peaks at 192, 205, and 370 m/z.

The utilization of fungicide and insecticide from medicinal plants for conservation of cultural properties (천연약재로부터 문화재보존용 방충방균제 개발연구)

  • Chung, Yong-Jae;Lee, Kyu-Shik;Han, Sung-Hee;Kang, Dai-Ill;Lee, Myeong-Hui
    • 보존과학연구
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    • s.22
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    • pp.5-25
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    • 2001
  • The germicidal and insecticidal properties of volatile components extracted from star anise(Illicium verum Hooker filius) and clove (Eugenia caryophyllata THUNBERG)were evaluated against five microorganisms and three insects for the purpose of developing biocidal active substances from medicinal plants. The volatile components of star anise and clove showed strong antimicrobial effect against Aspergillus niger, Penicillium funiculosum, Mucor hiemalis, Trichoderma viride, and Aureobasidium pullulans. The extracts of each medicine also showed insecticidal effects against Sitophilusoryzae L., Lyctus linearis GOZE, and Reticulitermes spertus kyushuensis Morimoto. Fumigant toxicities to adult insects were determined. In the case of fumiganttoxicity, the extract of star anise showed 100% mortality against R. spertus, S.oryzae, and L. linearis at rates of $2.5\mu\ell$, $50\mu\ell$, $250\mu\ell$/filter paper, respectively but showed no killing effects by clove. The volatile components of star anise and clove were investigated by means of GC/MS. The main constitute, anethole among 20components from star anise and eugenol among 9 components from clove were identified. The mixture of star anise and clove as the volume ratio of 2 : 1 showed higher properties for antimicrobial and insecticidal effect than each volatile component. A. niger was inhibited by the mixture(125ml/$m^3$) for up to 10 days of exposure. Also, from the result of observing state change of organic materials by volatile extracts of star anise and clove, volatile extracts effects have no effect on natural organic materials of organic cultural properties and can be used as biological control agent. As research contents as above, the insecticidal and germicidal agents from star anise and clove and the mixture of them were more efficient and high level to prevent biological damage for conservation of organic cultural properties. So they may be used in new development of biologicalinsecticidal and germicidal agents for conservation of cultural properties.

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