• Title/Summary/Keyword: metabolites analysis

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Proteomic analysis of serum proteins responsive to styrene exposure (Styrene 노출에 반응을 보이는 혈청 단백질에 대한 프로테오믹스 분석)

  • Kim, Ki-Woong;Heo, Kyung-Hwa;Won, Yong Lim;Jeong, Jin Wook;Kim, Tae Gyun;Park, Injeong
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.17 no.3
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    • pp.235-244
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    • 2007
  • By comparing the proteins from the workers exposed to styrene with the ones from controls, it may be possible to identify proteins that play a role in the occurrence and progress of occupational disease and thus to study the molecular mechanisms of occupational disease. In order to find the biomarkers for assessing the styrene effects early, before clinical symptoms develop and to understand the mechanisms of adverse health effects, we surveyed 134 employees, among whom 52 workers(30 male and 22 female) were chronically exposed to styrene in 10 glass-reinforced plastic boat manufacturing factories in Korea and 82 controls had never been occupationally exposed to hazardous chemicals including styrene. The age and drinking habits and serum biochemistry such as total protein, BUN and serum creatinine in both groups were significantly different. Exposed workers were divided into three groups according to exposure levels of styrene(G1, below 1/2 TLV; G2, 1/2 TLV to TLV; G3, above TLV). The mean concentration of airborne styrene in G1 group was $10.93{\pm}11.33ppm$, and those of urinary mandelic acid(MA) and phenylglyoxylic acid(PGA) were $0.17{\pm}0.21$ and $0.13{\pm}0.11g/g$ creatinine, respectively. The mean concentration of airborne styrene in G2 and G3 groups were $47.54{\pm}22.43$ and $65.33{\pm}33.47ppm$, respectively, and levels of urinary metabolites such as MA and PGA increased considerably as expected with the increase in exposure level of styrene. The airborne styrene concentration were significantly correlated to the urinary concentration of MA(r=0.784, p=0.000) and PGA(r=0.626, p<0.001). In the 2D electrophoresis, the concentration of five proteins including complement C3 precursor, alpha-1-antitrypsin(AAT), vitamin D binding protein precursor(DBP), alpha-1-B-glycoprotein(A1BG) and inter alpha trypsin inhibitor(ITI) heavy chain-related protein were significantly altered in workers exposed to styrene compared with controls. While expression of complement C3 precursor and AAT increased by exposure to styrene, expression of DBP, A1BG and ITI heavy chain-related protein decreased. These results suggest that the exposure of styrene might affects levels of plasma proteinase, carriers of endogenous substances and immune system. In particular, increasing of AAT with the increase in exposure level of styrene can explain the tissue damage and inflammation by the imbalance of proteinase/antiproteinase and decrease of DBP, A1BG and ITI heavy chain-related protein in workers exposed to styrene is associated with dysfunction and/or declination in immune system and signal transduction

Simultaneous Determination of Aflatoxins and Ochratoxin A in Pork by LC-MS/MS (LC-MS/MS를 이용한 돼지고기 중 총아플라톡신 및 오크라톡신 A 동시분석법 확립)

  • Paek, Ockjin;Park, Songyi;Park, Ki Hun;Kim, Sheen-Hee;Suh, Saejung;Yoon, Hae Jung
    • Journal of Food Hygiene and Safety
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    • v.31 no.3
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    • pp.194-200
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    • 2016
  • Aflatoxins and ochratoxin A (AFTs and OTA) are secondary fungal metabolites produced by several moulds, mainly by Aspergillus flavus by Aspergillus ochraceus and Penicillium verrucosum, and these toxins can be transferred to animals and humans through the ingestion of contaminated feed and food. This study was to develop the analytical method for determination the levels of AFTs ($B_1$, $B_2$, $G_1$ and $G_2$) and OTA in pork. The AFTs and OTA were analyzed simultaneously by electrospray ionization in positive ion mode and mass reaction monitoring (MRM) after solid phase extract (SPE) columns clean-up. Performance characteristics, such as accuracy, precision, linear range, limit of detection (LOD) and quantification (LOQ), were also determined. Matrix-matched standard calibration was used for quantification, obtaining the recoveries in the range of 67.3~108.2% with the relative standard deviations of < 20%. Limits of detection and quantification were also estimated, obtaining the limits of quantification ranged in $0.7{\sim}1.3{\mu}g/kg$. The results of the inter-day study, which was performed with pork samples for 3 days, showed an accuracy of 92.0~109.9%. The precisions (expressed as relative standard deviation values) for the inter day variation were 2.6~17.8%. The method developed in this study was able to carry out the analysis with the satisfactory intensity and accuracy.

Sensing the Stress: the Role of the Stress-activated p38/Hog1 MAPK Signalling Pathway in Human Pathogenic Fungus Cryptococcus neoformans

  • Bahn, Yong-Sun;Heitman, Joseph
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2007.05a
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    • pp.120-122
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    • 2007
  • All living organisms use numerous signal-transduction pathways to sense and respond to their environments and thereby survive and proliferate in a range of biological niches. Molecular dissection of these signalling networks has increased our understanding of these communication processes and provides a platform for therapeutic intervention when these pathways malfunction in disease states, including infection. Owing to the expanding availability of sequenced genomes, a wealth of genetic and molecular tools and the conservation of signalling networks, members of the fungal kingdom serve as excellent model systems for more complex, multicellular organisms. Here, we employed Cryptococcus neoformans as a model system to understand how fungal-signalling circuits operate at the molecular level to sense and respond to a plethora of environmental stresses, including osmoticshock, UV, high temperature, oxidative stress and toxic drugs/metabolites. The stress-activated p38/Hog1 MAPK pathway is structurally conserved in many organisms as diverse as yeast and mammals, but its regulation is uniquely specialized in a majority of clinical Cryptococcus neoformans serotype A and D strains to control differentiation and virulence factor regulation. C. neoformans Hog1 MAPK is controlled by Pbs2 MAPK kinase (MAPKK). The Pbs2-Hog1 MAPK cascade is controlled by the fungal "two-component" system that is composed of a response regulator, Ssk1, and multiple sensor kinases, including two-component.like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. We also identified and characterized the Ssk2 MAPKKK upstream of the MAPKK Pbs2 and the MAPK Hog1 in C. neoformans. The SSK2 gene was identified as a potential component responsible for differential Hog1 regulation between the serotype D sibling f1 strains B3501 and B3502 through comparative analysis of their meiotic map with the meiotic segregation of Hog1-dependent sensitivity to the fungicide fludioxonil. Ssk2 is the only polymorphic component in the Hog1 MAPK module, including two coding sequence changes between the SSK2 alleles in B3501 and B3502 strains. To further support this finding, the SSK2 allele exchange completely swapped Hog1-related phenotypes between B3501 and B3502 strains. In the serotype A strain H99, disruption of the SSK2 gene dramatically enhanced capsule biosynthesis and mating efficiency, similar to pbs2 and hog1 mutations. Furthermore, ssk2, pbs2, and hog1 mutants are all hypersensitive to a variety of stresses and completely resistant to fludioxonil. Taken together, these findings indicate that Ssk2 is the critical interface protein connecting the two-component system and the Pbs2-Hog1 pathway in C. neoformans.

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Cisplatin and Extract of Tissue Cultured Mountain Ginseng-Induced Apoptosis in Human Cervical Cancer Cells (인체 자궁암세포에서 cisplatin과 산삼배양근추출물에 의한 apoptosis유도)

  • Lee, Myeong-Seon
    • Applied Microscopy
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    • v.40 no.3
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    • pp.133-138
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    • 2010
  • Mountain ginseng is a perennial crop rarely found in the deep mountains of Korea. The medicinal effect of the mountain ginseng is well known as a panacea in traditional Chinese medicine for a long time. But scientific studies to elucidate the medicinal effect of the mountain ginseng have never been made on account of lack of sample. Recently an improved method of adventitious root culture system through the use of bioreactor has been developed in Panax ginseng that seems to be a reliable way of commercialization of root derived secondary metabolites. This experiment was conducted to evaluated chemotherapeutic effect against human cervical cancer cells by cisplatin (CDDP) and extract of tissue cultured mountain ginseng (ETCMG). CDDP and ETCMG-induced apoptotic cell death in human cervical cancer cell line, HeLa was confirmed by the analysis of cell growth, morphological changes, DNA fragmentation, flow cytometry showed that ETCMG is an inducer of apoptosis and synergizes with CDDP. These results suggest that ETCMG present evidence of anticancer effect and could have a possibly natural therapeutic potential in cervical cancer patients.

Quality Characteristics and Composition Profile of Traditional Doenjang and Manufactured Doenjang during Storage Time (저장기간에 따른 전통된장과 개량식된장의 이화학적 특성, 기능성분 및 대사체 분석)

  • Kang, Hee-Joo;Kim, Jin-Hee;Kim, Ri-Rang;Kim, Kang Sung;Hong, Sang-Phi;Kim, Min-Jung;Yang, Hye Jeong
    • The Korean Journal of Food And Nutrition
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    • v.29 no.5
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    • pp.785-794
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    • 2016
  • This research was conducted to evaluate quality changes in traditional Doenjang and manufactured Doenjang during a storage period of 8 weeks. Low-salt Doenjang and commercial Doenjang were purchased from different manufacturers and proximate analysis as well as changes in isoflavone, polyphenol, flavonoid contents of the samples were investigated using a mass spectrophotometer. The salinity of traditional Doenjang, low salt Doenjang, and commercial Doenjang were $13.2{\pm}1.15$, $7.17{\pm}2.74$, $10.67{\pm}0.35%$, respectively and the salt concentrations of the soybean pastes did not change during storage. After 8 weeks at $35^{\circ}C$, chromatic values of all the paste samples decreased somewhat, with traditional Doenjang exhibiting fewer changes as compared to manufactured Doenjang. Amino acid nitrogen, acidity, microbial population all tended to increase with time, although some samples showed fluctuations during the test period. Moreover, the total isoflavone contents of traditional Doenjang increased with storage time while that of manufactured Doenjang tended to decrease. The isoflavone aglycone was shown to be the highest in traditional Doenjang, while isoflavone glycoside was abundant in manufactured Doenjang. Total flavonoid contents showed similar trends regardless of samples; initial contents of total flavonoid was 0.6 mg/g which increased to more than twice to 1.4 mg/g at the end of storage period. Composition profile of Doenjang extracts was analyzed using UPLC-Q-ToF.

Correlation between Biosurfactants and Antifungal Activity of a Biocontrol Bacterium, Bacillus amyloliquefaciens LM11 (생물적 방제균 Bacillus amyloliquefaciens LM11의 유래 생물계면활성물질과 항균활성과의 상관관계)

  • Kang, Beom Ryong;Kim, Yong Hwan;Nam, Hyo Song;Kim, Young Cheol
    • Research in Plant Disease
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    • v.23 no.2
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    • pp.177-185
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    • 2017
  • Bacillus amyloliquefaciens LM11 was isolated from the feces of larvae of the rhino beetle and showed strong antifungal activities against various phytopathogenic fungi by producing biosurfactants. In this study, our overall goal was to determine relationship between biosurfactants produced from the LM11 strain and its role in growth inhibition of phytopathogenic fungi. Production and expression levels of B. amyloliquefaciens LM11 biosurfactants were significantly differed depending on growth phases. Transcriptional and biochemical analysis indicated that the biosurfactants of the LM11 strain were greatly enhanced in late log-phase to stationary phase. Inhibitions of phytopathogenic mycelial growth and spore germination were directly correlated (P<0.001, R=0.761) with concentrations of the LM11 cell-free culture filtrates. The minimum inhibitory surface tension of the culture filtrate of the B. amyloliquefaciens LM11 grown in stationary phase to inhibit mycelial growth of the phytopathogenic fungi was 38.5 mN/m (P<0.001, R=0.951-0.977). Our results indicated that the biosurfactants of B. amyloliquefaciens LM11 act as key antifungal metabolites in biocontrol of plant diseases, and measuring surface tension of the cell-free culture fluids can be used as an easy indicator for optimal usage of the biocontrol agents.

Assessment of Volatile Organic Compounds in Blood and Urine among Residents around Camp Carroll (캠프 캐럴 인근 주민의 혈중 및 요중 휘발성 유기화합물 평가)

  • Lim, Hyun-Sul;Yang, Wonho;Kim, Geun-Bae;Cho, Young-Sung;Min, Young-Sun;Lee, Kwan;Lee, Duk Hee;Ju, Young-Su;Kim, Sunshin;Heo, Jung;Jung, Dayoung
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.26 no.1
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    • pp.11-19
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    • 2016
  • Objectives: Exposure to volatile organic compounds such as trichloroethylene(TCE) and perchloroethylene(PCE), along with Agent Orange, that were issued around Camp Carroll US Army Base situated in Waegwan, Chilgok-gun, Gyeongsangbuk-do Province, Korea. The main objective of this study was to assess the exposure to TCE and PCE of residents of the area surrounding Camp Carroll. Methods: The TCE, PCE and trichloroethanol(TCEOH) concentrations in blood and trichlroroacetic acid(TCA) and TCEOH concentrations in urine were measured and analyzed in a total of 1,033 residents around Camp Carroll. TCA and TCEOH are metabolites of TCE and PCE, respectively. The information on demographic characteristics and exposure variables in relation to underground water were obtained through a questionnaire completed by the subjects. Results: TCE, PCE and TCEOH concentrations were not detected in blood. Detection rates of TCA and TECOH concentrations in urine were 98.5% and 36.6%, respectively. Creatinine-corrected average TCA and TCEOH concentrations were $12.23{\pm}23.81{\mu}g/g$ and $0.66{\pm}4.31{\mu}g/g$, respectively. A significant difference was not shown between the drinking group and no drinking group for underground water, which was assumed as a potential route of exposure to TCE and PCE through the consumption of ground water. However, females drinking ground water showed a significantly higher mean level of TCA in urine than did males. There was no significant difference according to drinking ground water as a potential source of exposure to TCE and PCE in residents around Camp Carroll. Conclusions: Considering the statistical analysis of factors affecting exposure to TCE and PCE in ground water along with previous reports, TCA in urine as exposure to TCE and PCE might not be appropriate because it is found in chlorinated drinking water. Therefore, TCA concentration in urine may be the result of drinking of chlorinated water.

Quantification of urea in serum by isotope dilution HPLC/MS (동위원소 희석 HPLC/MS에 의한 혈청 내 urea의 정량)

  • Lee, Hwashim;Park, Sangryoul
    • Analytical Science and Technology
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    • v.18 no.4
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    • pp.271-277
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    • 2005
  • Urea in blood has been measured as an effective marker for diagnosis of renal function. Urea which is e end-product of nitrogen containing metabolites such as proteins is filtered through glomeruli of kidneys and then excreted as urine. If the renal function is deteriorated, the urea concentration in blood will be increased, from which the healthiness of renal function is judged. In order to improve the confidence of diagnosis results, the results must keep traceability chain to certified reference materials, which was certified by primary reference method. In this study, we proposed isotope dilution-liquid chromatography/mass spectrometry (ID-LC/MS) as a candidate primary method, in which $15^N_2$-urea is used as an internal reference material. The developed method is highly accurate in principle and is convenient as it does not require cumbersome derivatization. 0.1 mmol/L ammonium chloride was selected as a mobile phase for HPLC because it provided low interference in MS analysis of relatively low molecular weighted urea. HPLC and MS were connected with an electrospray ionization (ESI) interface of positive mode, which provided high sensitivity and reproducibility. The developed method was validated with internationally recognized reference materials, and we have obtained satisfactory results in an international ring trial. The expanded uncertainty calculated according to ISO guide was 1.8% at 95% confidence interval. The developed method is being used as a primary reference measurement method such as for certification of serum certified reference materials (CRMs).

Identification and FT-IR Spectrum Analysis of Lichens on Flagpole Support in Beopjusa Temple (법주사 당간지주 지의류의 동정 및 FT-IR 스펙트럼 특성 분석)

  • Kim, Young Hee;Lee, Jeung Min;Choie, Myoungju;Hong, Jin Young;Jo, Chang Wook;Kim, Soo Ji;Jeong, So Young
    • Journal of Conservation Science
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    • v.33 no.5
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    • pp.391-398
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    • 2017
  • This study was conducted to investigate lichen as a typical biomass damage on the surfaces of flagpole-supporting stones in the Beopjusa temple. The lichens present on the flagpole-supporting stones were limited to five species. Two dominant lichen species were identified: Aspicilia sp. and Pertusaria flavicans. One foliose species and one fruticose species, which are rarely observed on crustose lichens, were identified as Xanthoparmelia conspersa and Ramalina sekika, respectively. The lichen inhabiting the black algae layer was confirmed as Leprocaulon textum. ATR-FTIR was performed to analyze the secondary metabolites synthesized by the lichens. By comparing the FTIR spectra of Xanthoparmelia conspersa and Ramalina sekika, the synthesized organic acids were confirmed to differ from each other. Furthermore, the spectral changes and characteristics due to functional groups in the molecules were confirmed.

The Algicidal Activity of Pseudoalteromonas sp. NH-12 against the Toxic Dinoflagellate Alexandrium catenella (유독성 와편모류 Alexandrium catenella에 대한 Pseudoalteromonas sp. NH-12의 살조능)

  • Jeoung, Nam-Ho;Son, Hong-Joo;Jeong, Seong-Yun
    • Korean Journal of Environmental Agriculture
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    • v.31 no.2
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    • pp.175-184
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    • 2012
  • BACKGROUND: The aim of this study was to isolate and identify algicidal bacterium that tends to kill the toxic dinoflagellate Alexandrium catenella, and to determine the algicidal activity. METHODS AND RESULTS: Among of four algicidal bacteria isolated in this study, NH-12 isolate was the strongest algicidal activity against A. catenella. NH-12 isolate was identified on the basis of biochemical characteristics and analysis of 16S rRNA gene sequences. The isolate showed 97.67% homology with Pseudoalteromonas prydzensis ACAM $620^T$ (U85855), and was designated Pseudoalteromonas sp. NH-12. The optimal culture conditions of this isolate were $25^{\circ}C$, initial pH 8.0, and 3.0% (w/v) NaCl concentration. The algicidal activity of NH-12 was significantly increased to maximum value in the late of logarithmic phase of bacterial culture. As a result of 'cell culture insert' experiment, NH-12 is assumed to produce secondary metabolites, as an indirect attacker. When 10% culture filtrate of NH-12 was applied to A. catenella, over 99% of algal cells were destroyed within 24 h. In addition, the killing effects were increased in dose and time dependent manners. CONCLUSION(S): Taken together, our results suggest that Pseudoalteromonas sp. NH-12 could be a candidate for controlling of toxic algal blooms.