• Journal of Plant Biotechnology
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• 제38권1호
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• pp.9-14
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• 2011

To determine whether pattern recognition based on metabolite fingerprinting for whole cell extracts can be used to discriminate cultivars metabolically, leaves of nine commercial Orostachys plants were subjected to Fourier transform infrared spectroscopy (FT-IR). FT-IR spectral data from leaves were analyzed by principal component analysis (PCA) and Partial least square discriminant analysis (PLS-DA). The dendrogram based on hierarchical clustering analysis of these PLS-DA data separated the nine Orostachys species into five major groups. The first group consisted of O. iwarenge 'Yimge', 'Jeju', 'Jeongsun' and O. margaritifolius 'Jinju' whereas in the second group, 'Sacheon' was clustered with 'Busan,' both of which belong to O. malacophylla species. However, 'Samchuk', belong to O. malacophylla was not clustered with the other O. malacophylla species. In addition, O. minuta and O. japonica were separated to the other Orostachys plants. Thus we suggested that the hierarchical dendrogram based on PLS-DA of FT-IR spectral data from leaves represented the most probable chemotaxonomical relationship between commercial Orostachys plants. Furthermore these metabolic discrimination systems could be applied for reestablishment of precise taxonomic classification of commercial Orostachys plants.

• Journal of Microbiology and Biotechnology
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• 제21권2호
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• pp.162-169
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• 2011

A dynamic model of lactic acid fermentation using Lactococcus lactis was constructed, and a metabolic flux analysis (MFA) and metabolic control analysis (MCA) were performed to reveal an intensive metabolic understanding of lactic acid bacteria (LAB). The parameter estimation was conducted with COPASI software to construct a more accurate metabolic model. The experimental data used in the parameter estimation were obtained from an LC-MS/MS analysis and time-course simulation study. The MFA results were a reasonable explanation of the experimental data. Through the parameter estimation, the metabolic system of lactic acid bacteria can be thoroughly understood through comparisons with the original parameters. The coefficients derived from the MCA indicated that the reaction rate of L-lactate dehydrogenase was activated by fructose 1,6-bisphosphate and pyruvate, and pyruvate appeared to be a stronger activator of L-lactate dehydrogenase than fructose 1,6-bisphosphate. Additionally, pyruvate acted as an inhibitor to pyruvate kinase and the phosphotransferase system. Glucose 6-phosphate and phosphoenolpyruvate showed activation effects on pyruvate kinase. Hexose transporter was the strongest effector on the flux through L-lactate dehydrogenase. The concentration control coefficient (CCC) showed similar results to the flux control coefficient (FCC).

• 분석과학
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• 제24권4호
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• pp.237-242
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• 2011

본 연구에서는 쇠고기 및 돼지고기 중 의약품인 메타미졸(metamizol)의 잔류량을 LC/MS/MS를 이용하여 정량하는 방법을 개발하였다. 분석은 그 주대사체인 4-methylaminoantipyrin (MAA)을 대상으로 하였으며, 식육에서 acetonitrile로 추출하여 간단한 정제를 거쳐 multiple reaction monitoring (MRM) 법으로 분석하였다. 정량을 위해 바탕시료에 MAA를 첨가한 식육을 추출하여 검정곡선을 작성하였다. MAA의 검량선의 결정계수는 위 두 가지의 시료에 대해 > 0.99 이었다. 개발된 시험법으로 본 실험실 내에서 6회 반복실험을 실시하였고 타 기관 두 곳에 의뢰하여 실험실간 비교 시험을 실시하였다. 쇠고기 분석의 실험실내 정확도는 78-102% (CV 5.5-9.1%), 실험실간 정확도 98% (CV 14%), 돼지고기 분석의 실험실내 정확도는 95-99% (CV 3.9-5.6%), 실험실간 정확도는 111% (CV 13%)를 보였다.

• 한국자기공명학회논문지
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• 제21권1호
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• pp.31-43
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• 2017

Together with radiotherapy, chemotherapy using cytotoxic agents is one of the most common therapies in cancer. Metabolic changes in cancer cells are drawing much attention recently, but the metabolic alterations by anticancer agents have not been much studied. Here, we investigated the effects of commonly used cytotoxic agents on lung normal cell MRC5 and lung cancer cell A549. We employed cis-plastin, doxorubicin, and 5-Fluorouracil and compared their effects on the viability and metabolism of the normal and cancer cell lines. We first established the concentration of the cytotoxic reagents that give differences in the viabilities of normal and cancer cell lines. In those conditions, the viability of A549 decreased significantly, whereas that of MRC5 remained unchanged. To study the metabolic alterations implicated in the viability differences, we obtained the metabolic profiles using $^1H$-NMR spectrometry. The $^1H$-NMR data showed that the metabolic changes of A549 cells are more remarkable than that of MRC5 cells and the effect of 5-FU on the A549 cells is the most distinct compared to other treatments. Heat map analysis showed that metabolic alterations under treatment of cytotoxic agents are totally different between normal and cancer cells. Multivariate analysis and weighted correlation network analysis (WGCNA) revealed a distinctive metabolite signature and hub metabolites. Two different analysis tools revealed that the changes of cell metabolism in response to cytotoxic agents were highly correlated with the Warburg effect and Reductive lipogenesis, two pathways having important effects on the cell survival. Taken together, our study addressed the correlation between the viability and metabolic profiles of MRC5 and A549 cells upon the treatment of cytotoxic anticancer agents.

• 식물조직배양학회지
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• 제28권6호
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• pp.347-351
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• 2001

인삼 (Panax ginseng C.A. Meyer)은 우리나라의 전통약재로 수 세기 동안 사용되어 왔으며, 약효 및 성분에 대한 연구가 매우 활발하게 이루어져 왔으나, 인삼의 분자생물학적, 단백질 화학적 측면에서 생리연구는 매우 미미하였다. 본 연구에서는 프로테옴믹스를 이용한 인삼 단백질 군을 연구하기 위한 첫 단계로 인삼 (천풍)및 모상근의 단백질 추출 방법을 확립하고, 이차원 전기영동을 통하여 추출방법의 유용성을 확인하였다. Homogenizer와 황산암모늄 침전을 통한 단백질 추출과 액체질소 및 TCA를 이용한 단백질 추출 방법을 수행하여 비교해 본 결과 추출 단백질 양에는 큰 차이가 없으나 이차원 전기영동 수행 시 액체질소 및 TCA를 통한 방법이 프로테옴 연구에 적합하여 훨씬 해상도가 높은 gel 이미지를 얻었을 뿐 아니라, gel당 660개 이상의 단백질 sopt을 확인하였다. 또한 인삼과 모상근의 이차원 전기영동을 비교한 결과 상당히 다른 발현 패턴을 보여, 생장조건 등 외부 환경과 생리상태의 차이에 따라, 같은 유전자를 가지고 있는 조직체라 할지라도, 매우 다른 프로테옴 (proteome)을 보일 수 있음을 보여주었다.

• 한국자원식물학회지
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• 제31권6호
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• pp.641-651
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• 2018

Floral scents and metabolites from cut flowers of 14 peony cultivars (Paeonia lactiflora Pall.) were analyzed to discriminate different cultivars and to compare the Korean cultivar with the other cut peonies imported to Korea using electronic nose (E-nose) and Fourier transform infrared (FT-IR) spectroscopy combined with multivariate analysis, respectively. Principal component analysis (PCA) and discriminant function analysis (DFA) dendrogram of peony floral scents were not precisely same but there were 3 groups including same cultivars. PCA and partial least squares-discriminant analysis (PLS-DA) dendrograms of peony metabolites showed that different cut peony cultivars were clustered into two major groups including same cultivars. Fragrance pattern of Korean 'Taebaek' was classified to same group with 'Jubilee' on the PCA and DFA results and its metabolite pattern was clearly discriminated by the PCA and PLS-DA compared to the other cultivars. These results show that the 14 peony cut flowers could be discriminated corresponding to their chemical relationship and the metabolic profile of Korean 'Taebaek' has distinctive characteristics. Furthermore, we suggest that these results could be used as the preliminary data for breeding new cut peony cultivars and for improving the availability of Korean cut peony in cosmetic industry.

• Toxicological Research
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• 제25권2호
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• pp.71-78
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• 2009

We investigated the pathogenesis of liver tissue damage during the lipid peroxidation and fibrogenesis with the observation of correlations between the parameters of collagen synthesis (and deposition) and lipid peroxidation in liver fibrosis (cirrhosis) rats. Rats were randomly divided into two groups, normal and $CCl_4$-thiopental sod. intoxicated group. And the one group was treated intragastrically with the mixture of $CCl_4$-thiopental sod. 3 times per week for 3 weeks. The liver tissue and sera were used for the measurement of hydroxyproline (HYP), malonedialdehyde (MDA) and superoxide dismutase (SOD). Biochemical parameters such as aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total-bilirubin and blood urea nitrogen (BUN) were measured. Additionally, the expression of collagen ${\alpha}1$(III) and $\beta$-actin mRNA was observed by RTPCR. The histological change in liver tissue was also observed by Masson's trichrome and H&E staining. Correlation analysis was carried by Spearman's rho method. All biochemical parameters except total-bilirubin were significantly higher in the $CCl_4$-thiopental sod. treated group than that of the normal group (p < 0.01). In the $CCl_4$-thiopental sod. treated group, Hyp as a parameter of collagen synthesis (deposition) and MDA as a metabolite of lipid peroxidation, were significantly elevated by 1.98 and 2.11 times higher than that of the normal group (p < 0.001) respectively. The activity of SOD in the $CCl_4$-thiopental sod. treated group is decreased significantly by 44.8% (p < 0.001). And collagen ${\alpha}1$(III) mRNA was more expressed in the $CCl_4$-thiopental sod. treated group than that of the normal group. However, the expression of $\beta$-actin mRNA is showed similar in both of groups. A good correlation was observed between the content of hyp and MDA concentration (r = 0.70, n = 40) in the two groups. And the correlation between the levels of hyp and SOD (r = -0.71, n = 25) is also reliable. However, no correlation were observed between MDA concentration and SOD (r = -0.40, n = 25) in the two groups. Elevated levels of MDA in $CCl_4$-thiopental sod. treated rats indicated enhancement of lipid peroxidation, which is accompanied by a decrease in SOD activity. Moreover, we could confirm that the parameters of collagen synthesis (and deposition) is in good correlation with the metabolite of lipid peroxidation (MDA) and the lipid peroxidation antagonizing enzyme (SOD). Hence, we propose that (1) lipid peroxidation and collagen synthesis (and deposition) could be enhanced by intragastrically application of $CCl_4$-thiopental sod. during a short terms. And (2) the intoxication of $CCl_4$-thiopental sod. could be used for monitoring of lipid peroxidation and collagen synthesis (and deposition) for test of antioxidant and antifibrotic agent.

• 농약과학회지
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• 제12권4호
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• pp.315-322
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• 2008

실험실 내 조건에서, 물과 토양 중 cyhalofop-butyl과 그 대사물질인 cyhalofop acid를 위한 잔류분석법이 고감도에서도 간단하고 매우 효과적으로 개발되었다. 물과 토양 중 cyhalofop-butyl과 cyhalofop acid를 분석하기 위하여 액액분별추출과 silica gel chromatographic 정제를 수행하였으며 HPLC-UV를 이용하여 정성/정량하였다. cyhalofop-butyl의 회수율은 2 가지 농도에서 3 반복 수행하여 각각 82.5-100.0%와 66.7-97.9%이었고, 검출한계와 최소검출량은 두 시료에서 모두 0.02 ppm과 10 ng이었다. Cyhalofop acid의 회수율은 물과 토양에서 각각 80.7-104.8%와 76.9-98.1%이 었으며, 검출한계는 각각 0.005 ppm과 0.01 ppm이었고 최소검출량은 두 시료에서 모두 2 ng이었다. Cyhalofop-butyl의 반감기는 물과 토양에서 각각 4.14와 6.6 day였다. 개발되어진 본 시험법은 cyhalofop-butyl의 30% 유탁제를 처리한 물과 토양에서 그 잔류량을 분석하기 위하여 성공적으로 적용되었다.

• Journal of Microbiology and Biotechnology
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• 제30권4호
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• pp.505-514
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• 2020

The symbiotic nature of the relationship between algae and marine bacteria is well-studied among the complex microbial interactions. The mutual profit between algae and bacteria occurs via nutrient and vitamin exchange. It is necessary to analyze the genome sequence of a bacterium to predict its symbiotic relationships. In this study, the genome of a marine bacterium, Pseudoruegeria sp. M32A2M, isolated from the south-eastern isles (GeoJe-Do) of South Korea, was sequenced and analyzed. A draft genome (91 scaffolds) of 5.5 Mb with a DNA G+C content of 62.4% was obtained. In total, 5,101 features were identified from gene annotation, and 4,927 genes were assigned to functional proteins. We also identified transcription core proteins, RNA polymerase subunits, and sigma factors. In addition, full flagella-related gene clusters involving the flagellar body, motor, regulator, and other accessory compartments were detected even though the genus Pseudoruegeria is known to comprise non-motile bacteria. Examination of annotated KEGG pathways revealed that Pseudoruegeria sp. M32A2M has the metabolic pathways for all seven vitamin Bs, including thiamin (vitamin B1), biotin (vitamin B7), and cobalamin (vitamin B12), which are necessary for symbiosis with vitamin B auxotroph algae. We also identified gene clusters for seven secondary metabolites including ectoine, homoserine lactone, beta-lactone, terpene, lasso peptide, bacteriocin, and non-ribosomal proteins.

• Toxicological Research
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• 제26권4호
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• pp.275-283
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• 2010

Placenta transfer study in non-human primate (NHP) is one of the crucial components in the assessment of developmental toxicity because of the similarity between NHP and humans. To establish the method to determine placenta transfer in non-human primate, toxicokinetics of valproic acid (VPA), a drug used to treat epilepsy in pregnant women, were determined in pregnant cynomolgus monkeys. After mating, pregnancy-proven females were daily administered with VPA at dose levels of 0, 20, 60 and 180 mg/kg by oral route during the organogenesis period from gestation day (GD) 20 to 50. Concentrations of VPA and its metabolite, 4-ene-VPA, in maternal plasma on GDs 20 and 50, and concentrations of VPA and 4-ene-VPA in placenta, amniotic fluid and fetus on GD 50 were analyzed using LC/MS/MS. Following single oral administration of VPA to pregnant monkeys, concentrations of VPA and 4-ene-VPA were generally quantifiable in the plasma from all treatment groups up to 4-24 hours post-dose, demonstrating that VPA was absorbed and the monkeys were systemically exposed to VPA and 4-ene-VPA. After repeated administration of VPA to the monkeys, VPA was detected in amniotic fluid, placenta and fetus from all treatment groups, demonstrating that VPA was transferred via placenta and the fetus was exposed to VPA, and the exposures were increased with increasing dose. Concentrations of 4-ene-VPA in amniotic fluid and fetus were below the limit of quantification, but small amount of 4-ene-VPA was detected in placenta. In conclusion, pregnant monkeys were exposed to VPA and 4-ene-VPA after oral administration of VPA at dose levels of 20, 60 and 180 mg/kg during the organogenesis period. VPA was transferred via placenta and the fetus was exposed to VPA with dose-dependent exposure. The metabolite, 4-ene VPA, was not detected in both amniotic fluid and fetus, but small amount of 4-ene-VPA was detected in placenta. These results demonstrated that proper procedures to investigate placenta transfer in NHP, such as mating and diagnosis of pregnancy via examining gestational sac with ultrasonography, collection of amniotic fluid, placenta and fetus after Caesarean section followed by adequate bioanalysis and toxicokinetic analysis, were established in this study using cynomolugus monkeys.