• Title/Summary/Keyword: 대사체 분석

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Urinary Profiles of the Endogenous Steroids in Pre-Menopausal Women with Uterine Leiomyoma (자궁근종 성장에 있어서 내인성 성호르몬의 역할)

  • Jung, Byung-Hwa;Bai, Sang-Wook;Chung, Bong-Chul;Kim, Sei-Kwang;Park, Ki-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.1
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    • pp.23-29
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    • 2003
  • 목 적: 본 연구의 목적은 내인성 성호르몬이 자궁근종의 성장에 미치는 영향을 알아보고자 연구를 시행하였다. 연구대상 및 방법: 폐경전 자궁근종 환자 27명과 같은 연령대의 정상여성 25명을 대상으로 하였다. 모든 대상군의 여성에서 24시간 소변을 모아서 소변내 estrogen, androgen의 대사체들을 GC-MS를 이용하여 측정하였으며 두 군에서의 차이를 비교분석 하였다. 결 과: 소변내 $17{\beta}$-estradiol, 5-androstene-$3{\beta}$, $16{\beta}$, $17{\beta}$-triol, 11-keto-ethiocholanolone, $11{\beta}$-hydroxy-androsterone, THS, THA, THE, a-cortolone, a-cortol 및 $\beta$-cortol가 환자군에서 의의있게 증가하였으며 $17{\beta}$estradiol/estrone 및 $11{\beta}$-hydroxy-ethiocholanolone/$11{\beta}$-hydroxy-androsterone도 환자군에서 의의있게 증가하였다. 결 론: 자궁근종의 성장은 요중 estrogen과 androgen의 농도와 밀접한 관련이 있으며 이는 환자의 스테로이드 호르몬 대사 감소에 기인한 것으로 사료된다.

Determination of alkylphenol ethoxylate in water by high performance liquid chromatography/electrospray ionization/mass spectrometry (HPLC/ESI/MS를 이용한 물 중의 알킬페놀에톡실레이트 분석)

  • Lee, Jeongae;Park, Song-Ja;Chung, Bong Chul
    • Analytical Science and Technology
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    • v.17 no.3
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    • pp.263-270
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    • 2004
  • A method is described for the analysis of short-chain alkylphenol ethoxylates (APEOs), 4-octylphenol-di-ethoxylate (OP2EO) and 4-nonylphenol-di-ethoxylate (NP2EO), in drinking water or wastewater using reversed phase high-performance liquid chromatography with electrospray ionization mass spectrometry. The solvent system was water and methanol containing $10{\mu}M$ trifluoroacetic acid as an ionization solvent. We acidified 1 L of water samples to less than pH 2 with concentrated $H_2SO_4$ and loaded onto Sep-Pak $C_{18}$, and eluted with acetone. The calibration of OP2EO and NP2EO was performed for the concentration range from 20 to 500 ng/L and the correlation coefficients were 0.999 and 0.990, respectively. The limits of detection were 20 ng/L (OP2EO) and 50 ng/L (NP2EO) at a signal-to-noise ratio of 3. Accuracy and precision of this analytical method were 85.8 ~ 122.1% and 8.2 ~ 18.8%, respectively. The proposed method allowed a sensitive and rapid detection of OP2EO and NP2EO and it could be applied for monitoring of APEOs from environmental samples.

Simultaneous GC/MS Analyses of Organic acids and Amino acids in Urine using TMS-TFA derivative (TMS-TFA 유도체화를 이용한 소변여지 중 유기산과 아미노산의 GC/MS 동시분석)

  • Yoon, Hye-Ran
    • Analytical Science and Technology
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    • v.19 no.1
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    • pp.107-114
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    • 2006
  • Early diagnosis and medical intervention are critical for the treatment of patients with metabolic disorders. A rapid analytical method was developed for simultaneous quantification of organic acids and amino acids in urine without labor-intensive pre-extraction procedure showing high sensitivity and specificity. A new method consisted of simple two-step trimethylsilyl (TMS)-trifluoroacetyl (TFA) derivatization using GC/MS-selective ion monitoring (SIM). Filter paper urine specimens were dried under nitrogen after being fortified with internal standard (tropate) in a mixture of distilled water and methanol. Methyl orange was added to the residue as indicator reagent. Silyl derivative of carboxylic functional group was followed by trifluoroacetyl derivative for amino functional group. N-methyl-N-(trimethylsilyl-trifluoroacetamide) and N-methyl-bistrifluoroacetamide were consecutively added and heated for 15-20 min at $65^{\circ}C-70^{\circ}C$, for TMS-TFA derivative, respectively. This reactant was analyzed by GC/MS-SIM. Linear dynamic range showed 0.001-50 mg with the detection limit of (S/N=3) 10-200 ng, and the quantification limit of 80-900 ng in urine. Correlation coefficient of regression line was 0.994-0.998. When the method was applied to the patients 'urine, it clearly differentiated the normal from the patient with metabolic disorder. The study showed that the developed method could be the method of choices in rapid and sensitive screening for organic aciduria and amino acidopathy.

DNA microarray analysis of RNAi plant regulated expression of NtROS2a gene encoding cytosine DNA demethylation (시토신 탈메틸화 관련 NtROS2a 유전자 발현을 제어한 RNAi 식물의 DNA microarray 분석)

  • Choi, Jang Sun;Lee, In Hye;Jung, Yu Jin;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.231-239
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    • 2016
  • To study the transcript levels of epigenetically regulated genes in tobacco, we have developed a transgenic line OX1 overexpressing NtROS2a gene encoding cytosine DNA demethylation and a RNAi plant line RNAi13. It has been reported that salt- and $H_2O_2$-stress tolerance of these transgenic lines are enhanced with various phenotypic characters (Lee et al. 2015). In this paper, we conducted microarray analysis with Agilent Tobacco 4 x 44K oligo chip by using overexpression line OX1, RNAi plant line RNAi 13, and wild type plant WT. Differentially expressed genes (DEGs) related to metabolism, nutrient supply, and various stressed were up-regulated by approximately 1.5- to 80- fold. DEGs related to co-enzymes, metabolism, and methylation functional genes were down-regulated by approximately 0.03- to 0.7- fold. qRT-PCR analysis showed that the transcript levels of several candidate genes in OX1 and RNAi lines were significantly (p < 0.05) higher than those in WT, such as genes encoding KH domain-containing protein, MADS-box protein, and Zinc phosphodiesterase ELAC protein. On the other hand, several genes such as those encoding pentatricopeptide (PPR) repeat-containing protein, histone deacetylase HDAC3 protein, and protein kinase were decreased by approximately 0.4- to 1.0- fold. This study showed that NtROS2a gene encoding DNA glycosylase related to demethylation could regulate adaptive response of tobacco at transcriptional level.

Analytical trends in mass spectrometry based metabolomics approaches of neurochemicals for diagnosis of neurodegenerative disorders (퇴행성신경질환의 진단을 위한 신경전달물질 대사체의 질량 분석법 동향)

  • Lee, Na-Kyeong;Jeon, Won-Jei;Jeong, Seung-Woo;Byun, Jae-Sung;Lee, Wonwoong;Hong, Jongki
    • Analytical Science and Technology
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    • v.30 no.6
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    • pp.355-378
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    • 2017
  • Because neurochemicals are related to homeostasis and cognitive and behavioral functions in human body and because they enable the diagnosis of numerous neurodegenerative disorders, there has been increasing interest in the development of analytical platforms for neurochemical profiling in biological samples. In particular, mass spectrometry (MS)-based analytical methods combined with chromatographic separation have been widely used to profile neurochemicals in metabolic pathways. However, development of delicate sample preparation procedures and highly sensitive instrumental detection is necessary considering the trace levels and chemical instabilities of neurochemicals in biological samples. Therefore, in this review, analytical trends in MS-based metabolomics approaches to neurochemicals in multiple biological samples, such as urine, blood, CSF, and biological tissues, are discussed. This paper is expected to contribute to the development of an analytical platform to discover biomarkers that will aid diagnosis, prognosis, and treatment of neurodegenerative disorders.

Astaxanthin Biosynthesis in Transgenic Arabidopsis by Using Chyb Gene Encoding β-Carotene Hydroxylase (β-Carotene Hydroxylase 관련 Chyb 유전자를 이용한 형질전환 Arabidopsis에서 Astaxanthin의 생합성)

  • Lee, Ho-Jae;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.231-237
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    • 2004
  • Oxycarotenoids are oxygenated carotenoids that perform critical roles in plants. $\beta$-Carotene hydroxylase adds hydroxyl groups to the $\beta$-rings of carotenes and has been cloned from several bacteria and plants including Arabidopsis. This study was carried out to investigate the effect of $\beta$-carotene hydroxylase gene (Chyb) on the oxycarotenoids biosynthesis in the transgenic Arabidopsis. Construct of pGCHYB containing Chyb was established onto Gateway vector system (pENTR3C gateway vector and pH2GW7 destination vector). Arabidopsis thaliana (cv. Columbia) was transformed with Agrobacterium tumerfacience GV3101 harboring pGCHYB construct driven by 35S promoter and hygromycin resistant gene. Seven hundred bases paired PCR products, indicating the presence of Chyb gene, were found in the transformants by PCR analysis using Chyb primers. Hygromycin resistance assay showed that transgenes were stably inherited to next generation. The overexpression of the Chyb gene resulted in the decrease carotenoid content. Especially, astaxanthin unusual oxycarotenoid in wild type Arabidopsis was detected in the transgenic plants. This means that decreased carotenoids might be converted into astaxanthin metabolism with the aid of silent gene in the host.

Transcriptome Profiling of Kidney Tissue from FGS/kist Mice, the Korean Animal Model of Focal Segmental Glomerulosclerosis (국소성 분절성 사구체 신병증의 동물 모델 (FGS/kist 생쥐) 신 조직의 유전자 발현 양상)

  • Kang, Hee-Gyung;Lee, Byong-Sop;Lee, Chul-Ho;Ha, Il-Soo;Cheong, Hae-Il;Choi, Yong
    • Childhood Kidney Diseases
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    • v.15 no.1
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    • pp.38-48
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    • 2011
  • Purpose: Focal segmental glomerulosclerosis (FSGS) is the most common glomerulopathy causing pediatric renal failure. Since specific treatment targeting the etiology and pathophysiology of primary FSGS is yet elusive, the authors explored the pathophysiology of FSGS by transcriptome analysis of the disease using an animal model. Methods: FGS/kist strain, a mouse model of primary FSGS, and RFM/kist strain, as control and the parent strain of FGS/kist, were used. Kidney tissues were harvested and isolated renal cortex was used to extract mRNA, which was run on AB 1700 mouse microarray chip after reverse transcription to get the transcriptome profile. Results: Sixty two genes were differentially expressed in FGS/kist kidney tissue compared to the control. Those genes were related to cell cycle/cell death, immune reaction, and lipid metabolism/vasculopathy, and the key molecules of their networks were TNF, IL-6/4, IFN${\gamma}$, TP53, and PPAR${\gamma}$. Conclusion: This study confirmed that renal cell death, immune system activation with subsequent fibrosis, and lipid metabolism-related early vasculopathy were involved in the pathophysiology of FSGS. In addition, the relevance of methodology used in this study, namely transcriptome profiling, and Korean animal model of FGS/kist was validated. Further study would reveal novel pathophysiology of FSGS for new therapeutic targets.

Effects of soy protein supplementation and treadmill running exercise on the changes of body composition, blood metabolic markers, estradiol, estrogen receptor gene expression in ovariectomized rats (콩 단백질 섭취 및 트레드밀 운동이 난소절제 흰쥐의 신체구성, 혈중 대사변인, 에스트라디올 농도 및 에스트로겐 수용체 유전자 발현에 미치는 영향)

  • Kim, K.J.
    • Exercise Science
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    • v.21 no.2
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    • pp.243-254
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    • 2012
  • This study tried to suggest the applicapability of soy protein supplementation and treadmill running exercise for the replacement theraphy on negative effects to estrogen metabolism in menopause. This study was analyzed the effects of 8 week soy protein supplementation and treadmill running exercise on the changes of body composition, blood concentrations of glucose, insulin, triglycerides, C-reactive protein (CRP) and estradiol, estrogen receptor gene expression of liver in ovariectomized rats. Ovariectomized groups showed the increasing responses of body weight, body fat percentage, and blood concentration of triglycerides, but these groups showed the decreasing responses of blood estradiol level and estrogen receptor gene expression in liver. Ovariectomized groups showed the positive responses of blood concentrations of lipid markers, insulin, estradiol, and estrogen receptor gene expression of liver except bone mineral contents after 8 week soy protein supplementation and treadmill running exercise. I could find the positive effects of 8 week soy protein supplementation and treadmill running exercise on the estrogen and lipid metabolism in ovariectomized rats, but this study could not confirmed the detailed replacement program of exercise intensity, duration, and soy protein volume for estrogen metabolism in ovariectomized rats.

Complete genome sequencing of Pseudomonas fluorescens NBC275, a biocontrol agent against fungal pathogens of plants and insects (식물 및 곤충의 곰팡이 병원균에 항균력을 가진 Pseudomonas fluorescens NBC275 균주의 유전체 염기서열)

  • Dutta, Swarnalee;Yu, Sang-Mi;Nagendran, Rajalingam;Jeong, Sang Chul;Lee, Yong Hoon
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.157-159
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    • 2019
  • Pseudomonas fluorescens NBC275 (Pf275) isolated from soil sample collected at riverside of Nakdonggang showed antagonistic activity against fungal pathogens of plants and insects. Here we present complete genome sequence of Pf275. The genome comprises of 6,610,362 bp with GC content of 60.9%, which includes 5,869 predicted protein-coding genes, 16 rRNAs, and 65 tRNAs. Genome analysis revealed gene clusters encoding antimicrobial secondary metabolites such as pyoverdine, 2, 4-diacetylphloroglucinol, and phenazine, which are known to play essential roles in biocontrol of diseases.

Complete genome sequencing of Pseudomonas parafulva PpaJBCS1880, a biocontrol and plant growth promoting agent (식물 병 방제 및 생육촉진 효과를 나타내는 Pseudomonas parafulva PpaJBCS1880균주의 유전체 염기서열)

  • Dutta, Swarnalee;Wabyona, Alex;Kakembo, David;Lee, Yong Hoon
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.286-288
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    • 2019
  • Pseudomonas parafulva PpaJBCS1880 (PpaJBCS1880) isolated from rice seeds showed strong antagonistic activity against bacterial plant pathogens by producing lipopeptide. Furthermore, the strain controlled the incidence of bacterial pustule in soybean plants and promoted the growth of rice plants. Here we present complete genome sequence of PpaJBCS1880. The genome comprises of 5,208,480 bp with GC content of 63.4%, which includes 4,487 predicted protein-coding genes, 19 rRNAs, and 74 tRNAs. Genome analysis revealed genes encoding antimicrobial secondary metabolites such as lipopeptide, pyoverdine, phenazine, and hydrogen cyanide, which are known to play essential roles in biocontrol of plant diseases.