• 한국의학물리학회지:의학물리
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• 제12권1호
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• pp.31-39
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• 2001

핵자기공명분광학(NMR Spectroscopy)을 이용해서 우리는 생물체의 조직안에 있는 각종 대사물질들을 측정할 수 있게 되었으며, 이것은 또 지금까지 알려진 여러 방법 중에서 생물체내의 신진대사 물질이나 생화학적인 변화를 비침습적(noninvasive)으로 알아낼 수 있는 유일한 방법에 속한다. 양성자 핵자기공명분광학은 최근들어 사람이나 흑은 동물뇌의 생화학적인 특성을 연구하는데 많이 사용되어오고 있다. 핵자기공명영상은 주로 물분자에 있는 양성자의 밀도와 그들의 이완현상(relaxation phenomena)을 측정하여 영상화하지만 핵자기공명분광학은 생체조직내의 신진대사에 관한 생화학적인 정보를 제공한다. 많은 경우에 있어서 핵자기공명분광학은 CT나 MRI 영상에서는 발견할 수 없는 정보를 제공함으로써 특정질병의 진단에 사용될 수 있으며, 따라서 환자들의 질병상태를 효과적으로 진단하는데 쓰여질 수 있다. 따라서 본 연구에서는 사람의 뇌에서 수소 핵자기공명분광학을 이용하여 대사물질들의 농도를 정량화하는 방법을 시도하였다. 이를 위해 펀텀을 이용하여 대사물질들 각각의 스팩트럼을 구했고, 성인 남녀 16명을 대상으로 insula gray matter 부위의 대사물질들의 농도를 측정하였다. 그리고 이 값들은 다른 연구자들에 의해 밝혀진 값들과 매우 근사한 값을 제공하였다.

• Journal of Animal Science and Technology
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• 제64권2호
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• pp.247-261
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• 2022

Ketosis is associated with high milk yield during lactating or insufficient feed intake in lactating dairy cows. However, few studies have been conducted on the metabolomics of ketosis in Korean lactating dairy cows. The present study aimed to investigate the serum and urine metabolites profiling of lactating dairy cows through proton nuclear magnetic resonance (¹H-NMR) spectroscopy and comparing those between healthy (CON) and subclinical ketosis (SCK) groups. Six lactating dairy cows were categorized into CON and SCK groups. All experimental Holstein cows were fed total mixed ration. Serum and urine samples were collected from the jugular vein of the neck and by hand sweeping the perineum, respectively. The metabolites in the serum and urine were determined using ¹H-NMR spectroscopy. Identification and quantification of metabolites was performed by Chenomx NMR Suite 8.4 software. Metabolites statistical analysis was performed by Metaboanalyst version 5.0 program. In the serum, the acetoacetate level was significantly (p < 0.05) higher in the SCK group than in the CON group, and whereas acetate, galactose and pyruvate levels tended to be higher. CON group had significantly (p < 0.05) higher levels of 5-aminolevulinate and betaine. Indole-3-acetate, theophylline, p-cresol, 3-hydroxymandelate, gentisate, N-acetylglucosamine, N-nitrosodimethylamine, xanthine and pyridoxine levels were significantly (p < 0.05) higher in the urine of the SCK group than that in the CON group, which had higher levels of homogentisate, ribose, gluconate, ethylene glycol, maltose, 3-methyl-2-oxovalerate and glycocholate. Some significantly (p < 0.05) different metabolites in the serum and urine were associated with ketosis diseases, inflammation, energy balance and body weight. This study will be contributed useful a future ketosis metabolomics studies in Korea.

• Investigative Magnetic Resonance Imaging
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• 제12권1호
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• pp.8-19
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• 2008

목적 : 본 연구에서는 인체 뇌 대사물질에 대하여 2D MR 기술인 correlation spectroscopy (COSY) 와 nuclear Overhauser effect/enhancement spectroscopy (NOESY)를 직접 적용하고, 데이터를 획득하여 인체 뇌대사물질들간의 스칼라 짝지움 (coupling)과 쌍극자 (dipolar) 상호작용- NOE에 대한 분석을 통하여 결합연결관계 및 공간연결관계에 대한 정보를 획득하고자 하였다. 대상 및 방법 : 모든 2-D (dimensional) MR 실험 (즉, COSY와 NOESY)은 z축 능동차폐 펄스경사자장, 삼중공명 동결탐침기가 장착된 Bruker Avance 500 (11.8 T) 장비에서 298 K에 수행되었다. MRS상에서 뇌 대사물질과 유사하도록만든 희석액을 만들었고, 최종 MRS 샘플은 10% $D_2O$를 이용하였다. 2-D 스펙트라는 2048 복합 (complex) 데이터 포인트로서 총 320개 의 free induction decay (FID)를 평균화 (averaging)하였고, $H_2O$에서 얻어진 스펙트라는 8012 Hz 였다. 반복지연 (repetition delay) 시간은 2초, 각각의 FID는 4개의 평균화를 선택하였다. 얻어진 2D-COSY, 2D-NOESY 데이터는 Top Spin 2.0 소프트웨어에서 후 처리기법 (post-processing)에 의해 분석되었다. 분석 대상 대사물질은 N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), glutamine (Gln), glutamate (Glu), myo-inositol (Ins), lactate (Lac)로서 총 7 가지 화학물로서 주요 목표 피크로 정했다. 결과 : 인체 뇌 대사물질에 대한 대칭형태의 2D-COSY와 2D-NOESY 스펙트럼을 획득하였고, COSY 스펙트럼상에서는 오직 1.0-4.5 ppm 사이에서만 교차피크들이 생성된 반면 NOESY 스펙트럼상에서는 1.0-4.5 ppm 외에도 7.9 ppm에서 공명 교차피크를 발견할 수 있었다. COSY 스펙트럼을 통하여 lactate에서 메틸 프로톤과 CH 프로톤의 COSY 크로스피크가 발견되었고, NAA에서 메틸렌 프로톤들간과 메틸렌 프로톤과 NH프로톤의 크로스피크가 발견되었고, Ins에서 CH 프로톤 들간의 크로스피크가 발견되었다. NOESY 스펙트럼을 통하여 NAA 분자내 NH 프로톤과 메틸 (-CH3) 프로톤과의 NOESY 크로스피크가 발견되었고, lactate에서 메틸 프로톤과 CH 프로톤과의 크로스피크가 발견되었고, Cr에서 메틸 프로톤과 메틸렌 프로톤과의 크로스피크가 발견되었고, Glu에서 메틸렌 프로톤 들간과 또한 메틸렌 프로톤과 CH 프로톤과의 크로스피크가 발견되었고, Gln에서 메틸렌 프로톤과 CH 프로톤과의 크로스피크가 발견되었고, Ins에서 CH 프로톤 들간의 크로스피크가 발견되었다. 결론 : 본 연구에서는 in vitro 상태의 인체 뇌 대사물질에 2D-COSY와 2D-NOESY 기술을 직접 적용하고, 결합연결관계 및 공간연결관계에 대한 정보를 성공적으로 획득하여 분석하여 보았다. 본 연구 결과물은 향후 인체 내 in vivo 2D-COSY를 이용한 뇌 대사물질 연구에 매우 유용하리라 사료된다.

• Animal Bioscience
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• 제34권2호
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• pp.213-222
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• 2021

Objective: The metabolites that constitute the rumen fluid and milk in dairy cattle were analyzed using proton nuclear magnetic resonance (1H-NMR) spectroscopy and compared with the results obtain for other dairy cattle herds worldwide. The aim was to provide basic dataset for facilitating research on metabolites in rumen fluid and milk. Methods: Six dairy cattle were used in this study. Rumen fluid was collected using a stomach tube, and milk was collected using a pipeline milking system. The metabolites were determined by 1H-NMR spectroscopy, and the obtained data were statistically analyzed by principal component analysis, partial least squares discriminant analysis, variable importance in projection scores, and metabolic pathway data using Metaboanalyst 4.0. Results: The total numbers of metabolites in rumen fluid and milk were measured to be 186 and 184, and quantified as 72 and 109, respectively. Organic acid and carbohydrate metabolites exhibited the highest concentrations in rumen fluid and milk, respectively. Some metabolites that have been associated with metabolic diseases (acidosis and ketosis) in cows were identified in rumen fluid, and metabolites associated with ketosis, somatic cell production, and coagulation properties were identified in milk. Conclusion: The metabolites measured in rumen fluid and milk could potentially be used to detect metabolic diseases and evaluate milk quality. The results could also be useful for metabolomic research on the biofluids of ruminants in Korea, while facilitating their metabolic research.

• Animal Bioscience
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• 제36권8호
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• pp.1199-1208
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• 2023

Objective: The purpose of this study was to perform a comparative analysis of metabolite levels in serum and milk obtained from cows fed on different concentrate to forage feed ratios. Methods: Eight lactating Holstein cows were divided into two groups: a high forage ratio diet (HF; 80% Italian ryegrass and 20% concentrate of daily intake of dry matter) group and a high concentrate diet (HC; 20% Italian ryegrass and 80% concentrate) group. Blood was collected from the jugular vein, and milk was sampled using a milking machine. Metabolite levels in serum and milk were estimated using proton nuclear magnetic resonance and subjected to qualitative and quantitative analyses performed using Chenomx 8.4. For statistical analysis, Student's t-test and multivariate analysis were performed using Metaboanalyst 4.0. Results: In the principal component analysis, a clear distinction between the two groups regarding milk metabolites while serum metabolites were shown in similar. In serum, 95 metabolites were identified, and 13 metabolites (include leucine, lactulose, glucose, betaine, etc.) showed significant differences between the two groups. In milk, 122 metabolites were identified, and 20 metabolites (include urea, carnitine, acetate, butyrate, arabinitol, etc.) showed significant differences. Conclusion: Our results show that different concentrate to forage feed ratios impact the metabolite levels in the serum and milk of lactating Holstein cows. A higher number of metabolites in milk, including those associated with milk fat synthesis and the presence of Escherichia coli in the rumen, differed between the two groups compared to that in the serum. The results of this study provide a useful insight into the metabolites associated with different concentrate to forge feed ratios in cows and may aid in the search for potential biomarkers for subacute ruminal acidosis.

• Animal Bioscience
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• 제34권12호
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• pp.1930-1939
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• 2021

Objective: The aim of the study was to conduct metabolic profiling of dairy cattle serum and urine using proton nuclear magnetic resonance (¹H-NMR) spectroscopy and to compare the results obtained with those of other dairy cattle herds worldwide so as to provide a basic dataset to facilitate research on metabolites in serum and urine. Methods: Six dairy cattle were used in this study; all animals were fed the same diet, which was composed of total mixed ration; the fed amounts were based on voluntary intake. Blood from the jugular neck vein of each steer was collected at the same time using a separate serum tube. Urine samples were collected by hand sweeping the perineum. The metabolites were determined by ¹H-NMR spectroscopy, and the obtained data were statistically analyzed by performing principal component analysis, partial least squares-discriminant analysis, variable importance in projection scores, and metabolic pathway data using Metaboanalyst 4.0. Results: The total number of metabolites in the serum and urine was measured to be 115 and 193, respectively, of which 47 and 81, respectively were quantified. Lactate (classified as an organic acid) and urea (classified as an aliphatic acylic compound) exhibited the highest concentrations in serum and urine, respectively. Some metabolites that have been associated with diseases such as ketosis, bovine respiratory disease, and metritis, and metabolites associated with heat stress were also found in the serum and urine samples. Conclusion: The metabolites measured in the serum and urine could potentially be used to detect diseases and heat stress in dairy cattle. The results could also be useful for metabolomic research on the serum and urine of ruminants in Korea.

• 한국자기공명학회논문지
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• 제12권1호
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• pp.14-25
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• 2008

To investigate the 3-bond connectivity of human brain metabolites by scalar coupling interaction through 2D-correlation spectroscopy (COSY) techniques using high field NMR spectroscopy. All NMR experiments were performed at 298K on Unity Inova 500 or 600 (Varian Inc.) equipped with a triple resonance probe head with z-shield gradient. Human brain metabolites were prepared with 10% $D_2O$. Two dimensional 2D COSY spectra were acquired with 4096 complex data points in $t_2$ and 128 or 256 increments in $t_1$ dimension. The spectral width was 9615.4 Hz and solvent suppression was achieved using presaturation using low power irradiation of the water resonance during 2s of relaxation delay. NMR data were processed using VNMRJ (Varian Instrument) software and all the chemical shifts were referenced to the methyl resonance of N-acetyl aspartate (NAA) peak at 2.0 ppm. Total 10 metabolites such as N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), glutamine (Gln), glutamate (Glu), myo-inositol (Ins), lactate (Lac), taurine (Tau), ${\gamma}$-aminobutyricacid (GABA), alanine (Ala) were included for major target metabolites. Symmetrical 2D-COSY spectra were successfully acquired. Total 14 COSY cross peaks were observed even though there were parallel/orthogonal noisy peaks induced by water suppression. Except for Cr, all of human brain metabolites produced COSY cross peaks. The spectra of NAA methyl proton at 2.02 ppm and Glu methylene proton ($CH_2(3)$) at 2.11 ppm and Gln methylene proton ($CH_2(3)$) at 2.14 ppm were overlapped in the similar resonance frequency between 2.00 ppm and 2.15 ppm. The present study demonstrated that in vitro 2D-COSY represented the 3-bond connectivity of human brain metabolites by scalar coupling interaction. This study could aid in better understanding the interactions between human brain metabolites in vivo 2D-COSY study. Also it would be helpful to determine the molecular stereochemistry in vivo by using two-dimensional MR spectroscopy.

• Animal Bioscience
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• 제36권1호
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• pp.53-62
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• 2023

Objective: In this study, metabolites that changed in the rumen fluid, urine and feces of dairy cows fed different feed ratios were investigated. Methods: Eight Holstein cows were used in this study. Rumen fluid, urine, and feces were collected from the normal concentrate diet (NCD) (Italian ryegrass 80%: concentrate 20% in the total feed) and high concentrate diet (HCD) groups (20%: 80%) of dairy cows. Metabolite analysis was performed using proton nuclear magnetic resonance (NMR) identification, and statistical analysis was performed using Chenomx NMR software 8.4 and Metaboanalyst 4.0. Results: The two groups of rumen fluid and urine samples were separated, and samples from the same group were aggregated together. On the other hand, the feces samples were not separated and showed similar tendencies between the two groups. In total, 160, 177, and 188 metabolites were identified in the rumen fluid, urine, and feces, respectively. The differential metabolites with low and high concentrations were 15 and 49, 14 and 16, and 2 and 2 in the rumen fluid, urine, and feces samples, in the NCD group. Conclusion: As HCD is related to rumen microbial changes, research on different metabolites such as glucuronate, acetylsalicylate, histidine, and O-Acetylcarnitine, which are related to bacterial degradation and metabolism, will need to be carried out in future studies along with microbial analysis. In urine, the identified metabolites, such as gallate, syringate, and vanillate can provide insight into microbial, metabolic, and feed parameters that cause changes depending on the feed rate. Additionally, it is thought that they can be used as potential biomarkers for further research on subacute ruminal acidosis.

• 한국자기공명학회논문지
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• 제22권1호
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• pp.10-17
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• 2018

Metabolite identification and quantification are one of the foremost important issues in metabolomics. In NMR based metabolomics, mainly one-dimensional proton NMR spectra of biofluids, such as urine and serum are measured. However, it is not always easy to identify and quantify metabolites in one-dimensional proton NMR spectra. This article introduces useful public metabolite databases, metabolic profiling software, and articles.

• 대한방사선기술학회지:방사선기술과학
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• 제44권4호
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• pp.315-325
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• 2021

The purpose of this study is to investigate the changes in lipid proton (LP) composition according to the induced obese fatty liver and to use it as basic data for treatment and diagnosis of fatty liver in the future. The phantom study was conducted to identify differences between STEAM and PRESS Pulse sequences in LP concentration. A high-fat diet (60%) was administered to eight Sprague-Dawley rats to induce obesity and fatty liver disease. Baseline magnetic resonance imaging /spectroscopy data were obtained prior to the introduction of high-fat diet, and data acquisition experiments were performed after eight weeks using procedures identical to those used for baseline studies. The six lipid proton metabolites were calculated using LCModel software. The correlation between the fat percentage and each LP, revealed that the methylene protons at 1.3 ppm showed the highest positive correlation. The α-methylene protons to carboxyl and diallylic protons showed negative correlation with fat percentage. The methylene proton showed the highest increase in the LP; however, it constituted only 71.86% of the total LP concentration. The methylene proton plays a leading role in fat accumulation in liver parenchyma.