• Title/Summary/Keyword: Glutamine

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In Vitro Nuclear Maturation of Canine Oocytes Obtained from Differents Stage of Estrus Cycle (개의 발정주기가 난자의 체외성숙에 미치는 영향)

  • Kim, Min-Kyu;Kim, Hye-Jin;Cho, Jong-Ki;Jang, Gu;Lee, Kyu-Seung;Kang, Sung-Geun;Lee, Byung-Chun;Hwang, Woo-Seok
    • Journal of Embryo Transfer
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    • v.17 no.2
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    • pp.145-151
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    • 2002
  • The aim of these experiments was to investigate in vitro nuclear maturation of canine oocyte collected from various stages of estrus cycles, Ovaries were obtained from 1 to 4 year-old mongrel bitch and minced for oocyte collection in phosphate buffered saline with 100 iu penicillin-G $m\ell$/sup -1/, 50 $\mu\textrm{g}$ streptomycin sulphate $m\ell$/sup -1/ and 0.1% (w/v) polyvinyl alcohol. Cumulus-oocyte-complexes (COCs) were washed in HEPES buffered tissue culture medium (TCM)199 and in vitro matured in TCM-199 culture medium supplemented with sodium pyruvate 0.028mg/$m\ell$, L-glutamine 0.146mg/$m\ell$, penicillin G 10,000 IU/$m\ell$, streptomycin 0.031mg/$m\ell$ and 10% (v/v) fatal calf serum. COCs were in vitro matured for 48~72 hrs at 39$^{\circ}C$ in humidified 5% $CO_2$ in air atmosphere. In vitro matured oocytes were remove the cumulus cells using 0.2% (v/v) hyaluronidase. After denuding, oocyte were placed in acetic acid : methanol : chlorform solusion (3:6 : 1.5 v/v) for 30 sec and acetic acid: ethanol(1:3 v/v) for 48hrs fixation. Nuclear maturation was classified to GV, GVBD, MI, MII and degenerate oocyte under microscopy after 1% aceto-orcein stain. In vitro maturation rates at 48hrs were not significantly difference among the oocytes collected from different stage of estrus at 15.9%, 16.3%, 23.7% and 18.2% for anestrus, proestrus, estrus and diestrus. However, the oocytes maturation(36.6%) of collected from estrus ovaries were significantly different from oocytes derived from proesturs, diestrus and anestrus ovaries(30.8%, 17.5% and 22.1%; p<0.05). The overall in vitro maturation rates were significantly higher (p<0.05) in 72hrs culture than 48hrs culture system. In summary, there was a tendency for higher in vitro maturation rates with the oocyte collected from estrus ovary than other stages of estrus. Also, for nuclear maturation, in vitro culture of oocyte for 72hrs was better than 48hrs culture.

Traumatic Contusion of ICR Mouse Brain by FPI : $^{1}\textrm{H}$ MR Spectroscopic Study (유체타진손상기법에 의한 ICR 쥐의 뇌손상: 자기공명분광법)

  • Park, Chi-Bong;Kim, Hwi-Yool;Jeun, Sin-Soo;Han, Young-Min;Han, Duk-Young;Kang, Young-Woon;Choe, Bo-Young
    • Progress in Medical Physics
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    • v.14 no.4
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    • pp.259-267
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    • 2003
  • In vivo $^1$H magnetic resonance spectroscopy (MRS) at 4.7 T was applied to investigate the cerebral metabolite changes of mice brain before and after experimental brain trauma. In vivo $^1$H MR spectra were acquired from a voxel covering right parietal cortex in normal brain, used as control subjects. After experimental brain trauma using the fluid percussion injury (FPI) method, $^1$H MR spectra were acquired from the same lesion three days after trauma. Metabolite ratios of the injured lesion were compared to those of controls. After trauma, N-acetylaspartate (NAA)/creatine (Cr) ratio, as a neuronal marker was decreased significantly versus controls, indicating neuronal loss. The ratio of NAA/Cr in traumatic brain contusion was 0.90$\pm$0.11, while that in normal control subjects was 1.13$\pm$0.12 (P=0.001). Choline (Cho)/Cr ratio had a tendency to rise in experimental brain contusion (P=0.02). Cho/Cr ratio after trauma was 0.91$\pm$0.17 while that before traumas was 0.76$\pm$0.15. Cho/Cr ratio was increased and this might indicate a inflammatory activity. However, no significant difference of [(glutamate+glutamine) (Glx)]/Cr was established between experimental traumatic brain injury models and normal controls. Lactate (Lac)/Cr ratio was appeared as a sign of shifted posttraumatic energy metabolism and increased versus controls. These findings strongly suggest that in vivo $^1$H MRS may be a useful modality for clinical evaluation of traumatic contusion and could aid in better understanding the neuropathologic process of traumatic contusion induced by FPI. In the present study, in vivo $^1$H MRS was proved to be a useful non-invasive method for in vivo diagnosis and monitoring of posttraumatic metabolism in models of brain contusion.

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Studies on the Chemical Constituents of the New Zealand Deer Velvet Antler Cervus elaphus var. scoticus-(I)

  • Lee, Nam Kyung;Shin, Hyun Jung;Kim, Wan Seok;Lee, Jong Tae;Park, Chae Kyu
    • Natural Product Sciences
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    • v.20 no.3
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    • pp.160-169
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    • 2014
  • 44 compounds and 9 minerals were isolated from and detected in the New Zealand deer velvet antler Cervus elaphus var. scoticus L$\ddot{o}$nnberg. The chemical structures of (1 - 26) were identified on the basis of the spectroscopic methods and comparisons with literature, respectively. The structures were identified as cholesterol (CS, 6), 7-keto-CS (7), $7{\beta}$-hydroxy-CS (8), and $7{\alpha}$-hydroxy-CS (9), and included 12 steroid $3{\beta}$-O-(palmitic/stearic/myristic acid esters; PM/SA/MS) [CS-$3{\beta}$-O-PM (1 - 1), CS-$3{\beta}$-O-SA (1 - 2), CS-$3{\beta}$-O-MR (1 - 3), 7-keto-CS-$3{\beta}$-O-PM (2 - 1), 7-keto-CS-$3{\beta}$-O-SA (2 - 2), 7-keto-CS-$3{\beta}$-O-MR (2 - 3), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-SA (3 -1), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-PM (3 - 2), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-MR (3 - 3), $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-SA (4 - 1), $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-PM (4 - 2), and $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-MR (4 - 3)], dinonyl phthalate (5), 8 nucleic acids analogues [uracil (10), deoxyguanosine (11), deoxyuridine (12), uridine (13), deoxyadenosine (14), adenosine (15), inosine (16), and guanosine (17)], and the 9 free amino acids [L-phenylalanine (18), L-isoleucine (19), L-leucine (20), L-tyrosine (21), L-valine (22), L-proline (23), L-threonine (24), L-alanine (25), and L-hydroxyproline (26)]. Also, there are 8 kinds of amino acids [asparagine, serine, glutamine, glycine, histidine, arginine, methionine, and lysine], 2 sialic acids [N-acetylneuraminic acid (27), ketodeoxynonulosonic acid (28)], and 9 minerals [Na > K > Ca > Mg > Fe > Zn > B > Al > Cu] were detected from the autoaminoacid analyzer and ICP spectrometer, HPAEC-PAD/HPLC-FLD, respectively. 9 kinds of oxycholesterol-$3{\beta}$-O-fatty acid ester (2 - 1, 2 - 2, 2 - 3, 3 - 1, 3 - 2, 3 - 3, 4 - 1, 4 - 2, and 4 - 3) and 3 nucleic acids (12, 14, and 15) were isolated from the velvet antler for the first time. 6 kinds of steroids (7, 8, 9, 2 - 1, 3 - 1, and 4 - 1) were examined for their anti-proliferative effects against L1210, P388D1, K562, MEG-01, KG-1, MOLT-4, A549, HepG2, MCF-7, SK-OV-3, and SW-620 cancer cell lines. They showed anti-proliferative effects with $IC_{50}$ values of 0.06, 2.16, 2.42, > 50.0, 1.66 and $8.31{\mu}M$ against L1210, while the values were 24.05, 9.44, 5.22, 0.25. 9.48 and $49.77{\mu}M$ against P388D1, respectively. The others were inactive.

Quality Changes in Tomato Fruits Caused by Genotype and Environment Interactions (재배환경과 유전형의 상호작용에 따른 토마토 과실 품질 변화)

  • Park, Minwoo;Chung, Yong Suk;Lee, Sanghyeob
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.361-372
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    • 2017
  • Bred and grown around the world, tomato (Solanum spp.) has highly valuable fruits containings various anti-oxidants such as lycopene, flavonoids, glutamine, and ${\beta}-carotene$. Several studies have explored, way in which to enhance the growth, management and quality of tomato, we focus on the management of growth for yield rather than quality. The expression of superior agronomic traits depends on where cultivars are grown. We evaluated 10 cultivars grown in three environment for their lycopene. HTL3137 ($70.48mg{\cdot}kg^{-1}$), which was grown in Yoeju in spring/summer, contained the highest lycopene content, while HTL10256 ($20.9mg{\cdot}kg^{-1}$), which was grown in Suwon in spring/summer, contain the least lycopene.Correlations between color components and lycopene content varied according to growing location and season. In spring/summer-grown tomatoes from Suwon, no significant correlation was observed between any color component (redness [R], greenness [G], blueness [B], luminosity, $L^*$, $a^*$, $b^*$, hue and chroma) and lycopene content. A correlation was observed between B and lycopene content in tomatoes grown in Yeoju during the same season. In tomatoes grown in Yeoju in fall/winter, significant correlations were found between lycopene content and G, luminosity, $L^*$, and hue. Variance in interactions between genotype, environment, and genotype ${\times}$ environment (G ${\times}$ E) using Minimum Norm Quadratic Unbiased Estimate (MINQUE) analysis indicated that lycopene content depends on genotype (51.33%), environment (49.13%), and G ${\times}$ E (21.43%). However, when the Additive Main Effects and Multiplicative Interaction (AMMI) was used, the G ${\times}$ E value was highest.

Active Aging: Roles of Physical Activity and Immunity (건강한 노후 : 운동활동과 면역반응을 중심으로)

  • Park, Chan Ho;Kim, Ji-Seok;Kwak, Yi Sub
    • Journal of Life Science
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    • v.28 no.5
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    • pp.621-626
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    • 2018
  • We introduced the physiological responses of aging, active aging and also suggest the impact of physical exercise on body health status and elderly immunity. In this purpose, we searched the Pub Med data base for the articles (include our experimental papers) and review papers having the terms 'Aging', 'Active aging' and 'Physical activity and elderly' in the title, published from 1999 until 2018. The results were as follows: Exercise training has been extensively studied about the reduction of inflammation, oxidative stress, disease, and aging in syndrome X patients and elderly. Combined and aerobic or resistance exercise training could reduce obesity, insulin resistance, type 2 diabetes and hypertension. Exercise training has been extensively studied in cancer settings as part of prevention or treatment strategies. From this research, regular exercise has the potential to target tumor growth through regulation of inflammation and immune responses such as lactate clearance, NK cell activation (innate immunity), activation of cytotoxic immune cells, T cell activation (adaptive immunity), and immune surveillance. However, Endurance physical activity not only induces thermogenesis and diverse sports injuries but also elicits mobilization and functional enhancement of monocytes, neutrophils (which is caused by the cytokine changes such as TNF-alpha, IL-1) whereas it suppresses cell mediated immunity causing to increased susceptibility to inflammation and infections like cough and URTIs (upper respiratory track infections) in young and especially in elderly people. Therefore, Strategies to prevent physical fatigue, sports injuries include avoid overtraining, Adequate recovery and various type of rest during and after physical activity and assuring adequate nutrition supplementation such as glutamine, vitamin B, vitamin C, carbohydrate, ion or berry-contain sports beverages is helpful in physically active elderly.

Chondrogenesis of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood (사람 제대혈 유래 간엽줄기세포로부터 연골세포 분화)

  • Koh, Phil-Ok;Cho, Jae-Hyun;Nho, Kyoung-Hwan;Cha, Yun-Im;Kim, Young-Ki;Cho, Eun-Hae;Lee, Hee-Chun;Jung, Tae-Sung;Yeon, Seong-Chan;Kang, Kyung-Sun;Lee, Hyo-Jong
    • Journal of Veterinary Clinics
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    • v.26 no.6
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    • pp.528-533
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    • 2009
  • In the current study, the mesenchymal stem cells (MSCs) isolated and propagated from the human umbilical cord blood (UCB) were tested for their capabilities of differentiation into chondrocytes in vitro. The mesenchymal progenitor cells (MPCs) collected from UCB were cultured in a low glucose DMEM medium with 10% FBS, L-glutamine and antibiotics. The human MSC colonies were positively stained by PAS reaction. When the immunophenotypes of surface antigens on the MSCs were analyzed by fluorescence-activated cell sorter (FACS) analysis, these cells expressed positively MSC-related antigens of CD 29, CD44, CD 90 and CD105, whereas they did not express antigens of CD14, CD31, CD34, CD45, CD133 and HLA-DR. Following induction these MSCs into chondrocytes in the chondrogenic differentiation medium for 3 weeks or more, the cells were stained positively with safranin O. We clearly confirmed that human MSCs were successfully differentiated into chondrocytes by RT-PCR and immunofluorescent stain of type-II collagen protein. These data also indicate that the isolation, proliferation and differentiation of the hUCB-derived MSCs in vitro can be used for elucidating the mechanisms involved in chondrogenesis. Moreover this differentiation technique can be applied to developing cell-based tissue regeneration or repair damaged tissues.

Comparison of Extractive Nitrogenous Constituents in Cultured and Wild Olive Flounder (Paralichthys olivaceus) Muscle (양식 및 자연산 넙치의 함질소엑스성분 비교)

  • 박춘규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.1
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    • pp.174-179
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    • 2000
  • The nitrogenous compounds in the muscle extracts of cultured and wild olive flounder, Paralichthys olivaceous, were analyzed. The analyzed coumpounds were extractive nitrogen, free amino acids, oligopeptides, ATP and its related ompounds, quaternary ammonium bases, and guanidino compounds. The distribution pattern of these compounds in cultured and wild fish was found to be very similar. Although the ATP and its related compounds and creatine in the muscle of cultured fish were slightly abundant than those in the muscle of wild one, the extractive nitrogen, total free amino acid, oligopeptides, and TMAO were found to be slightly rich in the muscle of wild fish than those in the muscle of cultrued one. The moisture content of cultured fish was relatively lower but the protein and fat contents of cultured one were higher than those of wild fish. However the differences in the proximate composition, extractive nitrogen and nitrogenous compounds between two fishes were not significantly different.

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Varietal Response of Tobacco Plants Through Tissue Culture to Butachlor and Bialaphos Herbicides (조직배양(組織培養)에 의한 제초제(除草劑) Butachlor와 Bialaphos에 대(對)한 담배의 품종간반응(品種間反應))

  • Bae, Y.Z.;Kim, K.U.;Jeong, H.J.
    • Korean Journal of Weed Science
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    • v.8 no.1
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    • pp.53-58
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    • 1988
  • This study was carried out to determine effect of butachlor [N.-(buthoxymethyl)-2-chloro-N-(2,6-diethylphenyl) acetamide] and bialaphos [2-amino-4(hydroxy)(methyl) phosphionyl] butyryl-alanylalanine sodium salt on the germination of tobacco seed, induction and growth of callus from tobacco. Further, fatty acids and ammonia content of tobacco calli were determined. Bialaphos had no effect on tobacco seed germination, but the growth of seedling was markedly affected by an application of 10 ppm bialaphos. However, regardless of varieties tested, tobacco seed germination was completely inhibited by $5{\times}10^{-5}M$ of butachlor. At an application of $5{\times}10^{-5}M$ butachlor, tobacco seeds were to some extent germinated and showed further growth. Hyangcho among varieties tested, showed the most tolerant response to butachlor. In induction of callus from various tobacco varieties and their growth, aromatic type of tobacco varieties exhibited the most tolerance against bialaphos. However, no distinct varietal differences were determined in the treatment of butachlor. The major fatty acids identified in tobacco calli were palmitic, oleic and linoleic acid. No marked difference in terms of fatty acids was observed among tobacco varieties used, but it was observed that there was the higher ratio of quantity in unsaturated fatty acids over saturated one, bialaphos treatment accumulated about 9 times higher ammonia content than that of the untreated control, giving an evidence that bialaphos might inhibit glutamine synthetase activity.

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Cloning of Low-molecular-weight Glutenin Subunit Genes and Identification of their Protein Products in Common Wheat (Triticum aestivum L.) (보통 밀에서 저분자글루테닌 유전자 클로닝 및 단백질 동정)

  • Lee, Jong-Yeol;Kim, Yeong-Tae;Kim, Bo-Mi;Lee, Jung-Hye;Lim, Sun-Hyung;Ha, Sun-Hwa;Ahn, Sang-Nag;Nam, Myung-Hee;Kim, Young-Mi
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.547-554
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    • 2010
  • Low-molecular-weight glutenin subunit (LMW-GS) in common wheat (Triticum aestivum L.) is important for quality processing of bread and noodles. The objectives of this study were to clarify the composition of LMW-GSs and to identify their corresponding proteins. Using LMW-GS specific primers we cloned and characterized 43 LMW-GS genes in the wheat cultivar 'Jokyoung'. Some of these genes contain polypeptides different in size due to the presence of various deletions or insertions within repetitive and glutamine-rich domains. The comparison of deduced amino acid sequence of the LMW-GS genes in Jokyoung with that of 12 groups LMW-GSs of wheat cultivar Norin 61 showed that the deduced amino acid sequences were nearly the same to LMW-GS groups of 1, 2, 3/4, 5, 7, 10 and 11. All LMW-GS genes contain eight cysteine residues, which are conserved among all of the typical LMW-GS sequences. The relative positions of cysteine residues are also conserved, except those of the first and seventh. Based on phylogenetic analysis, the 43 sequences with the same N-terminal and C-terminal amino acid sequences were clustered in the same group. To identify the proteins containing the corresponding amino acid sequences, we determined the N-terminal amino acid sequence of 7 spots of LMW-GSs of Jokyoung separated by two-dimensional gel electrophoresis (2DE). Of them, Glu-B3 (LMW-m and LMW-s) and Glu-D3 (LMW-m) were detected in two and three spots, respectively and the others were not clear. Collectively, we classified diverse LMW-GSs and identified their corresponding protein products. These results will be helpful in breeding programs for improvement of wheat flour quality.

Evaluations of Spectral Analysis of in vitro 2D-COSY and 2D-NOESY on Human Brain Metabolites (인체 뇌 대사물질에서의 In vitro 2D-COSY와 2D-NOESY 스펙트럼 분석 평가)

  • Choe, Bo-Young;Woo, Dong-Cheol;Kim, Sang-Young;Choi, Chi-Bong;Lee, Sung-Im;Kim, Eun-Hee;Hong, Kwan-Soo;Jeon, Young-Ho;Cheong, Chae-Joon;Kim, Sang-Soo;Lim, Hyang-Sook
    • Investigative Magnetic Resonance Imaging
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    • v.12 no.1
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    • pp.8-19
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    • 2008
  • Purpose : To investigate the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar nuclear Overhauser effect/enhancement (NOE) interaction through 2D- correlation spectroscopy (COSY) and 2D- NOE spectroscopy (NOESY) techniques. Materials and Methods : All 2D experiments were performed on Bruker Avance 500 (11.8 T) with the zshield gradient triple resonance cryoprobe at 298 K. Human brain metabolites were prepared with 10% $D_2O$. Two-dimensional spectra with 2048 data points contains 320 free induction decay (FID) averaging. Repetition delay was 2 sec. The Top Spin 2.0 software was used for post-processing. Total 7 metabolites such as N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), lutamine (Gln), glutamate (Glu), myo-inositol (Ins), and lactate (Lac) were included for major target metabolites. Results : Symmetrical 2D-COSY and 2D-NOESY pectra were successfully acquired: COSY cross peaks were observed in the only 1.0-4.5 ppm, however, NOESY cross peaks were observed in the 1.0-4.5 ppm and 7.9 ppm. From the result of the 2-D COSY data, cross peaks between the methyl protons ($CH_3$(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methylene protons (CH2(3,$H{\alpha}$)) at 2.50ppm and methylene protons ($CH_2$,(3,$H_B$)) at 2.70 ppm were observed in NAA. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. From the result of 2-D NOESY data, cross peaks between the NH proton at 8.00 ppm and methyl protons ($CH_3$) were observed in NAA. Cross peaks between the methyl protons ($CH_3$(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methyl protons (CH3) at 3.03 ppm and methylene protons (CH2) at 3.93 ppm were observed in Cr. Cross peaks between the methylene protons ($CH_2$(3)) at 2.11 ppm and methylene protons ($CH_2$(4)) at 2.35 ppm, and between the methylene protons($CH_2$ (3)) at 2.11 ppm and methine proton (CH(2)) at 3.76 ppm were observed in Glu. Cross peaks between the methylene protons (CH2 (3)) at 2.14 ppm and methine proton (CH(2)) at 3.79 ppm were observed in Gln. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. Conclusion : The present study demonstrated that in vitro 2D-COSY and NOESY represented the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar NOE interaction. This study could aid in better understanding the interactions between human brain metabolites in vivo 2DCOSY study.

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