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% D2O. 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), glutamine (Gln), glutamate (Glu), myo-inositol (Ins), and lactate (Lac) were included for major target metabolites. RESULTS: Symmetrical 2D-COSY and 2D-NOESY spectra 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 (CH3(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 (CH2,(3,HB)) 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 (CH3) were observed in NAA. Cross peaks between the methyl protons (CH3(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 (CH2(3)) at 2.11 ppm and methylene protons (CH2(4)) at 2.35 ppm, and between the methylene protons(CH2 (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.
Aspartic Acid ; Brain ; Choline ; Creatine ; Glutamic Acid ; Glutamine ; Humans ; Lactic Acid ; Protons ; Spectrum Analysis

BACKGROUND: ras gene mutations have been described in various human malignancies, suggesting that their activation may play a role in oncogenesis. However, there are few reports concerning ras gene alterations in malignant fibrous histiocytomas. We therefore designed a study to determine the prevalence and type of mutations in the first exons of H-ras and K-ras genes in these tumors. METHODS: Twenty-seven malignant fibrous histiocytomas were investigated by direct sequencing analysis with the automated DNA sequencing of polymerase chain reaction-amplified ras sequences. RESULTS: Twenty-four mutations were found in 18 (67%) of the tumors: GGC to GAC transition mutations at codon 13 of K-ras (coding for aspartic acid instead of glycine) in 18 of the samples and GGC to GTC transversions at codon 12 of H-ras (coding for valine instead of glycine) in six of the lesions. CONCLUSIONS: Our data suggest an involvement of the ras gene mutation in conjunction with other yet unknown events in the tumorigenesis and/or progression of malignant fibrous histiocytomas. The K-ras gene activation predominated in these tumors by a mutation at codon 13. It is noteworthy that H-ras mutations were detected only in association with the lesions containing K-ras mutated genes, the significance of which remains to be determined.
Aspartic Acid ; Carcinogenesis ; Codon ; Exons ; Genes, ras* ; Histiocytoma, Malignant Fibrous* ; Humans ; Prevalence ; Sequence Analysis, DNA ; Valine

Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality of AR. This study supplied a new method for the comparison of extract of herbal drugs.
Arginine ; analysis ; Aspartic Acid ; analysis ; Choline ; analysis ; Citric Acid ; analysis ; Drugs, Chinese Herbal ; analysis ; chemistry ; Fumarates ; analysis ; Glutamic Acid ; analysis ; Magnetic Resonance Spectroscopy ; Multivariate Analysis ; Phylogeography ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Succinic Acid ; analysis ; Sucrose ; analysis ; Taurine ; analysis

OBJECTIVETo explore the effects of 1,2-dichloroethane (1,2-DCE) on the behavior and the brain neurotransmitter levels in mice.

METHODSThirty mice were randomly divided into four groups, which were control group and groups of low, middle and high exposure (225, 450 and 900 mg/m3) to 1,2-DCE for 10 days (3.5 h a day) by inhalation. After the last exposure, the open field test was performed immediately. After exposure all mice were killed and the brain tissues were taken up rapidly. The levels of aspartate (Asp), glutamate (Glu) and gamma-aminobutyric acid (GABA) in the brain were detected by high performance liquid chromatography (HPLC).

RESULTSLevels of Asp and Glu in all exposure groups increased with doses. As compared to the control group, levels of Glu in all exposure groups increased significantly (P < 0.05). Levels of GABA in the low exposure group were significantly lower than those in control group, but those in the high exposure group were significantly higher than those in control group. The results of the open field test showed that effect of low exposure to 1,2-DCE on the behavior was stimulant, but the high exposure to 1,2-DCE inhibited behavior of exploration, excitement and sport.

CONCLUSIONSSubacute exposure to 1,2-DCE could result in the change of amino acid neurotransmitter content and ratio in the brain, thereby change the behavior of mice appeared, which might be the mechanism of neurotoxicity caused by 1,2-DCE in part.

Animals ; Aspartic Acid ; analysis ; Behavior, Animal ; drug effects ; Brain ; metabolism ; Ethylene Dichlorides ; toxicity ; Female ; Glutamic Acid ; analysis ; Mice ; Mice, Inbred Strains ; Neurotransmitter Agents ; metabolism ; gamma-Aminobutyric Acid ; analysis

OBJECTIVE: MR spectroscopy is a noninvasive method of monitoring in vivo metabolite concentration changes over time. In this study we evaluated the usefulness of combined magnetic resonance imaging and spectroscopy on the diagnosis of cerebral infarction. METHODS: Combined magnetic resonance imaging and spectroscopy investigations were carried out with 1.5-T system in 18 volunteers, 10 patients with acute infarction (< 8 hours). RESULTS: Acute ischemic infarctions were characterized by decreased N-acetyl aspartate and elevation of lactate. CONCLUSION: Metabolic alterations in ischemic tissue can be monitored.
Aspartic Acid ; Cerebral Infarction* ; Diagnosis ; Humans ; Infarction ; Lactic Acid ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy* ; Spectrum Analysis ; Volunteers

PURPOSE: The aim of this study is to evaluate stage IV breast cancer at the initial presentation by the review of a single institute' data. We also tried to figure out the factors to predict stage IV breast cancer. METHODS: We reviewed the prospectively collected database of 1,424 consecutive patients with primary breast cancer at the National Cancer Center in Korea from October 2000 to January 2005. RESULTS: The proportion of stage IV breast cancer was 2.7% (38/1,424). The median tumor size of the stage IV patients was 4.1 cm. The most common metastatic site was bone (47.4%) followed by lung (44.7%) and liver (36.8%). Metastases were found in 0.9% (6/672) of the T1 tumors, 2.4% (13/535) of the T2 tumors, 8.3% (4/48) of the T3 tumors, and 27.1% (13/48) of the T4 tumors (p<0.001). On multivariate analysis, the statistically significant predictors of distant metastasis were tumor size (> or =2 cm) (p=0.026), positive lymph node status (p<0.001), alkaline phosphatase (>104 IU/L) (p=0.013), aspartate transferase (>40 IU/L) (p=0.003) and CA15-3 (>32 U/mL) (p=0.025). CONCLUSION: Our study showed that the factors to predict distant metastasis of breast cancer were large size of tumor, positive lymph node status, elevated alkaline phosphatase, aspartate transferase and CA15-3. Therefore breast cancer patients with those clinical characteristics should be carefully evaluated to detect distant metastasis.
Alkaline Phosphatase ; Aspartic Acid ; Breast Neoplasms* ; Breast* ; Humans ; Korea ; Liver ; Lung ; Lymph Nodes ; Multivariate Analysis ; Neoplasm Metastasis ; Prospective Studies ; Transferases

PURPOSE: To determine the usefulness of in-vivo proton MR spectroscopy (MRS) with short TE for grading glial brain tumors. MATERIALS AND METHODS: For the purpose of tumor grading, 32 patients with pathologically confirmed glial tu-mors were examined by proton MRS. This and MRI were performed on a 1.5 T superconductive MR scanner. T2-weighted FSE images (TR/TE=4,000/100msec) were used to obtain anatomical reference images. The stim-ulated- echo acquisition mode (STEAM: TR/TE/MT=3000/30/13.7msec) was used to acquire MRS data from the localized single-voxel (2 x2 x2 cm 3 ) in both hemispheres. Residual water resonance in the spectra was re-moved using a CHESS pulse sequence. Prior to baseline correction, MRS raw-data, free induction decay sig-nals were zero-filled, apodized by an exponential function with 8Hz line-broadening, and fourier transformed. To normalize signal intensities of metabolites such as N-acetyl aspartate (NAA), total choline (Cho), myo-inosi-tol (mI), and lactate (Lac), the creatine(Cr) peak was used as a standard. RESULTS: The concentration ratios of Cho/Cr, mI/Cr, alpha-Glx, Lac, and NAA/Cr changed linearly according to tu-mor grade. Increased Cho, mI, alpha-Glx, and Lac levels were clearly seen in all grades. The most dramatic in-creases, observed in either Grade III or IV, were 78% and 228% for Cho(p<0.001), 106% and 61% for mI (p<0.001), 32% and 5% for alpha-Glx, and 727% and 450% for Lac (p<0.001), respectively. Increase of concen-tration ratio of Lac/Cr observed only in Grade III and Grade IV. The concentration ratios of NAA/Cr decreased gradually as tumor grade increased(p<0.001). CONCLUSION: The metabolic changes seen on proton MR spectroscopy using short TE might be useful for grad-ing glial brain tumors.
Aspartic Acid ; Brain Neoplasms ; Brain* ; Choline ; Fourier Analysis ; Humans ; Lactic Acid ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy* ; Neoplasm Grading ; Protons* ; Spectrum Analysis ; Water

OBJECTIVETo research the content changes of excitatory neurotransmitter and inhibitory neurotransmitter in corpus striatum of rats after single-used borneol and combining it with diazepam in hope of comprehending the activity of borneol on central nervous system and to observe whether borneol could increase the penetration of other drugs into the brain.

METHODThe content of four amino acids neurotransmitters in corpus striatum of rats were sampled by brain microdialysis technology at different time after administration and were determined by RP-HPLC which involved pre-column derivation with orthophthaladehyde (OPA), using phosphate gradient elution and fluorescence detection to detect the content of excitatory neurotransmitter aspartate (Asp), glutamate (Glu) and inhibitory neurotransmitter glycine (Gly), gamma-aminobutyric acid (GABA) in standards and samples and carry on statistical analysis.

RESULTThe content of both Gly and GABA in corpus striatum of rats with borneol increased significantly, compared with diazepam group (P < 0.05), while Asp and Glu showed no significant difference.

CONCLUSIONBorneol can improve permeability of diazepam through BBB.

Animals ; Aspartic Acid ; analysis ; Blood-Brain Barrier ; Bornanes ; administration & dosage ; pharmacology ; Corpus Striatum ; chemistry ; drug effects ; Diazepam ; administration & dosage ; pharmacology ; Glutamic Acid ; analysis ; Glycine ; analysis ; Male ; Neurotransmitter Agents ; analysis ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; analysis

OBJECTIVETo investigate the variety of vitro recovery of amino acids for microdialysis probe after different dialysis time in vivo.

METHODSProbes were dialyzed in the amino acids standard solutions with microdialysis system,amino acid standard solutions and the microdialysate of probe were detected by the method of precolumn derivation with HPLC-RF.

RESULTAfter using different time of probe made by regenerated cellulose membrane, the vitro recoveries of Asp, Glu and GABA were not completely same (Asp: F=19.669, P=0.000; Glu: F=103.955, P=0.000; GABA: F=3.454, P=0.040); while the vitro recovery of Tau had no obvious difference(F=2.001, P=0.152). After using 6 h in vivo, recovery remain percentage (RRP) of Asp, Glu,Tau and GABA was 64.34 %, 67.36%, 103.11 % and 98.23 %, respectively, the recoveries of Asp, Glu decreased obviously (Asp: P < 0.01,Glu: P <0.05). After using 12 h in vivo, the RRP of Asp, Glu, Tau and GABA was 43.44 %, 24.42%, 77.45 % and 67.36 %, respectively, the recoveries of Asp, Glu and GABA decreased obviously (Asp: P < 0.001, Glu: P < 0.001, GABA: P < 0.05). After using 24 h in vivo, the RRP of Asp, Glu,Tau and GABA was 36.26 %, 12.24 %, 89.48 % and 71.35 %, respectively, the recoveries of Asp, Glu, GABA decreased obviously (Asp: P < 0.0001, Glu: P < 0.0001, GABA: P < 0.01).

CONCLUSIONDialysis in vivo could lead to the decline of recovery of probe, the decline is more obvious after longer dialysis. So when making brain dialysis experiments, the use time of probe should not be too long. To improve the validity of data, some calibration should be made on the recoveries of probe.

Amino Acids ; analysis ; Animals ; Aspartic Acid ; analysis ; Brain Chemistry ; Chromatography, High Pressure Liquid ; methods ; Dialysis Solutions ; analysis ; Glutamic Acid ; analysis ; Microdialysis ; methods ; Rats ; Rats, Sprague-Dawley ; Time Factors ; gamma-Aminobutyric Acid ; analysis

OBJECTIVEThis study examined the biochemical metabolism by proton magnetic resonance spectroscopy ('H-MRS) in order to explore the value of 'H-MRS in idiopathic epilepsy in children.

METHODSThirty-three children with idiopathic epilepsy (14 cases with history of febrile seizures and 19 cases without) and six normal controls experienced MRI of the skull and brain and single-voxel 'H-MRS examinations of the hippocampi-temporal lobe. The signal intensities of N-acetylaspartate (NAA), eatine+phosphocreatine (Cr), choline-containing compounds (Cho) and lactate (Lac) and the ratios of NAA/ (Cho+Cr) and Lac/Cr were compared between the patients and normal controls.

RESULTSMRI examination showed that only one child with epilepsy had myelin dysplasia. 'H-MRS examination showed that the ratio of NAA/ (Cho+Cr) in the epilepsy group was lower than that in the control group (0.64+/-0.07 vs 0.73+/-0.05; P<0.01). The epileptic children with history of febrile seizures had a more decreased ratio of NAA/ (Cho+Cr) compared with those without the history (0.61+/-0.07 vs 0.66+/-0.06; P<0.05). There were no significant differences in the ratio of Lac/Cr between the epilepsy and the control groups.

CONCLUSIONS'H-MRS may provide early information on brain injury sensitively and non-invasively in children with epilepsy. It may be used for diagnosis and prognosis evaluation of epilepsy.

Aspartic Acid ; analogs & derivatives ; analysis ; Child ; Child, Preschool ; Choline ; analysis ; Epilepsy ; diagnosis ; metabolism ; Female ; Humans ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; methods ; Male ; Phosphocreatine ; analysis ; Protons