A study of patients with alcoholic liver disease by 31P nuclear magnetic resonance spectroscopy

1. Patients with a history of alcohol abuse were studied by 31P n.m.r. spectroscopy of the liver in vivo, and the results were related to the pattern of disease assessed by standard biochemical and histological techniques. 2. The ratios of metabolites measured from the 31P n.m.r. spectra were abnormal in patients with alcoholic hepatitis but not in those with fatty change or cirrhosis in the absence of hepatitis. In particular, the levels of phosphomonoesters were raised, with respect either to Pi, or to adenosine 5'-triphosphate. The level of phosphomonoesters showed a significant positive correlation with the severity of alcoholic hepatitis, assessed by histology. 3. The ratio of Pi to adenosine 5'-triphosphate was used as a measure of the energy status of the hepatocytes, and was unchanged between patients and controls.     

Muscle metabolism in patients with polymyositis simultaneously evaluated by using 31P-magnetic resonance spectroscopy and near-infrared spectroscopy

Simultaneous measurements of muscle energy metabolism using (31)P-magnetic resonance spectroscopy ((31)P-MRS) and the kinetics of muscular oxygen metabolism using near-infrared spectroscopy (NIRS) were conducted in polymyositis (PM) patients. The subjects were 12 PM patients (age 45 +/- 12 years) and 12 normal controls (age 41 +/- 12 years). The muscle phosphocreatine (PCr) index and intracellular pH (pHi) were determined with (31)P-MRS and the changes in intramuscular oxygenated (oxy-Hb), deoxygenated (deoxy-Hb), and total haemoglobin (total Hb) were evaluated with NIRS . The pHi and PCr index before steroid therapy in PM patients were significantly lower during exercise than in normal controls, and their recovery was statistically significantly delayed compared with the controls. The pattern of changes in NIRS over time before steroid therapy in PM patients differed from that in normal controls. There were smaller changes in deoxy-Hb and oxy-Hb during exercise, and total Hb decreased during exercise. In contrast, the kinetics of muscular metabolism after steroid therapy showed changes similar to those seen in normal controls. Simultaneous (31)P-MRS and NIRS measurements to determine the kinetics of muscular metabolism are expected to be useful as a noninvasive approach for the evaluation of treatment effects in PM patients.     

应用31P磁共振波谱对心肌梗死后能量代谢变化的评价

目的：探讨^31磷磁共振波谱（^31PMRS）在心肌梗死临床应用价值，评价心肌活力，为心肌梗死治疗方案决策提供依据。方法：健康志愿者12例行^31PMRS和共振成像（MRI）检查，结果作为正常对照组。选择10例心肌梗死病人，进行MRI和^31PMRS检查，所有病人经冠脉造影等检查证实。结果：与正常对照组比较，患者存活心肌PCr/ATP比值无显著差异；病变区无活力心肌r-ATP-SNR(the signal to noise ratio)与对照组和存活心肌比较明显降低。结论：①^31PMRS可以提供存活心肌和梗死（无活力）心肌能量代谢变化特征，为^31PMRS评价心肌活力的临床应用提供方法论。②顿抑心肌状态，高能磷酸盐代谢变化不是心肌顿抑心功能异常的直接病因。③^31PMRS显示人心肌梗死能量代谢的变化，用^31PMRS可以预测血管重建后心功能恢复情况。     

Skeletal muscle metabolism in heart failure: a 31P nuclear magnetic resonance spectroscopy study of leg muscle

1. The gastrocnemius muscle of seven patients with mild to moderate chronic heart failure and of five healthy control subjects was studied using 31P nuclear magnetic resonance spectroscopy. Spectra were collected at rest and during an incremental, symptom-limited, exercise protocol. Blood flow was measured in the same study during brief interruptions to exercise. 2. The phosphocreatine/(phosphocreatine plus inorganic phosphate) ratio was lower in patients with heart failure than in control subjects at an exercise rate of 1.5 W, although intracellular pH and blood flow were similar. 3. The cytosolic free adenosine 5'-diphosphate concentration was markedly increased in patients with heart failure exercising at 1.5 W compared with control subjects exercising at the same workload. 4. Although the maximum workload achieved by patients with heart failure was less than half of that reached by control subjects, the pH and the phosphocreatine/(phosphocreatine plus inorganic phosphate) ratio were lower in patients with heart failure at maximal load. Blood flow was less at maximal exercise in patients with heart failure than in control subjects in keeping with the reduced work load. 5. The phosphocreatine depletion induced in the gastrocnemius muscle by exercise was more severe than previously described in the forearm of patients with heart failure. 6. Metabolic abnormalities in skeletal muscle may contribute to exercise intolerance in heart failure, particularly during submaximal exercise.     

A new approach to the study of ifosfamide metabolism by the analysis of human body fluids with 31P nuclear magnetic resonance spectroscopy.

[31P] nuclear magnetic resonance spectroscopy was used to analyze body fluids from patients treated with ifosfamide (IF). This technique, which requires no labeled drug, allows a direct study of the biological sample with no need for extraction or derivatization and a simultaneous detection and quantification of all the different phosphorated metabolites in a single analysis. In urine, isophosphoramide mustard was detected in addition to the already known human urinary compounds [i.e., unchanged IF, carboxyifosfamide, 2-dechloroethylifosfamide, 3-dechloroethylifosfamide, ketoifosfamide]. 2,3-Didechloroethylifosfamide itself was not found, but two of its degradation compounds were detected, thus showing a minor route of didechloroethylation of IF in humans. Several other signals corresponding to unknown metabolites or to degradation compounds of IF metabolites were observed. None of them corresponded to IF-activated metabolites (4-hydroxyifosfamide, aldoifosfamide) or to conjugates of IF or its metabolites with mesna. The urinary excretion of IF and metabolites over 24 h amounted to 39 to 50% of the injected dose. Unmetabolized IF was the major compound in 0- to 8-h and 8- to 16-h fractions. 2-Dechloroethylifosfamide and 3-dechloroethylifosfamide were the main metabolites detected in each 8-h fraction. The two unknown compounds at 19.16 ppm and 16.06 ppm represented a non-negligible fraction of the excretion, above that of carboxyifosfamide. Only unchanged IF could be detected in plasma samples. Unmetabolized IF and 3-dechloroethylifosfamide were found in a cerebrospinal fluid sample. Neither IF nor IF metabolites could be observed in the corresponding plasma sample. This indicates a long persistence of these compounds in cerebrospinal fluid.     

In vivo 31P nuclear magnetic resonance spectroscopy of bone mineral for evaluation of osteoporosis

The mineral content of stationary bone samples can be quantified by 31P nuclear magnetic resonance (NMR) spectroscopy. The assay can be performed in regions of the anatomy that pose problems for absorptiometric techniques, because the mineral content is measured within a selected volume without concern for the geometry of the bone. In vivo 31P NMR spectra of the bones in human fingers and wrist are reported. Soft tissue such as marrow and skeletal muscle contributes little to the 31P NMR spectra of human fingers and wrist and thus should not seriously affect the accuracy of the mineral assay. 31P NMR spectrometry should prove helpful for confirming rapid bone mineral loss in those at risk and for monitoring response to treatment.     

Modification of intra-erythrocytic homeostasis in uremic patients, as studied with 31P nuclear magnetic resonance

Intra-erythrocytic pH, ATP concentrations, and 2,3-diphosphoglycerate relaxation times were studied in living erythrocytes by "high-resolution" 31P NMR spectroscopy to assess homeostasis within the cells. In uremic patients, intra-erythrocytic pH is significantly decreased before hemodialysis, but is corrected equally well by hemodialysis against either acetate or bicarbonate. This acidic pHi may be correlated with the increased concentration of ATP in erythrocytes in uremia, which is partly corrected by these two types of hemodialysis. Similarly, the significant decrease of spin-spin relaxation times in uremic patients is corrected by hemodialysis.     

Phospholipid and energy metabolism of cancer cells monitored by 31P magnetic resonance spectroscopy: possible clinical significance

31P magnetic resonance spectroscopy (MRS) has been established as an excellent noninvasive procedure for studying metabolism in cancer cells. For a full understanding of the results of in vivo MRS studies of metabolism, it is necessary to be able to conduct controlled experiments on the same intact functioning cells in a perfusion apparatus. The specific techniques used for this purpose are described, and the applications to cancer cell bioenergetics and phospholipid metabolism in drug-resistant and drug-sensitive breast cancer cells are illustrated. Spectra of the same cell line grown in a "model tumor" in natural basement membrane gel and in nude mice in vivo are compared. The results indicate a potential for the use of 31P MRS in a clinical setting for monitoring therapy and for diagnosis.     

Diagnosing inborn errors of lipid metabolism with proton nuclear magnetic resonance spectroscopy

BACKGROUND: Many severe diseases are caused by defects in lipid metabolism. As a result, patients often accumulate unusual lipids in their blood and tissues, and proper identification of these lipids is essential for correct diagnosis. In this study, we investigated the potential use of proton nuclear magnetic resonance (1H-NMR) spectroscopy to simultaneously identify and quantify (un)usual lipids present in the blood of patients with different inborn errors of lipid metabolism. METHODS: We extracted blood plasma or serum lipids in chloroform-methanol (2:1 by volume). After addition of the nonvolatile chemical shift and concentration reference compound octamethylcyclotetrasiloxane, we performed 1H-NMR measurements on a 500-MHz spectrometer. Assignments were based on the literature, computer simulations, and reference spectra of relevant authentic standards. RESULTS: Spectra of normal plasma samples allowed the identification of 9 lipid species. We found good correlation between conventional methods and 1H-NMR for cholesterol and triglyceride concentrations. We also investigated 4 inborn errors of lipid metabolism (3 in sterol metabolism and 1 in fatty acid metabolism). NMR analysis led to a correct diagnosis for all 4 diseases, whereas the concentration of the diagnostic metabolite could be determined for 3. CONCLUSIONS: 1H-NMR spectroscopy of blood plasma or serum lipid extracts can be used to accurately identify and quantify lipids. The method can also identify unusual lipids in the blood of patients with inborn errors of lipid metabolism. This technique may therefore be applicable in clinical diagnosis and follow-up.     

Lipoprotein particle analysis by nuclear magnetic resonance spectroscopy

Laboratory measurements of plasma lipids (principally cholesterol and triglycerides) and lipoprotein lipids (principally low-density lipoprotein [LDL] and low-density lipoprotein [HDL] cholesterol) are the cornerstone of the clinical assessment and management of atherosclerotic cardiovascular disease (CVD) risk. LDL particles, and to a lesser extent very-low-density lipoprotein [VLDL] particles, cause atherosclerosis, whereas HDL particles prevent or reverse this process through reverse cholesterol transport. The overall risk for CVD depends on the balance between the "bad" LDL (and VLDL) and "good" HDL particles. Direct assessment of lipoprotein particle numbers us now possible through nuclear magnetic resonance spectroscopic analysis.     

Alzheimer病和血管性痴呆的磁共振波谱分析

目的 探讨质子磁共振波谱(1H-MRS)对Alzheimer病(AD)和血管性痴呆(vaD)的诊断和鉴别诊断的作用。方法 采用点解析波谱法(PRESS),对24例AD患者、16例VaD患者和24例年龄匹配健康对照的双侧颞顶叶灰质区1H-MRS进行分析,比较各组间N-乙酰天门冬氨酸/肌酸(NAA/Cr)和胆碱/肌酸(Cho/Cr)比值的差别,并对AD和VaD组的简易智能精神状态量表(MMSE)评分与NAA/Cr、Cho/Cr进行相关分析。 结果 AD组和VaD组的NAA/Cr显著低于对照组(P<0.05),AD组的NAA/Cr显著低于VaD组(P<0.05),三组间的Cho/Cr无差异。AD组的MMSE评分与NAA/Cr明显正相关(P<0.05),与Cho/Cr无关。VaD组的MMSE评分与NAA/Cr、Cho/Cr均无关。结论 AD和VaD患者的双侧颞顶叶灰质区NAA/Cr比值的降低和AD患者NAA/Cr的变化与痴呆严重程度的相关性,对AD和VaD的诊断和鉴别诊断有一定参考作用。1H-MRS对AD的疾病监测也有重要作用。     

Metabolic abnormalities in skeletal muscle of patients receiving zidovudine therapy observed by 31P in vivo magnetic resonance spectroscopy.

Patients on long-term zidovudine (AZT) therapy experience muscle fatigue and weakness attributed to AZT-induced mitochondrial toxicity in skeletal muscle. To determine if the clinico-pathological abnormalities in these patients correspond to abnormal muscle energy metabolism, we used 31P in vivo magnetic resonance spectroscopy to follow phosphorylated metabolites during exercise. We studied 19 normal volunteers, 6 HIV-positive patients never treated with AZT, and 9 HIV-positive patients who had been treated with AZT for a mean period of 33 mo (range 12-48 mo) and had muscle biopsy-proven AZT-myopathy with abnormal mitochondria. Changes in phosphocreatine, ATP, and intracellular pH in the gastrocnemius muscle were followed during a graded steady state exercise protocol, and the recovery of phosphocreatine was followed on cessation of exercise. We found that graded steady state exercise produced a greater depletion of muscle phosphocreatine levels in the AZT-treated patients, compared to either HIV-positive patients who were not treated with AZT or normal controls. No differences in the effects of steady state exercise on muscle phosphocreatine levels were observed between the control group and the HIV-positive patients who had not been treated with AZT. The results suggest that the effect of AZT on muscle energy metabolism is significant, and similar to the effect observed in patients with known mitochondrial myopathies. Using a well-known model for control of mitochondrial metabolism, the observed differences in steady state phosphocreatine levels during exercise suggest that AZT treatment decreases the maximal work output and the maximal rate of muscle ATP synthesis.     

A study of intracellular orthophosphate concentration in human muscle and erythrocytes by 31P nuclear magnetic resonance spectroscopy and selective chemical assay

In order to study the relationship between extracellular and intracellular concentrations of orthophosphate (Pi), phosphorus nuclear magnetic resonance spectra were recorded, at rest, from the flexor digitorum superficialis muscle of hypophosphataemic patients with vitamin D-resistant rickets, and patients with Paget's disease of bone before and after they had been made hyperphosphataemic by treatment with the drug ethylidene-1-hydroxy-1,1-bisphosphonate. Changes in intramuscular P1 were estimated from the ratio of the areas of the Pi to adenosine 5'-triphosphate peaks. Even though the plasma Pi concentration in these patients spanned a fourfold range (0.5-2.0 mmol/l) the corresponding intramuscular Pi concentration increased by only 70%. A similar effect was observed in erythrocytes, from patients with these disorders, which were incubated in autologous plasma at 37 degrees C, under an atmosphere of O2 + CO2 (95:5, v/v). However, chloride ions, which are transported passively across the cell membrane, showed no change in distribution between cells and plasma, indicating that there was no general effect on passive anion distribution. When erythrocytes from normal subjects were incubated in autologous plasma (1.0 mmol of Pi/l) and in plasma supplemented with Pi (2.3 mmol of Pi/l), the Pi concentration in the cells, at steady state, increased only from 0.57 to 0.78 mmol/l cells, suggesting that the effect was not an artifact of disease or drug therapy. It is concluded that, in human skeletal myocytes and erythrocytes, the percentage change in the concentration of cytoplasmic Pi is lower than that in plasma. This implies that these cells can buffer or regulate cytoplasmic Pi when the extracellular concentration is disturbed.     

Relative metabolic efficiency of concentric and eccentric exercise determined by 31P magnetic resonance spectroscopy

To determine the relative metabolic efficiency (metabolic energy used per unit of mechanical energy output) of negative to positive muscular power, we used 31P magnetic resonance spectroscopy to monitor the cellular energy metabolism of limb muscles in eight healthy subjects during a nonfatiguing, mixed concentric-eccentric activity and during its concentric and eccentric components. We also studied isometric contractions. We found that in terms of the flow of metabolic energy through the muscle cells, the cost of concentric exercise at this intensity was proportional to the mechanical power generated, but the cost of eccentric and isometric exercise did not increase significantly as the apparent intensity of the exercise increased over the range studied. Although the pattern was similar in all subjects, the quantitative relationship between metabolic cost and mechanical output was different in subjects with different muscular strength. The qualitative results can be explained in the context of the known biochemistry and biophysics of the cellular contractile apparatus (sliding filament theory, with independent force generators).     

Study of the metabolism of flucytosine in Aspergillus species by 19F nuclear magnetic resonance spectroscopy

The metabolism of flucytosine (5FC) in two Aspergillus species (Aspergillus fumigatus and A. niger) was investigated by 19F nuclear magnetic resonance spectroscopy. In intact mycelia, 5FC was found to be deaminated to 5-fluorouracil and then transformed into fluoronucleotides; the catabolite alpha-fluoro-beta-alanine was also detected in A. fumigatus. Neither 5-fluoroorotic acid nor 5-fluoro-2'-deoxyuridine-5'-monophosphate was detected in perchloric acid extracts after any incubation with 5FC. 5FC, 5-fluorouracil, and the classical fluoronucleotides 5-fluorouridine-5'-mono-, di-, and triphosphates were identified in the acid-soluble pool. Two hydrolysis products of 5-fluorouracil incorporated into RNA, 5-fluorouridine-2'-monophosphate and 5-fluorouridine-3'-monophosphate, were found in the acid-insoluble pool. No significant differences in the metabolic transformation of 5FC were noted in the two species of Aspergillus. The main pathway of 5FC metabolism in the two species of Aspergillus studied is thus the biotransformation into ribofluoronucleotides and the subsequent incorporation of 5-fluorouridine-5'-triphosphate into RNA.     

原发性失眠患者脑磁共振波谱成像研究

目的研究原发性失眠(primary insomnia,PI)患者不同脑区(丘脑、海马、壳核)代谢特点.材料与方法采用氢质子磁共振波谱成像技术检测38例PI患者(PI组)和39例健康对照者(healthy control,HC)组丘脑、海马区、壳核区的氮-乙酰基天门冬氨酸(N-acetyl-aspartate,NAA)、...     

Quantification of plasma lipoproteins by proton nuclear magnetic resonance spectroscopy

A new analytical procedure for quantifying plasma lipoproteins by proton nuclear magnetic resonance (NMR) spectroscopy has been developed that potentially offers significant advantages over existing clinical methods used for assessing risk of coronary heart disease. Analysis of a single spectrum of a nonfasting plasma sample, acquired simply and rapidly at moderate magnetic field strength (250 MHz), yields a complete profile of lipoprotein concentrations: chylomicrons and very-low-, low-, and high-density lipoproteins. The method is based on curve-fitting (spectral deconvolution) of the plasma methyl lipid resonance envelope, the amplitude and shape of which depend directly on the amplitudes of the superimposed methyl resonances of the lipoprotein components. A linear least-squares curve-fitting algorithm was developed to efficiently extract the signal amplitudes (concentrations) of the lipoproteins from the plasma spectrum. These signal amplitudes correlate well with lipoprotein concentrations determined by triglyceride and cholesterol measurements.