Diagnosing diabetic nephropathy by 1H NMR metabonomics of serum

Object: The most severe complication of type 1 diabetes (T1DM) is diabetic nephropathy. It is associated with a high risk of cardiovascular complications and premature death and requires early detection to be efficiently treated. The clinical practice to diagnose diabetic nephropathy is also a non-optimal and tedious set up based on albumin excretion rate in multiple overnight or 24h urine samples. Conversely, in this study, these independent diagnostic data are used to provide a realistic testing case for applying ¹H NMR metabonomics of serum in a diagnostic fashion. Materials and Methods: 182 T1DM and 21 non-diabetic (non-T1DM) individuals were studied. The ¹H NMR of serum at 500 MHz was targeted at two molecular windows: lipoprotein lipids and low-molecular-weight metabolites. Results: T1DM and non-T1DM individuals were exclusively separated by ¹H NMR. For diabetic nephropathy diagnosis in the T1DM patients, ¹H NMR data (and clinical biochemistry data) gave a sensitivity of 87.1% (83.9%) and a specificity of 87.7% (95.9%). The predictive values of positive and negative tests were 89.0% (95.5%) and 83.6% (79.2%), respectively. Conclusions: ¹H NMR metabonomics clearly distinguishes metabolic characteristics of T1DM and appears approximately as good a means to diagnose diabetic nephropathy from serum as an advanced set of biochemical variables.     

Metabolic changes in the rat brain after a photochemical lesion treated by stem cell transplantation assessed by 1H MRS

Object  Metabolite changes in an experimental lesion in the rat cortex and in the contralateral hemisphere after the intravenous administration of mesenchymal stem cells (MSCs) were assessed by proton MR spectroscopy to verify the impact of the cell treatment on the brain tissue. Materials and methods  Wistar rats with a photochemical cortical lesion and transplanted MSCs or sham transplanted rats were examined. Proton spectra were obtained from the lesion and from the contralateral cortex. Results  Magnetic resonance spectroscopy revealed a gradual recovery of the damaged tissue; however, we found no significant differences in metabolite concentrations in the lesioned hemisphere between treated and untreated animals. Higher concentrations of glutamate and N-acetyl aspartate were found in the contralateral hemisphere in cell-treated animals compared to untreated ones. Lesioned animals showed neurogenesis in the contralateral hemisphere; the number of newly generated cells in stem cell-treated animals was 50% higher than those observed in untreated animals. Conclusion  No direct impact of cell transplantation was observed in the lesion. However, changes in the contralateral hemisphere suggest that the transplanted MSCs might stimulate repair processes and plasticity resulting in the generation of newborn cells, which might enable the faster adoption of the damaged tissue’s function by healthy tissue.     

Valproic acid intoxication identified by1H and1H-13C correlated NMR spectroscopy of urine samples

Analysis of biological fluids by proton and carbon nuclear magnetic resonance spectroscopy (¹H and¹³C NMR) is a promising tool in clinical biology. We used this method for rapid toxicological screening in the case of two suicide attempts. For each case, a urine sample was analysed at 300 MHz by 1D and 2D sequences (TOCSY and HMBC) in a short experimental time. Quantification was performed by peak integration on the 1D¹H NMR spectrum. For the two patients, results showed the same resonances of the major metabolite, valproyl-O-glucuronide at concentrations of 121 and 44 mmol/l.     

1H NMR analysis of choline metabolites in fine-needle-aspirate biopsies of breast cancer

Object

The relative amounts of choline (Cho), phosphocholine (PC), and glycerophosphocholine (GPC) may be sensitive indicators of breast cancer and the degree of malignancy. Here we implement some simple modifications to a previously developed ¹H NMR analysis of fine-needle-aspirate (FNA) biopsies designed to yield sufficient spectral resolution of Cho, PC, and GPC for usable relative quantitation of these metabolites.

Materials and methods

FNA biopsies of eighteen breast lesions were examined using our modified procedure for direct ¹H NMR at 400 MHz. Resonances of choline metabolites and potential interferences were fit using the computer program NUTS.

Results

Quantitation of PC, GPC, and Cho relative to each other and to (phospho)creatine was obtained for eleven confirmed cases of infiltrating ductal carcinoma. Reliable results could not be obtained for the remaining cases primarily due to interference from lidocaine anesthetic.

Conclusion

Some simple modifications of a previously developed ¹H NMR analysis of FNAs yielded sufficient spectral resolution of Cho, PC, and GPC to permit usable relative quantitation at 400 MHz. In 9 of the 11 quantified cases the sum of GPC and Cho exceeded 42 % of the total choline-metabolite peak area.     

Quantification of metabolites from single-voxelin vivo 1H NMR data of normal human brain by means of time-domain data analysis

We present here a combination of time-domain signal analysis procedures for quantification of human brainin vivo ¹H NMR spectroscopy (MRS) data. The method is based on a separate removal of a residual water resonance followed by a frequency-selective time-domain line-shape fitting analysis of metabolite signals. Calculation of absolute metabolite concentrations was based on the internal water concentration as a reference. The estimated average metabolite concentrations acquired from six regions of normal human brain with a single-voxel spin-echo technique for theN-acetylaspartate, creatine, and choline-containing compounds were 11.4±1.0,6.5±0.5, and 1.7±0.2 mmol kg^–1 wet weight, respectively. The time-domain analyses ofin vivo ¹H MRS data from different brain regions with their specific characteristics demonstrate a case in which the use of frequency-domain methods pose serious difficulties.     

1H and13C HR-MAS NMR investigations on native and enzymatically digested bovine nasal cartilage

Rheumatic diseases are accompanied by a progressive destruction of the cartilage layer of the joints. Despite the frequency of the disease, degradation mechanisms are not yet understood and methods for early diagnosis are not available. Although some information on pathogenesis could be obtained from the analysis of degradation products of cartilage supernatants, the most direct information on degradation processes would come from the native cartilage as such. We have used¹H as well as¹³C HR-MAS (high resolution magic angle spinning) NMR spectroscopy to obtain suitable line-widths of NMR resonances of native cartilage. ID and 2D NMR spectra of native cartilage were compared with those of enzymatically-treated (collagenase and pa pain) samples. In the¹H NMR spectra of native cartilage, resonances of polysaccharides, lipids and a few amino acids of collagen were detectable, whereas the¹³C NMR spectra primarily indicated the presence of chondroitin sulfate. Treatment with papain resulted only in small changes in the¹H NMR spectrum, whereas a clear diminution of all resonances was detectable in the¹³C NMR spectra. On the other hand, treatment with collagenase caused the formation of peptides with an amino acid composition typical for collagen (glycine, proline, hydroxyproline and lysine). It is concluded that the HR-MAS NMR spectra of cartilage may be of significance for the investigation of cartilage degradation since they allow the fast evaluation of cartilage composition and only very small amounts of sample are required. © 2001 Elsevier Science B.V. All rights reserved.     

The effect of decalcification on the microstructure of articular cartilage assessed by 2H double quantum filtered spectroscopic MRI

²H quadrupolar splitting of deuterated water molecules is a sensitive measure of the order and density of the collagen fibers in articular cartilage. In the calcified zone, near the bone, two pairs of quadrupolar split satellites were previously observed. To examine whether the large splitting observed originates from the presence of calcium ions and hydroxyapatite, one-dimensional ²H single and double quantum filtered spectroscopic imaging were performed on articular cartilage-bone plugs before and after decalcification. After decalcification, the magnitude splitting of the two pairs of satellites did not change and orientation dependency was kept. However, the intensity of the large splitting was greatly enhanced. According to these results the two pairs of satellites do not stem from the presence of calcium ions and hydroxyapatite but originate from the presence of two groups of collagen fibers with different degrees of hydration. The enhanced intensity of the large splitting is attributed to an increased amount of water molecules that fill the void, resulting from the removal of hydroxyapatite, which resides near the fibers responsible for the large splitting. The quadrupolar splitting observed in the trabecular bone was not orientation-dependent, indicating a random orientation of the collagen fibers in that tissue.     

1H spectroscopy in stroke

We used proton spectroscopy to compare metabolite levels in infarcted brain tissue with the levels in corresponding normal white matter. In a preliminary study of six patients, the ratios (mean ± standard deviation) were 0.72±0.25 (p=0.04) for choline, 0.41±0.24 (p=0.002) for creatine, and 0.21±0.22 (p<0.001) forN-acetylaspartate.     

Alleviating artifacts in 1H MRI thermometry by single scan spatiotemporal encoding

Objective

Recent years have seen an increased interest in combining MRI thermometry with devices capable of destroying malignancies by heat ablation. Expected from the MR protocols are accurate and fast thermal characterizations, providing real time feedback on restricted tissue volumes and/or rapidly moving organs like liver. This article explores the potential advantages of relying on spatiotemporally encoded (SPEN) sequences for retrieving real-time thermometric images based on the water’s proton resonance frequency (PRF) shifts.

Materials and methods

Hybrid spatiotemporal/k-space encoding single-scan MRI experiments were implemented on animal and human scanners, and their abilities to deliver single- and multi-slice real-time thermometric measurements based on PRF-derived phase maps in phantoms and in vivo, were compared against echo planar imaging (EPI) and gradient-echo counterparts.

Results

Under comparable acquisition conditions, SPEN exhibited advantages vis-à-vis EPI in terms of dealing with inhomogeneous magnetic field distortions, with shifts arising due to changes in the central frequency offsets, with PRF distributions, and for zooming into restricted fields-of-view without special pulse sequence provisions.

Conclusion

This work confirms the ability of SPEN sequences, particularly when implemented under fully-refocused conditions, to exploit their built-in robustness to shift- and field-derived inhomogeneities for monitoring thermal changes in real-time under in vitro and in vivo conditions.     

Detection and quantification of d-glucuronic acid in human bile using 1H NMR spectroscopy: relevance to the diagnosis of pancreatic cancer

Objective  

There are no specific biomarkers available for the definitive diagnosis of pancreatic cancer. Analysis of d-glucuronic acid (GlcUA) in bile could be valuable in this regard.     

Quantitative metabolic profiles of 2nd and 3rd trimester human amniotic fluid using 1H HR-MAS spectroscopy

Object

To establish and compare normative metabolite concentrations in 2nd and 3rd trimester human amniotic fluid samples in an effort to reveal metabolic biomarkers of fetal health and development.

Materials and methods

Twenty-one metabolite concentrations were compared between 2nd (15–27 weeks gestation, N = 23) and 3rd (29–39 weeks gestation, N = 27) trimester amniotic fluid samples using ¹H high resolution magic angle spinning (HR-MAS) spectroscopy. Data were acquired using the electronic reference to access in vivo concentrations method and quantified using a modified semi-parametric quantum estimation algorithm modified for high-resolution ex vivo data.

Results

Sixteen of 21 metabolite concentrations differed significantly between 2nd and 3rd trimester groups. Betaine (0.00846±0.00206 mmol/kg vs. 0.0133±0.0058 mmol/kg, P < 0.002) and creatinine (0.0124±0.0058 mmol/kg vs. 0.247±0.011 mmol/kg, P < 0.001) concentrations increased significantly, while glucose (5.96±1.66 mmol/kg vs. 2.41±1.69 mmol/kg, P < 0.001), citrate (0.740±0.217 mmol/kg vs. 0.399±0.137 mmol/kg, P < 0.001), pyruvate (0.0659±0.0103 mmol/kg vs. 0.0299±0.286 mmol/kg, P < 0.001), and numerous amino acid (e.g. alanine, glutamate, isoleucine, leucine, lysine, and valine) concentrations decreased significantly with advancing gestation. A stepwise multiple linear regression model applied to 50 samples showed that gestational age can be accurately predicted using combinations of alanine, glucose and creatinine concentrations.

Conclusion

These results provide key normative data for 2nd and 3rd trimester amniotic fluid metabolite concentrations and provide the foundation for future development of magnetic resonance spectroscopy (MRS) biomarkers to evaluate fetal health and development.     

Validation of glutathione quantitation from STEAM spectra against edited 1H NMR spectroscopy at 4T: application to schizophrenia

Objective: Quantitation of glutathione (GSH) in the human brain in vivo using short echo time ¹ H NMR spectroscopy is challenging because GSH resonances are not easily resolved. The main objective of this study was to validate such quantitation in a clinically relevant population using the resolved GSH resonances provided by edited spectroscopy. A secondary objective was to compare several of the neurochemical concentrations quantified along with GSH using LCModel analysis of short echo time spectra in schizophrenia versus control. Materials and Methods: GSH was quantified at 4T from short echo STEAM spectra and MEGA-PRESS edited spectra from identical volumes of interest (anterior cingulate) in ten volunteers. Neurochemical profiles were quantified in nine controls and 13 medicated schizophrenic patients. Results: GSH concentrations as quantified using STEAM, 1.6 ± 0.4 μmol/g (mean ± SD, n=10), were within error of those quantified using edited spectra, 1.4 ± 0.4 μmol/g, and were not different (p=0.4). None of the neurochemical measurements reached sufficient statistical power to detect differences smaller than 10% in schizophrenia versus control. As such, no differences were observed. Conclusions: Human brain GSH concentrations can be quantified in a clinical setting using short-echo time STEAM spectra at 4T.     

<Superscript>1</Superscript>H NMR relaxation times of skeletal muscle metabolites at 3 T

This study reports proton relaxation times of water and metabolites in soleus and tibialis anterior muscles of young healthy volunteers at 3 T. The results are in agreement with data reported for 1.5 and 4 T, showing a steady increase of spin-lattice relaxation times of water, creatine and lipids with B₀ and no effect of B₀ on spin–spin relaxation. Comparison between muscles revealed a longer spin–spin relaxation time of water in soleus than in tibialis anterior muscle (31±1 ms vs. 28±1 ms, p<0.05). These data can be applied to relaxation correction for the absolute quantification of skeletal muscle metabolite concentrations and further sequence optimization.     

The size of cytoplasmic lipid droplets varies between tumour cell lines of the nervous system: a 1H NMR spectroscopy study

Object

Cytoplasmic lipid droplets (LDs) are dynamic cellular organelles; their accumulation is associated with several cellular processes, such as cell proliferation, apoptosis and necrosis. ¹H Nuclear Magnetic Resonance (NMR) spectroscopy detects resonances from lipids present in cytoplasmic (LDs); an understanding of the relationship between LD characteristics and NMR lipid signals is important.

Materials and methods

In this study, five nervous system cancer cell lines were investigated. Nile red staining was used to measure the diameter of LDs. High-resolution magic angle spinning NMR (HR-MAS) was performed on harvested cell pellets to quantify the patterns of lipid signals.

Results

LDs were present in all five cell lines with different morphology. An average LD diameter of approximately 0.2???m was found in all cell types. Diameter of the largest LDs varied across the cell lines. The intensity of NMR lipid signals varied greatly between cell types, and a good correlation was found between total volume of LDs and the proton NMR lipid signal intensity at 0.9 and 1.3?ppm.

Conclusion

The correlation implied that little NMR signal is detected from LDs of diameters less than approximately 0.34???m, most likely due to restriction of rotational motion of the lipids.     

Magnetization transfer characteristics in atherosclerotic plaque components assessed by adapted binomial preparation pulses

Increasing the contrast between atheromatous plaque components is a major issue in cardiovascular MRI research. It would allow one to identify unstable plaque by differentiating the lipid core associated with vulnerability, from the fibrous cap, considered as a factor of stability. T₂ and diffusion-weighted imaging have already provided satisfying results. Magnetization transfer (MT) between restricted protons H_r and free-water protons H_f could achieve a different contrast related to collagen and lipoprotein macromolecules present in the fibrous cap and lipid core, respectively. The purpose of this work was to evaluate in vitro the MT effect produced by adapted T₂-selective 1-3-3-1 binomial pulses on isolated samples of atheromatous arteries at 3 T. A method based on simulation was used in order to improve the MT specificity: it is shown that 50% 1-3-3-1 pulses (the percentage indicating the level of H_r saturation) allow an estimation of T_2r, the H_r T₂. Using this technique, magnetization transfer was observed for the first time in atherosclerotic plaque components, an effect more pronounced for the fibrous cap and media than for the lipid core and advantitia. The T_2r estimation gave values ranging from 20 to 25 μs for the four samples. This preliminary study provides a basis for establishing an MT imaging sequence of atheromatous arteries, by using 50% 1-3-3-1 pulses calibrated for saturating protons with a 20 μs T₂. This MT protocol should be further compared to T₂ and diffusion-weighted imaging.     

氢冷发电机氢气品质监督的新手段--QQFX-Ⅰ型智能在线氢气分析仪

叙述了能够全面检测氢冷机组氢气品质参数的 QQFX- 型智能在线氢气分析仪研制的背景及其意义。介绍了该系统的组成部分 ,各自的功能、仪器的参数以及它的技术特点 ,该仪器除在电力系统的氢冷机组使用外 ,还可应用于石化、航天等领域     

Impact of prenatal stress on <Superscript>1</Superscript>H NMR-based metabolic profiling of rat amniotic fluid

Object  

The impact of inflammation in utero on amniotic fluid composition, the delivery term and the number of newborn rats per litter was investigated. The growth of newborns during the first fourteen days of life was analysed.     

In vivo1H MR spectra analysis by means of second derivative method

Short echo time (TE) in vivo PRESS¹H MR spectra (2 T. TE = 35 ms) of normal brain were fitted in the frequency domain using the second derivative method. In this approach, local maxima and hidden peaks are found as local minima of spectrum second derivative. The Lorentzian robust minimisation procedure (referred to as maximum likelihood or m-estimate fitting) using Levenburg -Marquardt non-linear fitting engine was applied. Spectral lines were approximated under the assumption of the mixed Lorentzian/Gaussian lineshapes. The same procedure was applied to 18 proton spectra. The number of peaks found within the range of 0.74/4.2 parts per million (ppm) was 52 ± 3 and their positions were almost the same. The fitted lines were assigned on the basis of the J-pattern recalculated for the field strength of 2 T and by comparing the chemical shifts with the shifts in the single compound spectra. The ratios of main metabolites, such as NAA/Cr, Cho/Cr, Cho/NAA and ml/Cr, are in accord with those obtained earlier using the software supplied with the MR imager and the absolute concentrations ofN-acetylaspartate (NAA). choline containing compounds (Cho),myo Inositol (ml), glucose (Gle) and glutamate (Glu) obtained from the fit agree with those reported in literature, which confirms the usefulness of the second derivative method in routine analyses of¹H MR brain spectra.     

Comparison of in vivo1H MRS of human brain tumours with1H HR-MAS spectroscopy of intact biopsy samples in vitro

High resolution magic angle spinning (MAS)¹H nuclear magnetic resonance (NMR) spectroscopy has been employed to study intact human brain tumour tissue and comparison with the corresponding in vivo spectrum has been made. Two dimensional¹H MAS-NMR measurements, including J-resolved and homonuclear shift correlation spectra, were obtained to aid metabolite signal assignment. MAS gave greatly improved line-shape and reduced line-width in comparison to conventional high resolution in vivo¹H MRS of intact tissue, permitting the simultaneous detection of cellular lipids and metabolites. The technique provides the most direct method for comparison of in vivo spectra with high resolution spectra in vitro and hence allows more reliable peak assignment of in vivo¹H MRS spectra.