Biomedical subjects

C Arús

Publications and source records attributed to C Arús.

At least 19 recordsLinked to original sources

PubMed2006

Analysis of the changes in the 1H NMR spectral pattern of perchloric acid extracts of C6 cells with growth.

The aim of this work was to identify spectral markers of cell proliferation that could be of use in clinical MRS. Cultured C6 ATCC rat glioma cells were used as models for this purpose and metabolites were extracted with perchloric acid at three different growth curve stages: log, confluence and post-confluence. 1D and 2D in vitro(1)H NMR spectra were recorded at 9.4 T. Statistically significant changes in myo-inositol and glutamine concentrations between log phase and post-confluence were found when normalized to the creatine ratio. The myo-inositol/creatine ratio was 2.76 +/- 0.82 at log phase increasing to 7.43 +/- 1.34 at post-confluence, while the glutamine/creatine ratio decreased from 0.22 +/- 0.03 to 0.10 +/- 0.02. No significant differences were recorded for other metabolites investigated. The fact that both myo-inositol and glutamine are detectable by in vivo MRS at clinical fields makes their changes relevant as potential astrocytic tumour proliferation rate markers in clinical MRS.

Animals↗

PubMed2004

Assignment of the 2.03 ppm resonance in in vivo 1H MRS of human brain tumour cystic fluid: contribution of macromolecules.

MRI and MRS are established techniques for the evaluation of intracranial mass lesions and cysts. The 2.03 ppm signal recorded in their (1)H-MRS spectra is often assigned to NAA from outer volume contamination, although it has also been detected in non-infiltrating tumours and large cysts. We have investigated the molecular origin of this resonance in ten samples of cystic fluids from human brain tumours. The NMR detected content of the 2.03 ppm resonance in 136 ms echo time spectra, assuming an N- CH(3) origin, was 3.19 +/- 1.01 mM. Only one third (34 +/- 12%) of the N-acetyl containing compound (NAC) signal could be extracted by perchloric acid (PCA) indicating that most of it originated in a macromolecular PCA-insoluble component. Chemical analysis of the cyst fluids showed that sialic acid bound to macromolecules would account for 64.3% and hexuronic containing compounds for 29.2% of the NMR-detectable ex vivo signal, 93.4% of the signal at TE 136 ms. Lactate content measured by NMR (6.4 +/- 4.4 mM) and the predominance of NAC originating in sialic acid point to a major origin from tumour rather than from plasma for this 2.03 ppm resonance.

Brain↗

PubMed2004

Brain tumor classification based on long echo proton MRS signals.

There has been a growing research interest in brain tumor classification based on proton magnetic resonance spectroscopy (1H MRS) signals. Four research centers within the EU funded INTERPRET project have acquired a significant number of long echo 1H MRS signals for brain tumor classification. In this paper, we present an objective comparison of several classification techniques applied to the discrimination of four types of brain tumors: meningiomas, glioblastomas, astrocytomas grade II and metastases. Linear and non-linear classifiers are compared: linear discriminant analysis (LDA), support vector machines (SVM) and least squares SVM (LS-SVM) with a linear kernel as linear techniques and LS-SVM with a radial basis function (RBF) kernel as a non-linear technique. Kernel-based methods can perform well in processing high dimensional data. This motivates the inclusion of SVM and LS-SVM in this study. The analysis includes optimal input variable selection, (hyper-) parameter estimation, followed by performance evaluation. The classification performance is evaluated over 200 stratified random samplings of the dataset into training and test sets. Receiver operating characteristic (ROC) curve analysis measures the performance of binary classification, while for multiclass classification, we consider the accuracy as performance measure. Based on the complete magnitude spectra, automated binary classifiers are able to reach an area under the ROC curve (AUC) of more than 0.9 except for the hard case glioblastomas versus metastases. Although, based on the available long echo 1H MRS data, we did not find any statistically significant difference between the performances of LDA and the kernel-based methods, the latter have the strength that no dimensionality reduction is required to obtain such a high performance.

Artificial Intelligence↗

PubMed2004

Classification of brain tumours using short echo time 1H MR spectra.

The purpose was to objectively compare the application of several techniques and the use of several input features for brain tumour classification using Magnetic Resonance Spectroscopy (MRS). Short echo time 1H MRS signals from patients with glioblastomas (n = 87), meningiomas (n = 57), metastases (n = 39), and astrocytomas grade II (n = 22) were provided by six centres in the European Union funded INTERPRET project. Linear discriminant analysis, least squares support vector machines (LS-SVM) with a linear kernel and LS-SVM with radial basis function kernel were applied and evaluated over 100 stratified random splittings of the dataset into training and test sets. The area under the receiver operating characteristic curve (AUC) was used to measure the performance of binary classifiers, while the percentage of correct classifications was used to evaluate the multiclass classifiers. The influence of several factors on the classification performance has been tested: L2- vs. water normalization, magnitude vs. real spectra and baseline correction. The effect of input feature reduction was also investigated by using only the selected frequency regions containing the most discriminatory information, and peak integrated values. Using L2-normalized complete spectra the automated binary classifiers reached a mean test AUC of more than 0.95, except for glioblastomas vs. metastases. Similar results were obtained for all classification techniques and input features except for water normalized spectra, where classification performance was lower. This indicates that data acquisition and processing can be simplified for classification purposes, excluding the need for separate water signal acquisition, baseline correction or phasing.

Brain Chemistry↗

PubMed2003

Utility of proton MR spectroscopy in the diagnosis of radiologically atypical intracranial meningiomas.

Our aim was to evaluate the usefulness of proton MR spectroscopy ((1)H MRS) in the diagnosis of radiologically atypical brain meningiomas. We studied 37 patients with intracranial meningiomas with MRI and (1)H MRS (TE 136 ms). Their spectra were quantitatively assessed and compared with those of 93 other intracranial brain neoplasms: 15 low-grade and 14 anaplastic astrocytomas, 30 glioblastomas and 34 metastases. The most characteristic features of meningiomas were the presence of alanine, high relative concentrations of choline and glutamine/glutamate and low concentrations of creatine-containing compounds, N-acetyl-containing compounds and lipids. These resonances were assembled in algorithms for two-way differentiation between meningioma and the other tumours. The performance of the algorithms was tested in the 130 patients using the leave-one-out method, with 94% success in differentiating between meningioma and other tumour. Of the 37 meningiomas, five (14%) were thought atypical on MRI, and in only one of these, found to be malignant on histology, was a diagnosis other than meningioma suggested by the algorithm. The other four were correctly classified. We suggest that (1)H MRS provides information on intracranial meningiomas which may be useful in diagnosis of radiologically atypical cases.

Adolescent↗

PubMed2001

Mobile lipid production after confluence and pH stress in perfused C6 cells.

NMR-visible mobile lipid (ML) has been observed in aggressive tumors and also in in vitro tumor cell models subjected to growth-inhibiting conditions, such as confluence or low-pH stress. The aim of the present study was to determine if ML production after confluence or low pH stress in a cultured cell model of brain tumor is due to growth arrest alone. ML was observed in situ by one- and two-dimensional (1)H NMR in viable but growth-arrested C6 glioma cells superfused for a period of 48 h after harvesting. The rate of ML production in cells harvested at subconfluence was compared to the rate in cells confluent for one cell cycle and to the rate in subconfluent-harvested cells superfused at low pH (pH 6.1). Confluent-harvested cells produced ML at a markedly greater rate than that of cells harvested at subconfluence, suggesting the involvement of prior cell-cell contact rather than simple growth arrest. A high rate was also observed in subconfluent-harvested cells subjected to low pH, indicating that ML in pH-stressed cells also does not arise from growth arrest alone. Furthermore, two-dimensional data on the degree of unsaturation of the ML fatty acyl chains and one-dimensional (31)P and two-dimensional (1)H NMR data on the GPC content of the cells suggest distinct metabolic pathways for the production of ML following confluence and low-pH stress.

Adenosine Triphosphate↗

PubMed2001

Magnetic resonance spectroscopy of brain hemangiopericytomas: high myoinositol concentrations and discrimination from meningiomas.

OBJECT: Hemangiopericytomas are a rare type of brain tumor that are very similar to meningiomas in appearance and symptoms but require different treatment. It is not normally possible to distinguish between them by using magnetic resonance (MR) imaging and computerized tomography studies. However, discrimination may be possible by using in vivo MR spectroscopy (MRS) because the biochemical composition of these two lesions is different. The goal of this study was to describe the use of MRS in discriminating between these similar tumor types. METHODS: In vivo MRS spectra were acquired in 27 patients (three with hemangiopericytomas and 24 with meningiomas) by using a single-voxel proton brain examination system at 1.5 teslas with short- (20-msec) and long- (135-msec) echo times. In addition, brain biopsy specimens obtained by open craniotomy were frozen within 5 minutes of resection and stored in liquid nitrogen until they were used. The specimens were powdered, extracted with perchloric acid, redissolved in 2H2O2 and high-resolution in vitro MRS was used at 9.4 teslas to record their spectra. CONCLUSIONS: In this study the authors show that hemangiopericytomas could be clearly distinguished from meningiomas because they have a larger peak at 3.56 ppm. Measurements of extracts of the tumors and comparison of spectra acquired with MRS at long- (135-msec) and short- (20-msec) echo times established that this was due to the much higher levels of myoinositol in the hemangiopericytomas.

Brain Neoplasms↗

PubMed2001

A prototype decision support system for MR spectroscopy-assisted diagnosis of brain tumours.

Our objective is to develop a decision support system that improves the accuracy of non-invasive brain tumour diagnosis and grading by enabling radiologists to use data from Magnetic Resonance Spectroscopy (MRS). The system, which uses pattern recognition techniques, is trained on a validated database of spectra and associated clinical information to provide automated classification of spectra from brain tumours. An innovative user-interface presents classification results as a two-dimensional overview plot in which points representing cases of different diseases form distinct clusters. Users can inspect any cases in these plots and compare them with the new, unknown spectrum. Hence, the overview plot can both communicate the classification of a case and help provide explanation for that classification in part by supporting human case-based reasoning. This paper describes the development of a prototype system implemented in JAVA.

Brain Neoplasms↗

PubMed1999

The relationship between nuclear magnetic resonance-visible lipids, lipid droplets, and cell proliferation in cultured C6 cells.

There is an ongoing controversy about the subcellular origin of the fatty acyl chains that give rise to the NMR visible mobile lipids (MLs) resonance at approximately 1.24 ppm in the 1H spectra of cells and solid tumors. Some groups have been supporting the hypothesis that triglycerides originating MLs are isotropically tumbling in small membrane microdomains, whereas other authors back the proposal that they are inside cytosolic or extracellular (necrotic areas) lipid droplets. Furthermore, MLs are frequently present in in vivo spectra recorded from human brain tumors, but the meaning of this detection is not fully clear. We have addressed the possible contribution of intracellular droplets to the ML pattern recorded from human brain tumors in vivo by studying cultured C6 rat glioma cells as a model system for astrocytic tumors. We show here that cultured C6 cells display ML resonances in high field (9.4 T) 1H NMR spectra recorded at 136 ms echo time when grown at saturation density conditions, but no MLs are visible for log-phase cells. Fluorescence microscopy analysis of cells stained with the lipophylic dye Nile red shows intracellular spherical yellow-gold droplets containing neutral lipids; cells at saturation density present lipid droplets of diameters about 1.6 microm in most cells (85%), whereas they are almost absent in log-phase cells (only 6% of the cells contain them). Furthermore, log-phase cells can be induced to display MLs and accumulate Nile red-positive droplets by culturing them for 24 h at pH 6.2. This acid pH effect can be fully reversed by 24 h of standard media incubation. Lipid droplet volume calculated from fluorescence microscopy preparations in an average cell is different for both culture conditions (2.2 times higher volume for saturation density than for pH-stressed cells). This difference in lipid droplet volume is reflected by a different ML peak height at 1.24 ppm (about 2 times higher for saturation density than for pH-stressed cells). Flow cytometry analysis shows that both culture conditions result in a slowing down of the proliferation rate of the cells. The fact that MLs are found to originate in lipid droplets inside cells that are growth compromised but still viable suggests that changes in the proliferative state of tumor cells, in the absence of necrosis, may be detected non invasively by in vivo NMR spectroscopy.

Animals↗

PubMed1998

1H MRS markers of tumour growth in intrasplenic tumours and liver metastasis induced by injection of HT-29 cells in nude mice spleen.

We have characterized, by in vitro magnetic resonance spectroscopy (MRS), the metabolite pattern of perchloric acid (PCA) extracts of intrasplenic tumours and hepatic metastasis, produced by intra-spleen injection of the human colorectal carcinoma cell line HT-29 and its metastatic variant HT-29 MMM into nude mice. Our aim was to gain further understanding of colorectal tumour metabolism as a basis for future in vivo studies of human colon cancer by 1H MRS. Metabolite PCA extract analysis showed a good reproduction of the spectral pattern observed in human primary colon tumours, while they were very different from the spectral pattern of the host tissues (spleen and liver). The main differences between host and tumour tissues involved taurine, phosphocholine (PC), phosphoethanolamine (PE), creatine, glycogen and glucose. Creatine is the most promising marker to follow tumour growth because of its practical absence in the nude mice host tissues. Detection of variable levels of this compound and of taurine in hepatic foci in man, are suggested as possible diagnostic markers. No correlation could be found between spectral pattern differences and the different ability to metastasize of the two HT-29 cell lines used. Furthermore, indirect evidence for a functional link between taurine and myo-inositol in colon tumour cells is presented. In summary, our data suggest that the nude mice model may be a suitable system for the MRS study of the changes taking place in host tissues upon tumour progression.

Animals↗

PubMed1998

Pattern recognition analysis of 1H NMR spectra from perchloric acid extracts of human brain tumor biopsies.

Pattern recognition techniques (factor analysis and neural networks) were used to investigate and classify human brain tumors based on the 1H NMR spectra of chemically extracted biopsies (n = 118). After removing information from lactate (because of variable ischemia times), unsupervised learning suggested that the spectra separated naturally into two groups: meningiomas and other tumors. Principal component analysis reduced the dimensionality of the data. A back-propagation neural network using the first 30 principal components gave 85% correct classification of meningiomas and nonmeningiomas. Simplification by vector rotation gave vectors that could be assigned to various metabolites, making it possible to use or to reject their information for neural network classification. Using scores calculated from the four rotated vectors due to creatine and glutamine gave the best classification into meningiomas and nonmeningiomas (89% correct). Classification of gliomas (n = 47) gave 62% correct within one grade. Only inositol showed a significant correlation with glioma grade.

Biomarkers, Tumor↗

PubMed1997

Evidence that mobile lipids detected in rat brain glioma by 1H nuclear magnetic resonance correspond to lipid droplets.

Mobile lipids have been detected by proton nuclear magnetic resonance (NMR) in animal and human tumors (cultured cells, biopsies, and in vivo), but their origin and subcellular location are still unclear. They have been associated with malignancy, metastatic ability, drug resistance, and necrosis. We wanted to determine whether these lipids are located within plasma membrane microdomains or in lipid droplets for a C6 cell-induced rat glioma. NMR-visible mobile lipids were found in all subcellular fractions isolated from the rat tumor, except in the cytosolic supernatants. Transmission electron microscopy showed that lipid droplets were present in all subcellular fractions containing NMR-visible lipids and in the necrotic and perinecrotic areas of the tumor. The mean diameter of droplets isolated by flotation in the subcellular fractionation protocol was 0.97 microm (n = 682; droplet profile diameter range between 0.2 and 5.0 microm). The apparent diffusion coefficient for these lipids (46 +/- 17 microm2 s(-1) measured in vivo by proton spectroscopy was four orders of magnitude higher than would be expected if mobile lipids were inside plasma membrane microdomains. The combined results demonstrated that mobile lipids detected in vivo by proton NMR in the C6 rat glioma are located in large lipid droplets, associated with the necrotic process.

Animals↗

PubMed1997

Diagnosis of brain abscess by magnetic resonance spectroscopy. Report of two cases.

Two cases of brain abscess were diagnosed by combining magnetic resonance spectroscopy (MRS) and magnetic resonance (MR) imaging. The resonances observed in vivo were assigned by means of an in vitro MRS study of the exudates extracted during surgical aspiration of the abscesses. The technique of MRS was demonstrated to be very powerful in the differential diagnosis of brain abscesses from other brain pathologies such as neoplasms. Amino acids, probably originating from extracellular proteolysis, and other compounds, such as acetate, arising from bacterial metabolism, were visible in the MRS spectra of the abscess, whereas they are not present in spectra of neoplasms. In this sense, MRS complemented the information provided by MR imaging to achieve a correct diagnosis of brain abscesses and could be added to routine MR examinations with only a small increase in cost and time.

Adult↗

PubMed1997

Measurement of intracellular pH of maize seeds (Zea mays) during germination by 31P nuclear magnetic resonance spectroscopy.

The 31P NMR spectra of germinating maize seeds showed a single broad resonance that shifted its position as germination proceeded (studied between 0 and 10 days). The resonance was shown to originate from the phosphate groups of phytine (Mg2+, Ca2+ and K+ salt of myoinositol hexakisphosphate) in a subcellular compartment of the embryo scutellar cells. A series of calibration curves for the chemical shift dependence of the phytate resonance in the presence of Mg2+ and Ca2+ were constructed. These calibration curves allowed us to determine that an acidification of the phytate containing compartment in the seed embryo takes place, reaching a minimum at about pH 4 after three days of germination. This acidification could be important in allowing phytate solubilization for export to growing parts of the maize seedling.

Cell Compartmentation↗

PubMed1996

Quantitative and qualitative characterization of 1H NMR spectra of colon tumors, normal mucosa and their perchloric acid extracts: decreased levels of myo-inositol in tumours can be detected in intact biopsies.

Sixteen colonic tumours and 10 normal mucosa biopsies have been examined by 1H NMR spectroscopy at 9.4 T. A complete characterization and quantification of the aliphatic region of PCA extract spectra and the analysis of the two-dimensional COSY spectra of five pairs of intact biopsies (tumor and control mucosa) has been carried out. The analysis of the PCA extracts demonstrated a significant increase in the concentration of the endogenous compounds: lactate, glutamate, aspartate, taurine, spermine, glutathione and glycerophosphoethanolamine, and a significant decrease of myo- and scyllo-inositol, in tumours with respect to mucosae. Among these metabolites, the high myo-inositol and taurine levels and the reciprocal changes found between them in tumours and mucosae make their resonances interesting as possible malignancy markers if they are detectable in vivo. In contrast to the easy observation of taurine in one-dimensional spectra of intact biopsies, the difficulty of observing myo-inositol prompted us to use two-dimensional COSY spectra for the detection and quantification of both these metabolites. In the two-dimensional spectra, the use of a ratio between the cross-peak volumes of both metabolites permits an excellent differentiation between tumours and normal mucosa and suggests its potential to detect malignant changes in the healthy tissue, provided a two-dimensional approach is used.

Biopsy↗

PubMed1995

A simple approach to the design of a shielded gradient probe for high-resolution in vivo spectroscopy.

A simple approach to the design of a shielded gradient coil set for a high-resolution NMR probe is described in detail. An Appendix is provided that shows the step-by-step Mathematica routine used to carry out the necessary calculations. The probe provided excellent water suppression and gradient-accelerated acquisition of one- and two-dimensional spectra from perfused bovine retina.

Algorithms↗

PubMed1994

In vivo, ex vivo, and in vitro one- and two-dimensional nuclear magnetic resonance spectroscopy of an intracerebral glioma in rat brain: assignment of resonances.

An in vivo study of intracerebral rat glioma using proton-localized NMR spectroscopy showed important modifications of the spectra in the tumor as compared with the contralateral brain. To carry out the assignment of the resonances of the glioma spectra, tumoral and normal rat brain tissues were studied in vivo, ex vivo, and in vitro by one-dimensional and two-dimensional proton spectroscopy. N-Acetylaspartate was found at an extremely low level in the glioma. The change of peak ratio total creatine/3.2 ppm peak was found to be due to a simultaneous decrease of the total creatine content and an increase of the 3.2 ppm peak. The 3.2 ppm resonance in the glioma spectra has been shown to originate from choline, phosphocholine, glycerophosphocholine, taurine, inositol, and phosphoethanolamine. The increase of the 3.2 ppm peak in the glioma was found to result from the increase of taurine and phosphoethanolamine contents. The peak in the 1.3 ppm region of the glioma spectra was due to both lactate and mobile fatty acids. Moreover, two-dimensional spectroscopy of excised tissues and extracts showed the presence of hypotaurine only in the tumor.

Amino Acids↗

PubMed1993

Development and characterization of an ergometer to study the bioenergetics of the human quadriceps muscle by 31P NMR spectroscopy inside a standard MR scanner.

A description is given of an ergometer made of nonmagnetic materials which fits into a standard whole body MR magnet. T2-weighted images show that exercise is highly specific for quadriceps muscles. The ergometer permits the noninvasive study of T2-related changes in the proton images of the leg as well as changes occurring in muscle bioenergetics during exercise and recovery.

Equipment Design↗