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At least 217 records · Page 12Linked to original sources

PubMed2004

Cerebral perfusion assessment by bolus tracking using hyperpolarized 13C.

Cerebral perfusion was assessed with 13C MRI in a rat model after intravenous injections of the 13C-labeled compound bis-1,1-(hydroxymethyl)-1-13C-cyclopropane-D8 in aqueous solutions hyperpolarized by dynamic nuclear polarization (DNP). Since the tracer acted as a direct signal source, several of the problems associated with techniques based on traditional dynamic susceptibility contrast (DSC) MRI contrast agents were avoided. Maps of cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT) were calculated. The MTT was determined to be 2.8 +/- 0.8 sec. However, arterial partial-volume effects in the animal model prevented accurate absolute quantification of CBF and CBV. It was demonstrated that depolarization of the hyperpolarized 13C tracer via relaxation and the imaging sequence had little influence on CBF assessment when the time resolution of the imaging sequence was short compared to the MTT. However, CBV and MTT were increasingly underestimated as MTT or the depolarization rate increased if depolarization was not taken into account. With a modified bolus-tracking theory depolarization could be compensated for, assuming that the depolarization rate was known. Three separate compensation methods were investigated experimentally and by numerical simulations.

Algorithms↗

PubMed2005

New method for the simultaneous detection of metabolites and water in localized in vivo 1H nuclear magnetic resonance spectroscopy.

A new two-scan method for localized 1H in vivo NMR spectroscopy (MRS) without water suppression (WS) is described. In one of the scans, two chemical shift selective 180 degrees pulses are applied prior to a standard localization sequence to invert all metabolite signals upfield and downfield from water, which remains unaffected. The difference spectrum records the metabolites whereas water and accompanying gradient induced artifacts are widely suppressed. The method was implemented on a 4.7-T system using point resolved spectroscopy with a short echo time of 18 ms. Phantom measurements proved the feasibility of absolute quantification using water as an internal reference. Measurements on healthy rat brain yielded comparable spectrum quality as measurements with water presaturation. The method does not require additional adjustments or sophisticated data postprocessing and scales favorably with increasing B(0) field. Therefore, the method should be useful for 1H MRS without WS. Although the two-step method doubles the minimum total measurement time, it may also be of interest for spectroscopic imaging (SI) without WS, in particular if fast SI techniques are applied.

Algorithms↗

PubMed1988

A critical assessment of methods of measuring metabolite concentrations by NMR spectroscopy.

Many NMR studies are made by scientists and clinicians in order to measure metabolite levels. There is a growing need to obtain absolute quantification of these metabolites. We have therefore reviewed the methods that are available and those which are being developed, for studies in vivo and on isolated tissues. We describe the following methods and assess their advantages and limitations: (1) Conventional spectroscopy and area ratios, (2) Tissue extracts, (3) External capillary, (4) Internal endogenous marker, (5) Internal exogenous marker, (6) External marker in bath, (7) Reference to internal 1H water concentration, (8) Gradient-localized spectroscopy and (9) Fourier series analysis localization. The relevant biological and technical aspects associated with quantification of metabolite concentrations are also discussed. This includes NMR visibility, origin of the NMR signals, definitions of concentration, influence of the RF coil, data collection and data analysis.

Humans↗

PubMedDate not supplied

Identification of Scyllo-inositol in proton NMR spectra of human brain in vivo.

Scyllo-inositol has been identified in proton NMR spectra of mammalian brain in vitro and in vivo. In contrast to myo-inositol this isomer comprises six equivalent CH protons that yield a singlet resonance at a chemical shift of 3.35 ppm. 1-D and 2-D J-resolved proton NMR studies (7.0 T) of perchloric acid extracts of brain tissues revealed different amounts of scyllo-inositol in man, sheep, cow and rat. Absolute quantification of localized short-echo time proton NMR spectra (2.0 T) of human brain in vivo resulted in scyllo-inositol concentrations of 0.35 +/- 0.06 mM for white matter (n = 25), 0.43 +/- 0.11 mM for grey matter (n = 23) and 0.57 +/- 0.14 mM for cerebellum (n = 10). Evidence for a tight metabolic link to myo-inositol was supported by a simultaneous variation of myo- and scyllo-inositol in patients with various brain diseases.

Animals↗

PubMed2005

FAWSETS perfusion measurements in exercising skeletal muscle.

Arterial spin labeling (ASL) techniques are now recognized as valid tools for providing accurate measurements of cerebral and cardiac perfusion. The labeling process used with most ASL techniques creates two problems, magnetization transfer (MT) effects and arterial transit time effects, that require compensation. The compensation process limits time resolution and hinders absolute quantification. MT effects are particularly problematic in skeletal muscle because they are large and change rapidly during exercise. The protocol presented here was developed specifically for quantification of perfusion in exercising skeletal muscle. The ASL technique that was implemented, FAWSETS, eliminates MT effects and arterial transit times. Localized, single-voxel perfusion measurements were acquired from rat hind limbs at rest, during ischemia and during three different levels of stimulated exercise. The results demonstrate sufficient sensitivity to determine the time constants for perfusion changes at onset of, and during recovery from, exercise and to distinguish the differences in the amplitude of the perfusion response to different levels of exercise. Additional measurements were conducted to demonstrate insensitivity to MT effects. The exercise protocol is easily adaptable to phosphorous magnetic resonance measurements, allowing the possibility to acquire local measurements of perfusion and metabolism from the same tissue in future experiments.

Animals↗

PubMed2004

Assessing factors for reliable quantitative proteomics based on two-dimensional gel electrophoresis.

We statistically analysed various factors to get accurate estimates of protein quantities from two-dimensional gels. Yeast proteins were labelled with (35)S or stained with Coomassie Brilliant Blue G-250, and spots were automatically quantified with software packages Kepler, ImageQuaNT, Melanie 3.0 and Progenesis. The different software packages proved to have very similar performances. With (35)S-labelled actin spot as a reference, we studied the staining efficiency of colloidal Coomassie blue as a function of amino acid composition of the protein, and derived an equation to estimate the number of molecules per cell from blue-stained proteins. Absolute quantification of most glycolytic enzymes was carried out in two yeast strains.

Actins↗

PubMed2006

Primary amine coding as a path to comparative proteomics.

Various isotope coding strategies are being used today in the field of comparative proteomics. This article specifically reviews the strengths and limitations of various N-termini-directing strategies. N-termini-directed coding strategy allows for use of different chromatographic enrichment techniques. Since N-termini-directed coding strategies are global in nature, they can be utilized in studying PTMs as well as protein expression. Using different N-termini-directed coding strategies, both relative and absolute quantification of proteins can be achieved either in the MS mode or in the MS/MS mode. The review ends with the conclusion that significant improvements have been made in the last decade. Among various issues, a need still exists for a better understanding of the kinetic issues in proteomics, relative protein pool sizes for different proteins and the issue of stimulus-induced changes in protein aggregation. Another critical issue that needs to be addressed in great detail is the role of PTMs in regulation.

Amines↗

PubMed2006

An integrated approach to mapping the proteome of the human bone marrow stromal cell.

Bone marrow stromal cells, also known as mesenchymal stem or progenitor cells, are thought to play a critical role in hematopoiesis. Because hematopoiesis is dysregulated in leukemic individuals, expression profiling in leukemic stromal cells might yield valuable insights into mechanisms of normal and dysregulated hematopoiesis. Here, proteomics was used to identify expressed proteins in normal and leukemic bone marrow stromal cells. Isobaric tags for relative and absolute quantification (iTRAQ) technology and MS/MS were used to identify and quantify proteins in bone marrow stromal cells from two normal and two leukemic individuals. Approximately 900 distinct proteins were identified with >95% confidence, 73 of which were differentially expressed in leukemic stromal cells. cDNA microarray analysis on Affymetrix oligonucleotide gene chips showed that transcripts correlated with approximately 90% of the expressed proteins. A novel integrated approach for mining and visualization of iTRAQ data is presented. The results provide an initial assessment of the proteome in human bone marrow stromal cells, laying the groundwork for comprehensive analysis of the proteome in these cells, and ultimately, for improved understanding of normal and dysregulated hematopoiesis.

Adult↗

PubMed1999

Evidence against the current hypothesis of "gene dosage effects" of trisomy 21: ets-2, encoded on chromosome 21" is not overexpressed in hearts of patients with Down Syndrome.

BACKGROUND: The major current concept for the pathogenesis of the Down Syndrome (DS) phenotype including congenital heart disease (CHD) is the so-called "gene dosage effect." According to this hypothesis, genes encoded by chromosome 21 at the "critical region" (which is thought to be crucial for the development of the DS phenotype) are overexpressed in the trisomic state, thus leading to an imbalance of genes as, e.g., the protooncogene ets-2, superoxide dismutase, etc. METHODS: We studied heart biopsies obtained at surgery from 6 patients with DS and 7 patients with congenital heart disease. ets-2-mRNA steady state levels were determined by a competitive reverse transcription-polymerase chain reaction (RT-PCR) technique which allowed the determination of this gene at the attomol level. RESULTS: ets-2 mRNA in total ventricular tissue of DS patients showed concentrations of 0.60 +/- 0.42 fg/10 ng total RNA (mean, +/- SD). When normalized versus the housekeeping gene beta-actin to rule out general transcriptional changes in that disorder, the ratio of 0.56 +/- 0.28 (mean, +/- SD) was calculated. ets-2 mRNA in total ventricular tissue of patients with non-DS CHD showed concentrations of 0.45 +/- 0.22 fg/10 ng total RNA (mean, +/-SD) and ratios of 0.48 +/- 0.35 (mean, +/-SD). No differences could be found at the p<0.05 level. CONCLUSION: No absolute quantification of a gene incriminated in the "gene dosage effect-hypothesis" was performed so far and the only approach to (semi-) quantitative determination of the ets-2 gene using northern blotting was published on one individual DS sample only. This is the first report to clearly show that no overexpression of ets-2 can be found in heart of patients with DS, thus providing evidence against the current gene dosage effect-hypothesis.

Chromosome Mapping↗

PubMed1997

SIMS microscopy: methodology, problems and perspectives in mapping drugs and nuclear medicine compounds.

Secondary ion mass spectrometry (SIMS) microscopy, a mass spectrometry method designed in the 1960s, offers new analytical capabilities, high sensitivity (ppm to ppb region), high specificity and improved lateral resolution, thus facilitating insight into many physiological and biomedical questions. Apart from the sample preparation and the physical characteristics of the detection, the biological model must also be considered. SIMS analysis of diffusible ions and molecules requires strict cryogenic procedures which always begin by a flash-freeze fixation. Cellular integrity can be checked by mapping the major element distributions since intra and extracellular ions are redistributed only in damaged cells. Cryofixing may be followed either by a freeze-fracture methodology or by cryoembedding and dry-cutting. Chemical sample preparation is only used for ions or molecules bound to fixed cell structures. The use of scanning procedures ameliorates the lateral resolution and chromosome imaging has been reported with probe size of below 50 nm. Absolute quantification can be derived for embedded specimen by using internal references included in tissue equivalent resins. The sensitivity is limited by the ionization yield of the tag element and may be further impaired when working at high mass resolution (> or = 5000) to eliminate interfering cluster ions. SIMS drug mapping is usually performed after in vitro administration of a molecule to cell culture systems. Drug detection is accomplished indirectly by detecting a tag isotope naturally present or introduced by labelling, mainly with halogens, 15N and 14C. Molecular imaging with TOF-SIMS is an appealing alternative especially for heavier compounds. We stress some biological problems through a critical review of published SIMS drug studies. SIMS proved useful in assessing the targeting specificity of nuclear medicine pharmaceutics, even after in vivo administration. The first microscopic evidence of a thionamide induced follicular blockade of the iodine organification process is presented in a human sample.

Animals↗

PubMed1992

Absolute quantitation of gallium-67 citrate accumulation in the lungs and its importance for the evaluation of disease activity in pulmonary sarcoidosis.

Our modification of a method for the absolute quantification of gallium-67 uptake in lungs with a scintillation camera and computer is described. The uptake of 67Ga in lungs, expressed in percentage of administered radioactivity, was determined by the transmission-emission method. We proved theoretically and experimentally that a 67Ga planar source could be replaced with a 57Co planar source. The performance of lung perfusion scans allows a more accurate delineation of the regions of interest on gallium scans. The method was applied to control subjects (n = 27) and to patients (n = 114) suffering from biopsy-proven pulmonary sarcoidosis (28 with inactive and 86 with active disease). The obtained results were compared with chest X-ray findings, the percentage of lymphocytes in the bronchoalveolar fluid (BAF-ly%), and serum angiotensin-converting enzyme (SACE) values. The method seems suitable for the assessment of disease activity in sarcoidosis. It is more accurate in detecting parenchymal involvement in lung sarcoidosis than the commonly used X-ray criteria. No correlation was found between 67Ga uptake and the BAF-ly% and SACE values.

Bronchoalveolar Lavage Fluid↗

PubMedDate not supplied

Ischaemic ATP degradation studied by HPLC and 31P-NMR spectroscopy: do the two techniques observe the same ATP pools?

31P-NMR spectroscopy has become the major tool for studying myocardial high energy phosphates. Conflicting results concerning NMR visibility of ATP in ischaemic myocardium were reported. A detailed study was undertaken to resolve this controversy. After cardioplegic arrest, canine hearts were excised and preserved for 24 h at 1 degree C (group 1) or for 6h at 23 degrees C (group 2). ATP breakdown was followed by 31P-NMR spectroscopy in a transmural piece of the anterior wall introduced in the NMR magnet, and by HPLC analysis using serial transmural biopsies from the rest of the anterior wall. At both temperatures, identical relative ATP decay curves were obtained, whether measured by NMR or by HPLC. Absolute quantification of ATP was carried out after varying periods of ischaemia at 1 degree C. The NMR-measured ATP concentration was 106 +/- 8% of the ATP concentration determined by HPLC. From our experiments, we conclude that ATP visibility for 31P-NMR spectroscopy is complete and constant during prolonged periods of hypothermic ischaemia in canine hearts.

Adenosine Triphosphate↗

PubMed1996

Radioimagers as an alternative to film autoradiography for in situ quantitative analysis of 125I-ligand receptor binding and pharmacological studies.

Three radioimagers, the mu-imager, the beta-imager and the phosphorimager, were tested as alternatives to quantitative autoradiography on film, for receptor imaging and pharmacological in situ quantitative analysis. Two iodinated ligands 125I-interleukin-1 alpha and 125I-gonadotropin releasing hormone agonist were used for receptor characterization in mouse brain and pituitary sections. Due to the high number of the agonist receptors in rat pituitary gland, this tissue was used to compare measurements obtained from digital autoradiograms with classical gamma detector determination. This permits the evaluation of radioimager efficiency and absolute quantification. Radioimagers represent an improvement in terms of time of image acquisition. All the radioimagers are more sensitive than film for the detection of low levels of radioactivity. The spatial resolution provided by the mu-imager compares favourably with that obtained on film autoradiograms while digital autoradiograms from the phosphorimager and beta-imager did not show precise definition under our experimental conditions. Superimposition of histological structures from the stained sections with radiolabelled areas in the autoradiograms remains, at this time, the unique advantage of film. In conclusion, radioimagers represent an alternative to autoradiography on film or emulsion for in situ quantitative studies on tissue sections. They combine precise imaging for in situ binding studies with easy and direct access to counts in cpm. The improvement in radioimaging technology has, therefore, brought in situ analysis of iodinated ligand binding to the level of accuracy that is obtained with classical detectors of radioactivity.

Animals↗

PubMed2000

Cardiac magnetic resonance spectroscopy: potential clinical applications.

MR spectroscopy is the only method for non-invasive detection of various aspects of cardiac metabolism in humans. While the 1H nucleus of water and fat molecules is the signal source for MR imaging, the MR spectroscopic technique allows for the study of a number of other nuclei, such as 13C, 19F, 23Na, 31P, 39K and 87Rb. Clinical applications presently are confined to the 31P nucleus. 31P-MR spectroscopy allows the non-invasive study of cardiac high-energy phosphate metabolites ATP and phosphocreatine. The phosphocreatine/ATP ratio is considered an index of the energetic state of the heart. Possible clinical indications include heart failure, valve disease and coronary artery disease. In heart failure, the phosphocreatine/ATP ratio is reduced and correlates with clinical severity, ejection fraction and prognosis. In mitral and aortic valve disease, a reduced phosphocreatine/ATP ratio may indicate the optimum timing for valve replacement. In coronary artery disease, a regional decrease of phosphocreatine during stress ("biochemical ergometry") may indicate local ischemia. Furthermore, absolute quantification of high-energy phosphates may allow diagnosis of myocardial viability. Major technical developments, leading to improved spatial and temporal resolution will be necessary to establish MR spectroscopy as a routine clinical tool.

Angioplasty, Balloon, Coronary↗

PubMed2000

[Nuclear medicine diagnosis of the kidneys].

BACKGROUND: Renal studies have a long tradition in nuclear medicine and are available for routine use for more than 30 years. Their high clinical acceptance is mainly based upon the fact that they allow for quantitative evaluation of different functional parameters such as glomerular filtration, effective renal plasma flow, or postrenal transport. The used methods are validated by experimental as well as numerous clinical studies and are performed throughout the world in a highly standardized way. INDICATIONS: Indications are verification/exclusion of a disturbed renal function, detection or evaluation of renal artery stenosis, and differential diagnosis of urinary tract obstruction. Further diagnostic improvement might be achieved by use of positron emission tomography which has the potential for absolute quantification of physiological parameters such as renal blood flow in ml.min-1.100 g-1 tissue.

Glomerular Filtration Rate↗

PubMed2001

Magnetic resonance imaging and 1H-magnetic resonance spectroscopy in amyotrophic lateral sclerosis.

We aimed to increase confidence in the combined use of MRI and proton MR spectroscopy (1H-MRS) in diagnosis of amyotrophic lateral sclerosis (ALS). We investigated 12 patients with ALS, seven definite and five probable, taking into account clinical measures of motor neuron function. On T2-weighted images we found high signal in the corticospinal tract in six and low signal in the primary motor cortex in seven of the 12 patients. Atrophy of the precentral gyrus was apparent in all the patients apart from one with probable ALS. Absolute quantification of cerebral metabolites using 1H-MRS demonstrated a significantly lower mean concentration of N-acetylaspartate (NAA) in the precentral gyrus of patients with probable and definite ALS (8.5 +/- 0.62) than in control subjects (10.4 +/- 0.71; P < 0.001). NAA concentration in primary motor cortex correlated with Norris scale scores (r = 0.30; P < 0.0001) but not with the ALS Functional Rating Scale score or disease duration. Significantly lower levels of NAA were detected in patients with low signal in the motor cortex than in those without (P < 0.01). Mean choline (Cho) and creatine (Cr) values did not differ between patients with ALS and controls.

Aged↗

PubMed2001

Non-uniform versus uniform attenuation correction in brain perfusion SPET of healthy volunteers.

Although non-uniform attenuation correction (NUAC) can supply more accurate absolute quantification, it is not entirely clear whether NUAC provides clear-cut benefits in the routine clinical practice of brain SPET imaging. The aim of this study was to compare the effect of NUAC versus uniform attenuation correction (UAC) on volume of interest (VOI)-based semi-quantification of a large age- and gender-stratified brain perfusion normal database. Eighty-nine healthy volunteers (46 females and 43 males, aged 20-81 years) underwent standardised high-resolution single-photon emission tomography (SPET) with 925 MBq 99mTc-ethyl cysteinate dimer (ECD) on a Toshiba GCA-9300A camera with 153Gd or 99mTc transmission CT scanning. Emission images were reconstructed by filtered back-projection and scatter corrected using the triple-energy window correction method. Both non-uniform Chang attenuation correction (one iteration) and uniform Sorenson correction (attenuation coefficient 0.09 cm(-1)) were applied. Images were automatically reoriented to a stereotactic template on which 35 predefined VOIs were defined for semi-quantification (normalisation on total VOI counts). Small but significant differences between relative VOI uptake values for NUAC versus UAC in the infratentorial region were found. VOI standard deviations were significantly smaller for UAC, 4.5% (range 2.6-7.5), than for NUAC, 5.0% (2.3-9.0) (P<0.01). Higher filter roll-off values of the transmission reconstruction filter increased noise in the emission images and altered estimated cortical attenuation coefficients as well as uptake values. In conclusion, semi-quantification based upon reconstruction with UAC results in very similar 99mTc-ECD uptake values in healthy volunteers to those obtained with NUAC, although values for the infratentorial region are slightly lower. NUAC produces a slight increase in inter-subject variability. Further study is necessary in various patient populations to establish the full clinical impact of NUAC in brain perfusion SPET.

Adult↗

PubMed1999

"Ecstasy"-induced changes of cerebral glucose metabolism and their correlation to acute psychopathology. An 18-FDG PET study.

The aim of this study was to determine the acute effects of the "Ecstasy" analogue MDE (3,4-methylene dioxyethamphetamine) on cerebral glucose metabolism (rMRGlu) of healthy volunteers and to correlate neurometabolism with acute psychopathology. In a randomized double-blind trial, 15 healthy volunteers without a history of drug abuse were examined with fluorine-18-deoxyglucose (18FDG) positron emission tomography (PET) 110-120 min after oral administration of 2 mg/kg MDE (n = 7) or placebo (n = 8). Two minutes prior to radiotracer injection, constant cognitive stimulation was started and maintained for 32 min using a word repetition paradigm to ensure constant and comparable mental conditions during cerebral glucose uptake. Individual brain anatomy was represented using Tl-weighted 3D flash magnetic resonance imaging (MRI), followed by manual regionalization into 108 regions of interest and PET/MRI overlay. After absolute quantification of rMRGlu and normalization to global metabolism, normalized rMRGlu under MDE was compared to placebo using the Mann-Whitney U-test. Acute psychopathology was assessed using the Positive and Negative Syndrome Scale (PANSS) and rMRGlu was correlated to PANSS scores according to Spearman. MDE subjects showed significantly decreased rMRGlu in the bilateral frontal cortex: left frontal posterior (-7.1%, P < 0.05) and right prefrontal superior (-4.6%, P < 0.05). On the other hand, rMRGlu was significantly increased in the bilateral cerebellum (right: +10.1%, P < 0.05; left: +7.6%, P < 0.05) and in the right putamen (+6.2%, P < 0.05). There were positive correlations between rMRGlu in the middle right cingulate and grandiosity (r = 0.87, P < 0.05), both the right amygdala (r = 0.90, P < 0.01) and the left posterior cingulate (r = 0.90, P < 0.01) to difficulties in abstract thinking, and the right frontal inferior (r = 0.85, P < 0.05), right anterior cingulate (r = 0.93, P < 0.01), and left anterior cingulate (r = 0.85, P < 0.05) to attentional deficits. A negative correlation was found between the left frontal operculum (Broca's area) and attentional deficits (r = -0.85, P < 0.05). The present study revealed acute neurometabolic changes under the "Ecstasy" analogue MDE, indicating a frontostriatocerebellar imbalance paralleling other psychotropic substances or various psychiatric disorders.

3,4-Methylenedioxyamphetamine↗