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PubMed1984

New approach to metabolism of 5'-deoxy-5-fluorouridine in humans with fluorine-19 NMR.

The metabolism of 5'-deoxy-5-fluorouridine (5'dFUrd), an antitumor fluoropyrimidine, has been investigated in human biofluids (blood, plasma, urine) using a new method: fluorine-19 NMR spectrometry. This method allows direct study of the biological sample and simultaneous identification of all the fluorinated metabolites. In the blood of a patient treated with 5'dFUrd during a 6-h continuous perfusion, we observed unmetabolized 5'dFUrd, 5-fluorouracil, 5,6-dihydrofluorouracil, and another metabolite which has not previously been reported alpha-fluoro-beta-alanine. The two major metabolites in urine are unmetabolized 5'dFUrd and alpha-fluoro-beta-alanine.

Antineoplastic Agents↗
PubMed2002

A mathematical model of flow in a liquid-filled visco-elastic tube.

In biofluid mechanics the fluid-solid interaction is important. To this aim the propagation of waves in a distensible tube filled with a viscous fluid was studied numerically. Based on the assumption of long wavelength and small amplitude of pressure waves, a quasi-1D differential model was adopted. The model accounted for vessel wall visco-elasticity and included the wall deformations in both radial and axial directions. The non-linear problem was solved in non-dimensional form by a finite difference method on a staggered grid. The boundary conditions were for two relevant cases: natural oscillations in a deformable tube fixed at the ends and persistent oscillations due to a periodical forcing pressure. The natural frequency St* was found to vary as the square root of the elasticity coefficient K, with 0 < or = K < or = 6000, and was not affected by the viscosity. These results highlight a strong influence of both wall visco-elasticity and blood viscosity. The natural oscillations are damped in a few time units and the damping time was found to be inversely proportional to the wall viscosity coefficient and the fluid viscosity provided an even larger damping factor.

Blood Viscosity↗
PubMedDate not supplied

Mesh updating in fluid-structure interactions in biomechanics: an iterative method based on an uncoupled approach.

In this study, a computational uncoupled approach to fluid-structure interaction problems in biofluid mechanics is presented. It is based on the finite element method and is applied to study the local fluid dynamics in two specific situations: the left ventricular ejection phase and the motion of an isolated red blood cell along a small artery. Particularly, the focus is on the algorithms developed to deal with mesh updating, because both examined districts are characterized by geometrical deformations of the fluid domain edges. This is currently a challenging issue in the application of computational fluid dynamics techniques to living systems, especially to the cardiovascular system. Although the chosen approach uses a commercial computational fluid dynamics package for the solution of the fluid domain, original algorithms have been developed to perform the boundary displacement calculations correctly, as well as the corresponding mesh updating. Results are reported and compared with available data in the literature pertinent to the two studied problems.

Algorithms↗
PubMed2006

A path or a new road in laboratory diagnostics? Biological mass spectrometry: facts and perspectives.

Proteins in tissues and biofluids and their many attributes define the proteome. Proteome can be directly correlated to known diseases and histological regions allowing the diagnosis and monitoring of disease progression as well predicting the patient's response to specific treatments. Proteomics performs large-scale, high-throughput characterization of the human proteome, among others by biological mass spectrometry. Proteinchip technology coupled with bioinformatics is able to screen any protein source for putative disease biomarkers from a small sample volume (microliter range) by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). This article discusses on a basic level both the technology and reliability of these methods.

Humans↗
PubMed2006

Genetic and epigenetic biomarkers in cancer : improving diagnosis, risk assessment, and disease stratification.

Gene expression patterns change during the initiation, progression, and development of cancer, as a result of both genetic and epigenetic mechanisms. Genetic changes arise due to irreversible changes in the nucleotide sequence, whereas epigenetic changes occur due to changes in chromatin conformation, histone acetylation, and methylation of the CpG islands located primarily in the promoter region of a gene. Both genetic and epigenetic markers can potentially be utilized to identify different stages of tumor development. Several such markers exhibit high sensitivity and specificity for different tumor types and can be assayed in biofluids and other specimens collected by noninvasive technologies. In spite of the availability of large numbers of diagnostic markers, only a few have been clinically validated so far. The current status and the challenges in the field of genetic and epigenetic markers in cancer diagnosis, risk assessment, and disease stratification are discussed.

Biomarkers, Tumor↗
PubMed1987

Stable isotope dilution analysis of pipecolic acid in cerebrospinal fluid, plasma, urine and amniotic fluid using electron capture negative ion mass fragmentography.

A sensitive and accurate stable isotope dilution assay was developed for the measurement of pipecolic acid in body fluids using electron capture negative ion mass fragmentography. The method utilizes [2H11]pipecolic acid as the internal standard. Sample preparation consisted of derivatization in aqueous solution (pH 11.5) of the amine moiety with methyl chloroformate to the N-methylcarbamate, followed by acidic ethyl acetate extraction (pH 2) and further derivatization of the carboxyl moiety to the pentafluorobenzyl ester. Normal values have been determined in cerebrospinal fluid (mean means = 0.041 mumol/l, range 0.010-0.120 mumol/l), in plasma of at term infants (age less than 1 wk, means = 5.73 mumol/l, range 3.75-10.8 mumol/l; age greater than 1 wk, means = 1.46 mumol/l, range 0.70-2.46 mumol/l), in urine of at term infants (age less than 6 mth, means = 32.5 mumol/g. creat., range 9.81-84.5 mumol/g. creat; age greater than 6 mth, means = 6.35 mumol/g. creat., range 0.15-13.6 mumol/g. creat.) and in amniotic fluid (means = 4.65 mumol/l, range 2.24-8.40 mumol/l). The utility of the method was demonstrated for the pipecolic acid quantification in these biofluids of patients with peroxisomal disorders. As affected fetuses with infantile Refsum's disease and Zellweger syndrome showed no significant elevation of pipecolic acid in their surrounding amniotic fluids, the measurement of pipecolic acid in amniotic fluid seemed not to be useful for prenatal diagnosis in these disorders.

Adult↗
PubMed1983

Velocity measurements in steady flow through axisymmetric stenoses at moderate Reynolds numbers.

The velocity field in the neighborhood of axisymmetric constrictions in rigid tubes was investigated using laser Doppler anemometry and flow visualization. Upstream flow conditions were steady; and Reynolds numbers were in the range 500-2000, values which are representative of the larger arteries in humans. Stenoses of 25, 50 and 75% area reduction were studied. Velocity profiles are presented in sufficient detail to allow comparison with computational biofluid dynamics models. Wall shear stresses were estimated from the near wall velocity gradient, and the nature of observed poststenotic flow disturbances is discussed. Results indicate that flow disturbances of discrete oscillation frequency may be more valuable than turbulence as an indicator of early stages of stenosis development. Additionally, despite the fact that poststenotic turbulence exists for the higher degrees of stenosis and Reynolds numbers, the resulting wall shear stresses are only three to four times greater than the Poiseuille value and are considerably less than the wall shear stress within the stenosis itself.

Aortic Valve Stenosis↗
PubMed1995

Use of high-performance liquid chromatography-diode array detection in forensic toxicology.

A comprehensive approach to the analysis for many drugs in postmortem blood and biological fluids using high-performance liquid chromatography and diode array detection has been developed. To reduce the likelihood of co-eluting interference components of postmortem blood or other drugs, selective back-extraction was also used to screen and quantitate drugs in blood and biofluids. An isocratic mobile phase (acetonitrile, phosphoric acid and triethylamine buffer, pH 3.4) was developed and found stable, reliable and convenient for general drug screening and quantitation. A library of drug spectra in the ultraviolet wavelength range (210-367 nm) was established for 272 drugs on two reversed-phase columns: Supelcosil (biphenyl) and LiChrospher RP-8. The application of several methods to whole blood, the analysis of complex cases and the use of multicomponent analysis for qualitative and quantitative analysis is discussed.

Chromatography, High Pressure Liquid↗
PubMed1994

Improved detection and derivatization in capillary electrophoresis.

Capillary electrophoresis is well known for its low mass detectabilities, but suffers from poor concentration detection limits. This review will discuss improvements in concentration detectability with an emphasis on derivatization methods. Sample concentration techniques and improved detector designs will also be discussed. Pre- and post-capillary derivatization methods for biofluid analytes such as amino acids, peptides, proteins, oligonucleotides, and oligosaccharides will be examined in detail.

Amino Acids↗
PubMed1996

Immunochromatographic analysis of bovine growth hormone releasing factor involving reversed-phase high-performance liquid chromatography-immunodetection.

We have developed high-performance immunoaffinity chromatography (HPIAC) methods for the detection and quantitation of bovine growth hormone releasing factor (GHRF), which could also be applicable to its metabolites in biofluids. These approaches have involved a combination of IAC using immobilized antibody (Ab) to GHRF, together with reversed-phase high-performance liquid chromatography (RP-HPLC) separations of initially isolated and concentrated protein, followed by selective detection, involving on-line immunodetection (ID) schemes. ID methods involved HPIAC supports of the Ab, together with synthesized Ab-fluorescein isothiocyanate conjugates. We have demonstrated optimization methods for each step of the entire hyphenated technique (IAC-HPLC-ID), and then actually quantitated GHRF using this overall system. The minimum detectable concentration was about 1 ng/5 ml (200 ppt) with fluorescence detection (excitation wavelength, 490 nm; emission wavelength, 510-650 nm). We have also tested a single blind, spiked biological sample (bovine plasma), spiked with a known level of GHRF. Accuracy (7.4%) and precision (S.D. = +/- 22%) were quite acceptable for a double immunoassay method.

Animals↗
PubMed1996

Determination of phase II drug metabolites in equine urine by micellar electrokinetic capillary chromatography.

Micellar electrokinetic capillary chromatography (MECC) using diode array detection has been investigated for the determination of phase I and phase II metabolites of drugs in biofluids. Methods were optimised for the determination of morphine, morphine-3-glucuronide, morphine-6-glucuronide, normorphine, meclofenamic acid and its metabolites in equine urine. Solid-phase extraction procedure were developed to concentrate and purify the analytes from spiked and post administration urines for MECC analysis. A simple on-line procedure for monitoring the kinetics of hydrolysis of morphine-glucuronide conjugates by beta-glucuronidase was demonstrated.

Analgesics, Opioid↗
PubMed1993

A new investigation of copper(II)-serine, copper(II)-histidine-serine, copper(II)-asparagine, and copper(II)-histidine-asparagine equilibria under physiological conditions, and implications for simulation models relative to blood plasma.

Some years ago, the application of computer modeling to metal speciation in biofluids was questioned based on the discrepancy between the simulated distribution of copper(II) in blood plasma and related experimental results obtained by Neumann and Sass-Kortsak in reconstituted serum. A recent investigation of the relevant copper(II)-amino acid equilibria reconciled these conflicting data, confirming that the reliability of computer models crucially depends on the data on which they are based. Since then, however, some of the constants of the copper-serine system used in that study have been suspected to be overestimated. This work thus reports the redetermination of copper-serine and copper-histidine-serine formation constants under physiological conditions. In addition, serine being close to asparagine in Neumann and Sass-Kortsak's classification, copper-asparagine and copper-histidine-asparagine equilibria have also been reinvestigated. For asparagine complexes, former constants have been basically confirmed. In contrast, all constants relative to serine have effectively been found lower than the previous ones. The effects of these new data on the stimulated distribution of plasma copper are only minor, but a better agreement is observed relative to Neumann and Sass-Kortsak's models in reconstituted serum.

Asparagine↗
PubMedDate not supplied

Magnetic resonance spectroscopy: a powerful tool for drug metabolism studies.

Studies on the metabolism and disposition of drugs using nuclear magnetic resonance spectroscopy (MRS) as the analytical technique are reviewed. An overview of the main studies classed in terms of the observed magnetic nucleus (1H, 2H, 7Li, 13C, 19F, 31P, 77Se) is followed by some typical examples of the way in which 19F and 31P MRS can be profitably employed to gain more understanding about the metabolism and disposition of the anticancer fluoropyrimidines (5-fluorouracil (FU) and its prodrugs) and ifosfamide (IF). The results of three recent studies carried out in our laboratory are developed. They concern the direct quantitative monitoring of the hepatic metabolism of FU in the isolated perfused mouse liver, the elucidation of the origin of the cardiotoxicity of FU and the metabolism of IF from an analysis of biofluids of patients. Finally, the advantages and limitations of MRS for investigations on drug metabolism are discussed.

Animals↗
PubMed1988

High-performance liquid chromatographic assay for dilevalol in human plasma and urine using a PRP-1 column and fluorimetric detection.

A single high-performance liquid chromatographic (HPLC) assay for the quantitative determination of dilevalol, the R,R isomer of labetalol, was developed for both plasma and urine. A significantly improved limit of detection for dilevalol in plasma was accomplished by extensive modification of an HPLC assay originally developed in our laboratory for labetalol. This simplified method is readily adaptable to urine and represents the first reported HPLC assay for the quantitative determination of dilevalol in this biofluid. Drug was recovered from plasma or urine by partition into diethyl ether under mildly alkaline conditions and back-extraction into dilute acid. Reversed-phase separation of dilevalol and the internal standard was accomplished on a 150 X 4.1 mm column commercially packed with a spherical (5 micron) macroporous copolymer (PRP-1). No interferences were observed in extracts obtained from drug-free plasma or urine. Selectivity for dilevalol in the presence of other beta-blockers was established. This method demonstrated a linear detector response to concentrations of unchanged drug typically observed in urine and plasma following once-a-day treatment with dilevalol hydrochloride (100-800 mg). The lowest limit of reliable quantitation was established at 1 ng/ml in plasma. The intra-assay precision (coefficient of variation) remained less than 6% at all concentrations evaluated from 1 to 800 ng/ml. In urine, the lowest limit of quantitation was validated to 20 ng/ml where the intra-assay precision (coefficient of variation) for unchanged drug was less than 4% at all concentrations evaluated up to 400 ng/ml. This method is suitable for routine quantitation of unchanged drug in human plasma and urine following the administration of therapeutically effective doses of dilevalol hydrochloride.

Adrenergic beta-Antagonists↗
PubMed1993

Determination of alclofenac in equine plasma and urine by high-performance liquid chromatography.

A high-performance liquid chromatographic method to measure plasma and urinary alclofenac levels in equine biofluids is described. Isolation of the drug from plasma is achieved using liquid-liquid extraction with diethyl ether. Reversed-phase C18 solid phase extraction is used for the extraction of free and conjugated alclofenac from urine. The reproducibility and accuracy of the method were well within acceptable limits over the concentration ranges 0-10 and 0-20 micrograms/ml, respectively, for plasma and urine. Starting with 2 ml of plasma, a concentration of 0.1 microgram/ml could easily be measured; the limit of quantification in urine (0.5 ml) was 1 microgram/ml. Hydrolysis of urine with strong alkali resulted in the decomposition of alclofenac. A pharmacokinetic profile of alclofenac in the horse is shown.

Animals↗
PubMed1995

Assay of the enantiomers of ibutilide and artilide using solid-phase extraction, derivatization, and achiral-chiral column-switching high-performance liquid chromatography.

Ibutilide fumarate and artilide fumarate, new drugs for the treatment of cardiac arrhythmias, each contain a stereogenic center bearing a secondary alcohol group. Reversed-phase achiral-chiral column-switching HPLC separations of the enantiomers of each compound were developed and validated for quantitation in plasma and other biofluids. The key component of the method was derivatization with 1-naphthyl isocyanate, which enhanced the sensitivity of fluorescence detection and enabled the enantiomers to be separated on a Pirkle column (covalent 3,5-dinitrobenzoyl-D-phenylglycine stationary phase). The lower limit of quantitation of ibutilide fumarate was typically 0.1 ng/ml or less of each enantiomer in 1 ml of plasma. Two of the special features of the column-switching system included operation with two samples in the system at one time, which reduced analysis time to 16 min/sample for ibutilide and 19 min/sample for artilide, and a relay-operated switching of detector outputs, which allowed achiral and chiral column chromatographic data to be gathered from two detectors into a single data acquisition channel.

Anti-Arrhythmia Agents↗
PubMed1993

Determination of peptide 520 in human plasma using post-column photolysis with electrochemical detection in liquid chromatography.

A simple LC method for the determination of peptide 520 in human plasma was developed. Based on micellar chromatography, sodium octyl sulphate (SOS) was added into the mobile phase in order to separate the peptide from human plasma components. The procedure was fast and sensitive for the determination of the peptide in untreated human plasma. The electrochemical (EC) detection limit for peptide 520 in human plasma was 0.5 microgram ml-1. Linearity of the calibration plot for peptide 520 in human plasma was 0.999. This approach represents a direct injection technique for the potential detection and analysis of numerous peptides in biofluids, besides just plasma, with absolute quantification.

Chromatography, Liquid↗