SEARCH · Search PubMed

Results for “phasing”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

SourceResearch connectionFrom yearThrough year

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,261 records · Page 70Linked to original sources

PubMed2005

Quantitative determination of pharmaceuticals using nano-electrospray ionization mass spectrometry after reversed phase mini-solid phase extraction.

The pre-concentration effect of solid phase microextraction (SPE) with nano-electrospray ionization mass spectrometry (nano-ESI MS) for selected pharmaceuticals is presented. An analytical method is developed for the quantitative determination of dicyclomine in serum with cyclopentolate as the internal standard by off-line nano-ESI ion-trap MS with reversed phase mini-SPE. Homemade C18 and C4 mini-SPE cartridges of 0.5 and 1cm in length and 1.55 mm i.d. have been tested for pre-concentration of samples originally 30 microL in volume. After SPE, the volume of the sample in methanol is about 1-2 microL and 0.5 microL can be injected into the nano-ESI MS instrument. Use of a 1cm C18 cartridge lowered the detection limit of dicyclomine 100 times to 16.1 fmole. Dicyclomine spiked in serum can be determined by nano-ESI MS after protein precipitation and further clean-up on the 1cm C18 cartridge. However, the slope of a calibration curve of dicyclomine standards spiked in serum is more than a factor of 10 less than that for a calibration curve of dicyclomine standards prepared in water indicating pre-concentration of pharmaceuticals could be compromised by a complex biological matrix.

Animals↗

PubMed2006

p38 mitogen-activated protein kinase mediates the sustained phase of hypoxic pulmonary vasoconstriction and plays a role in phase I vasodilation.

Hypoxic pulmonary vasoconstriction (HPV) and pulmonary hypertension present common and formidable clinical problems for thoracic, transplant, and trauma surgeons. We hypothesized that acute hypoxia causes pulmonary artery (PA) contraction and that p38 mitogen-activated protein (MAP) kinase is a key mediator in that process. To test this hypothesis, we measured isometric force displacement in isolated rat pulmonary artery rings during hypoxia in the presence and absence of the selective p38 MAP kinase inhibitor SB-20358, and stimulator anisomycin. In separate experiments, we measured the functional effects in isolated rat pulmonary artery rings of inhibiting p38 MAP kinase during normoxic conditions. p38 MAP kinase inhibition significantly attenuated the delayed, but not early, contractile phase of HPV. Additionally, stimulation of p38 MAP kinase significantly decreased the phase I vasodilation of HPV. Under normoxia conditions, there was no statistically significant difference in isometric force displacement between control and p38 MAPK inhibitor-treated pulmonary artery rings. We conclude that p38 MAP kinase may be a key mediator in the pathogenesis of HPV and that further understanding may lead to new therapies for HPV associated with acute lung injury.

Animals↗

PubMed2005

The effect of long-term depuration on phase I and phase II biotransformation in mullets (Mugil cephalus) chronically exposed to pollutants in River Douro Estuary, Portugal.

Pollutants such polycyclic aromatic hydrocarbons (PAHs) are released into the environment by urban communities and industries and the enzymes that catalyse the biotransformation of pollutants play a key role regarding the accumulation of these compounds in fish species inhabiting these areas. In this study the relationship between phase I (EROD activity) and phase II (GST activity) and PAH metabolites was measured in grey mullet (Mugil cephalus) after capture in the Douro estuary, and after long-term depuration in an unpolluted laboratory environment. The results showed a significant decrease in EROD activity after 1 month and in bile metabolites after 4 months in captivity, with both maintaining reduced levels at 4 and 8 months depuration. Liver GST activity did not showed significant changes. This study provides evidence that Douro estuary waters contain bioavailable PAHs that can be associated with the induction of cytochrome P450, and that mullets have the ability to metabolise and eliminate PAHs.

Animals↗

PubMed2006

Is P-glycoprotein (ABCB1) a phase 0 or a phase 3 colchicine transporter depending on colchicine exposure conditions?

This study investigates the P-glycoprotein (Pgp)-mediated transport of its substrates in accumulation or efflux modes under steady-state conditions. The kinetics of colchicine uptake and efflux, a substrate of both Pgp and intracellular tubulin, were studied in HL60 and HL60/DNR cells; HL60/DNR cells contain 25 times more Pgp than do HL60 cells. HL60/DNR cells in a medium containing 6.25 nM colchicine, which mimics therapeutic conditions, reached steady-state twice as rapidly as did HL60 cells, and accumulated 24-times less colchicine than did HL60 cells. The Pgp inhibitor GF120918, increased colchicine uptake by HL60 cells 1.2-fold and that of HL60/DNR cells 17-fold, while it had no effect on colchicine efflux from either cell line that had been incubated with colchicine for 24 h. Colchicine kinetics fitted well a two closed-compartment model, showing that the low intracellular accumulation of colchicine in HL60/DNR cells resulted from a 11-fold decrease in colchicine uptake and a 2.3-fold increase in colchicine efflux, that could be attributed to Pgp-mediated efflux activity in HL60/DNR cells. Intracellular colchicine was mainly and similarly distributed in the cytosol in both cell lines. These data demonstrate that the kinetics of the intracellular colchicine accumulation depend on the density of Pgp and that Pgp is more a phase 0 (preventing cellular uptake) than a phase 3 (effluxing intracellular substrate) transporter under steady-state conditions, although the situation is reversed after a short incubation time (30 min), when intracellular free colchicine concentration is probably high enough for it to be removed from the cell by Pgp.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1999

Exercise-induced myocardial ischemia triggers the early phase of preconditioning but not the late phase.

The aim of the present study was to establish whether exercise-induced ischemia triggers the second window of protection in 15 patients with coronary artery disease undergoing 2 consecutive treadmill exercise tests and a third test 24 hours later. Our findings confirm that a first exercise-induced ischemic challenge induces the early phase of preconditioning but not the late phase, thus suggesting that either a late protective effect of preconditioning does not exist in the setting of demand ischemia or, if it exists, it must be weaker than the early protective effect.

Aged↗

PubMed1992

Frequency domain impedance measurements of erythrocytes. Constant phase angle impedance characteristics and a phase transition.

We report measurements of the electrical impedance of human erythrocytes in the frequency range from 1 Hz to 10 MHz, and for temperatures from 4 to 40 degrees C. In order to achieve high sensitivity in this frequency range, we embedded the cells in the pores of a filter, which constrains the current to pass through the cells in the pores. Based on the geometry of the cells embedded in the filter a circuit model is proposed for the cell-filter saline system. A constant phase angle (CPA) element, i.e., an impedance of the form Z = A/(j omega)alpha, where A is a constant, j = square root of -1, omega is angular frequency, and 0 less than alpha less than 1 has been used to describe the ac response of the interface between the cell surface and the electrolyte solution, i.e., the electrical double layer. The CPA and other elements of the circuit model are determined by a complex nonlinear least squares (CNLS) fit, which simultaneously fits the real and imaginary parts of the experimental data to the circuit model. The specific membrane capacitance is determined to be 0.901 +/- 0.036 microF/cm2, and the specific cytoplasm conductivity to be 0.413 +/- 0.031 S/m at 26 degrees C. The temperature dependence of the cytoplasm conductivity, membrane capacitance, and CPA element has been obtained. The membrane capacitance increases markedly at approximately 37 degrees C, which suggests a phase transition in the cell membrane.

Biophysical Phenomena↗

PubMed1995

In vitro study of phase resetting and phase locking in a time-comparison circuit in the electric fish, Eigenmannia.

The electric fish Eigenmannia generates on oscillating weak electric field. The amplitude and timing information of this electric field is perceived by electroreceptors distributed on its skin. The pathway of timing information, consisting of spherical cells and giant cells, was studied in an in vitro preparation. The giant cells were identified to be endogenous oscillators and thus have the functional advantage of phase locking more easily to a periodic stimulus with a frequency in the range of the intrinsic frequency. Their spontaneous rhythmic activity was perturbed by delivering excitatory single pulses or periodic pulses via their synaptic inputs. The regular and irregular dynamics produced by periodic stimulation were discussed in the context of a mathematical analysis of the response to single pulses. Ambiguous representations of the timing of the stimulus pulse were observed and could be related to this analysis. Some spontaneously firing cells could be silenced with periodic excitatory stimulation in a narrow frequency and amplitude range. Some irregularly firing cells continued to fire periodically for several seconds after phase locking to a periodic stimulus. This study is the first description of an endogenous oscillator in a system devoted to the precise timing of sensory events.

Animals↗

PubMed1999

Characterization of Phase I and Phase II hepatic drug metabolism activities in a panel of human liver preparations.

The role of drug metabolism in drug discovery (lead compound selection) and the traditional role of identifying the enzymes involved in biotransformation pathways (reaction phenotyping) have both relied heavily on the availability and use of a human liver bank. The assessment of drug metabolizing enzyme activity and variability in a series of individual human livers is essential when characterizing the enzymes involved in metabolic pathways (i.e. correlation analysis). In this regard, a human liver bank of 21 samples (14 males, six females, and one unknown) was characterized with respect to the activity of several important drug metabolizing enzymes. The total CYP450 content of the livers ranged from 0.06 to 0.46 nmol/mg microsomal protein. The fold variations found in specific enzyme contents were as follows: CYP1A2 (3x), CYP2A6 (21x), CYP2C9 (8x), CYP2C19 (175x), CYP2D6 (18x), CYP2E1 (5x), CYP3A4 (18x), FMO (2.5x), UDPGT (4x), NAT (7x), COMT (5x), ST (5x), TPMT (3x), and GST (2.5x). In general, the fold variation of the Phase II enzymes was lower compared with the Phase I enzymes, with the exceptions of CYP1A2, CYP2E1, and FMO. Similar data were reviewed from other established liver banks and compared with regard to the relative variability observed in drug metabolizing capacities found in this study.

Arylamine N-Acetyltransferase↗

PubMed2000

Solid-phase extraction applied to the determination of ochratoxin A in wines by reversed-phase high-performance liquid chromatography.

A reversed-phase high-performance liquid chromatographic method is described for the analysis of Ochratoxin A at low microg l(-1) levels in samples of artificially contaminated wines. The method involves solid-phase extraction of samples using octadecylsilane cartridges and an additional preconcentration step prior to chromatography with isocratic elution and fluorimetric detection. The method was evaluated for accuracy and precision with relative standard deviations lower than 10%. Recoveries of ochratoxin A added to commercial wines over the range 0.1-3.0 microg l(-1) were higher than 80% in the assays. The performance of the octadecylsilane cartridge method tested compared very favourably with results of other published studies of ochratoxin A which use immunoaffinity columns or solvent extraction techniques.

Chromatography, High Pressure Liquid↗

PubMed1979

Combined reversed-phase and normal-phase high-performance liquid chromatography in the purification and identification of 7,12-dimethylbenz[a]anthracene metabolites.

A total of 37 compounds have been identified as rat liver microsomal metabolites of the potent carcinogen 7,12-dimethylbenz[a]anthracene and its hydroxymethyl derivatives 7-methyl-12-hydroxymethylbenz[a]anthracene, 7-hydroxymethyl-12-methylben[a]anthracene and 7,12-dihydroxymethylbenz[a]anthracene. The metabolites were characterized by: (i) retention times on reversed-phase (with a C18 column) and normal-phase (with a silica gel column) high-performance liquid chromatography (HPLC); (ii) ultraviolet absorption and fluorescence spectra; (iii) mass spectral analysis; (iv) optical activity; and (v) comparison of the physicochemical properties of the metabolites with those of some available synthetic standards. The 37 identified metabolites include four trans-3,4-dihydrodiols, four trans-5,6-dihydrodiols, four trans-8,9-dihydrodiols, four trans-10,11-dihydrodiols, two methyl carboxylic acids, two methyl aldehydes, two hydroxymethyl aldehydes, four 2-phenols, four 3-phenols, four 4-phenols and three hydroxymethyl derivatives. The trans configuration of the dihydrodiols was determined by their inability to form vicinal cisacetonides. Seven dihydrodiol metabolites were found to be optically active. Detailed physicochemical properties, such as ultraviolet absorption spectra, fluorescence spectra measured in methanol and in 0.1 N NaOH, major mass ions from mass spectral analysis and the retention times on two HPLC systems, are presented in support of the structural assignments.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1988

Polar contributions of the stationary phase to the reversed-phase ion-pair high-performance liquid chromatographic separation of quaternary ammonium drugs.

The chromatographic reproducibility of a methodology, developed for the separation and determination of quaternary ammonium drugs by reversed-phase ion-pair column liquid chromatography, was studied. The results in terms of retention dependence on the residual silanol content of the octadecyl stationary phase and column aging were compared with those obtained with conventional separation techniques. By on-column silylation with N-trimethylsilylimidazole, it was demonstrated that eluents containing both amines and alkanesulphonates, beside a higher resolving power, provide reproducible separations which are far less dependent on the residual and generated silanol groups compared to those obtained with eluents containing only an organic amine.

Chromatography, High Pressure Liquid↗

PubMed1988

Separation of proteins by reversed-phase high-performance liquid chromatography. II. Optimizing sample pretreatment and mobile phase conditions.

The effects of separation variables such as temperature, pH and composition of the mobile phase (including additives such as chaotropes, ion-pairing agents and surfactants), sample size and sample pretreatment for reversed-phase high-performance liquid chromatography (RP-HPLC) of proteins is examined. Experimental optimization of these parameters using the preferred instrumental and column conditions described previously lead to well behaved chromatographic performance for most proteins. This allowed us to achieve the required level of performance for the first dimension (RP-HPLC) separation of most protein samples by the chromatophoresis process.

Bacterial Proteins↗

PubMed1987

Partitioning behaviour of rat bone marrow cells in aqueous two-phase systems. Dependence of cell partition on the interfacial tension and electrical potential difference between the phases.

The partitioning of heterogeneous cell populations of rat bone marrow, in comparison with that of homogeneous populations of erythrocytes, has been studied in aqueous two-phase systems containing increasing concentrations of dextran-poly-(ethylene glycol) (to increase the interfacial tension) and/or decreasing phosphate concentrations (to decrease both the electrical potential difference between the phases and the interfacial tension). At any polymer and/or phosphate concentration tested the partition is lower for bone marrow cells, reflecting their lower surface charge and hydrophobicity.

Animals↗

PubMed2001

Characterisation of reversed-phase liquid chromatography stationary phases for the analysis of basic pharmaceuticals: eluent properties and comparison of empirical test methods.

The reversed-phase liquid chromatographic analysis of basic pharmaceuticals can be problematic. Both the properties of the eluent and the stationary phase can influence the chromatographic performance. Therefore selection of suitable experimental conditions for the analysis of basic compounds can be difficult. This paper shows that the organic modifier and the nature of the buffer influence the eluent properties. Moreover, the nature and amount of modifier also influence the basicity of the analytes. Investigations showed that the nature of the buffer can have a significant influence on retention and peak shape of basic compounds. Test procedures using basic analytes as test probes provided relevant information with respect to selecting columns to analyse basic pharmaceutical compounds. Test procedures using compounds like aniline, phenol and benzene were found to be less suitable.

Chromatography, High Pressure Liquid↗

PubMed1986

Reversed-phase determination of famotidine, potential degradates, and preservatives in pharmaceutical formulations by high-performance liquid chromatography using silica as a stationary phase.

An analytical method was developed for the determination of famotidine, potential degradates, and preservatives in several pharmaceutical formulations. The method employs reversed-phase chromatography on a silica column with a methanol-phosphate solution as the mobile phase. The influence of the concentration of phosphate and organic modifier are discussed. Accuracy and precision for this method along with assay data from different formulations of famotidine are presented. Two different commercial silica columns were tested with this method. Chromatographic differences related to the surface area and pore size of the silica are discussed.

Chemistry, Pharmaceutical↗

PubMed1990

Characterization of reversed-phase high-performance liquid chromatographic stationary phases using ribonuclease A.

The protein ribonuclease A (RNase A) represents a good model protein for studying reversible conformational refolding during gradient elution. Work is described utilizing RNase A under gradient conditions to evaluate several different reversed-phase materials. Columns (10 cm x 4.6 mm I.D.) were packed with Partisil C18, Vydac C18, Nucleosil C4, Nucleosil C18 and an adamantyl-modified Partisil silica. Measurements of the apparent first-order rate constant of refolding, as a function of temperature, are presented and compared for each stationary phase. Comparisons of peak shapes as functions of flow-rate and temperature are also discussed.

Chromatography, High Pressure Liquid↗

PubMed1987

Separation of basic drug substances by reversed-phase high-performance liquid chromatography on dynamically modified silica and on bonded-phase materials.

The chromatographic behaviour in the reversed-phase mode of various drug substances, which are nitrogen-containing bases, has been investigated. Three different types of column materials were used, ordinary chemically bonded ODS-silica, deactivated ODS-silica specifically designed for the separation of bases and bare silica dynamically modified by adding long-chain quaternary ammonium compounds to the eluent. Various brands were tested for all three types of column materials. When using ODS-silica as the column material, considerable tailing of peaks and prolonged retention was observed. Also the deactivated columns exhibited tailing problems, but less pronounced. For both types of bonded-phase material it was possible to reduce the tailing considerably by the addition of an anti-tailing agent to the eluent. On bare silica used in the dynamic modification mode the tailing was almost negligible.

Chromatography, High Pressure Liquid↗

PubMed1991

Use of polymeric reversed-phase columns for the characterization of polypeptides extracted from human pancreata. I. Effect of the mobile phase.

The high-performance liquid chromatographic (HPLC) behaviour of two different styrene-divinyl-benzene-based reversed-phase (RP) columns was evaluated using crude acetic acid extracts from normal and diabetic human pancreata as samples. Acetic acid gradients in water and acetonitrile gradients in triethylammonium phosphate (TEAP) and trifluoroacetic acid (TFA) were used as mobile phases, and comparisons were made with a silica-based C4 column. When two different polymeric RP columns were eluted with acetic acid gradients in water, surprisingly similar HPLC profiles of the pancreatic extracts were obtained. Elution of the polymer-based columns with acetonitrile gradients in TFA or TEAP resulted in changes in the polypeptide selectivity of these columns, in parallel with that of a silica-based C4 column eluted under similar conditions, indicating the general usability of polymeric columns for RP-HPLC of peptides and proteins. The pronounced difference in composition between normal and diabetic samples, which also was demonstrated after size-exclusion chromatography (SEC) on a silica-based and an agarose-based high-performance SEC column, was found to be related to the different ischaemia times for the two types of pancreata.

Acetates↗