Biomedical subjects

A D Jones

Publications and source records attributed to A D Jones.

At least 19 recordsLinked to original sources

PubMed2005

Thoracic size-selective sampling of fibres: performance of four types of thoracic sampler in laboratory tests.

The counting of fibres on membrane filters could be facilitated by using size-selective samplers to exclude coarse particulate and fibres that impede fibre counting. Furthermore, the use of thoracic size selection would also remove the present requirement to discriminate fibres by diameter during counting. However, before thoracic samplers become acceptable for sampling fibres, their performance with fibres needs to be determined. This study examines the performance of four thoracic samplers: the GK2.69 cyclone, a Modified SIMPEDS cyclone, the CATHIA sampler (inertial separation) and the IOM thoracic sampler (porous foam pre-selector). The uniformity of sample deposit on the filter samples, which is important when counts are taken on random fields, was examined with two sizes of spherical particles (1 and 10 microm) and a glass fibre aerosol with fibres spanning the aerodynamic size range of the thoracic convention. Counts by optical microscopy examined fields on a set scanning pattern. Hotspots of deposition were detected for one of the thoracic samplers (Modified SIMPEDS with the 10 microm particles and the fibres). These hotspots were attributed to the inertial flow pattern near the port from the cyclone pre-separator. For the other three thoracic samplers, the distribution was similar to that on a cowled sampler, the current standard sampler for fibres. Aerodynamic selection was examined by comparing fibre concentration on thoracic samples with those measured on semi-isokinetic samples, using fibre size (and hence calculated aerodynamic diameter) and number data obtained by scanning electron microscope evaluation in four laboratories. The size-selection characteristics of three thoracic samplers (GK2.69, Modified SIMPEDS and CATHIA) appeared very similar to the thoracic convention; there was a slight oversampling (relative to the convention) for d(ae) < 7 microm, but that would not be disadvantageous for comparability with the cowled sampler. Only the IOM thoracic sampler tended to undersample the fibres relative to the thoracic convention. With the data divided into four classes based on fibre length, the size-selection characteristics appeared to be unaffected by fibre length for GK2.69, Modified SIMPEDS and CATHIA. Only the IOM thoracic sampler (with the foam selector) showed slightly lower selection for longer length classes of fibres. These results indicate that the tested samplers follow the thoracic sampling convention for fibres, and may be used to improve the quality and reliability of samples that are taken when there is likely to be significant background dust.

Air Pollutants↗

PubMed2005

A comparison of fibre counting across three European national proficiency testing schemes.

Proficiency testing (PT) schemes seek to improve and maintain consistency in asbestos fibre counting by circulating mounted samples to laboratories, which return counts for comparison with consensus reference values. This study compares the level of these consensus values in three (Spanish, Belgian and UK) national PT schemes. It also assesses the effect of the imminent change in the European standard rules for counting asbestos fibres, to the new rules from the World Health Organisation (WHO), which will include more fibres. Forty samples from the three schemes were circulated to six laboratories, two in each national scheme. The UK and Spanish laboratories counted using both the ERM and the WHO method. Belgian laboratories already use the WHO method, and they counted only by this method. Densities from counts in this sample exchange served as a common basis against which we compared the national reference values (R). This produced a geometric overall mean ratio of R to exchange mean density of 0.94 for the UK scheme, 1.01 for the Spanish and 0.97 for the Belgian scheme, and thus indicated remarkably similar levels. Nevertheless, non-trivial systematic inter-laboratory differences confirmed the need for regular PT and international inter-laboratory comparisons. Examination of previous data from the national PT schemes and from an international scheme (AFRICA) provided further comparisons for the same laboratories (either comparing the laboratory's data with R values or making direct comparisons between laboratories) that are consistent with those from this special exchange. The change in fibre counting rule (to include fibres apparently touching particles with diameter >3 microm) produced mean increases in reported densities that ranged among samples from 0 to 70% (highest individual estimate 170%); the effect was broadly similar for the three schemes. The laboratories gave mostly similar estimates of percentage increase; however, one laboratory occasionally produced high estimates indicating a possible need for training for using the new method.

Air Pollutants, Occupational↗

PubMed2003

Modeling the retention and clearance of manmade vitreous fibers in the rat lung.

A mathematical model describing the dissolution and disintegration of long fibers and the clearance of short fibers is developed. For short fiber clearance, the model is based on previous modeling of the retention and clearance of particles, and most model parameters are taken from that particulate model. In addition to modeling the disappearance of long fibers, the present study includes a quantitative measure of goodness of fit of the model to observed data. Data from chronic inhalation experiments with insulation glass wools (MMVF10 and MMVF11) and rockwool (MMVF21) were provided for this study. These data comprised lung burdens at 10 time points at each of 3 concentrations for each fiber in inhalation experiments lasting up to 104 wk. At the two higher concentrations, the model had to take into account the effects of lung burden on macrophage-mediated clearance. The modeling shows that the overload dependence appears remarkably similar to that for low-toxicity particles in that the critical volumetric lung burden is similar to that for low toxicity dust. The model describes overload as leading to alveolar sequestration of short fibers or particles, and the estimated rate of alveolar sequestration for MMVF10 was similar to that for particles, but the estimated rate was lower for the other two fibers. Two alternative hypotheses to describe the process of the disappearance of longer fibers were tested by assessing their effect on a quantitative measure of fit of model predictions to the lung-burden data. These tests indicated that (a) dissolution leading to disintegration of long fibers into shorter fibers gave a much better fit than the alternative assumption that dissolution would leave only nonfibrous residue and (b) the relative rates of disintegration of the fibers in the lung appear to be directly dependent on their rates of in vitro dissolution and their diameters.

Administration, Inhalation↗

PubMed2001

Building a replisome solution structure by elucidation of protein-protein interactions in the bacteriophage T4 DNA polymerase holoenzyme.

Assembly of DNA replication systems requires the coordinated actions of many proteins. The multiprotein complexes formed as intermediates on the pathway to the final DNA polymerase holoenzyme have been shown to have distinct structures relative to the ground-state structures of the individual proteins. By using a variety of solution-phase techniques, we have elucidated additional information about the solution structure of the bacteriophage T4 holoenzyme. Photocross-linking and mass spectrometry were used to demonstrate interactions between I107C of the sliding clamp and the DNA polymerase. Fluorescence resonance energy transfer, analytical ultracentrifugation, and isothermal titration calorimetry measurements were used to demonstrate that the C terminus of the DNA polymerase can interact at two distinct locations on the sliding clamp. Both of these binding modes may be used during holoenzyme assembly, but only one of these binding modes is found in the final holoenzyme. Present and previous solution interaction data were used to build a model of the holoenzyme that is consistent with these data.

Biotin↗

PubMed2001

Recruitment of a foreign quinone into the A1 site of photosystem I. In vivo replacement of plastoquinone-9 by media-supplemented naphthoquinones in phylloquinone biosynthetic pathway mutants of Synechocystis sp. PCC 6803.

Interruption of the phylloquinone (PhQ) biosynthetic pathway by interposon mutagenesis of the menA and menB genes in Synechocystis sp. PCC 6803 results in plastoquinone-9 (PQ-9) occupying the A(1) site and functioning in electron transfer from A(0) to the FeS clusters in photosystem (PS) I (Johnson, T. W., Shen, G., Zybailov, B., Kolling, D., Reategui, R., Beauparlant, S., Vassiliev, I. R., Bryant, D. A., Jones, A. D., Golbeck, J. H., and Chitnis, P. R. (2000) J. Biol. Chem. 275, 8523-8530. We report here the isolation of menB26, a strain of the menB mutant that grows in high light by virtue of a higher PS I to PS II ratio. PhQ can be reincorporated into the A(1) site of the menB26 mutant strain by supplementing the growth medium with authentic PhQ. The reincorporation of PhQ also occurs in cells that have been treated with protein synthesis inhibitors, consistent with a displacement of PQ-9 from the A(1) site by mass action. The doubling time of the menB26 mutant cells, but not the menA mutant cells, approaches the wild type when the growth medium is supplemented with naphthoquinone (NQ) derivatives such as 2-CO(2)H-1,4-NQ and 2-CH(3)-1,4-NQ. Since PhQ replaces PQ-9 in the supplemented menB26 mutant cells, but not in the menA mutant cells, the phytyl tail accompanies the incorporation of these quinones into the A(1) site. Studies with menB26 mutant cells and perdeuterated 2-CH(3)-1,4-NQ shows that phytylation occurs at position 3 of the NQ ring because the deuterated 2-methyl group remains intact. Therefore, the specificity of the phytyltransferase enzyme is selective with respect to the group present at ring positions 2 and 3. Supplementing the growth medium of menB26 mutant cells with 1,4-NQ also leads to its incorporation into the A(1) site, but typically without either the phytyl tail or the methyl group. These findings open the possibility of biologically incorporating novel quinones into the A(1) site by supplementing the growth medium of menB26 mutant cells.

Alkyl and Aryl Transferases↗

PubMed2001

Oxidation of a tetrameric nonphenolic lignin model compound by lignin peroxidase.

The present study maps the active site of lignin peroxidase in respect to substrate size using either fungal or recombinant wild type, as well as mutated, recombinant lignin peroxidases. A nonphenolic tetrameric lignin model was synthesized that contains beta-O-4 linkages. The fungal and recombinant wild type lignin peroxidase both oxidized the tetrameric model forming four products. The four products were identified by mass spectral analyses and compared with synthetic standards. They were identified as tetrameric, trimeric, dimeric, and monomeric carbonyl compounds. All four of these products were also formed from single turnover experiments. This indicates that lignin peroxidase is able to attack any of the C(alpha)-C(beta) linkages in the tetrameric compound and that the substrate-binding site is well exposed. Mutation of the recombinant lignin peroxidase (isozyme H8) in the heme access channel, which is relatively restricted and was previously proposed to be the veratryl alcohol-binding site (E146S), had little effect on the oxidation of the tetramer. In contrast, mutation of a Trp residue (W171S) in the alternate proposed substrate-binding site completely inhibited the oxidation of the tetrameric model. These results are consistent with lignin peroxidase having an exposed active site capable of directly interacting with the lignin polymer without the advent of low molecular weight mediators.

Base Sequence↗

PubMed2001

Desorption-ionization mass spectrometry using deposited nanostructured silicon films.

We present a method for desorption ionization on silicon based on novel column/void-network-deposited silicon thin films. A number of different peptides and proteins in the < or = 6000 Daltons range are analyzed by time-of-flight mass spectrometry in this demonstration of our approach. A variety of sample preparation conditions, including the use of chemical additives, surface treatments, and sample purification are used to show the potential of mass analysis using deposited column/void-network silicon films for high throughput proteomic screening.

Citric Acid↗

PubMed2001

Structural requirements for the biosynthesis of backbone cyclic peptide libraries.

BACKGROUND: Combinatorial methods for the production of molecular libraries are an important source of ligand diversity for chemical biology. Synthetic methods focus on the production of small molecules that must traverse the cell membrane to elicit a response. Genetic methods enable intracellular ligand production, but products must typically be large molecules in order to withstand cellular catabolism. Here we describe an intein-based approach to biosynthesis of backbone cyclic peptide libraries that combines the strengths of synthetic and genetic methods. RESULTS: Through site-directed mutagenesis we show that the DnaE intein from Synechocystis sp. PCC6803 is very promiscuous with respect to peptide substrate composition, and can generate cyclic products ranging from four to nine amino acids. Libraries with five variable amino acids and either one or four fixed residues were prepared, yielding between 10(7) and 10(8) transformants. The majority of randomly selected clones from each library gave cyclic products. CONCLUSIONS: We have developed a versatile method for producing intracellular libraries of small, stable cyclic peptides. Genetic encoding enables facile manipulation of vast numbers of compounds, while low molecular weight ensures ready pharmacophore identification. The demonstrated flexibility of the method towards both peptide length and composition makes it a valuable addition to existing methods for generating ligand diversity.

Amino Acid Sequence↗

PubMed2001

Mechanism of pressure-induced gelation of milk.

The pressure-induced gelation of concentrated skimmed milk and milk-sugar mixtures was studied to discover the main components responsible for gelation. The major protein component responsible for gelation is micellar casein. Gelation occurs at similar pressures to casein micelle disintegration in dilute milk, and both can be prevented by inclusion of excess calcium chloride. Transmission electron micrographs show that the protein network is formed from particles with diameters approximately an order of magnitude smaller than those of intact casein micelles. Gelation occurs on decompression and is found to be baroreversible. Concentrations of sugar up to 30% reduce the critical concentration of casein required for gelation, but higher sugar concentrations inhibit gelation. A mechanism of gelation based on the aggregation of casein submicelles formed by pressure-induced disintegration of casein micelles is proposed. Observations on the effect of sucrose on gelation are discussed in terms of the influence of sugars on the solvent quality in aqueous casein systems.

Caseins↗

PubMed2001

Changes in myosin structure and function in response to glycation.

Nonenzymatic glycosylation (glycation) is recognized as an important post-translational modification underlying alterations of structure and function of extracellular proteins. The effect of glycation on intracellular proteins is, on the other hand, less well known despite the vital importance of intracellular proteins for cell, tissue, and organ function. The aim of this study was to explore the effects of glycation on the structure and function of skeletal muscle myosin. Myosin was incubated for up to 30 min with glucose and subsequently tested for structural and functional modifications by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and a single-fiber in vitro motility assay, respectively. MALDI spectra revealed glycation-related structural alterations as evidenced by the disappearance of specific Lys-C proteolysis products and the appearance of higher mass peaks that are attributed to cross-linking by glucose. This change was paralleled by a significant reduction in the in vitro motility speed, suggesting a structure-related decline in myosin mechanics in response to glucose exposure. Further evidence that early glycation products form in the regulatory regions of the myosin molecule is derived from the fact that there is complete reversal of motility speed after reaction with the Schiff base-cleaving agent hydroxylamine hydrochloride. Thus, glycation of skeletal muscle myosin has a significant effect on both the structural and functional properties of the protein, a finding that is important in understanding the mechanisms underlying the impairment in muscle function associated with aging and diabetes.

Actins↗

PubMed2001

Novel, wide bandwidth, micromachined ultrasonic transducers.

Surface micromachined, capacitive ultrasonic transducers have been fabricated using a low thermal budget, CMOS-compatible process. This process allows inherent control of parameters such as membrane size and thickness, cavity size and the intrinsic stress in the membrane to be achieved. Devices fabricated using this process exhibit interesting properties for transduction in air at frequencies in excess of 1 MHz when driven from a standard ultrasonic voltage source. Experiments have been performed with devices containing silicon nitride membranes of variable thicknesses over a 2 microm thick air cavity and with device dimensions of up to 5 mm square. This is much larger than has been reported for a device with a single membrane. Calibration measurements using 1/8 inch microphones in air, and miniature PVDF hyrdophones in water, have been performed. The dependence on d.c. bias voltage is examined, involving static membrane deflection measurements and received peak voltages. Pulse-echo and pitch-catch mode operation have been achieved. Interferometric measurements of membrane displacement have been performed in air to illustrate the membrane deflection characteristics. Operation in liquids is also discussed.

Equipment Design↗

PubMed2000

Protein targets of monocrotaline pyrrole in pulmonary artery endothelial cells.

A single administration of monocrotaline to rats results in pathologic alterations in the lung and heart similar to human pulmonary hypertension. In order to produce these lesions, monocrotaline is oxidized to monocrotaline pyrrole in the liver followed by hematogenous transport to the lung where it injures pulmonary endothelium. In this study, we determined specific endothelial targets for (14)C-monocrotaline pyrrole using two-dimensional gel electrophoresis and autoradiographic detection of protein metabolite adducts. Selective labeling of specific proteins was observed. Labeled proteins were digested with trypsin, and the resulting peptides were analyzed using matrix-assisted laser desorption ionization mass spectrometry. The results were searched against sequence data bases to identify the adducted proteins. Five abundant adducted proteins were identified as galectin-1, protein-disulfide isomerase, probable protein-disulfide isomerase (ER60), beta- or gamma-cytoplasmic actin, and cytoskeletal tropomyosin (TM30-NM). With the exception of actin, the proteins identified in this study have never been identified as potential targets for pyrroles, and the majority of these proteins have either received no or minimal attention as targets for other electrophilic compounds. The known functions of these proteins are discussed in terms of their potential for explaining the pulmonary toxicity of monocrotaline.

Actins↗

PubMed2000

Determination of blood folate using acid extraction and internally standardized gas chromatography-mass spectrometry detection.

Whole blood folate level is a superior indicator of folate nutritional status than serum/plasma level. Problems with and lack of confidence in results of current whole blood folate assays have limited its popularity for assessing folate nutritional status. Here, an acid extraction GCMS detection method that measures total folate whole blood is presented. Folates are released from the matrix of whole blood and cleaved to para-aminobenzoic acid (pABA) by acid hydrolysis in the presence of [(13)C(6)]pABA as internal standard (IS). The hydrolysate is passed over a C18 resin to remove heme. The pABA isotopomers are ethyl esterified, isolated on C18 resin, and trifluoroacetylated. Following normal-phase HPLC separation, the isotopomers are silylated to their tBDMS derivatives. The abundance of these derivatives are measured at m/z 324 for [(13)C(6)]pABA as IS and m/z 318 for pABA from whole blood folate. Our method uses readily available chemicals and our results agree well with those using Lactobacillus casei, the current gold standard reference assay. The presence of folate analogs (methotrexate) or antibacterials (sulfonamines) does not affect our method. This feature makes it useful in monitoring folate status of patients undergoing chemotherapy. Before using our method, pABA supplements must be discontinued for a few days.

Acids↗

PubMed2000

Inactivation of creatine kinase by S-glutathionylation of the active-site cysteine residue.

Protein S-thiolation, the formation of mixed disulphides of cysteine residues in proteins with low-molecular-mass thiols, occurs under conditions associated with oxidative stress and can lead to modification of protein function. In the present study, we examined the site of S-thiolation of the enzyme creatine kinase (CK), an important source of ATP in myocytes. Inactivation of this enzyme is thought to play a critical role in cardiac injury during oxidative stress, such as during reperfusion injury. Reaction of rabbit CK M isoenzyme with GSSG, used to model protein S-thiolation, was found to result in enzyme inactivation that could be reversed by GSH or dithiothreitol. Measurement of GSH that is released during the thiolation reaction indicated that the maximum extent of CK thiolation was approx. 1 mol of GSH/mol of protein, suggesting thiolation on one reactive cysteine residue. Accordingly, matrix-assisted laser-desorption ionization MS confirmed that the molecular mass of CK was increased, consistent with addition of one GSH molecule/molecule of CK. Using trypsin digestion, HPLC and MS analysis, the active-site cysteine residue (Cys(283)) was identified as the site of thiolation. Reversal of thiolation was shown to be rapid when GSH is abundant, rendering dethiolation of CK thermodynamically favoured within the cell. We conclude that S-glutathionylation of CK could be one mechanism to explain temporary reversible loss in activity of CK during ischaemic injury. The maintainance of GSH levels represents an important mechanism for regeneration of active CK from S-glutathionylated CK.

Amino Acid Sequence↗

PubMed2000

On-line coupling of subcritical water extraction with high-performance liquid chromatography via solid-phase trapping.

Although ambient water is very polar and cannot dissolve many organic species, water at elevated temperatures behaves like a polar organic solvent. Thus, subcritical water has been proven to be an effective extraction fluid for several classes of organic compounds. While solvent trapping was used to collect the extracted analytes in most of previous subcritical water extractions, sorbent trapping has also been developed for subcritical water extraction. In this study, an on-line system for subcritical water extraction and high-performance liquid chromatography (HPLC) was built and tested. A sorbent trap was used as the interface between subcritical water extraction and HPLC. Several shut-off valves have been utilized to switch the system from one mode to another (e.g., from the extraction mode to HPLC mode). The coupling technique of subcritical water extraction and HPLC eliminates the liquid-liquid extraction used in solvent trapping subcritical water extraction and provides higher sensitivity. Compared to the off-line system reported in an earlier work, the operation of this on-line system is even easier. Some peak broadening occurred after the coupling the water extraction with HPLC for the analytes studied. The performance of this on-line system was evaluated by the extraction and determination of caffeine, nitrotoluenes, polychlorinated biphenyls, chlorophenols and anilines.

Aniline Compounds↗

PubMed2000

Recruitment of a foreign quinone into the A(1) site of photosystem I. I. Genetic and physiological characterization of phylloquinone biosynthetic pathway mutants in Synechocystis sp. pcc 6803.

Genes encoding enzymes of the biosynthetic pathway leading to phylloquinone, the secondary electron acceptor of photosystem (PS) I, were identified in Synechocystis sp. PCC 6803 by comparison with genes encoding enzymes of the menaquinone biosynthetic pathway in Escherichia coli. Targeted inactivation of the menA and menB genes, which code for phytyl transferase and 1,4-dihydroxy-2-naphthoate synthase, respectively, prevented the synthesis of phylloquinone, thereby confirming the participation of these two gene products in the biosynthetic pathway. The menA and menB mutants grow photoautotrophically under low light conditions (20 microE m(-2) s(-1)), with doubling times twice that of the wild type, but they are unable to grow under high light conditions (120 microE m(-2) s(-1)). The menA and menB mutants grow photoheterotrophically on media supplemented with glucose under low light conditions, with doubling times similar to that of the wild type, but they are unable to grow under high light conditions unless atrazine is present to inhibit PS II activity. The level of active PS II per cell in the menA and menB mutant strains is identical to that of the wild type, but the level of active PS I is about 50-60% that of the wild type as assayed by low temperature fluorescence, P700 photoactivity, and electron transfer rates. PS I complexes isolated from the menA and menB mutant strains contain the full complement of polypeptides, show photoreduction of F(A) and F(B) at 15 K, and support 82-84% of the wild type rate of electron transfer from cytochrome c(6) to flavodoxin. HPLC analyses show high levels of plastoquinone-9 in PS I complexes from the menA and menB mutants but not from the wild type. We propose that in the absence of phylloquinone, PS I recruits plastoquinone-9 into the A(1) site, where it functions as an efficient cofactor in electron transfer from A(0) to the iron-sulfur clusters.

Alkyl and Aryl Transferases↗

PubMed2000

Quantification of individual glutathione S-transferase isozymes in hepatic and pulmonary tissues of naphthalene-tolerant mice.

Acute exposure to naphthalene produces severe bronchiolar epithelial cell necrosis in mice, whereas subchronic exposure to naphthalene (200 mg/kg/7 days) fails to produce epithelial necrosis and renders the animals tolerant to subsequent challenge doses of naphthalene. Mechanisms responsible for the development of tolerance have not been delineated. The few studies exploring naphthalene tolerance focus on expression of microsomal enzymes and have yet to delve into expression of the hepatic detoxification enzymes such as glutathione S-transferases (GSTs; EC 2.5.1.18). Glutathione conjugation catalyzed by GSTs accounts for one of the two primary routes of naphthalene detoxification. In this study, we rigorously quantify levels of individual GST isozymes expressed within the livers and lungs of mice with acquired tolerance to naphthalene. Subchronic exposure to naphthalene increases the abundance of some hepatic GSTs to levels as much as 68% greater than controls. Naphthalene-tolerant mice displayed increases in mGSTM1 (51%), mGSTM2 (58%), and mGSTP1 (66%), whereas no significant difference in mGSTA3 was observed between exposed and control mice. Extracts of pulmonary tissues from naphthalene-tolerant mice showed minor increases in levels of mGSTP1 (7%) and Peak 8 isozyme (27%) and decreases in levels of mGSTM1 (31%), mGSTM2 (17%), and mGSTA3 (8%). The total enzymatic activity for the conjugation of 1-chloro-2,4-dinitrobenzene (CDNB) was 22% lower in lung extracts from naphthalene-tolerant animals than in controls. These results indicate that induction of hepatic GSTs is substantial and may be an important factor in the development of tolerance to naphthalene.

Animals↗

PubMed2000

An economic analysis of an aggressive diagnostic strategy with single photon emission computed tomography myocardial perfusion imaging and early exercise stress testing in emergency department patients who present with chest pain but nondiagnostic electrocardiograms: results from a randomized trial.

STUDY OBJECTIVE: Conventional emergency department testing strategies for patients with chest pain often do not provide unequivocal diagnosis of acute coronary syndromes. This study was conducted to determine whether the routine use of single photon emission computed tomography (SPECT) imaging at rest and early exercise stress testing to assess intermediate-risk patients with chest pain and no ECG evidence of acute ischemia will lead to earlier discharges, more discriminate use of coronary angiography, and an overall reduction in average costs of care with no adverse clinical outcomes. METHODS: All patients in this study had technetium 99m tetrofosmin SPECT imaging at rest and were randomly assigned to either a conventional (results of the imaging test blinded to the physician) or perfusion imaging-guided (results of the imaging test unblinded to the physician) strategy. Patients in the conventional arm were treated at their physician's discretion. Patients in the perfusion imaging-guided arm were treated according to a predefined protocol based on SPECT imaging test results: coronary angiography after a positive scan result and exercise treadmill testing after a negative scan result. Study endpoints consisted of total in-hospital costs and length of stay. Hospital costs were calculated using hospital department-specific Medicare cost/charge ratios. Length of stay was calculated as total hospital room days billed (regular and intensive care). RESULTS: We enrolled 46 patients, 9 with acute myocardial infarctions. Patients randomly assigned to the perfusion imaging-guided arm had $1,843 (95% confidence interval [CI] $431 to $6,171) lower median in-hospital costs and 2.0-day (95% CI 1.0 to 3.0 days) shorter median lengths of stay but similar rates of in-hospital and 30-day follow up events as patients in the conventional arm. CONCLUSION: An ED chest pain diagnostic strategy incorporating acute resting (99m)Tc tetrofosmin SPECT imaging and early exercise stress testing may lead to reduced in-hospital costs and decreased length of stay for patients with acute chest pain and nondiagnostic ECGs.

Aged↗