Triglyceride Lenses at the Air-Water Interface as a Model System for Studying the Initial Stage in the Biogenesis of Lipid Droplets.

Lipid droplets (LD) are intracellular structures consisting of an apolar lipid core, composed mainly of triglycerides (TG) and steryl esters, coated by a lipid-protein mixed monolayer. The mechanisms underlying LD biogenesis at the endoplasmic reticulum membrane are a matter of many current investigations. Although models explaining the budding-off of protuberances of phase-segregated TG inside bilayers have been proposed recently, the assumption of such initial blisters needs further empirical support. Here, we study mixtures of egg phosphatidylcholine (EPC) and TG at the air-water interface in order to describe some physical properties and topographic stability of TG bulk structures in contact with interfaces. Brewster angle microscopy images revealed the appearance of microscopic collapsed structures (CS) with highly reproducible lateral size (∼1 µm lateral radius) not varying with lateral packing changes and being highly stable at surface pressures (π) beyond collapse. By surface spectral fluorescence microscopy, we were able to characterize the solvatochromism of Nile Red both in monolayers and inside CS. This allowed to conclude that CS corresponded to a phase of liquid TG and to characterize them as lenses forming a three-phase (oil-water-air) system. Thereby, the thicknesses of the lenses could be determined, observing that they were dramatically flattened when EPC was present (6-12 nm compared to 30-50 nm for lenses on EPC/TG and TG films, respectively). Considering the shape of lenses, the interfacial tensions, and the Neumann's triangle, this experimental approach allows one to estimate the oil-water interfacial tension acting at each individual microscopic lens and at varying compression states of the surrounding monolayer. Thus, lenses formed on air-water Langmuir films can serve to assess variables of relevance to the initial step of LD biogenesis, such as the degree of dispersion of excluded-TG phase and shape, spatial distribution, and oil-water interfacial tension of lenses.

The challenge of post-mortem GHB analysis: storage conditions and specimen types are both important.

BACKGROUND: For the interpretation of concentrations of gamma-hydroxybutyrate (GHB) in post-mortem specimens, a possible increase due to post-mortem generation in the body and in vitro has to be considered. The influence of different storage conditions and the specimen type was investigated. METHOD AND MATERIAL: Post-mortem GHB concentrations in femoral venous blood (VB), heart blood (HB), serum (S) from VB, urine (U), cerebrospinal fluid (CSF) and vitreous humour (VH) were determined by gas chromatography-mass spectrometry after derivatisation. Various storage conditions, that is 4 °C or room temperature (RT) and the addition of sodium fluoride (NaF), were compared during storage up to 30 days. Additionally, bacterial colonisation was determined by mass spectrometry fingerprinting. RESULTS: Twenty-six cases without involvement of exogenous GHB were examined. GHB concentrations (by specimen) at day 0 were 3.9-22.1 mg/L (VB), 6.6-33.3 mg/L (HB), < 0.5-18.1 mg/L (U), 1.1-10.4 mg/L (CSF) and 1.7-22.0 mg/L (VH). At 4 °C, concentrations increased at day 30 to 5.6-74.5 mg/L (VB), 4.6-76.5 mg/L (HB) and < 0.5-21.3 mg/L (U). At RT, concentrations rose to < 0.5-38.5 mg/L (VB), 1.2-94.6 mg/L (HB) and < 0.5-37.5 mg/L (U) at day 30. In CSF, at RT, an increase up to < 0.5-21.2 mg/L was measured, and at 4 °C, a decrease occurred (< 0.5-6.5 mg/L). GHB concentrations in VH remained stable at both temperatures (1.2-20.9 mg/L and < 0.5-26.2 mg/L). The increase of GHB in HB samples with NaF was significantly lower than that without preservation. No correlation was found between the bacterial colonisation and extent of GHB concentration changes. CONCLUSION: GHB concentrations can significantly increase in post-mortem HB, VB and U samples, depending on storage time, temperature and inter-individual differences. Results in CSF, VH, S and/or specimens with NaF are less affected.

Endogenous gamma-hydroxybutyric acid (GHB) concentrations in post-mortem specimens and further recommendation for interpretative cut-offs.

When interpreting gamma-hydroxybutyric acid (GHB) concentrations in post-mortem specimens, a possible increase in GHB concentrations because of post-mortem generation must be considered. In this study, endogenous GHB concentrations in post-mortem biological fluids were investigated. Additionally, we review post-mortem GHB concentrations already published in the literature. Heart and peripheral blood samples, cerebrospinal fluid, urine, and vitreous humor were collected from 64 autopsies in subjects where the cause of death excluded GHB exposure. Sample analysis was carried out either on the day of autopsy or later after immediate freezing and storage at -20 °C. GHB concentrations in venous blood samples (n = 61) were <0.6-28.7 mg/L (mean 11.9 mg/L; median 10.6 mg/L), <0.6-65.3 mg/L (mean 15.2 mg/L; median 12.8 mg/L) in heart blood (n = 56), <0.6-25.1 mg/L (mean 6.0 mg/L; median 3.8 mg/L) in urine (n = 50), <0.6-39.0 mg/L (mean 9.6 mg/L; median 7.5 mg/L), in vitreous humor (n = 54), and <0.6-24.0 mg/L (mean 4.2 mg/L; median 3.2 mg/L) in cerebrospinal fluid (n = 52). There was no significant difference between GHB concentrations in cases where there were signs of beginning putrefaction at the time of autopsy (n = 9) and cases without obvious signs of putrefaction. In one case with advanced putrefaction, the GHB concentration in venous blood was 32.7 mg/L. In conclusion, for post-mortem venous blood, urine, and cerebrospinal fluid, an interpretative cut-off of 30 mg/L for GHB concentrations is suggested in cases where GHB analysis is conducted on the day of sample collection at autopsy or if samples have been stored at -20 °C immediately after collection.

Dilatational yielding of solid Langmuir monolayers.

In a previous work, Muruganathan and Fischer observed laser-induced local collapse of a methyl stearate monolayer. These experiments opened the possibility of studying the collapse mechanism in a highly controlled manner, since the laser intensity can be easily varied and collapse happens in a definite place (the laser focus). In this paper we extended the work presented by Muruganathan et al., describing the local yielding as an alternative pathway toward monolayer collapse competing with the global collapse of the monolayer. We first corroborated that the laser-induced collapse is a thermocapillary effect and afterward determined the threshold laser power necessary for the local pathway to win over the global collapse. We show that the laser threshold is determined more by the gradients in temperature and pressure than by the global pressure and temperature. We propose that the flow of material into the focus of the laser is observed after the yield stress of the monolayer is overcome. The higher the yield stress, the higher the temperature gradient that is necessary for the monolayer to yield. The local pathway opens only when the derivative of surface pressure with temperature is negative such that stress gradients point toward the laser focus and a sink of material is generated. In such a case we are able to give estimates of the dilatational yield pressure of the solid monolayer.

Rheological properties of a two phase lipid monolayer at the air/water interface: effect of the composition of the mixture.

Many biologically relevant monolayers show coexistence of discrete domains of a long-range ordered condensed phase dispersed in a continuous, disordered, liquid-expanded phase. In this work, we determined the viscous and elastic components of the compressibility modulus and the shear viscosity of monolayers exhibiting phase coexistence with the aim at elucidating the contribution of each phase to the observed monolayer mechanical properties. To this purpose, mixed monolayers with different proportions of distearoylphosphatidylcholine (DSPC) and dimyristoylphosphatidylcholine (DMPC) were prepared and their rheological properties were analyzed. The relationship between the phase diagram of the mixture at 10 mN m(-1) and the rheological properties was studied. We found that the monolayer shear viscosity is highly dependent on the presence of domains and on the domain density. In turn, the monolayer compressibility is only influenced by the presence of domains for high domain densities. For monolayers that look homogeneous on the micrometer scale (DSPC amount lower that 23 mol %), all the analyzed rheological properties remain similar to those observed for pure DMPC monolayers, indicating that in this proportion range the DSPC molecules contribute as DMPC to the surface rheology in spite of having hydrocarbon chains four carbons longer.

Micron-scale phase segregation in lipid monolayers induced by myelin basic protein in the presence of a cholesterol analog.

It was previously shown that myelin basic protein (MBP) can induce phase segregation in whole myelin monolayers and myelin lipid films, which leads to the accumulation of proteins into a separate phase, segregated from a cholesterol-enriched lipid phase. In this work we investigated some factors regulating the phase segregation induced by MBP using fluorescent microscopy of monolayers formed with binary and ternary lipid mixtures of dihydrocholesterol (a less-oxidable cholesterol analog) and phospholipids. The influence of the addition of salts to the subphase and of varying the lipid composition was analyzed. Our results show that MBP can induce a dihydrocholesterol-dependent segregation of phases that can be further regulated by the electrolyte concentration in the subphase and the composition (type and proportion) of non-sterol lipids. In this way, changes of the lipid composition of the film or the ionic strength in the aqueous media modify the local surface density of MBP and the properties (phase state and composition) of the protein environment.

The influence of domain crowding on the lateral diffusion of ceramide-enriched domains in a sphingomyelin monolayer.

In this paper, we analyze the Brownian motion of ceramide-enriched condensed domains immersed in a fluid sphingomyelin-enriched monolayer at the air-water interface. The diffusion coefficient of the domains is determined under different molecular packings and domain arrays, and the monolayer viscosity is calculated. With this approach, the effect of domain crowding and of intrinsic monolayer viscosity can be split. We found that, for mixed monolayers of palmitoylated sphingomyelin and ceramide, the monolayer viscosity depends on the lateral pressure and on the domain-domain distance. In a 9:1 proportion of sphingomyelin:ceramide, the viscosity is about 4x10(-9) N s m(-1) below 15 mN m(-1) (continuum sphingomyelin phase in a liquid expanded state) and increases at higher lateral pressures (continuum phase in a condensed state). The viscosity change with pressure is caused by both an increase of intrinsic viscosity and an increase in domain crowding. At very high domain crowding, the monolayer viscosity increases because the domains are held in place by steric hindrance generated by the other condensed domains of the array. These are short-range forces, effective when the domains are close together. As the domain array is relaxed and the domain-domain distance increases, these forces become negligible, and repulsive dipolar interactions appear to acquire importance. For the lipid mixture analyzed, the dipolar repulsion is more noticeable on subphases of 0.15 M NaCl than on pure water, revealing the influence of surface electrostatics.

[Alcohol, the aged and traffic]. / Alkohol, Alter und Verkehr.

The roadworthiness of older people is affected by internal, neurological or orthopedic diseases and by the medication. These factors have an effect on alcohol tolerance. Therefore, alcoholization in elderly drivers needs specific attention. A retrospective evaluation of all alcoholized traffic participants > 60 years of age between 2003-2008 in Hamburg is described. The data analysis gives information about the alcohol level in various age groups, the resulting neurological/physiological deficits, chronic illnesses and the medications taken. The proportion of elderly people involved in a traffic delinquency has increased. However, the absolute number of old drunk drivers has decreased. Seniors are represented significantly less often concerning alcohol in traffic than other age groups.

Lysophosphatidic acid-induced platelet shape change revealed through LPA(1-5) receptor-selective probes and albumin.

Lysophosphatidic acid (LPA), a component of mildly-oxidized LDL and the lipid rich core of atherosclerotic plaques, elicits platelet activation. LPA is the ligand of G protein-coupled receptors (GPCR) of the EDG family (LPA(1-3)) and the newly identified LPA(4-7) subcluster. LPA(4), LPA(5) and LPA(7) increase cellular cAMP levels that would induce platelet inhibition rather than activation. In the present study we quantified the mRNA levels of the LPA(1-7) GPCR in human platelets and found a rank order LPA(4) = LPA(5) > LPA(7) > LPA(6) = LPA(2) >> LPA(1) > LPA(3). We examined platelet shape change using a panel of LPA receptor subtype-selective agonists and antagonists and compared them with their pharmacological profiles obtained in heterologous LPA(1-5) receptor expression systems. Responses to different natural acyl and alkyl species of LPA, and octyl phosphatidic acid analogs, alpha-substituted phosphonate analogs, N-palmitoyl-tyrosine phosphoric acid, N-palmitoyl-serine phosphoric acid were tested. All of these compounds elicited platelet activation and also inhibited LPA-induced platelet shape change after pre-incubation, suggesting that receptor desensitization is likely responsible for the inhibition of this response. Fatty acid free albumin (10 microM) lacking platelet activity completely inhibited platelet shape change induced by LPA with an IC(50) of 1.1 microM but had no effect on the activation of LPA(1,2,3,&5) expressed in endogenously non-LPA-responsive RH7777 cells. However, albumin reduced LPA(4) activation and shifted the dose-response curve to the right. LPA(5) transiently expressed in RH7777 cells showed preference to alkyl-LPA over acyl-LPA that is similar to that in platelets. LPA did not increase cAMP levels in platelets. In conclusion, our results with the pharmacological compounds and albumin demonstrate that LPA does not induce platelet shape change simply through activation of LPA(1-5), and the receptor(s) mediating LPA-induced platelet activation remains elusive.

Usefulness of cardiac magnetic resonance imaging for coronary artery disease detection.

Cardiac magnetic resonance imaging (cMRI) is a promising non-invasive technique to assess the presence of coronary artery disease (CAD), which is free of ionizing radiation and iodine contrast. cMRI can detect CAD by angiographic methods or indirectly by perfusion stress techniques. While coronary angiography by cMRI remains limited to research protocols, stress perfusion cMRI is currently being applied worldwide in the clinical setting. Studies have shown good correlation between adenosine-induced stress myocardial perfusion cMRI and single-photon-emission computed tomography or positron emission tomography to detect CAD. Quantitative methods to analyze cMRI perfusion data have been developed in an attempt to provide a more objective imaging interpretation. Standardization of such quantitative methods, with minimal operator dependency, would be useful for clinical and research applications. Myocardial perfusion reserve (MPR), calculated using Fermi deconvolution technique, has been compared with well established anatomical and physiological CAD detection techniques. MPR appears to be the most accurate quantitative index to detect anatomical and hemodynamically significant CAD. Beyond physiological assessment of CAD, cMRI provides information regarding regional and global left ventricular function and morphology, myocardial infarction size, transmurality and viability. Such comprehensive information would require the performance of multiple tests if other modalities were used. This article describes current applications of cMRI for evaluation of patients with CAD.

Postmortem determination of concentrations of stress hormones in various body fluids--is there a dependency between adrenaline/noradrenaline quotient, cause of death and agony time?

To find out whether a certain cause of death or a certain length of an agonal period shows specific adrenaline or noradrenaline profiles, heart blood, femoral vein blood, liquor, urine and vitreous humour were taken from corpses (n = 98) at the Medical School Hannover, and noradrenaline and adrenaline were determined using high-performance liquid chromatography (HPLC). Corpses were classified according to the following five categories: short agony, long agony, state after hanging, state after asphyxiation and state after CPR with documented administration of epinephrine. Once results were collected the adrenaline/noradrenaline quotient was determined. It became clear that there were no significant differences regarding the concentration of adrenaline and noradrenaline in the various body fluids in relation to the above-mentioned categories. The means adrenaline/noradrenaline quotients in femoral vein blood were 0.21 +/- 0.29 for hanged persons, 0.38 +/- 0.47 for asphyxiated persons, 0.17 +/- 0.19 for those with short agony and 0.42 +/- 0.43 for those with long agony, significantly below 1 (p < 0.001; p = 0.001; p = 0.003). For condition after CPR we found an adrenaline/noradrenaline quotient of 2.81 +/- 5.8. In liquor the adrenaline/noradrenaline quotients for short agony was 0.17 +/- 0.17, for hanged persons 0.18 +/- 0.19 and for asphyxiated ones 0.30 +/- 0.38, significantly lower than 1 (p < 0.001). In urine the adrenaline/noradrenaline quotients for all categories are lower than 1 (p < 0.001); short agony (0.13 +/- 0.09), long agony (0.21 +/- 0.16), hanged (0.15 +/- 0.16), asphyxiated (0.14 +/- 0.08) and CPR (0.14 +/- 0.06). In vitreous humour the quotients for short agony (0.14 +/- 0.28), long agony (0.13 +/- 0.12), hanged (0.07 +/- 0.09) and asphyxiated (0.09 +/- 0.11) are lower than 1 (p < 0.001). The spread of data for the adrenaline/noradrenaline quotient did not allow for any conclusions about cause of death and length of agony in individual cases.

Externally applied electric fields on immiscible lipid monolayers: repulsion between condensed domains precludes domain migration.

Lipid and protein molecules anisotropically oriented at a hydrocarbon-aqueous interface configure a dynamic array of self-organized molecular dipoles. Electrostatic fields applied to lipid monolayers have been shown to induce in-plane migration of domains or phase separation in a homogeneous system. In this work, we have investigated the effect of externally applied electrostatic fields on different lipid monolayers exhibiting surface immiscibility. In the monolayers studied, lipids in the condensed state segregate in discontinuous round-shaped domains, with the lipid in the liquid-expanded state forming the continuous phase. The use of fluorescent probes with selective phase partitioning allows analyzing by epifluorescence microscopy the migrations of the domains under the influence of inhomogeneous electric fields applied to the surface. Our observations indicate that a positive potential applied to an electrode placed over the monolayer promotes a repulsion of the domains until a steady state is reached, indicating the presence of a force opposed to the externally applied electric force. The experimental results were modeled by considering that the opposing force is generated by the dipole-dipole repulsion between the domains.

Biophysics of sphingolipids II. Glycosphingolipids: an assortment of multiple structural information transducers at the membrane surface.

Glycosphingolipids are ubiquitous components of animal cell membranes. They are constituted by the basic structure of ceramide with its hydroxyl group linked to single carbohydrates or oligosaccharide chains of different complexity. The combination of the properties of their hydrocarbon moiety with those derived from the variety and complexity of their hydrophilic polar head groups confers to these lipids an extraordinary capacity for molecular-to-supramolecular transduction across the lateral/transverse planes in biomembranes and beyond. In our opinion, most of the advances made over the last decade on the biophysical behavior of glycosphingolipids can be organized into three related aspects of increasing structural complexity: (1) intrinsic codes: local molecular interactions of glycosphingolipids translated into structural self-organization. (2) Surface topography: projection of molecular shape and miscibility of glycosphingolipids into formation of coexisting membrane domains. (3) Beyond the membrane interface: glycosphingolipid as modulators of structural topology, bilayer recombination and surface biocatalysis.

Properties of galactocerebroside layers transferred to glassy carbon electrodes: effect of an applied electric field.

Galactocerebroside films deposited onto glassy carbon electrodes have been previously studied through the electrochemical response of a redox couple present in solution. Those experiments indicated that the film is inhomogeneous and that there are lipid-free places. In this work, we present experimental results indicating that those bare regions are formed when the electrode is introduced in an aqueous solution, and that the size and/or amount of uncovered domains increase when negative potentials are applied to the film. The experimental techniques employed for these findings are epifluorescence microscopy and ellipsometry.

[Cricothyroidotomy training on cadavers - experiences in the education of medical students, anaesthetists, and emergency physicians]. / Koniotomieübungen an der Leiche - Erfahrungen in der Ausbildung mit Medizinstudenten, Anästhesisten und Notärzten.

OBJECTIVE: Should the technique of surgical cricothyroidotomy be practiced on cadavers and should it be a compulsory part of the teaching curriculum? Is it wise to use a speculum for the insertion of the endotracheal tube? What is the optimum size of the tube? METHODS: A surgical cricothyroidotomy with a speculum was carried out on 30 cadavers from the Institute of Legal Medicine, Medical School Hannover. This took place as part of a official and voluntary course for students of advanced semesters, anaesthetists and emergency doctors with the subjects "cricothyroidotomy, chest drainage and venous cut-down". The surgical cricothyroidotomy without the use of a speculum was carried out on 5 cadavers by two clinicians well practiced in this technique. The elapsed time between skin incision and the insertion of the endotracheal tube was measured on all five subjects. After the course the participants were asked if they were able to carry out a cricothyroidotomy in an emergency. They were also asked whether this course should be a compulsory part of their curriculum and whether practical sessions should take place. During autopsies at the Institute of Legal Medicine the length of the ligamentum conicum was measured on 40 corpses with reclined and non-reclined heads. RESULTS: The average time of storage of the cadavers was 4.2 days +/- 1.9 days. The cricothyroidotomy was possible on all 35 cadavers. In one case (3,3 %) the result was a complete rupture of the cricoid cartilage. In 5 cases (16.7 %) the horizontal incision was torn due to prising with the speculum. Difficult situations always occured when the skin incision was not exactly in the midline. The average time to place the endotracheal tube into the trachea by the surgical procedure of cricothyroidotomy was 22.4 seconds +/- 3.1 seconds (minimum 18 seconds, maximum 26 seconds). 10 % of the medical students and 50 % of the anaesthetists and emergency doctors felt they would be prepared to carry out a cricothyroidotomy in an emergency. 90 % of the students and respectively 80 % of the anaesthetists and emergency doctors stated that they would like to practice the technique on a cadaver again. Almost all participants were of the opinion that the course should be integrated as a compulsory course in a future educational curriculum. The average distance between the thyroid cartilage and the cricoid cartilage was 9.5 mm +/- 1.9 mm with non-reclined head (minimum 6 mm, maximum 14 mm) and 11.9 mm +/- 2.5 mm with reclined head (minimum 7 mm, maximum 18 mm). The average difference of distances was 2.4 mm +/- 1.2 mm (minimum 1 mm, maximum 6 mm) in reclined and non-reclined heads. CONCLUSIONS: In our opinion it is highly recommended that the technique of cricothyroidotomy should be practiced on cadavers and that the course should become a compulsory part in a future educational curriculum. In addition the incision of the ligamentum conicum using dilators or a speculum is not to be recommended from the point of view of this study. The tracheal tube used in this study (reinforced wire tube, ID 6.0) was best suited for surgical cricothyroidotomy.

Magnetic resonance first-pass perfusion imaging: overview and perspectives.

The data from clinical studies with quantitative MR first-pass perfusion imaging suggests that this technique outperforms SPECT--widely available clinical imaging tool--in sensitivity and specificity. Moreover, MRFP imaging may be combined with the assessment of global and segmental function of the heart and regional wall thickening, and in addition, performed with pharmacological stress agents. The inter- and intra-observer reproducibility of quantitative MRFP is comparable with clinically used nuclear medicine techniques. MRFP measurements can discern collateral myocardium and are able to identify small changes in myocardial blood flow and myocardial perfusion reserve (the ratio of stress blood flow over resting). MRFP imaging has been mainly used in context of coronary artery disease but many other exciting areas in clinical cardiology are awaiting of new insights that can be accomplished with this technique. Trials are needed to obtain the approval of the contrast agent (Gd-DTPA) and perfusion sequences by the Food and Drug Administration and to establish reimbursement procedures with the third-party insurance companies and health maintenance organizations.

Amplification of growth regulatory genes in intraductal breast cancer is associated with higher nuclear grade but not with the progression to invasiveness.

Ductal carcinoma in situ (DCIS), as an identifiable progenitor lesion of invasive breast cancer, represents a morphologically, biologically, and prognostically heterogeneous disease. It is not clear which molecular mechanisms are involved in progression to infiltrative growth. In this study, 83 DCIS classified according to the Van Nuys grading scheme were examined for amplification of growth regulatory genes that have been found to be amplified in invasive breast cancer (c-erbB2, topoisomerase IIalpha, c-myc, and cyclinD1 genes). Exact quantification of gene amplification was enabled by a combination of laser microdissection of paraffin-embedded tissue with real-time PCR. In DCIS, gene amplifications of all tested genes were found. The most frequently amplified gene was c-erbB2 found in 21 of 83 (25%) cases. Amplification of the other genes under investigation was observed in 4% to 6% of cases, high-grade DCIS being predominantly affected. High-grade DCIS differed significantly from low- and intermediate-grade DCIS in frequency and level of c-erbB2 amplification. In addition, high-grade DCIS revealed an accumulation of genetic aberrations. Amplification status in pure in situ lesions did not differ from intraductal carcinoma with an infiltrative component, indicating that although associated with a higher nuclear grade gene amplification might not represent an independent prognostic marker of disease progression.

Echo-planar magnetic resonance myocardial perfusion imaging: parametric map analysis and comparison with thallium SPECT.

Magnetic resonance (MR) perfusion FLASH imaging has been used for assessing coronary artery disease (CAD). Echo-planar MR techniques have advantages in speed and in making MR perfusion imaging results more clinically accessible through parametric maps, but have not been previously assessed. We implemented a spin-echo, echo-planar MR technique and applied it at rest and during adenosine stress in 26 patients with CAD and abnormal thallium single-photon-emission computed tomography (SPECT), and analyzed the results by using a newly developed parametric map analysis of time to peak, peak intensity, and slope of contrast washin. The results were compared with the results of conventional visual analysis of the perfusion cine series. For detecting abnormal coronary territories, MR and SPECT were comparable for sensitivity, specificity, and accuracy (thallium, 70%, 78%, and 73%; MR, 79% 83%, and 80%; P = NS). There was good agreement between thallium and MR during stress (kappa = 0.49), but defects were larger by MR (2.4 vs. 3.1 segments for slope; P < 0.01). Additional segments were detected at rest by MR (58 for slope vs. 25 for thallium), which correlated with areas that became abnormal with stress in the thallium (sensitivity, 100%; specificity, 63%). The parametric maps were easier and faster to interpret than review of the original first-pass series of images (chi2 = 10.8; P < 0.04). The diagnostic performance of echo-planar perfusion MR and SPECT was similar, and combining the results with parametric mapping was useful for interpretation and considerably improved data display for clinical interpretation. MR, however, was faster and yielded images of higher resolution with no radiation burden. In multislice mode, these new MR techniques may have clinical value.

Quantitative magnetic resonance first-pass perfusion analysis: inter- and intraobserver agreement.

Magnetic resonance first-pass (MRFP) imaging awaits longitudinal clinical trials for quantification of myocardial perfusion. The purpose of this study was to assess inter- and intraobserver agreement of this method. Seventeen MRFP studies (14 rest and 3 under adenosine-induced hyperemia) from 14 patients were acquired. Two observers visually graded study quality. Each study was subdivided into eight regions. Both observers analyzed all 17 studies (8 x 17 = 136 regions) for interobserver agreement. Each observer then analyzed 10 of the 17 studies a second time (2 x 8 x 10 = 160 regions) for intraobserver agreement. Signal intensity curves were obtained with Argus software (Siemens, Iselin, NJ). The maximum amplitude of the impulse response function (Rmax) and the change of signal intensity (deltaSImax) of the contrast bolus were determined. Intraclass correlation coefficient was used to determine intra- and interobserver agreement. The quality was good or excellent in 14 studies. Intraobserver agreement of Rmax and deltaSImax were good (0.85 and 0.80, n = 160). Interobserver agreement of Rmax was fair (0.55, n = 136) but improved after exclusion of poor-quality studies (0.88, n = 112). Interobserver agreement of deltaSImax was good (0.73) and improved less than Rmax with study quality (0.83). Interobserver agreement for Rmax in individual myocardial regions before and after exclusion of studies with poor quality changed most markedly in lateral and posterior regions (0.69 and 0.65 vs. 0.97 and 0.94), where signal-to-noise ratios were reduced compared with anteroseptal regions (p < 0.01). Analysis of MRFP images provides good intraobserver agreement. Interobserver agreement of the quantitative perfusion analysis is good under the premise of good image quality.