Body weight-dependent Rubidium-82 activity results in constant image quality in myocardial perfusion imaging with PET.

BACKGROUND: Clinical practice shows degrading image quality in heavier patients who undergo myocardial perfusion imaging (MPI) with Rubidium-82 (Rb-82) PET when using a fixed tracer activity. Our aim was to derive and validate a patient-specific activity protocol resulting in a constant image quality in PET MPI. METHODS: We included 251 patients who underwent rest MPI with Rb-82 PET (Discovery 670, GE Healthcare). 132 patients were included retrospectively and were scanned using a fixed activity of 740 MBq. The total number of measured prompts was normalized to activity and correlated to body weight, mass per body length and body mass index to find the best predicting parameter. Next, a patient-specific activity was derived and subsequently validated in 119 additional patients. Image quality was scored by three experts on a four-point scale. RESULTS: Both image quality and prompts decreased in heavier patients when using a fixed activity (p < .005). Body weight was used to derive a new activity formula: Activity = 8.3 MBq/kg. When applying this formula, both measured prompts and scored image quality became independent of body weight (p > .60). CONCLUSION: Administrating a Rb-82 activity that linearly depends on body weight resulted in a constant image quality across all patients and is recommended.

Effect of proton pump inhibitors on Rubidium-82 gastric uptake using positron emission tomography myocardial perfusion imaging.

BACKGROUND: Rb-82 positron emission tomography (PET) myocardial perfusion imaging (MPI) is a robust tool for the evaluation of coronary artery disease (CAD). However, gastric uptake and spillover can be seen in 10% of Rb-82 PET MPI studies, commonly affecting the inferior wall, and can preclude the accurate identification of myocardial ischemia. We sought to understand the relationship between Rb-82 gastric uptake and the use of proton pump inhibitors (PPI). METHODS: 600 consecutive patients who presented for a clinically indicated Rb-82 PET MPI study were prospectively enrolled. In addition to the clinical history, PPI use was ascertained (medication, dose, frequency and duration of use, and time of last dose). Patients were categorized as PPI and non-PPI users. Rb-82 uptake in the gastrium, myocardium, and liver were measured at rest. Absolute uptake values and gastric:hepatic ratios were compared in PPI and non-PPI users. RESULT: Of 600 enrolled patients, 181 (30.2%) patients were using PPI. The gastric Rb-82 uptake in PPI users was 23% higher than non-PPI users (146 ± 52 kBq/cc vs 119 ± 40 kBq/cc, respectively; P < 0.001). The resting gastric:hepatic Rb-82 uptake ratio was also 23% higher in PPI vs non-PPI users (2.7 ± 1.0 vs 2.2 ± 0.8, respectively; P < 0.001). CONCLUSION: The gastric uptake of Rb-82 appears to be greater in patients actively using PPI and may identify a group who might be at greater risk of non-diagnostic Rb-82 PET MPI.

The human brain somatostatin interactome: SST binds selectively to P-type family ATPases.

Somatostatin (SST) is a cyclic peptide that is understood to inhibit the release of hormones and neurotransmitters from a variety of cells by binding to one of five canonical G protein-coupled SST receptors (SSTR1 to SSTR5). Recently, SST was also observed to interact with the amyloid beta (Aß) peptide and affect its aggregation kinetics, raising the possibility that it may bind other brain proteins. Here we report on an SST interactome analysis that made use of human brain extracts as biological source material and incorporated advanced mass spectrometry workflows for the relative quantitation of SST binding proteins. The analysis revealed SST to predominantly bind several members of the P-type family of ATPases. Subsequent validation experiments confirmed an interaction between SST and the sodium-potassium pump (Na+/K+-ATPase) and identified a tryptophan residue within SST as critical for binding. Functional analyses in three different cell lines indicated that SST might negatively modulate the K+ uptake rate of the Na+/K+-ATPase.

Biodistribution and radiation dosimetry of (82)Rb at rest and during peak pharmacological stress in patients referred for myocardial perfusion imaging.

PURPOSE: (82)Rb is an ultra-short-lived positron emitter used for myocardial blood flow quantification with PET imaging. The aim of this study was to quantify the biodistribution and radiation dosimetry in patients with coronary disease and in healthy normal volunteers. METHODS: A total of 30 subjects, 26 patients with known or suspected coronary artery disease (CAD) and four healthy volunteers were injected with (82)Rb chloride at 10 MBq/kg followed by a 10-min dynamic PET scan. Chest scans at rest were acquired in all subjects, as well as one additional biodistribution scan of the head, neck, abdomen, pelvis or thighs. Chest scans under stress were acquired in 25 of the CAD patients. (82)Rb time-integrated activity coefficients were determined in 22 source organs using volume of interest analysis, including corrections for partial-volume losses. The mean time-integrated activity coefficients were used to calculate the whole-body effective dose using tissue weighting factors from the International Commission on Radiological Protection (ICRP) Publications 60 and 103. RESULTS: A total of 283 organ time-integrated activity coefficients were calculated, with a minimum of four values per source organ. The rest and stress mean effective dose was 0.8 mSv/GBq, according to the most recent ICRP definition. Using 10 MBq/kg for 3D PET imaging, the effective dose to a gender-averaged reference person (60 kg female and 73 kg male) is 1.1 mSv for a complete rest and stress perfusion study. For 2D PET using a typical injected activity of 1.1 to 2.2 GBq each for rest and stress, the effective dose for a complete study is 1.8 to 3.5 mSv. CONCLUSION: The current effective dose estimate in CAD patients is four times lower than the values reported previously by the ICRP, and about 35% lower than previous in vivo studies in young healthy subjects.

Quantification of myocardial blood flow in absolute terms using (82)Rb PET imaging: the RUBY-10 Study.

OBJECTIVES: The purpose of this study was to compare myocardial blood flow (MBF) and myocardial flow reserve (MFR) estimates from rubidium-82 positron emission tomography ((82)Rb PET) data using 10 software packages (SPs) based on 8 tracer kinetic models. BACKGROUND: It is unknown how MBF and MFR values from existing SPs agree for (82)Rb PET. METHODS: Rest and stress (82)Rb PET scans of 48 patients with suspected or known coronary artery disease were analyzed in 10 centers. Each center used 1 of 10 SPs to analyze global and regional MBF using the different kinetic models implemented. Values were considered to agree if they simultaneously had an intraclass correlation coefficient >0.75 and a difference <20% of the median across all programs. RESULTS: The most common model evaluated was the Ottawa Heart Institute 1-tissue compartment model (OHI-1-TCM). MBF values from 7 of 8 SPs implementing this model agreed best. Values from 2 other models (alternative 1-TCM and Axially distributed) also agreed well, with occasional differences. The MBF results from other models (e.g., 2-TCM and retention) were less in agreement with values from OHI-1-TCM. CONCLUSIONS: SPs using the most common kinetic model-OHI-1-TCM-provided consistent results in measuring global and regional MBF values, suggesting that they may be used interchangeably to process data acquired with a common imaging protocol.

Quantification of myocardial perfusion using cardiac magnetic resonance imaging correlates significantly to rubidium-82 positron emission tomography in patients with severe coronary artery disease: a preliminary study.

INTRODUCTION: Aim was to compare absolute myocardial perfusion using cardiac magnetic resonance imaging (CMRI) based on Tikhonov's procedure of deconvolution and rubidium-82 positron emission tomography (Rb-82 PET). MATERIALS AND METHODS: Fourteen patients with coronary artery stenosis underwent rest and adenosine stress imaging by 1.5-Tesla MR Scanner and a mCT/PET 64-slice Scanner. CMRI were analyzed based on Tikhonov's procedure of deconvolution without specifying an explicit compartment model using our own software. PET images were analyzed using standard clinical software. CMRI and PET data was compared with Spearman's rho and Bland-Altman analysis. RESULTS: CMRI results were strongly and significantly correlated with PET results for the absolute global myocardial perfusion differences (r=0.805, p=0.001) and for global myocardial perfusion reserve (MPR) (r=0.886, p<0.001). At vessel territorial level, CMRI results were also significantly correlated with absolute PET myocardial perfusion differences (r=0.737, p<0.001) and MPR (r=0.818, p<0.001). Each vessel territory had similar strong correlation for absolute myocardial perfusion differences (right coronary artery (RCA): r=0.787, p=0.001; left anterior descending artery (LAD): r=0.796, p=0.001; left circumflex artery (LCX): r=0.880, p<0.001) and for MPR (RCA: r=0.895, p<0.001; LAD: r=0.886, p<0.001; LCX: r=0.886, p<0.001). CONCLUSION: On a global and vessel territorial basis, CMRI-measured absolute myocardial perfusion differences and MPR were strongly and significantly correlated with the Rb-82 PET findings.

Initial human experience with Rubidium-82 renal PET/CT imaging.

INTRODUCTION: Preclinical data have shown that Rubidium-82 chloride ((82)Rb) is a radiotracer with high first pass extraction and slow washout in the kidneys. The goal of this study was to investigate the feasibility of human kidney imaging with (82)Rb positron emission tomography (PET) and obtain quantitative data of its uptake non-invasively. METHODS: Eight healthy volunteers underwent dynamic PET/CT imaging with (82)Rb. A preprogrammed pump was used to insure reproducible injections. Tissue time activity curves were generated from the renal cortex. An input function was derived from the left ventricular blood pool (LVBP), the descending thoracic aorta and the abdominal aorta. Renal blood flow was estimated by applying a two-compartment kinetic model. Results obtained with different input functions were compared. RESULTS: Radiotracer accumulation was rapid and reached a plateau within 15-30 s after the bolus entered the kidneys. The derived K1 and k2 parameters were reproducible using input functions obtained from diverse vascular locations. K1 averaged 1.98 ± 0.14 mL/min/g. The average k2 was 0.35 ± 0.11/min. Correlation between K1 values obtained from the LVBP from different bed positions when the kidneys and abdominal aorta were in the same field of view was excellent (R = 0.95). CONCLUSIONS: Non-invasive quantitative human kidney imaging with (82)Rb PET is feasible. Advantages of renal PET with (82)Rb include excellent image quality with high image resolution and contrast. (82)Rb has potential as a clinical renal imaging agent in humans.

Impact of point spread function modeling and time-of-flight on myocardial blood flow and myocardial flow reserve measurements for rubidium-82 cardiac PET.

BACKGROUND: Myocardial flow reserve (MFR) obtained from dynamic cardiac positron emission tomography (PET) with rubidium-82 (Rb-82) has been shown to be a useful measurement in assessing coronary artery disease. Advanced PET reconstructions with point spread function modeling and time-of-flight have been shown to improve image quality but also have an impact on kinetic analysis of dynamic data. This study aims to determine the impact of these algorithms on MFR data. METHODS: Dynamic Rb-82 cardiac PET images from 37 patients were reconstructed with standard and advanced reconstructions. Area under curve (AUC) of the blood input function (BIF), myocardial blood flow (MBF) and MFR were compared with each reconstruction. RESULTS: No significant differences were seen in MFR for the two reconstructions. A relatively small mean difference in MBF data of +11.9% was observed with advanced reconstruction compared with the standard reconstruction but there was considerable variability in the degree of change (95% confidence intervals of -16.2% to +40.0%). Small systematic relative differences were seen for AUC BIF (mean difference of -6.3%; 95% CI -17.5% to +5.4%). CONCLUSION: MFR results from Rb-82 dynamic PET appear to be robust when generated by standard or advanced PET reconstructions. Considerable increases in MBF values may occur with advanced reconstructions, and further work is required to fully understand this.

Vascular physiology and protein disposition in a preclinical model of neurodegeneration.

The development of clinically relevant preclinical models that mimic the hallmarks of neurodegenerative disease is an ongoing pursuit in early drug development. In particular, robust physiological characterization of central nervous system (CNS) disease models is necessary to predict drug delivery to target tissues and to correctly interpret pharmacodynamic responses to disease-modifying therapeutic candidates. Efficient drug delivery across the blood-CNS barrier is a particularly daunting task, prompting our strategy to evaluate the biodistribution of five distinct molecular probes in a well-characterized mouse model of neurodegeneration. A transgenic mouse model of amyotrophic lateral sclerosis was selected based on a phenotype resembling clinical symptoms, including loss of motor neurons from the spinal cord and paralysis in one or more limbs, due to expression of a G93A mutant form of human superoxide dismutase (SOD1). The tissue distributions of two proteins, albumin and a representative immunoglobulin G antibody, as well as two blood flow markers, the lipophilic blood flow marker Ceretec (i.e., (99m)Tc-HMPAO) and the polar ionic tracer, rubidium-86 chloride ((86)RbCl), were measured following intravenous injection in SOD1(G93A) and age-matched control mice. The radiopharmaceutical TechneScan PYP was also used to measure the distribution of (99m)Tc-labeled red blood cells as a blood pool marker. Both the antibody and (86)Rb were able to cross the blood-spinal cord barrier in SOD1(G93A) mice to a greater extent than in control mice. Although the biodistribution patterns of antibody, albumin, and RBCs were largely similar, notable differences were detected in muscle and skin. Moreover, vastly different biodistribution patterns were observed for a lipophilic and polar perfusion agent, with SOD1(G93A) mutation resulting in reduced renal filtration rates for the former but not the latter. Overall, the multiprobe strategy provided an opportunity to efficiently collect an abundance of physiological information, including the degree and regional extent of blood-CNS barrier permeability, in a preclinical model of neurodegeneration.

Washout of 8²Rb as a marker of impaired tissue integrity, obtained by list-mode cardiac PET/CT: relationship with perfusion/metabolism patterns of myocardial viability.

PURPOSE: Myocardial washout of the potassium analogue (82)Rb may indicate tissue impairment. Few studies have evaluated its usefulness for viability assessment, and controversial results were reported. We revisited this topic using list-mode positron emission tomography (PET)/CT. METHODS: A total of 22 patients with chronic ischemic cardiomyopathy (ICM) and 11 control subjects with normal CT coronary angiogram were studied. Rest (82)Rb PET/CT studies were acquired in list mode and resampled to static, gated, and dynamic images. Using a 17-segment model, (82)Rb washout was determined by monoexponential fitting of myocardial time-activity curves. In ICM patients, (18)F-fluorodeoxyglucose (FDG) studies were obtained in the same session and segments were classified as normally perfused, mismatch, or matched defect. RESULTS: (82)Rb washout was minimal and homogeneous in control subjects. Normally perfused segments of ICM did not differ (p = 0.33). ICM patients had a left ventricular ejection fraction (LVEF) of 25 ± 12%, 25/353 mismatched, and 46/353 matched defect segments. (82)Rb washout was higher in hypoperfused vs normal segments (p < 0.05), but not different between mismatch and matched defect (p = 0.18). Intraindividual analysis in nine patients showing both FDG mismatch and matched defect confirmed absence of differences. Overall, segmental (82)Rb washout correlated inversely with (82)Rb uptake (r = -0.70; p < 0.05) and less well with FDG uptake (r = -0.31; p < 0.05). CONCLUSION: Using state-of-the-art PET/CT technology for myocardial viability assessment, (82)Rb washout does not distinguish between perfusion/metabolism patterns of hibernating myocardium and scar. Tissue integrity may be at least partially impaired in hibernation.

Radiation dosimetry of 82Rb in humans under pharmacologic stress.

UNLABELLED: (82)Rb is used with PET for cardiac perfusion studies. Using human biokinetic measurements, in vivo, we recently reported on the resting-state dosimetry of this agent. The objective of this study was to obtain (82)Rb dose estimates under stress. METHODS: (82)Rb biokinetics were obtained in 10 healthy volunteers (5 male, 5 female; mean age ± SD, 33 ± 10 y; age range, 18-50 y) using whole-body PET/CT. The 76-s half-life of (82)Rb and the corresponding need for pharmacologic vasodilation require that all imaging be completed within 10 min. To accommodate these constraints, while acquiring the data needed for dosimetry we used the following protocol. First, a whole-body attenuation correction CT scan was obtained. Then, a series of 3 whole-body PET scans was acquired after a single infusion of 1.53 ± 0.12 GBq of (82)Rb at rest. Four minutes after the infusion of a 0.56 mg/kg dose of the vasodilator, dipyridamole, 3 serial whole-body PET scans were acquired after a single infusion of 1.50 ± 0.16 GBq of (82)Rb under stress. The time-integrated activity coefficient (TIAC) for stress was obtained by scaling the mean rest TIAC obtained from our previous rest study by the stress-to-rest TIAC ratio obtained from the rest-stress measurements described in this report. RESULTS: The highest mean organ-absorbed doses under stress were as follows: heart wall, 5.1, kidneys, 5.0, lungs, 2.8, and pancreas, 2.4 µGy/MBq (19, 19, 10.4, and 8.9 mrad/mCi, respectively). The mean effective doses under stress were 1.14 ± 0.10 and 1.28 ± 0.10 µSv/MBq using the tissue-weighting factors of the International Commission on Radiological Protection, publications 60 and 103, respectively. CONCLUSION: Appreciable differences in source-organ biokinetics were observed for heart wall and kidneys during stress when compared with the previously reported rest study. The organ receiving the highest dose during stress was the heart wall. The mean effective dose calculated during stress was not significantly different from that obtained at rest.

Pathophysiologic correlates of 82Rb biodistribution in cardiac PET/CT.

PURPOSE: PET perfusion imaging with (82)Rb is a powerful tool for evaluating coronary artery disease (CAD). Little is known about normal patterns or significance of (82)Rb lung distribution in the setting of heart disease. Herein, PET/CT hybrid imaging was used to obtain insights into the frequency and potential radiomorphologic correlates of altered (82)Rb distribution. METHODS: Myocardial perfusion PET/CT studies of 58 patients referred for workup of CAD were analyzed [28 normal and 30 patients with low left ventricular ejection fraction (LVEF)]. Organ regions of interest were placed on PET images, and (82)Rb uptake was measured and compared under resting and stress conditions. RESULTS: Qualitatively increased lung uptake was observed in 13 patients-5 with normal LVEF(R) and 8 with reduced LVEF(R); 12 of 13 had lung infiltrates/atelectasis on CT. Lung to heart ratios in the normal and low EF groups were (mean ± SD) 0.168 ± 0.047 and 0.171 ± 0.075 at rest, and 0.128 ± 0.035 and 0.147 ± 0.067 during stress (p = 0.87 and 0.18, respectively). Lung to liver ratios were not significantly different between the two groups under stress or rest conditions. CONCLUSION: Increased lung uptake of (82)Rb occurs in a subset of patients referred for workup of CAD by PET/CT and may be influenced by primary parenchymal abnormalities and LV dysfunction. Thus, the relevance of pulmonary (82)Rb uptake as a marker of cardiac outcome may be limited. Larger studies are needed to determine how non-cardiac (82)Rb uptake and CT findings may be integrated to increase the diagnostic and prognostic value of cardiac PET/CT.

Human biodistribution and radiation dosimetry of 82Rb.

UNLABELLED: Prior estimates of radiation-absorbed doses from (82)Rb, a frequently used PET perfusion tracer, yielded discrepant results. We reevaluated (82)Rb dosimetry using human in vivo biokinetic measurements. METHODS: Ten healthy volunteers underwent dynamic PET/CT (6 contiguous table positions, each with separate (82)Rb infusion). Source organ volumes of interest were delineated on the CT images and transferred to the PET images to obtain time-integrated activity coefficients. Radiation doses were estimated using OLINDA/EXM 1.0. RESULTS: The highest mean absorbed organ doses (µGy/MBq) were observed for the kidneys (5.81), heart wall (3.86), and lungs (2.96). Mean effective doses were 1.11 ± 0.22 and 1.26 ± 0.20 µSv/MBq using the tissue-weighting factors of the International Commission on Radiological Protection (ICRP), publications 60 and 103, respectively. CONCLUSION: Our current (82)Rb dosimetry suggests reasonably low radiation exposure. On the basis of this study, a clinical (82)Rb injection of 2 × 1,480 MBq (80 mCi) would result in a mean effective dose of 3.7 mSv using the weighting factors of the ICRP 103-only slightly above the average annual natural background exposure in the United States (3.1 mSv).

Right and left ventricular uptake with Rb-82 PET myocardial perfusion imaging: markers of left main or 3 vessel disease.

BACKGROUND: Relative myocardial perfusion imaging may underestimate severity of coronary disease (CAD), particularly in cases of balanced ischemia. Can quantification of peak left (LV) and right (RV) ventricular Rb-82 uptake measurements identify patients with left main or 3 vessel disease? METHODS: Patients (N = 169) who underwent Rb-82 PET MPI and coronary angiography were categorized as having no significant coronary stenosis (n = 60), 1 or 2 vessel disease (n = 81), or left main disease/3 vessel disease (n = 28), based on angiography. Maximal LV and RV ventricular myocardial Rb-82 uptake was measured during stress and rest. RESULTS: Failure to augment LV uptake by >or= 8500 Bq/cc at stress, predicted left main or 3 vessel disease with a sensitivity of 93% and specificity of 61% (area under curve = 0.83). A >or=10% increase in RV: LV uptake ratios with stress over rest was 93% specific (area under curve = 0.74) for left main or 3 vessel disease. These indices incrementally predicted left main or 3 vessel disease compared to models including age, gender, cardiac risk factors, and summed stress and difference scores. CONCLUSION: Quantifying maximal rest and stress LV and RV uptake with PET myocardial perfusion imaging may independently and incrementally identify patients with left main or 3 vessel disease.

Direct regulation of prokaryotic Kir channel by cholesterol.

Our earlier studies have shown that channel activity of Kir2 subfamily of inward rectifiers is strongly suppressed by the elevation of cellular cholesterol. The goal of this study is to determine whether cholesterol suppresses Kir channels directly. To achieve this goal, purified prokaryotic Kir (KirBac1.1) channels were incorporated into liposomes of defined lipid composition, and channel activity was assayed by (86)Rb(+) uptake. Our results show that (86)Rb(+) flux through KirBac1.1 is strongly inhibited by cholesterol. Incorporation of 5% (mass cholesterol/phospholipid) cholesterol into the liposome suppresses (86)Rb(+) flux by >50%, and activity is completely inhibited at 12-15%. However, epicholesterol, a stereoisomer of cholesterol with similar physical properties, has significantly less effect on KirBac-mediated (86)Rb(+) uptake than cholesterol. Furthermore, analysis of multiple sterols suggests that cholesterol-induced inhibition of KirBac1.1 channels is mediated by specific interactions rather than by changes in the physical properties of the lipid bilayer. In contrast to the inhibition of KirBac1.1 activity, cholesterol had no effect on the activity of reconstituted KscA channels (at up to 250 microg/mg of phospholipid). Taken together, these observations demonstrate that cholesterol suppresses Kir channels in a pure protein-lipid environment and suggest that the interaction is direct and specific.

Direct effects of eicosapentaenoic and docosahexaenoic acids on phospholipid and triglyceride fatty acid pattern, glucose metabolism, 86rubidium net uptake and insulin release in BRIN-BD11 cells.

The long-term metabolic and functional effects of a dietary deprivation of long-chain polyunsaturated omega3 fatty acids were recently investigated in second-generation omega3-depleted rats. This study represents the first attempt to explore the direct, but not immediate, effects of omega3 fatty acids on insulin-producing cells. For this purpose, BRIN-BD11 cells were cultured for 24 h in the absence or presence of both C20:5omega3 and C22:6omega3 (50 microM each) and, thereafter, examined for their phospholipid and triglyceride fatty acid pattern, and their metabolic, ionic, and secretory responses to D: -glucose and/or non-nutrient insulinotropic agents. The prior culture in the presence of the two omega3 fatty acids provoked an enrichment of cell lipids in such omega3 fatty acids, changes in the phospholipid fatty acid pattern of long-chain polyunsaturated omega6 fatty acids as well as saturated and monodesaturated fatty acids, and cell steatosis. It minimized the relative increase in D: -[5-(3)H]glucose utilization and D: -[U-(14)C]glucose oxidation otherwise resulting from an increase in the concentration of the hexose from 1.1 to 11.1 mM. It also minimized the changes in (86)Rb(+) net uptake otherwise provoked by rises in D: -glucose concentration and decreased the absolute values for insulin output. It is concluded that the major changes in metabolic, cationic, and secretory behavior of the omega3-enriched BRIN-BD11 cells are paradoxically similar to those encountered in pancreatic islets from omega3-depleted rats and, in both cases, possibly attributable to a phenomenon of lipotoxicity.

CT-based attenuation correction in (82)Rb-myocardial perfusion PET-CT: incidence of misalignment and effect on regional tracer distribution.

PURPOSE: Misalignment of low-dose-CT used for attenuation correction (AC) may cause artifacts in cardiac-PET-CT. The aim was to evaluate incidence and severity of misalignment and its quantitative effects on regional myocardial (82)Rb-distribution. METHODS: Rest/dipyridamole (82)Rb-perfusion-PET-CT studies of 92 consecutive patients were analyzed for misalignment. Two different scanning protocols were employed: the first 57 patients had separate CTs for rest and stress PET. The following 35 patients had one CT at rest, used for AC of rest and stress PET. Misalignment was visually scored on a five-point scale (0 = no, 1 = minimal, 2 = mild, 3 = moderate, and 4 = severe). In five representative patients with normal perfusion and low probability of disease, 95 polarmaps were created by shifting CT vs PET prior to reconstruction of attenuation-corrected data sets using dedicated software (three dimensions of space; magnitude of shifts, 5, 10, 14 mm). RESULTS: PET/CT -misalignment was detected in 60% of rest and 67% of stress studies. Alignment for rest was better than that for stress (0.7 +/- 0.7 vs 1.0 +/- 0.9, P = 0.03). Comparison of the two protocols revealed no effect on the alignment of the stress study (1.0 +/- 0.9 vs 1.0 +/- 0.9, P = 0.9). Quantitatively, the largest individual effect of any artificial misalignment was a 25% reduction of relative (82)Rb uptake. With a shift of 1 cm, the largest effect in an individual was a 19% decrease. Anterior wall was most frequently influenced by misalignment, but changes of uptake also occurred in all other segments. CONCLUSIONS: Misalignment between CT and PET in cardiac-PET-CT influences regional tracer distribution in multiple segments. Repeated CT imaging after dipyridamole does not improve alignment. These results emphasize the need for strategies to improve coregistration in clinical imaging protocols.

Role of homologous ASP334 and GLU319 in human non-gastric H,K- and Na,K-ATPases in cardiac glycoside binding.

Cardiac steroids inhibit Na,K-ATPase and the related non-gastric H,K-ATPase, while they do not interact with gastric H,K-ATPase. Introducing an arginine, the residue present in the gastric H,K-ATPase, in the second extracellular loop at the corresponding position 334 in the human non-gastric H,K-ATPase (D334R mutation) rendered it completely resistant to 2mM ouabain. The corresponding mutation (E319R) in alpha1 Na,K-ATPase produced a approximately 2-fold increase of the ouabain IC(50) in the ouabain-resistant rat alpha1 Na,K-ATPase and a large decrease of the ouabain affinity of human alpha1 Na,K-ATPase, on the other hand this mutation had no effect on the affinity for the aglycone ouabagenin. These results provide a strong support for the orientation of ouabain in its biding site with its sugar moiety interacting directly with the second extracellular loop.

Erythrocyte sodium pump stimulation by ouabain and an endogenous ouabain-like factor.

Cardiac glycosides inhibit the sodium pump. However, some studies suggest that nanomolar ouabain concentrations can stimulate the activity of the sodium pump. In this study, using the Na(+)/K(+)-ATPase of human erythrocytes, we compared the effect of digoxin, ouabain and an ouabain like-factor (OLF), on (86)Rb uptake. Ouabain concentrations below 10(-9) M significantly stimulate Rb(+) uptake, and the maximal increase above base-line values is 18 +/- 5% at 10(-10) M ouabain. No stimulation is observed in the same conditions by digoxin. OLF behaved like ouabain, producing an activation of Rb(+) flux at concentrations lower than 10(-9) M ouabain equivalents (14 +/- 3% at 10(-10) M). Western blot analysis revealed the presence of both alpha(1) and alpha(3) pump isoforms in human erythrocytes. Our data confirm the analogies between OLF and ouabain and suggest that Na(+)/K(+)-ATPase activation may be related to the alpha(3) isoform. In addition, we investigated whether ouabain at different concentrations was effective in altering the intracellular calcium concentration of erythrocytes. We found that ouabain at concentration lower than 10(-9) M did not affect this homeostasis.