Multiple-pinhole collimators improve intra- and between-rater agreement and the certainty of the visual interpretation in dopamine transporter SPECT.

BACKGROUND: Multiple-pinhole (MPH) collimators improve the resolution-sensitivity trade-off compared to parallel-hole collimators. This study evaluated the impact of MPH collimators on intra- and between-rater agreement, and on the certainty of visual interpretation in dopamine transporter (DAT)-SPECT. METHODS: The study included 71 patients (62.1 ± 12.7 y). Two SPECT acquisitions were performed in randomized order after a single injection of 182 ± 9 MBq 123I-FP-CIT, one with MPH and one with low-energy-high-resolution-high-sensitivity (LEHRHS) collimators. MPH projections were reconstructed with an iterative 3d Monte Carlo algorithm. LEHRHS projections were reconstructed with filtered backprojection (FBP) or with ordered-subsets expectation-maximization and resolution recovery (OSEM). Images were visually evaluated twice by three independent raters with respect to presence/absence of Parkinson-typical reduction of striatal 123I-FP-CIT uptake using a Likert 6-score (- 3 = clearly normal, …, 3 = clearly reduced). In case of intra-rater discrepancy, an intra-rater consensus was obtained. Intra- and between-rater agreement with respect to the Likert score (6-score and dichotomized score) was characterized by Cohen's kappa. RESULTS: Intra-rater kappa of visual scoring of MPH/LEHRHS-OSEM/LEHRHS-FBP images was 0.84 ± 0.12/0.73 ± 0.06/0.73 ± 0.08 (6-score, mean of three raters) and 1.00 ± 0.00/0.96 ± 0.04/0.97 ± 0.03 (dichotomized score). Between-rater kappa of visual scoring (intra-rater consensus) of MPH/LEHRHS-OSEM/LEHRHS-FBP images was 0.70 ± 0.06/0.63 ± 0.08/0.48 ± 0.05 (6-score, mean of three pairs of raters) and 1.00 ± 0.00/0.92 ± 0.04/0.90 ± 0.06 (dichotomized score). There was a decrease of (negative) Likert scores in normal DAT-SPECT by 0.87 ± 0.18 points from the LEHRHS-OSEM to the MPH setting. The (positive) Likert scores of reduced DAT-SPECT did not change on average. CONCLUSIONS: MPH collimators improve intra- and between-rater agreement as well as the certainty of the visual interpretation of DAT-SPECT.

Impact of Valve Plane Alignment on the Repeatability of Left Ventricular Ejection Fraction in ECG-gatedMyocard ial SPECT Using Corridor 4DM.

Background: In myocardial gated single-photon emission computed tomography (GSPECT), to differentiate true changes of left ventricular ejection fraction (LVEF) from inherent methodical variability is clinically relevant; however, data about repeatability of GSPECT LVEF in the same patients are rather inconsistent in literature. The aim of this study was therefore to determine repeatability coefficient (RC) of GSPECT LVEF at rest and to investigate the effect of the introduction of processing constraints in left ventricular edge detection. Methods: Thirty-five patients referred for one-day myocardial GSPECT stress-rest scan were included. After the routine stress-rest study, patients were completely repositioned on the imaging table for a second rest acquisition using the same acquisition parameters. LVEF was computed using Corridor 4DM software without and with manual alignment of valve plane. Repeatability was assessed using the Bland-Altman method. Results: RC of LVEF from unaligned datasets was 7.6% with upper and lower limits of agreement of 7.4% to -7.8%. After valve plane and ventricular long-axis length alignment, RC improved to 3.6% with upper and lower limits of agreement of 3.4% to -3.8%. Conclusions: RC using unaligned determination of GSPECT LVEF was comparable to that from previous publications. However, RC using valve plane alignment could be improved to below 4% on 95% confidence level.

Stimulation of erythrocyte cell membrane scrambling by mitotane.

UNLABELLED: background: Mitotane (1,1-dichloro-2-[o-chlorophenyl]-2-[p-chlorophenyl]ethane), a cytostatic drug used for the treatment of adrenocortical carcinomas, is effective by triggering tumor cell apoptosis. In analogy to apoptosis of nucleated cells, eryptosis is the suicidal death of erythrocytes, which is typically paralleled by cell shrinkage and breakdown of cell membrane phosphatidylserine asymmetry with subsequent phosphatidylserine exposure at the erythrocyte surface. Eryptosis may be triggered by increase of cytosolic Ca(2+) concentration ([Ca(2+)]i). The present study tested, whether treatment of human erythrocytes with mitotane is followed by eryptosis. METHODS: [Ca(2+)]i was estimated from Fluo3 fluorescence, cell volume from forward scatter, phosphatidylserine exposure from annexin V binding, and hemolysis from hemoglobin release. RESULTS: Exposure to mitotane (≥ 5 µg/ml ≈ 16 µM) significantly increased [Ca(2+)]i, increased annexin V binding and triggered hemolysis, but did not significantly modify forward scatter. The effect on annexin V binding was significantly blunted in the absence of extracellular Ca(2+). Within 30 min Ca(2+) ionophore ionomycin (1 µM) decreased forward scatter, an effect virtually abolished in the presence of mitotane (15 µg/ml). CONCLUSIONS: Mitotane increases [Ca(2+)]i with subsequent phosphatidylserine translocation. By the same token mitotane inhibits Ca(2+) induced cell shrinkage.

Estramustine-induced suicidal erythrocyte death.

BACKGROUND: The nitrogen mustard derivative of estradiol-17ß-phosphate estramustine is used for the treatment of prostate cancer. Estramustine may trigger suicidal death of cancer cells. Side effects of estramustine include anemia. At least in theory, estramustine could cause anemia by stimulation of eryptosis, the suicidal death of erythrocytes. Hallmarks of eryptosis include cell shrinkage, increased cytosolic Ca2+ activity ([Ca2+]), ceramide formation and phosphatidylserine translocation to the outer leaflet of the cell membrane with phosphatidylserine exposure at the erythrocyte surface. Eryptosis is stimulated by increase of cytosolic Ca2+ activity ([Ca2+]i). The present study explored whether estramustine triggers eryptosis. METHODS: [Ca2+]i was estimated from Fluo3 fluorescence, cell volume from forward scatter, phosphatidylserine exposure from annexin V binding, and hemolysis from hemoglobin release. RESULTS: A 24 h exposure to estramustine (≤ 100 µM) significantly increased [Ca2+]i, increased annexin V binding and increased hemoglobin release. The effect of estramustine on annexin V binding was significantly blunted by removal of extracellular Ca2+. CONCLUSIONS: Estramustine stimulates both, eryptosis and hemolysis. The estramustine induced translocation of phosphatidylserine to the cell surface is at least partially due to increase of cytosolic Ca2+ activity.

Effect of miltefosine on erythrocytes.

BACKGROUND: Miltefosine, an alkylphosphocholine drug with antiparasite, antibacterial, antifungal and antineoplastic potency, is the only oral drug that can be used to treat visceral and cutaneous leishmaniasis. The effect of miltefosine is at least partially due to triggering of apoptosis. Similar to apoptosis of nucleated cells, erythrocytes may enter suicidal death or eryptosis, which is characterized by cell shrinkage and by cell membrane scrambling with phosphatidylserine-exposure at the erythrocyte surface. Eryptosis may be triggered following increase of cytosolic Ca(2+)-level ([Ca(2+)]i). The present study explored, whether miltefosine elicits eryptosis. METHODS: Cell volume has been estimated from forward scatter, phosphatidylserine-exposure from annexin-V-binding, hemolysis from hemoglobin release, [Ca(2+)]i from Fluo3-fluorescence. RESULTS: A 48 h exposure to miltefosine (≥ 4.9 µM) was followed by significant decrease of forward scatter and significant increase of annexin-V-binding. The effect was paralleled by significant increase of [Ca(2+)]i. The annexin-V-binding following miltefosine treatment was significantly blunted in the nominal absence of extracellular Ca(2+). CONCLUSION: Miltefosine stimulates eryptosis, an effect at least partially due to stimulation of Ca(2+) entry.