Beta-ray imaging system with γ-ray coincidence for multiple-tracer imaging.

PURPOSE: Beta-ray imaging systems are widely used for various biological objects to obtain a two-dimensional (2D) distribution of ß-ray emitting radioisotopes. However, a conventional ß-ray imaging system is unsuitable for multiple-tracer imaging, because the continuous energy distribution of ß-rays complicates distinguishing among different tracers by energy information. Therefore, we developed a new type of ß-ray imaging system, which is useful for multiple tracers by detecting coincidence γ-rays with ß-rays, and evaluated its imaging performance. METHODS: Our system is composed of position-sensitive ß-ray and γ-ray detectors. The former is a 35 × 35 × 1-mm3 Ce-Doped((La, Gd)2 Si2 O7 ) (La-GPS) scintillation detector, which has a 300-µm pitch of pixels. The latter is a 43 × 43 × 16-mm3 bismuth germanium oxide (BGO) scintillation detector. Both detectors are mounted on a flexible frame and placed in a user-selectable position. We experimentally evaluated the performance of the ß-ray detector and the γ-ray efficiencies of the γ-ray detector with different energies, positions, and distances. We also conducted point sources and phantom measurements with dual isotopes to evaluate the system performance of multiple-tracer imaging. RESULTS: For the ß-ray detector, the ß-ray detection efficiencies for 45 Ca (245-keV maximum energy) and 90 Sr/90 Y (545 and 2280-keV maximum energy) were 14.3% and 21.9%, respectively. The total γ-ray detection efficiency of the γ-ray detector for all γ-rays from 22 Na (511-keV annihilation γ-rays and a 1275-keV γ-ray) in the center position with a detector distance of 20 mm was 17.5%. From a point-source measurement using 22 Na and 90 Sr/90 Y, we successfully extracted the position of a positron-γ emitter 22 Na. Furthermore, for a phantom experiment using 45 Ca and 18 F or 18 F and 22 Na, we successfully extracted the distribution of the second tracer using the annihilation γ-ray or de-excitation γ-ray coincidence. In all the imaging experiments, the event counts of the extracted images were consistent with the counts estimated by the measured γ-ray efficiencies. CONCLUSIONS: We successfully demonstrated the feasibility of our ß-ray autoradiography system for imaging multiple isotopes. Since our system can identify not only a ß-γ emitter but also a positron emitter using the coincidence detection of annihilation γ-rays, it is useful for PET tracers and various new applications that are otherwise impractical.

Measurement of the 43Sc production cross-section with a deuteron beam.

43,44Sc/47Sc is one of the most promising theranostic pairs in nuclear medicine. The co-emission of 1157 keV γ-rays with 99.9% branching ratio by 44Sc and the presence of its metastable state 44 mSc push to favour the adoption of 43Sc for Positron Emission Tomography (PET) diagnostic procedures to lighten the dose to the patient and to the personnel. The ß+ emitter 43Sc can be produced at a medical cyclotron by proton bombardment of an enriched 43Ca or 46Ti oxide target. 43Sc can be also produced by deuteron bombardment of an enriched 42Ca oxide target. Only a few medical cyclotrons currently in operation offer deuteron beams. Some can be adapted to operate both a proton or a deuteron source. To compare these three production routes, an accurate knowledge of the cross-sections is essential. In this paper, we report on the cross-section measurement of the reaction 42Ca(d,n)43Sc performed at the 6 MV HVEC EN-Tandem of the Ion Beam Physics group at ETH in Zürich. A study of the production yield by using commercially available enriched target materials is also presented.

Drug-loadable Mesoporous Bioactive Glass Nanospheres: Biodistribution, Clearance, BRL Cellular Location and Systemic Risk Assessment via (45)Ca Labelling and Histological Analysis.

Mesoporous bioactive glass (MBG) nanospheres with excellent drug loading property have attracted significant attention in the field of nano-medicine. However, systemic metabolism and biosafety of MBG nanospheres which are crucial issues for clinical application are yet to be fully understood. Isotope quantitative tracing combined with biochemical parameters and histopatological changes were used to analyze biodistribution, excretion path and the effect on metabolism and major organs, and then we focused on the hepatocellular location and damaging effect of MBG. The results indicated MBG possessed a longer residence time in blood. After being cleared from circulation, nanospheres were mainly distributed in the liver and were slightly internalized in the form of exogenous phagosome by hepatocyte, whereby more than 96% of nanospheres were located in the cytoplasm (nearly no nuclear involvement). A little MBG was transferred into the mitochondria, but did not cause ROS reaction. Furthermore, no abnormal metabolism and histopathological changes was observed. The accumulation of MBG nanospheres in various organs were excreted mainly through feces. This study revealed comprehensively the systemic metabolism of drug-loadable MBG nanospheres and showed nanospheres have no obvious biological risk, which provides a scientific basis for developing MBG nanospheres as a new drug delivery in clinical application.

Palliative care of bone pain due to skeletal metastases: Exploring newer avenues using neutron activated (45)Ca.

INTRODUCTION: With an objective to develop a cost-effective radiochemical formulation for palliation of pain due to skeletal metastases, we have demonstrated a viable method for large-scale production of (45)Ca (t½=163 days, Eßmax=0.3MeV) using moderate flux research reactor, its purification from radionuclidic impurities adopting electrochemical approach and preclinical evaluation of (45)CaCl2. METHODS: Irradiation parameters were optimized by theoretical calculations for production of (45)Ca with highest possible specific activity along with minimum radionuclidic impurity burden. Based on this, the radioisotope was produced in reactor by irradiation of isotopically enriched (98% in (44)Ca) CaO target at a thermal neutron flux of ~1 × 10(14) n.cm(-2).s(-1) for 4 months. Scandium-46 impurity co-produced along with (45)Ca was efficiently removed adopting an electrochemical separation approach. The bone specificity of (45)CaCl2 was established by in vitro studies involving its uptake in hydroxyapatite (HA) particles and also evaluating its biodistribution pattern over a period of 2 weeks after in vivo administration in Wistar rats. RESULTS: Thermal neutron irradiation of 100mg of enriched (98% in (44)Ca) CaO target followed by radiochemical processing and electrochemical purification procedure yielded ~37 GBq of (45)Ca with a specific activity of ~370 MBq/mg and radionuclidic purity>99.99%. The reliability and reproducibility of this approach were amply demonstrated by process demonstration in several batches. In vitro studies indicated significant uptake of (45)CaCl2 (up to 65%) in HA particles. In vivo biodistribution studies in Wistar rats showed specific skeletal accumulation (40-46%ID) with good retention over a period of 2 weeks. CONCLUSIONS: To the best of our knowledge, this is the first study on utilization of (45)CaCl2 in the context of nuclear medicine. The results obtained in this study hold promise and warrant further investigations for future translation of (45)CaCl2 to the clinics, thereby potentially enabling a cost-effective approach for metastatic bone pain palliation especially in developing countries.

Calcium isolation from large-volume human urine samples for 41Ca analysis by accelerator mass spectrometry.

Calcium oxalate precipitation is the first step in preparation of biological samples for (41)Ca analysis by accelerator mass spectrometry. A simplified protocol for large-volume human urine samples was characterized, with statistically significant increases in ion current and decreases in interference. This large-volume assay minimizes cost and effort and maximizes time after (41)Ca administration during which human samples, collected over a lifetime, provide (41)Ca:Ca ratios that are significantly above background.

Transport and distribution of (45)Ca(2+) in the perfused rat liver and the influence of adjuvant-induced arthritis.

The purpose of the present work was to investigate Ca(2+) transport and distribution under the conditions of the intact rat liver in health and disease (adjuvant-induced arthritis). The multiple-indicator dilution technique was used with the simultaneous injection of (45)Ca(2+) and indicators into the portal vein under defined conditions and analysis of the outflow profiles by means of a space-distributed variable transit time model. The best description of the (45)Ca(2+) outflow profiles corresponds to a model that assumes rapid distribution of (45)Ca(2+) between the vascular space and the cell surface and a slower transfer into the hepatocytes. In kinetic terms two distinct cellular pools were distinguishable, the cytosol and the endoplasmic reticulum. The concentration of Ca(2+) in the cytosol was much lower than in the vascular space and in the endoplasmic reticulum. The most prominent modification observed in the livers of arthritic rats was the increased Ca(2+) concentration in the hormone-sensitive cellular pool. Furthermore, reduced rates of Ca(2+) influx and efflux between the hormone-sensitive cellular pool and the cytosolic space were also detected in combination with a significantly reduced expression of the sarco-endoplasmic reticulum Ca(2+)-ATPase (SERCA2) protein. All these observations mean that in livers from arthritic rats more time is required to replenish the hormone sensitive Ca(2+) stores.

In vivo stability of hydroxyapatite nanoparticles coated on titanium implant surfaces.

PURPOSE: Nanotechnology has been employed in attempts to enhance bone incorporation of dental implants. Often, nanoparticles are applied to the implant surface as particle coatings. However, the same properties that may increase the functionality may also lead to undiscovered negative effects, such as instability of the nanocoating. The aim of this study was to investigate the stability/instability of the nanoparticles using a radiolabeling technique. MATERIALS AND METHODS: Twenty threaded and turned titanium microimplants were inserted in 10 rats. All 20 implants were coated with nanometer-sized hydroxyapatite (HA) particles. In order to trace the HA nanoparticles, the particles for 16 implants were labeled with calcium 45 (45Ca). After 1, 2, 4, and 8 weeks, the implants and surrounding bone were retrieved and analyzed using autoradiography with respect to particle migration from the implant surface. Samples from the brain, liver, thymus, kidney, and blood, as well as wooden shavings from the rats' cages, were also retrieved and analyzed using liquid scintillation counting. RESULTS: The radioactivity representing the localization of 45Ca decreased over time from the vicinity of the implant. The amounts of 45Ca found in the blood and in the rats' excretions decreased with time and corresponded well to each other. After 8 weeks, the only trace of 45Ca was found in the liver. CONCLUSION: The results indicated that released particles leave the body through the natural cleaning system, and the probability that the nanocoating will assemble in vital organs and thus become a potential biologic risk factor is unlikely.

Structural insight into Ca2+ specificity in tetrameric cation channels.

Apparent blockage of monovalent cation currents by the permeating blocker Ca(2+) is a physiologically essential phenomenon relevant to cyclic nucleotide-gated (CNG) channels. The recently determined crystal structure of a bacterial homolog of CNG channel pores, the NaK channel, revealed a Ca(2+) binding site at the extracellular entrance to the selectivity filter. This site is not formed by the side-chain carboxylate groups from the conserved acidic residue, Asp-66 in NaK, conventionally thought to directly chelate Ca(2+) in CNG channels, but rather by the backbone carbonyl groups of residue Gly-67. Here we present a detailed structural analysis of the NaK channel with a focus on Ca(2+) permeability and blockage. Our results confirm that the Asp-66 residue, although not involved in direct chelation of Ca(2+), plays an essential role in external Ca(2+) binding. Furthermore, we give evidence for the presence of a second Ca(2+) binding site within the NaK selectivity filter where monovalent cations also bind, providing a structural basis for Ca(2+) permeation through the NaK pore. Compared with other Ca(2+)-binding proteins, both sites in NaK present a novel mode of Ca(2+) chelation, using only backbone carbonyl oxygen atoms from residues in the selectivity filter. The external site is under indirect control by an acidic residue (Asp-66), making it Ca(2+)-specific. These findings give us a glimpse of the possible underlying mechanisms allowing Ca(2+) to act both as a permeating ion and blocker of CNG channels and raise the possibility of a similar chemistry governing Ca(2+) chelation in Ca(2+) channels.

Determination of Vitamin D-dependent calcium absorption by 45Ca gavage in the rat.

Precise determination of Vitamin D-dependent intestinal calcium absorption in longitudinal studies is problematic. We have assessed Vitamin D-dependent intestinal calcium absorption by (45)Ca gavage. Rats were gavaged with a 1mL solution containing (45)Ca (CaCl(2), 9.3MBq/mL) maintained at 37 degrees C. Total Ca concentration of the gavage fluid was optimised by comparing the absorption curves for fluids made up to 0.025, 2.025, 4.025 and 40.025 mmol/L with (40)CaCl(2). The effect of varying dietary Ca on fractional Ca absorption was determined in rats fed semi-synthetic diets containing either 0.05%, 0.2%, 0.4% or 1.0% Ca for 50 days. Serum 1,25 dihydroxyvitamin D (1,25D) was determined by radioimmunoassay. Total gavage Ca of 0.025 mmol/L achieved the highest peak fractional absorption and was adopted for all future experiments. Fifty days after allocation to the diets both fractional Ca absorption and 1,25D were highest in rats fed 0.05% Ca and lowest in those fed 1.0% Ca (absorption, P<0.05 and 1,25D, P<0.05). There was a strong logarithmic relationship between 1,25D and fractional Ca absorption (R(2) 0.69, P<0.001). Weekly repetition of the procedure did not cause a fall in haematocrit over 7 weeks. Radiocalcium ((45)Ca) absorption by gavage provides a simple measure of Vitamin D-dependent Ca absorption for repetitive use in longitudinal studies.

Counting individual 41Ca atoms with a magneto-optical trap.

Atom trap trace analysis, a novel method based upon laser trapping and cooling, is used to count individual atoms of 41Ca present in biomedical samples with isotopic abundance levels between 10(-8) and 10(-10). The method is calibrated against resonance ionization mass spectrometry, demonstrating good agreement between the two methods. The present system has a counting efficiency of 2x10(-7). Within 1 h of observation time, its 3-sigma detection limit on the isotopic abundance of 41Ca reaches 4.5x10(-10).

Influence of monovalent cation identity on parvalbumin divalent ion-binding properties.

Rat alpha- and beta-parvalbumins have distinct monovalent cation-binding properties [Henzl et al. (2000) Biochemistry 39, 5859-5867]. Beta binds two Na(+) or one K(+), and alpha binds one Na(+) and no K(+). Ca(2+) abolishes these binding events, suggesting that the monovalent ions occupy the EF-hand motifs. This study compares alpha and beta divalent ion affinities in Na(+) and K(+) solutions. Solvent cation identity seriously affects alpha. In Hepes-buffered NaCl, at 5 degrees C, the macroscopic Ca(2+)-binding constants are 2.6 x 10(8) and 6.4 x 10(7) M(-1) and the Mg(2+) constants, 1.8 x 10(4) and 4.3 x 10(3) M(-1). In Hepes-buffered KCl, the Ca(2+) values increase to 2.9 x 10(9) and 6.6 x 10(8) M(-1) and the Mg(2+) values to 2.2 x 10(5) and 3.7 x 10(4) M(-1). Monte Carlo simulation of alpha binding data-employing site-specific constants and explicitly considering Na(+) binding-yields a K(Na) of 630 M(-1) and indicates that divalent ion-binding is positively cooperative. NMR data suggest that the lone Na(+) ion occupies the CD loop. Solvent cation identity has a smaller impact on beta. In Na(+), the Ca(2+) constants for the EF and CD sites are 2.3 x 10(7) and 1.5 x 10(6) M(-1), respectively; the Mg(2+) constants are 9.2 x 10(3) and 1.7 x 10(2) M(-1). In K(+), these values shift to 3.1 x 10(7) and 3.8 x 10(6) M(-1) and the latter to 1.4 x 10(4) and 2.9 x 10(2) M(-1). These data suggest that parvalbumin divalent ion affinity, particularly that of rat alpha, can be significantly attenuated by increased intracellular Na(+) levels.

Bioavailability of the calcium in fortified soy imitation milk, with some observations on method.

BACKGROUND: Calcium-fortified soy milk is growing in popularity, particularly among vegetarians, but the bioavailability of its calcium was not previously known. Additionally, the validity of isotopic labeling methods for fortified liquid products had not been established. OBJECTIVES: The objectives of this study were to compare the bioavailability of the calcium in fortified soy milk with that of calcium in cow milk and to evaluate the method of labeling soy milk for bioavailability testing. DESIGN: A within-subject comparison of extrinsically labeled cow milk with intrinsically and extrinsically labeled soy milks was undertaken in 16 healthy men. In all tests, 300-mg Ca loads were given as a part of a light breakfast after an overnight fast. The milks were physically partitioned into liquid and solid phases to enable evaluation of tracer distribution. RESULTS: Calcium from intrinsically labeled soy milk was absorbed at only 75% the efficiency of calcium from cow milk. Extrinsic labeling of soy milk did not produce uniform tracer distribution throughout the liquid and solid phases and resulted in a 50% overestimate of true absorbability. CONCLUSION: Calcium-fortified soy milk does not constitute a calcium source comparable to cow milk, and extrinsic labeling of such calcium particulate suspensions does not produce the uniform tracer distribution needed for bioavailability testing. Hence, intrinsic labeling of the fortificant is required for such liquid suspensions.