G. E. Meinken

Biodistribution of Radioiodinated Adenovirus Fiber Protein Knob Domain after Intravenous Injection in Mice

ABSTRACT The knob domains from the fiber proteins of adenovirus serotypes 2 and 12 were labeled with radioiodine and then injected into the bloodstreams of mice. Knob proteins with functional binding sites for the coxsackie and adenovirus receptor (CAR) were cleared rapidly from the circulation, with radioactivity appearing predominantly in the stomach, while knob mutants unable to bind to CAR remained in the blood circulation for a prolonged period. The clearance of radiolabeled wild-type knob from the blood was slowed by coinjecting an excess of unlabeled wild-type knob protein. An earlier study showed that 99mTc-labeled knob protein with intact CAR-binding activity also cleared rapidly from the blood circulation of mice, with radioactivity accumulating predominantly in the liver (K. R. Zinn et al., Gene Ther. 5:798-808, 1998). Together these results suggest that rapid clearance of knob protein from the blood results from specific binding to CAR in the liver and that the bound knob then enters a degradative pathway. The elevated levels of radioiodine in the stomach observed in our experiments are consistent with deiodination of labeled knob by dehalogenases in hepatocyte microsomes and uptake of the resultant free radioiodine by Na/I symporters in the gastric mucosa. Although CAR has been shown to localize in tight junctions of polarized epithelial cells, where it functions in intercellular adhesion, the results of our study suggest that a subset of CAR molecules in the liver is highly accessible to ligands in the blood and able to rapidly deliver bound ligand to an intracellular degradative compartment.

Development of a large scale production of 67Cu from 68Zn at the high energy proton accelerator: closing the 68Zn cycle.

A number of research irradiations of (68)Zn was carried out at Brookhaven Linac Isotope Producer aiming to develop a practical approach to produce the radioisotope (67)Cu through the high energy (68)Zn(p,2p)(67)Cu reaction. Disks of enriched zinc were prepared by electrodeposition of (68)Zn on aluminum or titanium substrate and isolated in the aluminum capsule for irradition. Irradiations were carried out with 128, 105 and 92 MeV protons for at least 24h. After irradiation the disk was chemically processed to measure production yield and specific activity of (67)Cu and to reclaim the target material. The recovered (68)Zn was irradiated and processed again. The chemical procedure comprised BioRad cation exchange, Chelex-100 and anion exchange columns. Reduction of the oxidation degree of copper allowed for more efficient Cu/Co/Zn separation on the anion exchange column. No radionuclides other than copper isotopes were detected in the final product. The chemical yield of (67)Cu reached 92-95% under remote handling conditions in a hot box. Production yield of (67)Cu averaged 29.2 μCi/[μA-h×g (68)Zn] (1.08MBq/[μA-h×g (68)Zn]) in 24h irradiations. The best specific activity achieved was 18.6 mCi/μg (688.2 MBq/μg).

The reduction of pertechnetate-99 by stannous chloride—II. The stoichiometry of the reaction in aqueous solutions of several phosphorus(V) compounds

Abstract The reduction of pertechnetate-99 by excess stannous chloride in 1 M sodium phosphate at pH 7 produced a mixed-metal complex of Tc(III), Tc(IV), and Sn(II). This was shown by polarography and spectrophotometry. Coulometry at fixed potential produced a Tc(III) complex which was air-oxidized to a Tc(IV) complex, in agreement with other reports. In 0.2 M sodium pyrophosphate at pH 7, both coulometry at fixed potential and reduction by excess SnCl 2 produced compounds which slowly turned deep blue. From their identical visible absorption spectra, it was tentatively concluded that they were the same, as were their oxidation products (pink), and from the coulometry it was concluded that Tc(III) has been formed. In 1 M potassium pyrophosphate at pH 7, excess SnCl 2 and coulometry produced almost colorless solutions; dilution to 0.2 M pyrophosphate did not cause the blue complex to appear. Qualitatively, it was shown that the pyrophosphate complex was more stable than one formed in a sodium polyphosphate solution. The close resemblance between the spectrum of Tc(III) phosphate and that of Tc(III) pyrophosphate was explained by assuming that the phosphate complex was formed from a hydrogen-bonded dimer of phosphoric acid.

Biotransformation of pertechnetate by Clostridia

Summary Clostridia are strict anaerobic, spore-forming, fermentative bacteria commonly present in soils, sediments, and wastes; and, they play a major role in the decomposition of a wide variety of organic compounds. They also are involved in the reduction of iron, manganese, and uranium, thereby affecting their solubility. However, little is known of the ability of Clostridia to reduce technetium (Tc). We investigated the reduction and precipitation of pertechnetate by Clostridium sphenoides able to metabolize citrate as its sole carbon source, and Clostridium sp. capable of fermenting glucose but not citric acid. Both species reduced Tc(VII) to Tc(IV), although C. sphenoides did so at a greater rate and extent than Clostridium sp. The reduced Tc was predominantly associated with the cell biomass. It also was present in solution complexed with bacterial metabolic products (MW>5000). Adding diethylenetriaminepentaacetic acid (DTPA) to Clostridium sp. resulted in the formation of a soluble Tc(IV)-DTPA complex, whereas with C. sphenoides only a small amount of Tc was present in solution, indicating that insoluble Tc species were formed. These results suggest that Clostridia may play a major role in regulating the mobility of Tc under anaerobic conditions in wastes and subsurface environments.

Correlating labeling chemistry and in-vitro test results with the biological behavior of radiolabeled proteins

A study of the effect of various rediochemical labeling parameters on the in-vivo behavior of proteins, in particular of monoclonal antibodies, was carried out. Both radioiodination, and radiometal labeling (using protein-chelating agent conjugates), of antimelanoma, antiplatelet, and anticolon carcinoma monoclonal antibodies (222.28s, 7E3, and GA-733 respectively), as well as the direct labeling of human serum albumin with 99m Tc, were investigated. Different aspects of the biological behavior are affected in relation to the labeling chemistry involved. These include the immunoreactivity, blood clearance and tissue uptake kinetics, and rates and routes of excretion. Individual radionuclide effects have often to be addressed separately. Some antibodies are more susceptable to alteration from labeling conditions than others. Careful optimization of labeling and purification procedures is thus necessary for particular radionuclide/antibody combinations in order to obtain predictable and reproducible in-vivo results for both immunoscintigraphy and immunotherapy applications.

AN IODINE-123 GENERATOR/IODINATION KIT: A PRELIMINARY REPORT*

Preliminary results are described of a123Xe filled device to serve as a combination123I generator/iodination kit.123X was produced in the Brookhaven Linac Isotope Producer (BLIP) by the reaction127I(p, 5n)123Xe. The device consists of a small glass ampoule containing an internal glass breakseal and a flanged neck on which was crimped a multi-injection type septum. The ampoule contained a hydrogen sulfide atmosphere to assure that the iodine generated from the decay of the xenon was in the form of iodide. Following an adequate period for123Xe to decay (this period can be used for shipment), a needle is forced through the septum breaking the seal and residual gases are pumped off. The123I in the form of iodide can then be rinsed from the ampoule with any desired solvent or reagent added directly to the device to carry out an iodination in an enclosed environment. Preliminary results of both iodine recovery and iodinations have been promising.

123I research and production at Brookhaven National Laboratory

Abstract The procedures for preparing high purity 123 I at the Brookhaven Linac Isotope Producer using the 127 I(p, 5n) 123 Xe reaction on a NaI target are described. The activity is supplied in a glass ampoule with anhydrous 123 I deposited on the interior walls, allowing maximum flexibility in subsequent iodinations. Preliminary experience with a continuous flow target is also described. The results of a series of measurements of specific activity by neutron activation, x-ray fluorescence, u.v. absorption, and wet chemistry generally showed no detectable carrier. HPLC methods to analyze the chemical form of radioiodine and to characterize various iodinated radiopharmaceuticals have been developed. These methods provide higher sensitivity, speed and resolution than commonly used techniques.

Compact CdZnTe-based gamma camera for prostate cancer imaging

In this paper, we discuss the design of a compact gamma camera for high-resolution prostate cancer imaging using Cadmium Zinc Telluride (CdZnTe or CZT) radiation detectors. Prostate cancer is a common disease in men. Nowadays, a blood test measuring the level of prostate specific antigen (PSA) is widely used for screening for the disease in males over 50, followed by (ultrasound) imaging-guided biopsy. However, PSA tests have a high falsepositive rate and ultrasound-guided biopsy has a high likelihood of missing small cancerous tissues. Commercial methods of nuclear medical imaging, e.g. PET and SPECT, can functionally image the organs, and potentially find cancer tissues at early stages, but their applications in diagnosing prostate cancer has been limited by the smallness of the prostate gland and the long working distance between the organ and the detectors comprising these imaging systems. CZT is a semiconductor material with wide band-gap and relatively high electron mobility, and thus can operate at room temperature without additional cooling. CZT detectors are photon-electron direct-conversion devices, thus offering high energy-resolution in detecting gamma rays, enabling energy-resolved imaging, and reducing the background of Compton-scattering events. In addition, CZT material has high stopping power for gamma rays; for medical imaging, a few-mm-thick CZT material provides adequate detection efficiency for many SPECT radiotracers. Because of these advantages, CZT detectors are becoming popular for several SPECT medical-imaging applications. Most recently, we designed a compact gamma camera using CZT detectors coupled to an application-specific-integratedcircuit (ASIC). This camera functions as a trans-rectal probe to image the prostate gland from a distance of only 1-5 cm, thus offering higher detection efficiency and higher spatial resolution. Hence, it potentially can detect prostate cancers at their early stages. The performance tests of this camera have been completed. The results show better than 6-mm resolution at a distance of 1 cm. Details of the test results are discussed in this paper.

Synthesis and biological properties of the lipophilic technetium-99m complex 99mTc(acac)3

In the development of technetium-99m radiopharmaceuticals for the evaluation of regional cerebral perfusion, one series of complexes that has remained unexplored is the neutral lipophilic tris complexes formed with beta-diketonato ligands. The prototype complex of this series, tris(2,4-pentanedionato) technetium(III), has been prepared via a new synthetic route and chemically characterized using 99Tc and the biodistribution of the no-carrier-added 99mTc complex has been determined. The 99mTc complex was found to be distributed throughout the body with persistent high blood levels indicative of a high degree of protein binding. The primary route of excretion was the hepatobiliary system as indicated by the appearance of 99mTc in the gut and feces at longer sample times post-injection. Although this complex was not retained by the brain, it does provide a starting point from which a more effective agent might be developed.

Radiolabeled Antiplatelet Monoclonal Antibodies for the Scintigraphic Localization of In-vivo Thrombi and Vascular Lesions

Detection of thrombi in the extremities and in the trunk, including coronary and carotid thrombi, is extremely important. of equal importance is the ability to visualize the interaction between blood platelets and the arterial wall in thrombosis and intimai damage where platelet deposition occurs. In-vivo imaging would allow: (i) thrombus detection before thrombolytic therapy is instituted, and the monitoring of thrombosis once therapy is discontinued; (ii) monitoring platelet deposition on vessel walls following balloon angioplasty; and (iii) monitoring for acute occlusion of coronary artery bypass grafts. Quantitative imaging could permit visualization of coronary and carotid artery occlusion and thrombogenic and thrombolytic activity within the arterial circulation to the heart, lung, and the brain with better effectiveness.