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Indium-111-labeled antibody heavy metal chelate conjugates: a potential alternative to radioiodination.

An alternative method to radioiodination is outlined for labeling antibodies with a radioactive marker. The method requires the conjugation of the antibody molecule to chelating agents that contain a free amino group and are capable of binding heavy metal ions. Glutaraldehyde is the coupling agent used to bridge the free amino groups on both the chelate and the antibody molecules. 111In is then added to the antibody-chelate conjugate and is bound instantaneously by the chelating portion. Electrophoretic, autoradiographic, immunodiffusion, and hemagglutination techniques were used to confirm the integrity of the 111In-labeled antibodies.

Antigen-Antibody Reactions↗

Significance of skin metallization in the diagnosis of electrocution.

The differentiation of minute current marks from the faint traces of burns may offer some difficulties. Marcinkowski and Wojciechowski (1973) treated the skin of corpses with the same metal objects either heated to a high temperature or exposed to 250 V of alternating current, and determined (by an electrographic method) that metallization appeared only after applying the electric current. The continuation of these observations is linked with the actual experimental studies of Pankowski. By treating the skin of corpses with alternating and direct current of 10, 50, 100 and 250 V for 0.3 sec, 1 sec, 30 sec and 1 min passing through a radioactive electrode of 60Co, he has shown that the radioactivity of the skin at the site of electrode contact increases with the elevation of the voltage and its duration. In the case of direct current the rise was 630-54000-fold at the site of the positive electrode. Using electrodes of copper, aluminium and iron (not radioactive) it has also been shown (by an electrographic method) that metallization intensifies under the same conditions of time and voltage. Metallization could be detected even when no current marks on the skin were evident. Electrography appears to be extremely useful in the detection of metallization. No metallization was detected at the site of the negative electrode (as refers to direct current).

Electric Injuries↗

[Initial results in tumor diagnosis with radioactive lanthanide complexes].

Clinical investigations have been made in 70 patients to determine whether 169Yb-labelled metal ligand complexes accumulate in various tumors as was shown by previous animal experiments. The time course of these radiopharmaceuticals in tumors, the skeleton and the liver permitted a good to excellent tumor imaging in 61/70 cases. Even 2-5 h after intravenous injection of the radioactive compound, a meaningful diagnosis could be made in the majority of cases. A number of lesions were found for which no equivalent findings were available with other methods. They were therefore interpreted as metastases.

Colorectal Neoplasms↗

Radiocesium contamination at a steel plant in Ireland.

Radioactive sources have been inadvertently incorporated into consignments of scrap metal in various locations throughout the world. In 1990, a 3.7 GBq 137 Cs source, due for transfer from a Scottish industrial establishment to one in England, was mistakenly included in a scrap consignment destined for Irish Steel, a metal recycling plant in County Cork in the Republic of Ireland. Unaware of the presence of the source, Irish Steel smelted this consignment in the usual manner. This involved separation of some non-ferrous materials which were then exported, in pellet form, To Pasminco Europe (now Britannia Zinc Ltd.) in Avonmouth, U.K. The presence of 137 Cs contamination in these pellets was detected by the U.K. company in the course of a routine radiation survey. Irish Steel carried out extensive decontamination of its plant, placing the contaminated dust in secure storage. The company has equipped itself with radiation detection devices which monitor incoming scrap. Outgoing products and furnace dust are also monitored on a routine basis. While this incident was of negligible radiological significance as far as personnel were concerned, the financial cost to Irish Steel have been substantial. It highlights the need for surveillance, by national competent authorities, of the movement of radioactive sources from production through use to final disposal.

Cesium Radioisotopes↗

DNA-scission activities of ascorbate in the presence of metal chelates.

A systematic comparison of the DNA-scission activities of various transition metal ions and their metal chelates in the presence of reducing agents has been made. L-Ascorbate in combination with metal ions possesses a much higher scission activity as compared to sulfhydryl-containing reductants despite its low reduction potential. It is demonstrated that DNA-scission activities of metal complexes in the presence of various reducing agents is a general phenomenon for all transition metal ions albeit with different degrees of efficiency. The different reactivities of various metal complexes appear to be attributed, in part, to the differences in their redox electrochemical potentials of the reactions between metal chelates and the reducing agents. Agarose gel electrophoresis coupled with the determination of acid-soluble radioactivity from 3H-labeled DNA provides a rapid qualitative and quantitative assay for the comparison of the reactivities of different metal ions or metal chelates in combination with reductants on the DNA-scission reactions.

Antioxidants↗

"High specific activity" radiotracers for metallo-toxicological studies: cyclotron and nuclear reactor production, radiochemical separation and "quality control": platinum, iridium, gold, copper and gallium.

Very High Specific Activity RadioNuclides, HSARN, are a powerful tool to label a wide variety of chemical elements and compounds present in the biosphere in ultra-trace amounts. Medium and high Z radionuclides, can be produced by irradiation in light-ions accelerator and sometimes nuclear reactor. If the nuclear reaction product has atomic number different from irradiated target, it is possible separating the radioactive nuclide from irradiated target, without addition of isotopic carrier. These kinds of radionuclides are named No Carrier Added, NCA, and their specific activity is very high and can reach values close to the theoretical Carrier Free one. The true specific activity must be determined by use of very sensitive radioanalytical techniques. If a low isotopic dilution factor is obtained, these radiotracers are used to label inorganic species and complexes of elements, which are presently introduced into the echo-systems by human activities. New production methods for NCA Pt, Ir, Au, Cu and Ga radiotracers are presented, with some details on radiochemistry and quality controls.

Copper↗

DTPA-coupled proteins--procedures and precautions.

While demonstrating that the cyclic anhydride of diethylenetriaminepentaacetic acid (DTPA) may be used to covalently attach this chelator to proteins, it was necessary to develop several new procedures and techniques. We were able to show that coupled proteins may be labeled with indium-111 (111In) simply by transcomplexation from a weaker complex such as the acetate to avoid subjecting the protein, even momentarily, to the acidity of the chloride (however, we have also observed that too high a concentration of acetate and other complexing agents such as citrate will interfere with protein labeling by competing with DTPA for the radioactivity). In agreement with other investigators using bifunctional chelate methodology we have observed that trace metals may interfere with the labeling. Accordingly, we developed a simple paper chromatographic assay which may be used to establish whether trace metals are present at interfering concentrations and to identify their sources. We have also employed a hydrolyzed control assay to determine whether and to what degree the radioactivity is bound to protein other than at the DTPA groups. We have developed a simple method of measuring the average number of DTPA groups per protein molecule which involves labeling the products of coupling with 111In. Procedures used and precautions observed in the radiolabeling of proteins by these methods are discussed.

Immunoglobulin G↗

[Examination of gastric juice in animals fed with fodder containing power plant ashes and loess dusts].

Normal metabolism, regulation of stimulation and checking of electrolyte secretion in the gastric juice depend on the way of feeding and its nutritional ingredients. Feeding experimental animals with fodder containing power plant ashes or loess dusts causes a change in the gastric juice reaction, thus resulting in a different assimilation of elements contained in the fodder. There was shown an increase of some elements content (Mg, Na, Cu, Fe), a decrease in the content of some others (Pb,Zn) in the gastric juice and a positive tendency for blocking by calcium ions of heavy metals contained in the fodder by the mucous membrane of the stomach.

Animals↗

Axonal transport of manganese and its relevance to selective neurotoxicity in the rat basal ganglia.

The present study provides evidence for anterograde axonal transport of manganese (Mn) in the basal ganglia. Microinjections of 54Mn into rat substantia nigra or striatum revealed region-specific accumulation and retention of the isotope in globus pallidus, striatum, thalamus and substantia nigra for up to at least 48 or 72 h respectively. Within 4 h after intrastriatal injection of 54Mn, radioactivity accumulated in the substantia nigra, suggesting axonal transport of the metal. Subsequent studies using bilateral 54Mn injections into striatum or substantia nigra and unilateral colchicine injections into or transection of the medial forebrain bundle confirmed axonal transport of Mn through these fibres. Selective destruction of the striatonigral or nigrostriatal pathways using quinolinic acid or 6-hydroxydopamine 2 weeks before injection of the isotope, revealed uptake of 54Mn by cell bodies of both gamma-aminobutyric acidergic striatal and dopaminergic nigral neurons and subsequent anterograde transport through striatonigral or nigrostriatal fibres. In addition, the quinolinic acid-lesioned striatum retained three times more radioactivity than the intact striatum. In conclusion, the present data suggest that both glial cells and striatonigral and nigrostriatal neurons are potential targets for Mn toxicity. These results and the selective neurotoxicity of Mn are discussed with respect to the iron transport protein transferrin, transferrin receptors, the iron storage protein ferritin, and mitochondrial dysfunction.

Animals↗

Altering pharmacokinetics of radiolabeled antibodies by the interposition of metabolizable linkages. Metabolizable linkers and pharmacokinetics of monoclonal antibodies.

Monoclonal antibodies, their fragments and low molecular weight oncophilic molecules such as synthetic somatostatin derivatives have been used to deliver radioactivity to target cells for both diagnostic and therapeutic purposes. Clinical studies demonstrated high abilities of the radiolabeled antibodies and peptides for nuclear medicine applications. However, high and persistent localization of radioactivity was observed in the liver or kidney especially when these molecules are labeled with metallic radionuclides, which reduce diagnostic accuracy and compromise therapeutic effectiveness. Thus, radiolabeled antibodies and peptides would become much more useful in both targeted imaging and radiotherapy if the undesirable radioactivity localization can be diminished. As a means to reduce the undesirable radioactivity, interposition of a metabolizable linkage between an antibody and a radiolabel was proposed to generate radiolabeled small molecules of urinary excretion from the parental antibody by enzymatic cleavage of the linkage. In this paper, after indicating the rationale behind the radiopharmaceutical design, a significant role played by the interposition of the metabolizable linkage in altering pharmacokinetics of radiolabeled antibodies is described from a variety of studies so far reported with an emphasis being laid on the importance of radiometabolite-based design of metabolizable linkages.

Antibodies, Monoclonal↗

Radiation protection and radioactive scales in oil and gas production.

Low specific-activity scales consisting of alkaline earth metal carbonates and sulfates are often present in some gaseous and liquid hydrocarbon plants. These scales contain a certain concentration of radium, uranium, and thorium which can cause a risk of gamma irradiation and internal radiocontamination when they must be mechanically removed. The gamma dose rates and the 238U, 232Th, 226Ra concentrations were determined in sludges, scales, and waters of some gas and oil hydrocarbon plants located in Italy, Congo, and Tunisia. 238U and 232Th concentrations were were low. The isotopes 238U and 234U resulted in radioactive equilibrium, while 232Th and 228Th were not always equilibrium. A rough correlation was found between the gamma dose rate and the 226Ra concentration. Some considerations and conclusions about radiation protection problems are pointed out.

Fossil Fuels↗

Radiative decay engineering: biophysical and biomedical applications.

Fluorescence spectroscopy is a widely used research tool in biochemistry and molecular biology. Fluorescence has also become the dominant method enabling the revolution in medical diagnostics, DNA sequencing, and genomics. To date all the fluorescence observables, including spectral shifts, anisotropies, quantum yields, and lifetimes, have all been utilized in basic and applied uses of fluorescence. In this forward-looking article we describe a new opportunity in fluorescence, radiative decay engineering (RDE). By RDE we mean modifying the emission of fluorophores or chromophores by increasing or decreasing their radiative decay rates. In most fluorescence experiments the radiative rates are not changed because these rates depend on the extinction coefficient of the fluorophore. This intrinsic rate is not changed by quenching and is only weakly dependent on environmental effects. Spectral changes are usually caused by changes in the nonradiative rates resulting from quenching or resonance energy transfer. These processes affect the emission by providing additional routes for decay of the excited states without emission. In contrast to the relatively constant radiative rates in free solution, it is known that the radiative rates can be modified by placing the fluorophores at suitable distances from metallic surfaces and particles. This Review summarizes results from the physics literature which demonstrate the effects of metallic surfaces, colloids, or islands on increasing or decreasing emissive rates, increasing the quantum yields of low quantum yield chromophores, decreasing the lifetimes, and directing the typically isotropic emission in specific directions. These effects are not due to reflection of the emitted photons, but rather as the result of the fluorophore dipole interacting with free electrons in the metal. These interactions change the intensity and temporal and spatial distribution of the radiation. We describe the unusual effects expected from increases in the radiative rates with reference to intrinsic and extrinsic biochemical fluorophores. For instance, the decreased lifetime can result in an effective increase in photostability. Proximity to nearby metallic surfaces can also increase the local field and modify the rate of excitation. We predict that the appropriate localization of fluorophores near particles can result in usefully high emission from "nonfluorescent" molecules and million-fold increases in the number of photons observable from each fluorophore. We also describe how RDE can be applied to medical testing and biotechnology. As one example we predict that nearby metal surfaces can be used to increase the low intrinsic quantum yields of nucleic acids and make unlabeled DNA detectable using its intrinsic metal-enhanced fluorescence.

Energy Transfer↗

Stability of a metabolizable ester bond in radioimmunoconjugates.

Ester bonds have been used as metabolizable linkages to reduce radioactivity levels in non-target tissues following the administration of antibodies labeled with metallic radionuclides. In this radiochemical design of antibodies, while the ester bonds should be cleaved rapidly in non-target tissues, high stability of the ester bonds in plasma is also required to preserve target radioactivity levels. To assess the structural requirements to stabilize the ester bond, a new benzyl-EDTA-derived bifunctional chelating agent with an ester bond, (1-[4-[4-(2- maleimidoethoxy)succinamido]benzyl]ethylenediamine-N,N,N',N' -tetraacetic acid; MESS-Bz-EDTA), was developed. MESS-Bz-EDTA was coupled with a thiolated monoclonal antibody (OST7, IgG1) prepared by reducing its disulfide bonds to introduce the ester bond close and proximal to the antibody molecule. For comparison, 1-[4-(5- maleimidopentyl)aminobenzyl]ethylenediamine-N,N,N',N'-tetraacetic acid (EMCS-Bz-EDTA) and meleimidoethyl 3-[131I]iodohippurate (MIH) was coupled to OST7 under the same conjunction chemistry. When incubated in 50% murine plasma or a buffered-solution of neutral pH, OST7-MESS-Bz-EDTA-111In rapidly released the radioactivity, and more than 95% of the initial radioactivity was liberated after a 24 h incubation in both solutions, due to a cleavage of the ester bond. On the other hand, only about 20% of the radioactivity was released from OST7-MIH-131I in both solutions during the same incubation period. In mice biodistribution studies, while a slightly faster radioactivity clearance from the blood with less radioactivity levels in the liver and kidneys was observed with OST7-MIH-131I than with OST7-EMCS-Bz-EDTA-111In, OST7-MESS-Bz-EDTA-111In indicated radioactivity clearance from the blood much faster than and almost comparable to that of OST7-MIH-131I and succinamidobenzyl-EDTA-111In, respectively. These findings as well as previous findings on radiolabeled antibodies with ester bonds suggested that while an introduction of an ester bond close to an antibody molecule stabilized the ester bond against esterase access, chemical structures of the linkages and radiolabels attached to the ester bonds play a significant role in the chemical stability of the ester bond. This may explain the different stability of the ester bonds in radioimmunoconjugates so far reported.

Animals↗

Recent climate change in the Arctic and its impact on contaminant pathways and interpretation of temporal trend data.

The Arctic has undergone dramatic change during the past decade. The observed changes include atmospheric sea-level pressure, wind fields, sea-ice drift, ice cover, length of melt season, change in precipitation patterns, change in hydrology and change in ocean currents and watermass distribution. It is likely that these primary changes have altered the carbon cycle and biological systems, but the difficulty of observing these together with sporadic, incomplete time series makes it difficult to evaluate what the changes have been. Because contaminants enter global systems and transport through air and water, the changes listed above will clearly alter contaminant pathways. Here, we review what is known about recent changes using the Arctic Oscillation as a proxy to help us understand the forms under which global change will be manifest in the Arctic. For Pb, Cd and Zn, the Arctic is likely to become a more effective trap because precipitation is likely to increase. In the case of Cd, the natural cycle in the ocean appears to have a much greater potential to alter exposure than do human releases of this metal. Mercury has an especially complex cycle in the Arctic including a unique scavenging process (mercury depletion events), biomagnifying foodwebs, and chemical transformations such as methylation. The observation that mercury seems to be increasing in a number of aquatic species whereas atmospheric gaseous mercury shows little sign of change suggests that factors related to change in the physical system (ice cover, permafrost degradation, organic carbon cycling) may be more important than human activities. Organochlorine contaminants offer a surprising array of possibilities for changed pathways. To change in precipitation patterns can be added change in ice cover (air-water exchange), change in food webs either from the top down or from the bottom up (biomagnification), change in the organic carbon cycle and change in diets. Perhaps the most interesting possibility, presently difficult to predict, is combination of immune suppression together with expanding ranges of disease vectors. Finally, biotransport through migratory species is exceptionally vulnerable to changes in migration strength or in migration pathway-in the Arctic, change in the distribution of ice and temperature may already have caused such changes. Hydrocarbons, which tend to impact surfaces, will be mostly affected by change in the ice climate (distribution and drift tracks). Perhaps the most dramatic changes will occur because our view of the Arctic Ocean will change as it becomes more amenable to transport, tourism and mineral exploration on the shelves. Radionuclides have tended not to produce a radiological problem in the Arctic; nevertheless one pathway, the ice, remains a risk because it can accrue, concentrate and transport radio-contaminated sediments. This pathway is sensitive to where ice is produced, what the transport pathways of ice are, and where ice is finally melted-all strong candidates for change during the coming century. The changes that have already occurred in the Arctic and those that are projected to occur have an effect on contaminant time series including direct measurements (air, water, biota) or proxies (sediment cores, ice cores, archive material). Although these 'system' changes can alter the flux and concentrations at given sites in a number of obvious ways, they have been all but ignored in the interpretation of such time series. To understand properly what trends mean, especially in complex 'recorders' such as seals, walrus and polar bears, demands a more thorough approach to time series by collecting data in a number of media coherently. Presently, a major reservoir for contaminants and the one most directly connected to biological uptake in species at greatest risk-the ocean-practically lacks such time series.

Air Movements↗

Modelling the closure-related geochemical evolution of groundwater at a former uranium mine.

A newly developed reactive transport model was used to evaluate the potential effects of mine closure on the geochemical evolution in the aquifer downgradient from a mine site. The simulations were conducted for the Königstein uranium mine located in Saxony, Germany. During decades of operation, uranium at the former mine site had been extracted by in situ acid leaching of the ore underground, while the mine was maintained in a dewatered condition. One option for decommissioning is to allow the groundwater level to rise to its natural level, flooding the mine workings. As a result, pore water containing high concentrations of dissolved metals, radionuclides, and sulfate may be released. Additional contamination may arise due to the dissolution of minerals contained in the aquifer downgradient of the mine. On the other hand, dissolved metals may be attenuated by reactions within the aquifer. The geochemical processes and interactions involved are highly non-linear and their impact on the quality of the groundwater and surface water downstream of the mine is not always intuitive. The multicomponent reactive transport model MIN3P, which can describe mineral dissolution-precipitation reactions, aqueous complexation, and oxidation-reduction reactions, is shown to be a powerful tool for investigating these processes. The predictive capabilities of the model are, however, limited by the availability of key geochemical parameters such as the presence and quantities of primary and secondary mineral phases. Under these conditions, the model can provide valuable insight by means of sensitivity analyses.

Forecasting↗

Radiochemical investigations of 177Lu-DOTA-8-Aoc-BBN[7-14]NH2: an in vitro/in vivo assessment of the targeting ability of this new radiopharmaceutical for PC-3 human prostate cancer cells.

Bombesin (BBN), a 14 amino acid peptide, is an analogue of human gastrin releasing peptide (GRP) that binds to GRP receptors (GRPr) with high affinity and specificity. The GRPr is over expressed on a variety of human cancer cells including prostate, breast, lung, and pancreatic cancers. The specific aim of this study was to identify a BBN analogue that can be radiolabeled with (177)Lu and maintains high specificity for GRPr positive prostate cancer tumors in vivo. A preselected synthetic sequence via solid phase peptide synthesis (SPPS) was designed to produce a DOTA-BBN (DOTA = 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid) conjugate with the following general structure: DOTA-X-Q-W-A-V-G-H-L-M-(NH(2)), where the spacer group, X = omega-NH(2)(CH(2))(7)COOH (8-Aoc). The BBN-construct was purified by reversed phase-HPLC (RP-HPLC). Electrospray Mass Spectrometry (ES-MS) was used to characterize both metallated and non-metallated BBN-conjugates. The new DOTA-conjugate was metallated with (177)Lu(III)Cl(3) or non-radioactive Lu(III)Cl(3). The (177)Lu(III)- and non-radiolabeled Lu(III)-conjugates exhibit the same retention times under identical RP-HPLC conditions. The (177)Lu-DOTA-8-Aoc-BBN[7-14]NH(2) conjugate was found to exhibit optimal pharmacokinetic properties in CF-1 normal mice. In vitro and in vivo models demonstrated the ability of the (177)Lu-DOTA-8-Aoc-BBN[7-14]NH(2) conjugate to specifically target GRP receptors expressed on PC-3 human prostate cancer cells.

Animals↗

Synthesis and characterization of rhenium-complexed alpha-melanotropin analogs.

Receptor binding peptides labeled with medically important radionuclides such as technetium and rhenium are an important tool for the imaging and treatment of many forms of cancer. This paper describes a method of labeling peptides with rhenium using a natural amino acid chelating moiety. The structural characteristics of this chelate moiety, N-acetyl-cysteine-glycine-cysteine-glycine (NAc-CGCG) complexed with nonradioactive rhenium, have been investigated. The stability of this peptide-metal complex has been evaluated on the tracer level using radioactive rhenium-186. The rhenium-bound peptide has been appended to the N termini of receptor binding alpha-melanocyte stimulating hormone (alpha-MSH, NAc-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-NH2) fragments via solid phase peptide synthesis. Bioassays and receptor binding studies of the resulting complexes demonstrate that the fragments retained biological activity and exhibited receptor binding constants ranging from 0.3 to 1.1 nM. This method could provide a general means of labeling bioactive peptide fragments that would simplify product purification and characterization.

Animals↗