Technetium-99m distribution into Klebsiella pneumoniae.

Bacteria labelled with radionuclide has been the subject of much investigation and has been applied in microbiological research. Technetium-99m (99mTc) may be an alternative radionuclide for the labelling of bacteria employed in various microbiological procedures. This radionuclide is easily available, is not expensive and presents important physical and biological characteristics. 99mTc-labelled bacteria are stable and their cell viability and biological properties are not modified. Study of the distribution of radioactivity in 99mTc-labelled Klebsiella pneumoniae cultures, after homogenization and differential centrifugation of the cells fractions, showed that this radionuclide was present inside the cell, mainly in a ribosomal fraction. Treatment of these fractions with enzymes and detergent revealed a high sensitivity to pronase and Triton X-100. After phenol extraction, a large percentage of radioactivity was detected in the phenol phase. Treatment of the soluble fraction with trichloroacetic acid at different temperatures showed that the concentration of 99mTc in the precipitate was lower at 100 degrees than at 4 degrees C. These results suggest that 99mTc binds mainly to the proteins in Klebsiella pneumoniae.     

A new method for labelling of monoclonal antibodies and their fragments with technetium-99m

A new method of labelling of antibodies with technetium-99m is described. The antibody or fragment is derivatized by iminothiolane. Ligand exchange from a labile technetium complex such as TcO(GLU)₂⁻ results in >90% of the technetium labelling the antibody. This method appears to be equally applicable to antibodies and fragments.     

Uptake kinetics of 99Tc in common duckweed

The uptake of the nuclear waste product technetium-99 was studied in common duckweed (Lemna minor). In addition to measurements, a model involving two compartments in duckweed with different chemical forms of technetium was derived. The model was tested by chemical speciation, i.e. differentiating between reduced Tc-compounds and Tc(VII)O(4)(-). The TcO(4)(-) concentrations measured were in good agreement with those predicted by the model. Two processes determine technetium uptake: (1) transport of Tc(VII)O(4)(-) across the cell membrane, and (2) reduction of Tc(VII). The TcO(4)(-) concentration in duckweed reaches a steady state within 2 h while reduced Tc-compounds are stored, as a result of absence of release or re-oxidation processes. Bioaccumulation kinetic properties were derived by varying 99Tc concentration, temperature, nutrient concentrations, and light intensity. The reduction of technetium in duckweed was highly correlated with light intensity and temperature. At 25 degrees C the maximum reduction rate was observed at light intensities above 200 μmol m(-2) s(-1) while half of the maximum transformation rate was reached at 41 μmol m(-2) s(-1). Transport of TcO(4)(-) over the cell membrane requires about 9.4 kJ mol(-1), indicating an active transport mechanism. However, this mechanism behaved as first-order kinetics instead of Michaelis-Menten kinetics between 1x10(-14) and 2.5x10(-5) mol l(-1) TcO(4)(-). Tc uptake could not be inhibited by 10(-3) mol l(-1) nitrate, phosphate, sulphate or chloride.     

Development of 99mTc radiolabeled A85380 derivatives targeting cerebral nicotinic acetylcholine receptor: Novel radiopharmaceutical ligand 99mTc-A-YN-IDA-C4

Nicotinic acetylcholine receptors (nAChRs) are pentameric ligand-gated ion channels that have been implicated in higher brain functions. To elucidate the functional mechanisms underlying nAChRs and contribute significantly to development of drugs targeting neurological and neuropsychiatric diseases, non-invasive nuclear medical imaging can be used for evaluation. In addition, technetium-99m (^99mTc) is a versatile radionuclide used clinically as a tracer in single-photon emission computed tomography. Because A85380 is known as a potent α4β2-nAChR agonist, we prepared A85380 derivatives labeled with ^99mTc using a bifunctional chelate system. A computational scientific approach was used to design the probe efficiently. We used non-radioactive rhenium (Re) for a ^99mTc analog and found that one of the derivatives, Re-A-YN-IDA-C4, exhibited high binding affinity at α4β2-nAChR in both the docking simulation (?19.3 kcal/mol) and binding assay (Ki = 0.4 ± 0.04 nM). Further, ^99mTc-A-YN-IDA-C4 was synthesized using microwaves, and its properties were examined. Consequently, we found that ^99mTc-A-YN-IDA-C4, with a structure optimized by using computational chemistry techniques, maintained affinity and selectivity for nAChR in vitro and possessed efficient characteristics as a nuclear medicine molecular imaging probe, demonstrated usefulness of computational scientific approach for molecular improvement strategy.     

Single isomer technetium-99m tamoxifen conjugates

To produce an imaging agent for breast cancer using a technetium-99m-labeled agent specific for estrogen receptors, an N(2)S(2) bifunctional chelator was conjugated to Z- and E-aminotamoxifens through an amide linker. These bioconjugates have been chelated with both technetium-99m and rhenium. For the Z-isomer, chelation with rhenium in the presence of sodium acetate yields a mixture of two isomers, anti and syn, in a 1:1 ratio and in the presence of hydroxide results in only the anti isomer. Both the Z- and E-tamoxifen conjugates have been chelated with technetium-99m at the tracer level yielding a single isomer product, which is assigned as anti based on chromatographic comparison to the rhenium complexes. Radiochemical yields were consistently greater than 80%, with Sep-Pak column purification yielding a final product with >99% radiochemical purity and no residual starting material. Both in vitro and in vivo biological evaluation of the tamoxifen chelates indicated very limited estrogen receptor binding.     

An N2S2 bifunctional chelator for technetium-99m and rhenium: complexation, conjugation, and epimerization to a single isomer.

A bifunctional chelator 6 was prepared bearing an N2S2 core for binding rhenium or technetium and a carboxylic acid group for conjugation to amino groups of biomolecules. Complexation of 6 with rhenium(V) resulted in two kinetic isomers, anti-7 and syn-7, being formed in approximately equal amounts. Epimerization with 0.5 M NaOH yields a single isomer anti-7, as determined by NMR spectroscopy and single-crystal X-ray analysis. The 99mTc complex was prepared at the tracer level by reaction of the ligand with 99mTcO4-, tin(II) chloride and sodium gluconate giving a mixture of two isomers, but showing a preference for the anti isomer. Chelation in the presence of 1 M NaOH results in anti-8 being formed as the sole product. The bifunctional ability of the ligand was explored by amide formation with (S)-alpha-phenethylamine, either by direct DCC coupling or through the N-hydroxy succinimidyl ester 9 intermediate. The deprotected bioconjugate 11 was complexed with rhenium, yielding similar amounts of two isomeric rhenium complexes, anti-12 and syn-12, which were isolated and characterized by NMR spectroscopy. Treatment of the kinetic mixture of anti-12 and syn-12 with 1 M NaOH resulted in quantitative conversion to a single rhenium complex anti-12. With technetium-99m in 0.1 M sodium acetate, bioconjugate 11 yielded both technetium-99m complexes anti-13 and syn-13, in a 2:1 ratio, respectively. In contrast, complexation in the presence of 1 M NaOH gave only one technetium-99m complex, assigned the structure anti-13.     

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 β-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 persistant 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.     

Predicting the behaviour of pesticides in soil from their physical and chemical properties

Increasingly stringent environmental requirements for pesticides mean that both biological activity and favourable environmental behaviour must be assessed at an early stage in pesticide discovery. Soil behaviour is governed by the physical properties of the molecule: partition coefficient, dissociation constant, vapour pressure and melting point, which control potential movement under particular soil and environmental conditions and the soil persistence. Established chemical structure-physical property correlations generally allow physical properties to be estimated for the large number of compounds in a synthesis programme with adequate precision. Stability to chemical or biological transformations in soil is more difficult to estimate but a combination of measurement for a few compounds and analogy with known chemical and biological transformation rates for various functional groups can give useful structure-stability correlations.     

Radiolabeling of HYNIC-annexin V with technetium-99m for in vivo imaging of apoptosis

Apoptosis is a critical factor in AIDS and other viral illnesses, cerebral and myocardial ischemia, autoimmune and neurodegenerative states, organ and bone marrow transplant rejection, and tumor response to chemotherapy and radiation. Improved methods to identify sites of apoptosis are increasing our understanding of the pathophysiology and treatment of these and numerous other human disorders. Here we describe the most used method for labeling annexin V, a protein with a high affinity for apoptotic cells in vitro, with technetium-99m (99mTc) as a radionuclide imaging agent that can localize and non-invasively quantify apoptosis in vivo when coupled with single-photon emission tomography. In this method, annexin V is first attached to the bifunctional chelator molecule hydrazino nicotinate (HYNIC). Once prepared, HYNIC-annexin V can be labeled with 99mTc, a widely available gamma-radiation-emitting radionuclide, for intravenous injection in as little as 30 min without the need for specialized reagents or equipment.     

Bioremediation of radioactive waste: radionuclide-microbe interactions in laboratory and field-scale studies

Given the scale of the contamination associated with 60 years of global nuclear activity, and the inherent high financial and environmental costs associated with invasive physical and chemical clean-up strategies, there is an unparalleled interest in new passive in situ bioremediation processes for sites contaminated with nuclear waste. Many of these processes rely on successfully harnessing newly discovered natural biogeochemical cycles for key radionuclides and fission products. Recent advances have been made in understanding the microbial colonization of radioactive environments and the biological basis of microbial transformations of radioactive waste in these settings.     

A new method for protein labeling with 99mTc

A method is described for labeling proteins with ^99mTc, the radionuclide of choice in diagnostic nuclear medicine. Labeling efficiency, stability of label attachment and retained biological behaviour, e.g. immunoreactivity of monoclonal antibodies after radiolabeling are demonstrated. An application of a ^99mTc-labeled anti-fibrin monoclonal antibody in radioimmunoimaging of thrombi is presented.     

In vivo exposure of human lymphocytes to technetium-99m in nuclear medicine patients does not induce detectable genetic effects.

Recent reports have demonstrated that exposure of nuclear medicine patients to thallium-201 does not result in a detectable increase in mutation at the hprt locus in human lymphocytes. In an effort to study further the potential genetic effects of medical exposures to low dose radiation, we have examined chromosome aberrations and mutations in peripheral blood lymphocytes from nuclear medicine patients exposed to clinical doses of technetium-99m. Our results show that there is no exposure-related increase in chromosomal damage; furthermore, the data do not confirm earlier reports of exposure-related increases in mutations induced by technetium-99m.     

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 99mTc, 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 susceptible 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.     

Microbial detoxification of metals and radionuclides

Microorganisms have important roles in the biogeochemical cycling of toxic metals and radionuclides. Recent advances have been made in understanding metal-microbe interactions and new applications of these processes to the detoxification of metal and radionuclide contamination have been developed.     

Chemical and physical properties of radionuclides

There are many radionuclides with a wide range of energies and half-lives available for use as non-sealed radiotherapeutic agents. To date, no single radionuclide has emerged as being clearly superior to all others, in the way that ^99mTc predominates in diagnostic imaging. It is unlikely that one will emerge. Instead, if a particular application demands certain decay properties, the radionuclide which will be used will be the one for which appropriate chemistry can be developed and which can be produced and distributed most economically.     

Geochemical ecology and problems of health.

The state of health or disease is determined by the nature of the organism, the properties of the biosphere, the heterogeneity of its natural geochemical composition and changes brought about by technology (technogenic changes). For a systematic study of the conditions of health and endemic diseases we have suggested a system of biogeochemical regionalizing of the biosphere with the aid of biospheric taxa: regions of the biosphere, subregions of the biosphere, biogeochemical provinces. The main criteria of the regionalizing are biogenous cycles of chemical elements (links of the biogeochemical food chain from soil-forming rocks to man). An important criterion of the biogeochemical regionalizing is threshold concentrations of chemical elements. The organism regulates its metabolism within the ranges of chemical element concentration between the upper and lower thresholds (necessity range). When chemical elements are present in concentrations above the upper threshold and below the lower threshold, dysfunctions and endemic diseases are observed. Hence, the biogeochemical food chain allows us to establish critical links responsible for the state of health or endemic disease. Principles of optimizing the conditions of the environment and life have been worked out. The creation by us in the U.S.S.R. of biogeochemical maps relating conditions of the environment to biological reactions of organisms has proved a useful method of studying the ecological structure of the biosphere.     

The use of scintigraphic imaging as a tool in the development of liposome formulations

Scintigraphic imaging is a valuable tool that can be used during the development of liposome-based therapeutic agents. It provides the ability to non-invasively track and quantitate the distribution of liposomes in the body. This review article provides a general overview of the methods involved in producing scintigraphic images as well as methods of radiolabeling liposomes. Liposomes labeled with technetium-99m ((99m)Tc) are particularly useful for scintigraphic imaging due to the physical characteristics of (99m)Tc, which provides a high quality image. Examples of how scintigraphic imaging studies have contributed to the development of a variety of liposome-based formulations are covered in this article. These liposome formulations include long-circulating liposome-based oxygen carriers, liposome-based anti-cancer drugs, liposomes encapsulating antibiotics and anti-fungals, and liposomes targeted to lymph nodes. Studies using scintigraphic imaging for the investigation of immune responses to liposomes are also discussed. These examples demonstrate the usefulness of scintigraphic imaging for the development of novel liposome formulations.     

Chemical taphonomy of biomineralized tissues

Biomineralized tissues are chemically altered after death, and this diagenetic alteration can obscure original biological chemical features or provide new chemical information about the depositional environment. To use the chemistry of fossil biominerals to reconstruct biological, environmental or taphonomic information, a solid appreciation of biomineralization, mineral diagenesis and biomineral–water interaction is needed. Here, I summarize the key recent developments in the fields of biomineralization and post‐mortem trace element exchange that have significant implications for our understanding of the diagenetic behaviour of biominerals and the ways in which biomineral chemistry can be used in palaeontological and taphonomic research.     

Bioinspired polymeric materials: in-between proteins and plastics

Chemical and biological researchers are making rapid progress in the design and synthesis of non-natural oligomers and polymers that emulate the properties of natural proteins. Whereas molecular biologists are exploring biosynthetic routes to non-natural proteins with controlled material properties, synthetic polymer chemists are developing bioinspired materials with well-defined chemical and physical properties that function or self-organize according to defined molecular architectures. Bioorganic chemists, on the other hand, are developing several new classes of non-natural oligomers that are bridging the gap between molecular biology and polymer chemistry. These synthetic oligomers have both sidechain and length specificity, and, in some cases, demonstrate capability for folding, self-assembly, and specific biorecognition. Continued active exploration of diverse backbone and sidechain chemistries and connectivities in bioinspired oligomers will offer the potential for self-organized materials with greater chemical diversity and biostability than natural peptides. Taken together, advances in molecular bioengineering, polymer chemistry, and bioorganic chemistry are converging towards the creation of useful bioinspired materials with defined molecular properties.     

An introduction to peptide nucleic acid

Peptide Nucleic Acid (PNA) is a powerful new biomolecular tool with a wide range of important applications. PNA mimics the behaviour of DNA and binds complementary nucleic acid strands. The unique chemical, physical and biological properties of PNA have been exploited to produce powerful biomolecular tools, antisense and antigene agents, molecular probes and biosensors.