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241Am, 137Cs, Sr and Pb uptake by tobacco as influenced by application of Fe chelators to soil.

To determine the potential for phytoextraction of 241Am and other contaminants from soil, accumulation of 241Am, 137Cs, Sr, Fe, Al, Pb, and Mg by tobacco was determined for soil applications of two concentrations of ethylenediaminetetraacetic acid (EDTA), citric acid, and ascorbic acid. In tobacco receiving EDTA at 3.1 mmol/kg of soil, 241Am content of plants averaged 15 Bq/kg (ranging up to 26 Bq/kg) while Fe concentrations became constant at 4.5 mmol/kg. Soil treatment with 18.8 mmol/kg EDTA resulted in average 241Am concentrations of 29 Bq/kg (19 times higher than controls). Uptake of Pb was similar to 241Am. In these samples, Fe increased to a maximum of almost 18 mmol/kg and 241Am content increased linearly with both Fe and Al. Plants receiving ascorbic and citric acids took up smaller quantities of 241Am, Pb, and Fe, even though these reagents were able to elute about as much Fe from the soil as EDTA. Synchrotron microbeam X-ray fluorescence (SXRF) was used to determine radial distributions of elements in roots and stems with and without EDTA treatment. SXRF maps indicate differences in behavior between Fe and Pb that are consistent with the bulk plant observations and provide insight into changes in metal content of the roots in the presence of EDTA.

Americium↗

Human immunodeficiency virus type 1 Pr55gag and Pr160gag-pol expressed from a simian virus 40 late replacement vector are efficiently processed and assembled into viruslike particles.

Human immunodeficiency virus type 1 (HIV-1) gag and pol genes were expressed by using fragments of the BH10 clone of HIV inserted into a simian virus 40 late replacement vector. An initial construct containing the entire coding regions of gag, pol, and vif produced only minute amounts of the gag precursor, Pr55gag. However, high-level expression was obtained when an additional sequence from the env gene (the rev-responsive element) was inserted 3' of vif in the correct orientation, and rev was provided in trans from a second vector. Western immunoblot analysis of transfected cells showed the presence of large amounts of both Pr55gag and Pr160gag-pol as well as all of the expected cleavage products. Electron microscopy of thin sections of transfected cells showed a multitude of viruslike particles. Both immature particles in the process of budding and particles containing the condensed core characteristic of HIV were observed. Analysis of the released viruslike particles showed the presence of active reverse transcriptase. Sucrose gradient analysis of particles produced from [3H]uridine-labeled cells indicated a peak of radioactivity which cosedimented with a peak of p24, suggesting that the particles contained RNA.

Animals↗

[Defense mechanisms of the respiratory system].

The air we breathe contains certain gases and particles, some of which may be a threat to man. The respiratory tract is their first target because it comes into direct contact with them, and to protect the body itself it mobilizes defense mechanisms of high complexity. To understand these it is necessary to analyze the toxic modalities of these air contaminants: composition, deposition and transit through the respiratory tract. As far as the particles are concerned, the aerodynamic diameter is an important factor in determining the manner of deposition. Also of importance are shape, density, degree of humidity and radioactivity. Some particles are infectious, some are antigenic, and some are toxic due to their physical structure or chemical composition. Emphasis is placed on the role of aerosols, particularly tobacco smoke, as pollution factors or co-factors with other pollutants. The characteristics of the exposed subject capable of defining a risk profile varying from person to person are dwelt on. This approach presupposes a separation, within the cleaning and defense mechanisms, of the mechanical means which bring physical phenomena into play on the basis of fixed structures in the respiratory tract (cilium, mucous cells, surfactant). This is followed by study of the immunologic means of defense essentially involving cellular elements residing temporarily in the lung (T or B lymphocytes, plurinuclears, alveolar histiocytes in particular). Many enzyme systems are also involved in this pulmonary defense and in keeping homeostasis working. Particular attention is focused on some of them, such as the antiproteases and cytochrome P450.(ABSTRACT TRUNCATED AT 250 WORDS)

Air Pollutants↗

Implementation of a radiochemical procedure for the analysis of isotopes of Pu, Am and Cm in food and environmental samples.

A radiochemical procedure for the analysis of isotopes of Pu, Am and Cm was implemented during an expert mission to Thailand sponsored by the International Atomic Energy Agency. This method was to be used for the analysis of food and environmental samples, particularly those of marine origin. The project was initiated by evaluating a procedure which had recently been published by the IAEA. This procedure could not handle the wide range of matrix materials used in the testing and a program was undertaken to develop a modified procedure. Plutonium was radiochemically separated using an anion exchange procedure which gives clean separations from the matrix. Americium and curium came off the column unseparated from most of the other elements in the matrix and they were separated by using a number of steps, which were selected to handle the specific chemical and radiochemical interferences in a particular matrix material. Sources were prepared for alpha spectrometry by coprecipitation with CeF3. The procedure was tested with a wide variety of sample types and good chemical yields and separations were obtained in most cases.

Americium↗

A novel 111In-labeled antisense DNA probe with multi-chelating sites (MCS-probe) showing high specific radioactivity and labeling efficiency.

A multi-aminolinked oligodeoxynucleotide (ODN) was synthesized by substitution of dT with aminolinked dU in the sequence, following conjugation with isothiocyanobenzyl-EDTA (IBE) for 111In labeling. As a model target gene, the c-erbB-2 protooncogene was used. The probability of the number of aminolinked dU in the 20mer ODN was 5, but there were actually 3 and 4 in the selected antisense and sense ODNs, respectively. The IBE/ODN conjugation levels of probes with multi-chelating sites (MCS-probe) were 1.6 (antisense) and 2.4 (sense), more than 50 times higher than those of our previous studies using 5'-end aminolinked ODNs (IBE/ODN = 0.03). Labeling studies using the MCS-probe and 111In indicated that specific radioactivity as high as 48 MBq/nmol could be obtained with a labeling efficiency of over 90%. The 111In-antisense-MCS-probe could bound to sense ODN under physiological conditions, but the 111In-sense-MCA-probe could not. Thus, side-chain modification of ODN for metal labeling is considered to be useful for antisense techniques.

Antisense Elements (Genetics)↗

Spontaneous and induced minisatellite instability in the human genome.

1. Minisatellites, originally developed for forensic DNA fingerprinting and profiling, have also provided extremely informative systems for analysing processes of tandem repeat DNA turnover in the human genome. 2. Minisatellite instability involves distinct mutation processes in somatic and germline cells. In sperm, tandem repeat instability most likely arises at meiosis as a by-product of high-frequency meiotic recombination events occurring in and near minisatellites. Instability is not an intrinsic property of minisatellite repeat DNA, but instead appears to be controlled by flanking recombination initiation elements. 3. Minisatellites not only show high-frequency spontaneous mutation in the germline, but also appear to be very sensitive to mutation induction by ionizing radiation, both in experimentally irradiated mice and in human populations exposed following the Chernobyl disaster; the mechanisms of mutation induction by radiation remain enigmatic.

Animals↗

Lipopolysaccharide-induced methylation of HuR, an mRNA-stabilizing protein, by CARM1. Coactivator-associated arginine methyltransferase.

The RNA-binding protein HuR stabilizes labile mRNAs carrying AU-rich instability elements. This mRNA stabilization can be induced by hypoxia, lipopolysaccharide, and UV light. The mechanism by which these stimuli activate HuR is unclear and might be related to post-translational modification of this protein. Here we show that HuR can be methylated on arginine. However, HuR is not a substrate for PRMT1, the most prominent protein-arginine methyltransferase in mammalian cells, which methylates a number of heterogeneous nuclear ribonucleoproteins. Instead, HuR is specifically methylated by coactivator-associated arginine methyltransferase 1 (CARM1), a protein-arginine methyltransferase previously shown to serve as a transcriptional coactivator. By analyzing methylation of specific HuR arginine-to-lysine mutants and by sequencing radioactively methylated HuR peptides, Arg(217) was identified as the major HuR methylation site. Arg(217) is located in the hinge region between the second and third of the three HuR RNA recognition motif domains. Antibodies against a methylated HuR peptide were used to demonstrate in vivo methylation of HuR. HuR methylation increased in cells that overexpressed CARM1. Importantly, lipopolysaccharide stimulation of macrophages, which leads to HuR-mediated stabilization of tumor necrosis factor alpha mRNA in these cells, caused increased methylation of endogenous HuR. Thus, CARM1, which plays a role in transcriptional activation through histone H3 methylation, may also play a role in post-transcriptional gene regulation by methylating HuR.

Animals↗

Overview of tritium: characteristics, sources, and problems.

Tritium has certain characteristics that present unique challenges for dosimetry and health-risk assessment. For example, in the gas form, tritium can diffuse through almost any container, including those made of steel, aluminum, and plastics. In the oxide form, tritium can generally not be detected by commonly used survey instruments. In the environment, tritium can be taken up by all hydrogen-containing molecules, distributing widely on a global scale. Tritium can be incorporated into humans through respiration, ingestion, and diffusion through skin. Its harmful effects are observed only when it is incorporated into the body. Several sources contribute to the inventory of tritium in our environment. These are 1) cosmic ray interaction with atmospheric molecules; 2) nuclear reactions in the earth's crust; 3) nuclear testing in the atmosphere during the 1950s and 1960s; 4) continuous release of tritium from nuclear power plants and tritium production facilities under normal operation; 5) incidental releases from these facilities; and 6) consumer products. An important future source will be nuclear fusion facilities expected to be developed for the purpose of electricity generation. The principal health physics problems associated with tritium are 1) the determination of the parameters for risk estimation with further reduction of their uncertainties (e.g., relative biological effectiveness and dose-rate dependency); 2) risk estimation from complex exposures to tritium in gas form, tritium in oxide form, tritium surface contamination, and other tritium-contaminated forms, with or without other ionizing radiations and/or nonionizing radiations; 3) the dose contributions of elemental tritium in the lung and from its oxidized tritium in the gastrointestinal tract; 4) prevention of tritium (in oxide form) intake and enhancement of tritium (oxide form) excretion from the human body; 5) precise health effects information for low-level tritium exposure; and 6) public acceptance of tritium leakage and waste disposal from reactors and fuel reprocessing plants.

Environmental Pollutants↗

Transfer factors of some selected radionuclides (radioactive Cs, Sr, Mn, Co and Zn) from soil to leaf vegetables.

Transfer factors of radionuclides from soil to leaf vegetables (cabbage, Chinese cabbage, komatsuna, spinach and lettuce) have been studied by radiotracer experiments using Andosol as a representative of Japanese soils. The transfer factors of radioactive Cs, Sr, Mn, Co and Zn for edible parts of vegetables (average of five vegetables) were 0.11, 0.24, 0.61, 0.05 and 0.52, respectively. These values should be used in safety assessment for Japanese agricultural environment. The transfer factors of Mn, Co and Zn for spinach were higher than those for the other vegetables. The transfer factors of Cs for different organs of the leaf vegetables were rather homogeneous. The transfer factors of Sr and Mn were higher for older (outer) leaves than younger (inner) ones. In contrast to Sr and Mn, transfer factors of Zn for younger leaves were higher than those for older ones. The distribution ratios of the elements between soil-solution and soil were in the order Sr > Mn > Cs > Co > Zn, whereas the distribution ratios of the elements between plant and soil-solution were in the order Zn > Cs > Mn > Co > Sr. These results indicate that the selectivity for Sr by plants from the soil-solution was low and that for Zn was very high.

Biological Availability↗

[Bacteriological diagnosis of tuberculosis].

Microscopic examination and culture are still today essential elements of the bacteriological diagnosis of tuberculosis. Microscopic examination of a Ziehl fuchsin or auramine stained specimen allows detection of most strains in less than an hour. Culture on Löwenstein-Jensen medium is more sensitive than the microscopic examination and is required for identification and to measure sensitivity to antibiotics. Mycobacterium colonies, generally the causal agent in tuberculosis, usually grow within 28 days and are easily recognized by their "cauliflower" aspect. The niacin test is used for formal identification. Currently, radiometric respirometry allows detection of M. tuberculosis growth and provides antibiotic sensitivity results more rapidly, usually within 10 days. Use of this technique is however limited because the culture medium contains radioactive carbon. Genetic probes are on the other hand quite easy to use and allow identification of cultured bacteria in only a few hours. After polymerization chain reaction gene amplification, M. tuberculosis strains can be detected directly in the specimen within 2 or 3 hours, but in practice, this method has not become a routine laboratory technique, particularly due to lack of sufficient specificity and sensitivity. No other serologic tests are currently reliable enough for the diagnosis of tuberculosis. For cases with low-count specimens, there still is no reliable "on-the-spot" diagnostic test.

Bacteriological Techniques↗

Ultrafiltration of large volumes for the determination of colloidally bound artificial radionuclides in natural waters.

A methodology based on large volume ultrafiltration (> 1000 l) is developed in order to characterize the artificial radionuclides associated with the colloidal populations in natural waters. "Inorganic elements" and organic carbon sinks or sources due to sorption or post desorption within the ultrafiltration assembly/membranes of the Sartorius system are underlined and discussed from methodological step experiments as well as recoveries obtained for artificial radionuclides such as 106Ru, 125Sb, 137Cs and 239+240Pu fractionating natural waters from the Rhône River (France), its estuary and the Mediterranean Sea (Gulf of Lions).

Colloids↗

Tissue uptake of circulating hyaluronic acid. A whole body autoradiographic study.

Previous work in the rabbit has shown that there is a significant flux of plasma hyaluronic acid (HA) which is taken up and degraded mainly in the liver but also concentrated in the spleen. Purified 14C-labelled HA of high average molecular wt prepared by biosynthesis from D-[U-14C] glucose was injected i.v. in mice and its tissue distribution was determined by whole-body autoradiography during the next 24 h. As blood levels declined, radioactivity was concentrated in the liver and spleen as found in the rabbit, and also in bone marrow and lymph nodes. Distribution was uniform in liver tissue, concentrated and relatively persistent in the periphery of lymph nodes, and distinctly nodular within the spleen. Analysis of an aqueous liver extract taken 4 h after injection identified 14C in HA, in a macromolecular fraction resistant to fungal hyaluronidase, and in metabolites of low molecular wt. These findings confirm and extend observations based on tissue extraction in rabbits. The pattern of distribution through the body and the restricted localization within spleen and lymph nodes further suggest that HA is absorbed from plasma and tissue fluids by elements of the reticuloendothelial system.

Animals↗

Maintenance by nerve growth factor of the intracellular sodium environment in spinal sensory and sympathetic ganglionic cells.

A promising advance in our understanding of the mode of action of nerve growth factor (NGF) has been provided by our recent finding that dissociated cells from 8-day embryonic chick dorsal root ganglia (DRG) lose their competence to maintain a low intracellular Na+ when incubated without NGF for 6 h, and promptly recover it on delayed presentation of NGF. To ascertain whether control of intracellular Na+ is a general feature of the NGF action on its target neurons, we have now tested several NGF-sensitive tissues, both as intact ganglia and dissociates, for their ability to (i) accumulate large amounts of 22Na+ over a period of hours in the absence of NGF, and (ii) rapidly extrude the accumulated radioactivity upon delayed presentation of NGF. Intact and dissociated chick embryo DRG and sympathetic ganglia, as well as dissociated mouse DRG, displayed the expected Na+ responses to the lack or the administration of NGF. No such responses were observed with intact mouse DRG, which do not require NGF in explant cultures for neuritic outgrowth, or with intact and dissociated chick embryo ciliary ganglia, which are not sensitive to NGF in either explant or dissociated cell cultures. Thus, in all ganglionic preparations examined, the occurrence of Na+ responses to exogenous NGF correlated with traditional responses to NGF in culture. This together with other recent data, reinforces the view that ionic control may be a critical element in the mechanism of action of NGF.

Animals↗

ADD1/SREBP1 activates PPARgamma through the production of endogenous ligand.

Adipose differentiation is an important part of the energy homeostasis system of higher organisms. Recent data have suggested that this process is controlled by an interplay of transcription factors including PPARgamma, the C/EBPs, and ADD1/SREBP1. Although these factors interact functionally to initiate the program of differentiation, there are no data concerning specific mechanisms of interaction. We show here that the expression of ADD1/SREBP1 specifically increases the activity of PPARgamma but not other isoforms, PPARalpha, or PPARdelta. This activation occurs through the ligand-binding domain of PPARgamma when it is fused to the DNA-binding domain of Gal4. The stimulation of PPARgamma by ADD1/SREBP1 does not require coexpression in the same cells; supernatants from cultures that express ADD1/SREBP1 augment the transcriptional activity of PPARgamma. Finally, we demonstrate directly that cells expressing ADD1/SREBP1 produce and secrete lipid molecule(s) that bind directly to PPARgamma, displacing the binding of radioactive thiazolidinedione ligands. These data establish that ADD1/SREBP1 can control the production of endogenous ligand(s) for PPARgamma and suggest a mechanism for coordinating the actions of these adipogenic factors.

3T3 Cells↗

Is the transfer factor a relevant tool to assess the soil-to-plant transfer of radionuclides under field conditions?

The radiological impact of radionuclides released to the terrestrial environment is usually predicted with mathematical models in which the transfer of radionuclides from soil to the plant is described with the transfer factor (TF). This paper questions the validity of the protocols proposed by the International Atomic Energy Agency to measure TF in the field and in greenhouses conditions. We grew maize (Zea mays L.) both in the field after a surface application of radionuclides ((54)Mn, (57)Co, (65)Zn, and (134)Cs) and in a greenhouse with the same soil that has received the same fertilization and that had been previously sieved and homogeneously labeled with the same radionuclides before being repacked in pots. The analysis of the displacement of radionuclides in the field soil profile showed a higher concentration of the surface-applied radionuclides in the preferential flow path (PFP) in comparison to the soil matrix indicating that they infiltrated heterogeneously in the soil profile due to the structure-induced non-uniform water flow. A significantly higher recovery of (57)Co and (134)Cs was observed in the plants grown in the field soil, whereas no differences in the recovery of (54)Mn and (65)Zn between the two experiments were detected. These results suggest that (i) under field conditions the soil-to-plant transfer of radionuclides that co-exist as stable elements present at low concentrations in the soil and in the plant is higher than that measured under greenhouse conditions and (ii) the implicit assumption made when calculating the TF (that radionuclides are homogeneously distributed in the soil profile) is not valid, thereby preventing the calculation of an average concentration to obtain the TF parameter.

Models, Theoretical↗

Can angiotensin receptor antagonists prevent restenosis after stent placement?

Restenosis rates after coronary stent implantation in complex lesions are between 30 and 50%. Neointimal hyperplasia promoted by complex interaction between cellular and acellular elements, such as cytokines and growth factors, is thought to be the primary process responsible for restenosis. The risk of in-stent restenosis is increased in patients with a history of restenosis after percutaneous transluminal coronary angioplasty, in long lesions, in total occlusions, in patients with diabetes mellitus, in small vessels, in the proximal parts of the left anterior descending coronary artery and in cases of stent oversizing. In-stent restenosis represents a serious economic burden on society because treatment strategies include expensive approaches such as cutting-balloon angioplasty, rotational atherectomy and brachytherapy. A number of pharmacological agents, including ACE inhibitors, have been unsuccessful in preventing restenosis. Alternative procedures such as brachytherapy, radioactive stents and drug-eluting stents are under evaluation. Although sirolimus- or paclitaxel-eluting stents have been associated with very low restenosis rates over durations of 6 to 12 months, the long-term efficacy and tolerability of this approach is currently being investigated. Although ACE inhibitors have failed in reducing restenosis rates, the selective angiotensin II type 1 (AT(1)) receptor antagonist valsartan has shown encouraging results in the single-center Valsartan for Prevention of Restenosis after Stenting of Type B2/C lesions trial (ValPREST). The ValPREST trial is the first randomized, placebo-controlled study to have evaluated the effect of an angiotensin receptor antagonist on in-stent restenosis in a moderate number of patients. Compared with ACE inhibitors, angiotensin receptor blockers exert additional effects on the pathophysiological processes which lead to restenosis. Angiotensin receptor antagonists may affect several mechanisms involved in neointimal hyperplasia such as decreasing circulating cytokine and growth factor levels and reducing neutrophil activation, especially after stenting in acute coronary syndromes, but the results need to be confirmed in a large multicenter trial. The question whether long-term therapy, with an oral angiotensin receptor antagonist, is cost-effective and whether angiotensin receptor antagonists should be used as an add-on therapy to drug-eluting stents, requires clarification.

Angiotensin-Converting Enzyme Inhibitors↗

Accelerator mass spectrometry in pharmaceutical research and development--a new ultrasensitive analytical method for isotope measurement.

Accelerator mass spectrometry (AMS) permits the measurement of elemental isotopes at the individual atom level. The main application of AMS in drug discovery and development will be in the analysis of 14-carbon (14C). The principle behind AMS is the separation of individual positively charged atoms through mass, charge and momentum differences. In order to obtain the high-energy charge state required for separation, negative atoms are accelerated through a high voltage field (up to 10 million volts) generated by a tandem Van de Graaff accelerator. In the middle of the accelerator, the outer valency electrons are stripped from the atom and the resulting charged species are separated and counted. For 14C, AMS counts the number of individual atoms rather than measuring radioactive decays. The result is that AMS is up to one million times more sensitive than decay counting. Radioactivity levels as low 0.0001 dpm can be detected using AMS. The exquisite sensitivity of AMS analysis means that much lower amounts of 14C can be used than for conventional counting methods. This makes it easier to use 14C for in vitro, preclinical and clinical research programmes. As 14C poses both a biological and environmental hazard, AMS permits much lower doses to be used. Human drug mass balance studies have been conducted with doses of 50 nanoCuries and below. Radioactive HPLC metabolite profiles of plasma extracts from subjects given nanoCurie doses of 14C-labelled drug have been obtained by injecting as little as 0.25 dpm onto an HPLC column. In studies of biologics, biosynthetically 14C-labelled recombinant protein has been produced with a specific radioactivity sufficient to conduct human clinical studies with AMS analysis. For one human recombinant protein an increase in sensitivity of 2,000-fold over ELISA was obtained with AMS measurement. AMS is an enabling technology that should prove of value in increasing human and environmental safety as well as allowing new research directions to be followed.

Animal Testing Alternatives↗

The delayed respiratory consequences of inhaled radionuclides in population exposed to nuclear catastrophe.

Cytological analysis of bronchoalveolar lavage (BAL) and x-ray energy dispersive spectrometry of intercellular compounds were evaluated in 19 workers ("liquidators") commissioned to clean and contain the nuclear waste after the Chernobyl catastrophe in comparison with ten control subjects. Results showed a significant increase of neutrophils, small monocyte-like alveolar macrophages (AM) and AM with large black cytoplasmic inclusions. In addition, such mineral elements as Zr, Ce, Pr, Sm, Pa, PU and Cs were detected in AM phagosomes in liquidators. These elements were reported as markers of Chernobyl-type nuclear fuel. It is recognized that BAL gives access to particulate matter present in lung parenchyma. Thus, spectrometric analysis of AM inclusions can provide reliable information about the exposure to inhaled radionuclides seven years after the catastrophe.

Administration, Inhalation↗