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PubMed1991

Determination of tin in human blood serum by radiochemical neutron activation analysis.

A method was developed for the determination of tin in human serum by radiochemical neutron activation analysis, using the long-lived radioisotope Sn(T1/2 = 115.09 days). This radioisotope decays to a daughter isotope 113mIn, the most suitable nuclide for counting (T1/2 = 1.658 h, gamma-ray of 391.7 keV). Experience showed that, with the exception of the serum samples with the lowest tin levels, in the experimental conditions of the present study tin could mostly also be determined by using its radioisotope 117mSn(T1/2 = 13.61 days, gamma-ray of 158.5 keV). Samples were collected and prepared by using the procedure elaborated by the authors, which proved its effectiveness in preventing significant sample contamination on several occasions. Because samples had to be irradiated at 10(14) n.cm-2.s-1, dry ashing was necessary. After irradiation, tin was separated by solvent extraction of tin(IV) iodide from a sulfuric acid-ammonium iodide solution with toluene. The dry ashing and solvent extraction steps were exhaustively tested by means of radioactive tracer experiments whereas the accuracy and precision of the analytical method were thoroughly checked by analyzing biological reference materials (Bowen's kale powder, the NBS' bovine liver, the NBS' nonfat milk powder, and the "second-generation" biological reference material--freeze-dried human serum--for trace element determinations, developed by the authors).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1997

Sphingolipid synthesis as a target for antifungal drugs. Complementation of the inositol phosphorylceramide synthase defect in a mutant strain of Saccharomyces cerevisiae by the AUR1 gene.

We have identified a Saccharomyces cerevisiae gene necessary for the step in sphingolipid synthesis in which inositol phosphate is added to ceramide to form inositol-P-ceramide, a reaction catalyzed by phosphatidylinositol:ceramide phosphoinositol transferase (IPC synthase). This step should be an effective target for antifungal drugs. A key element in our experiments was the development of a procedure for isolating mutants defective in steps in sphingolipid synthesis downstream from the first step including a mutant defective in IPC synthase. An IPC synthase defect is supported by data showing a failure of the mutant strain to incorporate radioactive inositol or N-acetylsphinganine into sphingolipids and, by using an improved assay, a demonstration that the mutant strain lacks enzyme activity. Furthermore, the mutant accumulates ceramide when fed exogenous phytosphingosine as expected for a strain lacking IPC synthase activity. Ceramide accumulation is accompanied by cell death, suggesting the presence of a ceramide-activated death response in yeast. A gene, AUR1 (YKL004w), that complements the IPC synthase defect and restores enzyme activity and sphingolipid synthesis was isolated. Mutations in AUR1 had been shown previously to give resistance to the antifungal drug aureobasidin A, leading us to predict that the drug should inhibit IPC synthase activity. Our data show that the drug is a potent inhibitor of IPC synthase with an IC50 of about 0.2 nM. Fungal pathogens are an increasing threat to human health. Now that IPC synthase has been shown to be the target for aureobasidin A, it should be possible to develop high throughput screens to identify new inhibitors of IPC synthase to combat fungal diseases.

Amino Acid Sequence↗

PubMed2004

A modified ICRP 66 iodine gas uptake model and its parametric uncertainty.

Intakes via inhalation may occur from radionuclides released in the form of a gas. The chemical characteristics pertaining to the release influence the intake and subsequent dose to an exposed individual. Gases are taken up or absorbed in the entire respiratory tract and the associated uptake mechanisms are quite different from deposition of particulates. Gaseous iodine can exist in various chemical forms, e.g., elemental iodine, inorganic, and organic iodine compounds. These different chemical species play an integral role in the gaseous uptake o f iodine in t he respiratory tract. Gas uptake in the various regions of the respiratory tract results in the intake of iodinated material into the body. The radioactive iodine taken up in the gas-exchange tissues is absorbed into the bloodstream of an individual and subsequently transferred to other organs. Iodine in the circulatory system can then be taken up by the thyroid gland, with resulting dose to the thyroid. The magnitude and uncertainty in regional gas uptake is important in the assessment of individuals exposed to airborne releases of radioiodine. The current ICRP 66 model is rudimentary and estimates regional gas uptake based on solubility and reactivity of the different radionuclides entering the respiratory tract. The modified model proposed here employs methodology and a mathematical structure to determine estimates of fractional gas uptake rather than defaulting to literature values, as in the current ICRP model. Model parameters have been assigned input distributions and estimates of uncertainty have been determined. A sensitivity analysis of these parameters has been performed to demonstrate the importance of each of these parameters. The sensitivity analysis ranks the model-input parameters by their importance to estimates of regional gas uptake. The model developed herein may be used for improved estimation of gas uptake in the respiratory tract and subsequent dose estimates from the different chemical forms of radioiodine.

Adult↗

PubMed1998

Controlled introduction of selenium into Chlorella cells.

Selenium (Se) is an important element in the antioxidant system of the human body, and Chlorella, well-known for its therapeutic effects, is the ideal carrier to offer it in the wanted organic form. The kinetics of Se absorption by growing algal cells and its distribution in the cells are studied using radioactive 75Se labelled solutions. There is a rapid Se absorption within the first few minutes at the cell surfaces where it is irreversibly fixed and cannot be absorbed by the human body. In the final state, reached after 24-48 hr, about 40% of the total fixed Se is inside the cells in the wanted organic-bound form.

Chlorella↗

PubMed1997

[Molecular typing of Moroccan strains of Mycobacterium tuberculosis].

The insertion sequence IS 6110 was used to differentiate clinical Moroccan isolates of Mycobacterium tuberculosis by using two non radioactive probes. Among 16 strains isolated from patients clinically related, 10 had similar IS 6110 restriction fragment length polymorphism (RFLP) patterns, confirming that they were derived from a common source. Two strains were isolated from the same patient (sputum, lymph node) showed identical profiles hybridized with IS 6110 element. Four sequential strains isolated from the same patients before treatment and after one year had identical IS 6110 RFLP patterns suggesting relapse and not reinfection. Twenty-one strains with identical drug susceptibility showed different IS 6110 RFLP profiles confirming no correlation between antibiotic resistance profiles and IS 6110 RFLP patterns. Since RFLP analysis by using IS 6110 element is a useful tool for the epidemiological survey, of tuberculosis.

DNA, Viral↗

PubMed1979

Mechanistic and quantitative evaluation of precorneal pilocarpine disposition in albino rabbits.

The low ocular bioavailability of topically applied pilocarpine is attributed to extensive precorneal drug loss in conjection with the resistance to corneal penetration. Several elements of precorneal loss were reported earlier, but a complete mechanistic understanding has not been available. The present study was designed to gain a better understanding of the mechanisms governing pilocarpine disposition in the precorneal areas as well as the relative influence of these parameters on ocular drug bioavailability. Radioactive pilocarpine and glycerin solutions were instilled into the precorneal area of the albino rabbit eye under various experimental conditions, and the drug concentration in the lacrimal lake was monitored as a function of time. The results demonstrated that nonconjunctival loss of pilocarpine, vasodilation due to the drug, and lacrimation due to vehicle formulation are additional aspects of precorneal drug disposition. The individual influence of all precorneal loss parameters on drug bioavailability was then assessed using a mathematical model formulated from experimental findings on both precorneal and intraocular drug disposition. Drainage and vasodilation, as well as nonconjunctival pilocarpine loss, exerted major influences on drug loss at the absorption site.

Administration, Topical↗

PubMed1986

Studies on rat cytomegalovirus induced structural and non-structural proteins present at (immediate-)early and late times of infection.

Radioactive-labelled virions and nucleocapsids of rat cytomegalovirus (RCMV) were purified from the supernatant and subcellular fractions of infected rat embryo fibroblasts (REF) and analyzed by SDS-polyacrylamide gel electrophoresis. Nuclear nucleocapsids contain one major protein of 138 kD, which is considered to be the basic invariant structural element of RCMV. Enveloped virions consist of 28 protein species, five of which were clearly identified as glycoproteins (58, 64, 76, 112 and 118 kD). Using pulse labelling procedures on RCMV-infected REF cells, after removal of a previously established translation block by cycloheximide, two RCMV-induced immediate-early (IE) protein species (71 and 85 kD) were both detected in the nuclear and cytoplasmic cell fractions. When pulse-labelling was performed for 16 hours in presence of Actinomycin D, only the 85 kD IE protein was detected in the nucleus. The results indicate that the 85 kD IE polypeptide is of importance in early transcriptional events of viral genes. Protein metabolism studies revealed that late in the course of RCMV infection (at 3 days p.i.) protein synthesis has been dramatically changed. Some cellular proteins are greatly suppressed while other cellular proteins are clearly enhanced. Moreover, active synthesis of 8 new cytoplasmic proteins and 9 new nuclear proteins occurs. Most of these proteins were identified as structural constituents of virions.

Amino Acids↗

PubMed2000

Improved target system for production of high purity [18F]fluorine via the 18O(p,n)18F reaction.

An improved aluminium target system for production of elemental fluorine via the 18O(p,n)18F reaction using a two-step irradiation protocol is described. In the first step highly enriched gaseous oxygen-18 is irradiated with protons to form fluorine-18 which gets deposited on the inner target surface. In the second step, after cryogenic recovery of oxygen-18 target gas, a mixture of elemental 'cold' fluorine and krypton is introduced and a short proton irradiation is done, whereby an isotopic exchange between the gaseous fluorine and the deposited radiofluorine occurs. The second step leads to the recovery of the radiofluorine as [18F]fluorine. Optimisation studies were performed regarding the yield and specific radioactivity of [18F]fluorine. Furthermore, some irradiation parameters relevant to the recovery step were investigated. It was found that a 15 to 20 min irradiation with a beam current of 20 microA is sufficient for the isotopic exchange between the fluorine-carrier and the 18F-radioactivity deposited on the inner wall of the target. The distribution of the 18F-radioactivity deposited on the inner target surface is inhomogeneous, probably due to convection effects. Extensive radioanalytical techniques were applied to characterise the reactivity of [18F]fluorine and to identify undesired nonreactive 18F-compounds, mainly [18F]tetrafluoromethane and [18F]nitrogentrifluoride. The [18F]fluorine produced in the system used has the distinction of having a negligible contamination from those inert 18F-compounds. This is a combined effect of the use of highest purity gases and a welded target construction, which avoids any contact of the gases with organic material during irradiations. The target has proved to be very reliable for production of [18F]fluorine in high yields of up to 34 GBq and specific activities of 350-600 GBq/mmol, both at 30 min after end of activation bombardment.

Aluminum↗

PubMed2005

Radiation protection system at the RIKEN RI beam factory.

The RIKEN RI (radioactive isotope) Beam Factory is scheduled to commence operations in 2006, and its maximum energy will be 400 MeV u(-1) for ions lighter than Ar and 350 MeV u(-1) for uranium. The beam intensity will be 1 pmicroA (6 x 10(12) particles s(-1)) for any element at the goal. For the hands-on-maintenance and the rational shield thickness of the building, the beam loss must be controlled with several kinds of monitors. Three types of radiation monitors will be installed. The first one consists of a neutron dose equivalent monitor and an ionisation chamber, which are commercially available area monitors. The second one is a conventional hand-held dose equivalent monitor wherein the logarithmic signal is read by a programmable logic controller based on the radiation safety interlock system (HIS). The third one is a simple plastic scintillator called a beam loss monitor. All the monitors have threshold levels for alarm and beam stop, and HIS reads all these signals.

Equipment Design↗

PubMed1988

Selenium increases hydrogenase expression in autotrophically cultured Bradyrhizobium japonicum and is a constituent of the purified enzyme.

We have investigated the effect of added selenite on autotrophic growth and the time course of hydrogen oxidation derepression in Bradyrhizobium japonicum 122DES cultured in a medium purified to remove selenium compounds. In addition, hydrogenase was purified to near homogeneity and examined for the specific incorporation of Se into the enzyme. The addition of Se at 0.1 microM significantly increased total cell protein and hydrogenase specific activity of harvested cells. Also, the addition of SeO3(2-) enhanced the time course of hydrogenase derepression by 133%, whereas VO3, AsO2(2-), SO2(2-), and TeO3(2-) failed to substantially affect hydrogenase derepression. During the final chromatographic purification of hydrogenase, a striking coincidence in peaks of protein content, Se radioactivity, and hydrogenase activity of fractions was obtained. The total Se content expressed per milligram of protein increased manyfold during the purification procedure. The mean Se content of the purified hydrogenase was 0.56 +/- 0.13 mol of Se per mol of enzyme. These results indicate that Se is an important element in the H2 metabolism of B. japonicum and that hydrogenase from B. japonicum is a seleno protein.

Culture Media↗

PubMed1988

Translational control in early sea urchin embryogenesis: initiation factor eIF4F stimulates protein synthesis in lysates from unfertilized eggs of Strongylocentrotus purpuratus.

We have used cell-free translation systems from unfertilized eggs and embryos of the sea urchin Strongylocentrotus purpuratus to analyze the mechanisms limiting protein synthesis in early embryogenesis. Unfertilized egg lysates supplemented with nuclease-treated reticulocyte lysate were stimulated 2-4-fold in incorporation of radioactive amino acid into protein. Thirty-minute zygote lysates supplemented in this way were not stimulated. These results suggested that a component limiting translation in the unfertilized egg lysate was provided by the nuclease-treated lysate and that this component was no longer limiting protein synthesis following fertilization. In view of these results, partially fractionated lysates and individual purified translational components from mammalian cells were tested for stimulation of the unfertilized egg lysate. A 1000000g supernatant devoid of ribosomal subunits also stimulated the unfertilized egg lysate. Thus, the stimulation was not due to the addition of active ribosomal subunits but to soluble elements in the reticulocyte lysate. Of the soluble components tested, only the cap-binding protein complex eIF4F caused a dramatic stimulation of the unfertilized egg lysate (2-3.5-fold). The 30-min zygote lysate was not stimulated by eIF4F or by any of the other components tested, supporting the hypothesis that a block in the translational machinery is removed at fertilization. A rabbit reticulocyte shift assay was used to analyze whether mRNA is limiting in early development. When unfertilized egg lysate was added to the shift assay, there was no shift in radioactivity from 43S to 80S complexes, indicating the unfertilized egg mRNA is not available for translation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2005

Retention and translocation of foliar applied 239,240Pu and 241Am, as compared to 137Cs and 85Sr, into bean plants (Phaseolus vulgaris).

Foliar transfer of 241Am, 239,240Pu, 137Cs and 85Sr was evaluated after contamination of bean plants (Phaseolus vulgaris) at the flowering development stage, by soaking their first two trifoliate leaves into contaminated solutions. Initial retentions of 241Am (27%) and 239,240Pu (37%) were higher than those of 137Cs and 85Sr (10-15%). Mean fraction of retained activity redistributed among bean organs was higher for 137Cs (20.3%) than for 239,240Pu (2.2%), 241Am (1%) or 85Sr (0.1%). Mean leaf-to-pod translocation factors (Bq kg(-1) dry weight pod/Bq kg(-1) dry weight contaminated leaves) were 5.0 x 10(-4) for 241Am, 2.7 x 10(-6) for 239,240Pu, 5.4 x 10(-2) for 137Cs and 3.6 x 10(-4) for 85Sr. Caesium was mainly recovered in pods (12.8%). Americium and strontium were uniformly redistributed among leaves, stems and pods. Plutonium showed preferential redistribution in oldest bean organs, leaves and stems, and very little redistribution in forming pods. Results for americium and plutonium were compared to those of strontium and caesium to evaluate the consistency of the attribution of behaviour of strontium to transuranium elements towards foliar transfer, based on translocation factors, as stated in two radioecological models, ECOSYS-87 and ASTRAL.

Americium↗

PubMed2002

Distribution of long-lived radionuclides of the 238U series in the sediments of a small river in a uranium mineralized region of Spain.

A study is presented on the distribution and mobilization of the natural U isotopes (238U and 234U), 230Th, and 226Ra in the sediments of a small river crossing an uranium mineralized zone where a disused uranium mine is located. Due to the preferential directions for surface run-off waters and to the mine's situation, one sampling point along the river bed was identified as a point of accumulation of radionuclides. The average values of the activity concentrations (Bq/kg) in this sediment sample were 5,025, 5,055, 5,915 and 1,694 for 238U, 234U, 230Th and 226Ra, respectively, while the respective average values of the activity concentrations (Bq/kg) for the sediment sample considered to give the background level were 125, 124, 131 and 370. Isotopic ratios between the descendants of 238U served to clarify some paths of distribution, involving the soils nearest to the sampling points and the location of these points with respect to the disused mine. The differences in behaviour found between the uranium, thorium and radium isotopes were associated to the mobility of these radionuclides in the fluvial system studied. Correlations between the radionuclide activity concentration ratios and stable element concentrations in the sediment samples were also investigated.

Fresh Water↗

PubMed2006

Mixing in a square and a rectangular duct regarding selection of locations for extractive sampling of gaseous contaminants.

Tests were conducted to characterize the uniformity of velocity and tracer gas profiles in a square and a rectangular duct with respect to defining the suitability of locations for single point sampling of gaseous contaminants. Several configurations, such as a straight duct with unidirectional flow at the entrance section and straight ducts preceded by mixing elements (a 90 degrees mitered bend and double 90 degrees bends in S- and U-type configurations) were tested. Results are compared with those from circular ducts. For a straight duct of square cross section, which is not preceded by a mixing element, the coefficients of variation (COV) of tracer gas concentration at 19 duct diameters downstream of the gas release location is 143% (center release of tracer gas). COVs of velocity and tracer gas concentration in a straight square duct 9.5 duct diameters downstream of a 90 degrees mitered bend are 6% and 24.3% (top inside release), respectively, which does not meet the ANSI N13.1 limit of 20% for the tracer gas COV. In case of the rectangular duct with a 3:1 (width to height) aspect ratio, COVs of velocity and tracer gas concentration at 9 duct diameters downstream of a 90 degrees mitered bend are 29% and 62% (bottom inside release), respectively. A mixing element in a square duct comprised of two 90 degrees mitered bends in a U-configuration produces results similar to those obtained with a single 90 degrees bend. However, COVs of velocity and tracer gas concentration in a square duct 6 duct diameters downstream of an S-type double bend are 10.6% and 8.3% (top inside release), respectively, which comply with the ANSI tracer gas and velocity criteria for single point representative sampling. When mixing elements were employed in square ducts, the COV results were comparable with those of other researchers for circular ducts.

Air Pollutants, Radioactive↗

PubMed1961

Ability of some Black Sea organisms to accumulate fission products.

The coefficients of accumulation of strontium-90, cesium-137, and cerium-144 in seaweeds, eelgrass, actinia, mollusks, and crustaceans are presented. The discharge of strontium-90 into sea water from decomposing seaweed and the retention and additional absorption of cesium-137 and cerium-144 onto organic debris is discussed. Some observations are made about the ability of these elements to diffuse into sea water and about the relative hazard to man from strontium-90 and cerium-144 in marine life.

Animals↗

PubMed1975

Accumulation of 42K and 86Rb in the organs of the rat.

42K and 86Rb were administered in combination to anaesthetized rats by the intravenous route, and accumulation of the isotopes in the organs during a few cirulatory cycles (90 sec) and after 24 hours has been studied in the rat. a) Accumulation of the two elements was identical in skeletal and heart muscle at both points of time. b) Acute (90 sec) accumulation of 86Rb slightly exceeded that of 42K in the skin, liver, brain, external and internal medulla of the kidney. Therefore, the cardiac output fractions obtained with 42K and 86Rb according to Sapirstein's method must be compared with certain corrections, and they cannot be evaluated in a combined form. c) Considerable differences were found in the accumulation of the two elements after 24 hours. The differences were ascribed to the different intra- and extracellular concentrations of the natural isotopes of the two elements and, to a lesser extent to their presumbly different "lag phenomenon".

Animals↗

PubMed2002

Genome-wide expression profiling of 8-chloroadenosine- and 8-chloro-cAMP-treated human neuroblastoma cells using radioactive human cDNA microarray.

Previous reports raised question as to whether 8-chloro-cyclic adenosine 3,5-monophosphate (8-Cl-cAMP) is a prodrug for its metabolite, 8-Cl-adenosine which exerts growth inhibition in a broad spectrum of cancer cells. The present study was carried out to clarify overall cellular affects of 8-Cl-cAMP and 8-Cl-adenosine on SK-N-DZ human neuroblastoma cells by systematically characterizing gene expression using radioactive human cDNA microarray. Microarray was prepared with PCR-amplified cDNA of 2,304 known genes spotted on nylon membranes, employing (33)P-labeled cDNAs of SK-N-DZ cells as a probe. The expression levels of approximately 100 cDNAs, representing about 8% of the total DNA elements on the array, were altered in 8-Cl-adenosine- or 8-Cl-cAMP-treated cells, respectively. The genome-wide expression of the two samples exhibited partial overlaps; different sets of up-regulated genes but the same set of down-regulated genes. 8-Cl-adenosine treatment up-regulated genes involved in differentiation and development (LIM protein, connexin 26, neogenin, neurofilament triplet L protein and p21(WAF1/CIP1)) and immune response such as natural killer cells protein 4, and down-regulated ones involved in proliferation and transformation (transforming growth factor-beta, DYRK2, urokinase-type plasminogen activator and proteins involved in transcription and translation) which were in close parallel with those by 8-Cl-cAMP. Our results indicated that the two drugs shared common genomic pathways for the down-regulation of certain genes, but used distinct pathways for the up-regulation of different gene clusters. Based on the findings, we suggest that the anti-cancer activity of 8-Cl-cAMP results at least in part through 8-Cl-adenosine. Thus, the systematic use of DNA arrays can provide insight into the dynamic cellular pathways involved in anticancer activities of chemotherapeutics.

2-Chloroadenosine↗

PubMed2003

The methionine requirement is lower in neonatal piglets fed parenterally than in those fed enterally.

The requirements for the sulfur amino acids (SAA), methionine (Met) and cysteine (Cys), have seldom been determined in neonates and to our knowledge have not previously been determined directly in parenterally fed neonates. The sulfur amino acids are catabolized largely in the liver and kidney, and their metabolism by the gut has been studied less frequently. In the present research, the enteral and parenteral Met requirement was determined, without dietary Cys, using the indicator amino acid oxidation (IAAO) technique. Piglets [n = 32, 2 d old, 1.66 +/- 0.13 kg (SD)] received elemental diets containing adequate energy, phenylalanine (Phe) and excess tyrosine, with varied Met concentrations and no Cys. Diets were infused continuously via intravenous or intragastric catheters. Phenylalanine oxidation was determined during a primed, constant infusion of L-[1-(14)C]-Phe, by measuring expired (14)CO(2) and plasma specific radioactivity of Phe. For both parenteral and enteral groups, Phe oxidation (% of dose) decreased linearly (P < 0.01) as Met intake increased, then became low and unchanging. Using breakpoint analysis, the Met requirement was estimated to be 0.42 and 0.29 g/(kg. d) for enteral and parenteral feeding, respectively. Breakpoint analysis using absolute phenylalanine oxidation [ micro mol/(kg. h)] resulted in an estimation of the Met requirement of 0.44 and 0.26 g/(kg. d) for enteral and parenteral feeding, respectively. These data show that the parenteral Met requirement is approximately 69% of the enteral requirement and suggest that extraction of SAA by first-pass splanchnic metabolism may be responsible for this difference.

Amino Acids↗