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Non-specific binding of transferrin and lactoferrin to polystyrene culture tubes: role of the radioligand.

While evaluating the role of iron-binding glycoproteins on the in vitro uptake of 67Ga and 59Fe by tumor cells, it was observed that these radiometals bind to polystyrene culture tubes in the presence of transferrin or lactoferrin. The amount of 67Ga or 59Fe bound to the tube increases with glycoprotein concentrations up to 20 microgram/ml and decreases thereafter. This biphasic response is a reflection of metal-protein affinity and is greatest for 59Fe-lactoferrin. With 125I-labeled transferrin and lactoferrin, the amount of tube-bound radioactivity was inversely dependent on the glycoprotein concentration suggesting that glycoproteins bind to a limited number of binding sites on the tube wall. Approximately 10(13) glycoprotein molecules were bound per tube with an affinity constant of 1.89 X 10(7) 1/M for transferrin and 1.08 X 10(7) l/M for lactoferrin. These sites are not specific since addition of albumin inhibited the binding of radiolabeled glycoproteins to the tube. In the light of these observations, caution is required in interpreting results of cell culture experiments which have not directed attention to protein-plastic interaction.

Albumins↗

Response of the bronchial mucociliary clearance system in rabbits to inhaled sulfite and sulfuric acid aerosols.

A group of eight rabbits was exposed (orally) for 1 hr to a distilled water aerosol (sham control) or to submicrometer aerosols of either a transition metal sulfite complex (Fe(III)-S(IV], sodium sulfite (Na2SO3), or sulfuric acid (H2SO4). Mucociliary clearance was evaluated by external retention measurements of radioactively tagged tracer particles from the bronchial tree. Fe(III)-S(IV) in the range of 238 to 1227 micrograms/m3 (as SO2-3) produced no significant change from sham control in the mean residence time (MRT) of the tracer, indicating no effect upon mucociliary clearance rate. On the other hand, Na2SO3 at levels greater than or equal to 1200 micrograms/m3 (as SO2-3) resulted in clearance acceleration; the lack of effect of Fe(III)-S(IV) is possibly due to its stability. H2SO4 at 260 to 2155 micrograms/m3 produced a significant dose-related response, indicating clearance acceleration at low concentrations and a depression at higher levels.

Aerosols↗

Dissociation of human choriogonadotropin from the pseudopregnant rat ovary as a complex to a receptor fragment during perifusion and incubation.

Pseudopregnant rats were injected intravenously with radioactively-labelled human choriogonadotropin (CG). The animals were killed 2 h after the injection and the ovaries, liver and kidney were subjected to perifusion. Radioactivity was released from the ovaries at an increasing rate during perifusion, mainly in a complex form with a molecular size between human CG and the solubilized receptor-human CG complex. The increase in the rate of radioactivity release was inhibited by N-ethylmaleimide and CuCl2, but not by MgCl2, Trasylol, N alpha-tosyl-L phenylalanine chloromethyl ketone or N alpha-p-tosyl-L-lysine and CuCl2 chloromethyl ketone. Intact hormone dissociation from the complex at pH 3. After perifusion the ovarian tissue radioactivity only as receptor-hormone complexes. Only free radioiodine released from the control tissues, liver and kidney during perifusion. The low molecular weight hormone complex was also released from a homogenate of pseudopregnant rat ovaries prelabelled in vivo with radioactivity-labelled human CG during incubation in a hypotonic medium. The release of this complex was likewise inhibited by alkylating agents and heavy metals, and intact hormone dissociated from the complex at pH 3. A similar human CG complex was released also from purified receptor-human CG complex during incubation with ovarian homogenate, and presence of N-ethylmaleimide or use of heat inactivated ovarian homogenate inhibited this process. The present results indicate that the in vivo bound human CG sheds from the luteal tissue in perifusion and incubation as a low molecular weight complex. This may be a facet in the processing and elimination of occupied LH receptors from the ovary.

Animals↗

Minor burns lead to major changes in synthesis rates of plasma proteins in the liver.

The effect of minor burns on the rates of synthesis of plasma proteins in the liver was studied in white Buffalo rats. Burns of second to third degree, covering 0.8% of total body surface, were produced by short application of a hot piece of metal to the skin under ether anesthesia. Levels of mRNA in extracts from liver removed after 24 hr were measured by hybridization to radioactively labeled specific cDNA probes. The level of mRNA for major acute phase alpha 1-protein (also called cysteine proteinase inhibitor or T1-kininogen) increased 20-fold, that of fibrinogen mRNA 8-fold, and that of alpha 1-acid glycoprotein mRNA about 9-fold. The levels of albumin mRNA and transthyretin mRNA (also called prealbumin) decreased to about 80% of normal and the level of transferrin mRNA did not change significantly. Thus, although the percentage of burnt skin was only very small, a typical acute phase response of plasma protein synthesis in liver was observed.

Acute-Phase Proteins↗

Discovery of ancient silicate stardust in a meteorite.

We have discovered nine presolar silicate grains from the carbonaceous chondrite Acfer 094. Their anomalous oxygen isotopic compositions indicate formation in the atmospheres of evolved stars. Two grains are identified as pyroxene, two as olivine, one as a glass with embedded metal and sulfides (GEMS), and one as an Al-rich silicate. One grain is enriched in 26Mg, which is attributed to the radioactive decay of 26Al and provides information about mixing processes in the parent star. This discovery opens new means for studying stellar processes and conditions in various solar system environments.

Aluminum↗

Placental permeability of lead.

The detection of lead in fetal tissues by chemical analysis has long been accepted as prima facie evidence for the permeability of the placenta to this nonessential trace metal. However, only a few investigations, all on lower mammalian species, have contributed any direct experimental data bearing on this physiological process. Recent radioactive tracer and radioautographic studies on rodents have shown that lead crosses the placental membranes rapidly and in significant amounts even at relatively low maternal blood levels. While it is not possible to extrapolate directly the results of these experiments to humans because of differences in placental structure and other factors, the results do serve as a warning of the possible hazard to the human embryo and fetus of even low levels of lead in the maternal system.

Abortion, Spontaneous↗

Evidence for an inducible cadmium binding protein metallothionein in chick embryo.

Cadmium binding protein metallothionein was isolated in a partially purified form from the liver of cadmium treated chick embryos. This binding protein was totally absent in normal chick embryo, though the induction occurred with cadmium exposure in the embryonic liver. Incorporation studies with 3H-leucine or 3H-lysine indicated the active synthesis of metallothionein after 10 hrs of cadmium injection. Since the characteristic of metallothionein is by its metal content, we estimated the cadmium concentration in all fractions and it was evident that cadmium peak corresponded only with the peak of radioactive amino acid incorporation to metallothionein. This result suggests that binding protein is specific for cadmium. Further experiments have been conducted with actinomycin D treatment to cadmium treated embryo to understand the mechanism of metallothionein induction. The incorporation of 3H-leucine into hepatic metallothionein of cadmium exposed embryos was not inhibited by actinomycin D treatment. suggesting that cadmium does not act at the level of transcription in the metallothionein biosynthesis, occurred in chick embryo.

Animals↗

Ferritin. Binding of beryllium and other divalent metal ions.

Rat liver homogenates in 0.1 M Tris, pH 7.5, were heated to 80 degrees C, cooled immediately, and centrifuged at 24,000 X g, and 7Be2+ was added to the supernatant. Twenty-five per cent of the radioactivity was bound to a single protein. It was purified to homogeneity and identified to be ferritin as judged by different criteria. These were sucrose density gradient centrifugation, electrophoresis in polyacrylamide gel of the native or sodium dodecyl sulfate-treated protein, reactivity to antibodies, isoelectric focusing, and total amino acid composition. Comparative study of the ability of ferritin or apoferritin to bind Cd2+, Zn2+, Cu2+, and Be2+ was conducted by using a gel equilibrium technique, Centifree micropartition technique, and microcentrifuge desalting technique. Ferritin could be saturated with Cd2+ or Zn2+ or Cu2+ but not with Be2+ even after 800 g atoms of Be2+ were bound. None of the bound Be2+ was dialyzable at 4 degrees C in 0.05 Tris acetate buffer, pH 8.5, but at pH 6.5 over 80% of the bound metal ion was dialyzed after 72 h. By contrast, apoferritin bound similar amounts of all four metal ions, some of which were dialyzable. By spectrophotometric titrations at pH 6.5 of Be2+ with sulfosalicylic acid (SSA), BeKDSSA was calculated to be 5.0 X 10(-6) M and by competition of sulfosalicyclic acid and ferritin for Be2+ the BeKDferritin was calculated to be 6.8 X 10(-6) M.

Animals↗

Radionuclide applications in laboratory studies of environmental surface reactions.

The advantages of using radionuclides for laboratory studies of environmental processes include the wide range of element concentrations that can be studied, the capability to simultaneously study several isotopes in a single experiment, the direct applicability to the behaviour of radioactive waste or fallout, and the ability to study the mechanisms, reversibility and kinetics of environmental reactions under controlled conditions. These attributes are demonstrated using specific examples drawn from case studies in Australia, including radionuclide fallout onto tropical soils, the association of trace metals with harbour sediments and the behaviour of uranium in natural and contaminated systems.

Adsorption↗

[Study on air-powder abrasive system for prosthetic clinical application. 3. Bacterial adherence to the air-powder abraded metal surface].

The purpose of this study is to investigate the relation between the metal surface texture and bacterial adherence. The surface of Au-Ag-Pd alloy was treated with or without air-powder abrasion. Seven standard strains and two clinical isolates were used. The alloys were mixed with [6-3H]-thimidine labelled bacteria. The radioactivity was measured with a scintilation counter. Streptococcus sanguis and clinical isolated Bacteroides gingivalis were adhered in a larger amount to the air-powder abraded alloy surface than to nonabraded alloy surface. These results indicated that careful consideration is needed for application of air-powder abrasive system on metal crown with secondary caries and periodontal disease.

Bacterial Adhesion↗

The existence of specific antibodies to cobalt in hard metal asthma.

Twelve workers with hard metal asthma diagnosed on the basis of peak flow diaries and positive bronchial reactions to cobalt chloride (CoCl2) were studied for sensitization by detection of specific antibodies to radioactive cobalt (57Co), cobalt-conjugated human serum albumin (Co-HSA) and cobalt-conjugated exchange resin (Co-resin). Their IgE titres ranged from 73 to 1500 IU/ml and eight were atopic individuals. Sixty serum samples from asthmatic patients with IgE titres of 14-4300 IU/ml were studied as controls in all tests. Eleven of twelve subject sera that selectively bound to 57Co after incubation with saturated ammonium sulphate (greater than 232 c.p.m., P less than 0.01) were divided into three groups: (1) six sera showing evidence of specific IgE antibodies to Co-HSA (greater than 673 c.p.m., P less than 0.01) without those to Co-resin; (2) one serum giving a positive radio-allergosorbent test (RAST) only to Co-resin (greater than 417 c.p.m., P less than 0.01), and (3) four sera that were negative for two antigenic agents (Co-HSA, Co-resin). These results suggest that the subjects had occupational asthma due to hard metal exposure from cobalt sensitivity. An immuno-allergic mechanism mediated by specific IgE antibodies to cobalt was confirmed to be responsible for the development of hard metal asthma, with the possibility of some role of the reaction without reagins.

Adult↗

Distribution of heavy metals and radionuclides in sediments, water, and fish in an area of Great Bear Lake contaminated with mine wastes.

The concentrations of heavy metals and radionuclides in the sediments and water of Great Bear Lake were determined during 1978 near an operating silver mine and an abandoned uranium mine. Additional information on the level of mercury in fish tissues were also collected. The mines, situated on the same site, deposited tailings and other waste material directly into the lake. The concentrations of mercury, lead, manganese, and nickel in the sediments were highest near the tailings deposit and decreased significantly as the distance from the mine increased. Although there were also significant positive correlations between these metals and the organic content of the sediments, water depth and slope of the bottom had no impact on metal distribution. Since the concentrations of arsenic, cobalt, copper, 226radium, 210lead and 230thorium varied inconsistently throughout the study area, the distribution of these substances could not be related to any of the environmental factors that were measured. There were, however, significant negative correlations between the concentrations of 232thorium and 228thorium and distance from the mine and organic content of the sediments. Heavy metal and radionuclide levels in water were generally below detectable limits, reflecting the strong chemical bonding characteristics of the sediments. The low concentrations of mercury in the tissues of lake trout Salvelinus namaycush were probably related to low uptake rates and the ability of this species to move into uncontaminated areas of the lake.

Animals↗

Detection of zirconium in organic and aqueous phases using an inductively coupled plasma-mass spectrometer and the same aqueous standards.

Solvent extraction requires measurement of both aqueous and organic phases, and common techniques of such measurements are tedious and time consuming. This paper outlines an inductively coupled plasma-mass spectrometric technique in which both aqueous and organic phases (both contained in a carrier phase, ethanol) can be measured using the same calibration curve. The method has proved to be successful as long as the organic and aqueous content in the carrier phase is kept below 10%. The developed technique is a more convenient tool for solvent extraction chemists who have often used radioactive tracers within their experimental techniques. In addition, this technique may be used in environmental studies involving the analysis of organic fractions, such as metal fingerprinting of petroleum fluids. In such analyses, the ability to use more available and traceable aqueous standards will lead to significantly improved confidence in the analytical results.

Journal Article↗

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↗

Vertical profiles and enrichment pattern of natural radionuclides in monazite areas of coastal Kerala.

Detailed studies on radionuclides concentration in different environmental matrices of high background areas were undertaken in the coastal areas of Karunagapalli, Chavara, Neendakara and Kollam to study the distribution and enrichment of the radionuclides in the region. The sand samples collected at different distances from sea waterline and at different depths, were analysed for primordial radionuclides by gamma spectrometry. The activity of primordial radionuclides was determined for the different size fractions of sand to study the enrichment pattern. The highest activity was found confined in 125-63 microm particle size fraction in sand. The minimum (232)Th activity was 9.4 Bq kg(-1), found in Kollam at a depth of 10-20 cm, 40 m away from waterline in 500-250 microm particle size fraction and maximum activity of 136,811.2 Bq kg(-1) was observed in Chavara in grains of size 125-63 microm at a depth of 0-10 cm for a sample collected 20 m away from waterline. The lowest (226)Ra activity observed was 29.6 Bq kg(-1) at Kollam beach for a sample 40 m away from waterline in grains of size 1000-500 microm and at a depth of 20-30 cm and the highest activity observed was 10,309 Bq kg(-1) in grains of size 125-63 microm for a sample collected at a distance 20 m away from waterline and at a depth of 0-10 cm. The activity of (40)K was below detectable level in most of the samples collected from the high background monazite area. The (232)Th, (226)Ra activities decrease with depth for the samples collected 20 m away from the waterline and increase with depth for the samples collected 40 m away from the waterline at Chavara and Kollam beaches. No definite correlation was found between variation of the concentrations of (232)Th and (226)Ra with depth at Karunagapalli and Neendakara beach sands. There exists a strong correlation between (226)Ra and (232)Th activities in the region. The results of these investigations are presented and discussed in this paper.

Background Radiation↗

Novel rhenium complexes derived from alpha-tropanol as potential ligands for the dopamine transporter.

A series of rhenium complexes was synthesized as model compounds for the corresponding radioactive technetium-99m complexes for preliminary biological investigations. In a '3 + 1' mixed-ligand approach the tropanol molecule was linked with the metal core through a omega-mercaptoester group as monodentate ligand. Bis(thioethyl)sulphide was used as the tridentate dithiol ligand to block the remaining free coordination sites. The omega-mercaptoesters were synthesized via the trityl-protected precursors. Binding tests on the cloned human dopamine transporter revealed moderate binding affinities for some of the prepared compounds. The complexes were characterised by X-ray and the lipophilicity as well as pKa values were determined.

Carrier Proteins↗

Norwegian monitoring programme on the inorganic and organic contaminants in fish caught in the Barents Sea, Norwegian Sea and North Sea, 1994-2001.

The results from part of a monitoring programme of contaminant levels in fish and other seafood products initiated by the Directorate of Fisheries in Norway in 1994 are presented. Concentrations of 22 elements (four are presented here: As, Cd, Hg and Pb) and HCB, HCH, PCB 28, 52, 101, 105, 118, 138, 153, 156, 180, pp-DDD, p-DDE, p-DDT, sum DDT and (137)Cs were determined in 17 species of fish caught in three sampling locations: the Barents Sea, the Norwegian Sea and the North Sea. The fish species analysed in the survey were limited to species of commercial importance for Norway with catching volumes of at least 10,000 metric tons year(-1). The survey started in 1994 and is expected to continue beyond 2010. The analyses were carried out on 25 individual fish from each species and each sampling location, and the locations were representative of commercial fishing grounds for the species in question. The concentrations of contaminants found were considerably lower than the maximum levels permissible in fish set by CODEX and the European Union for contaminants in seafood products.

Animals↗

Ribonucleotide reductase of Brevibacterium ammoniagenes is a manganese enzyme.

Ribonucleotide reduction and not DNA replication is the site for the specific manganese requirement of DNA synthesis and cell growth in the coryneform bacterium Brevibacterium ammoniagenes. To characterize the metal effect we have isolated and purified ribonucleoside-diphosphate reductase from overproducing bacteria that were first deprived of and then reactivated by manganese ions. Purification on columns of Sephacryl S400, DEAE-cellulose and hydroxyapatite provided an apparently homogeneous enzyme consisting of two protein subunits. These were characterized by affinity chromatography on 2',5'-ADP-Sepharose as nucleotide-binding protein B1 (Mr = 80,000) and catalytic protein B2 (Mr = 100,000, composed of two Mr = 50,000 polypeptides), which were both necessary for activity. In vitro the purified enzyme does not require added metal ions except for an unspecific, twofold activity increase observed in the presence of Mg2+ and other divalent cations. Enzyme activity is inhibited by hydroxyurea (I50 = 2.5 mM). The electronic spectrum with maxima around 455 nm and 485 nm closely resembles that of manganese(III)-containing pseudocatalase and of oxo-bridged binuclear Mn(III) model complexes. Denaturation of the enzyme in trichloroacetic acid liberated an equimolar amount of Mn(II) which was detected by EPR spectroscopy. It was not possible to remove and reintroduce metal ions without loss of enzyme activity. Manganese-deficient cell cultures were also grown in the presence of 54MnCl2. Ribonucleotide reductase activity and radioactivity cochromatographed in several systems. Non-denaturing polyacrylamide gel electrophoresis showed that protein subunit B2 was specifically 54Mn-labeled. All these properties suggest that the ribonucleotide reductase of B. ammoniagenes is a manganese-containing analog of the non-heme-iron-containing reductases of Escherichia coli and eukaryotes.

Brevibacterium↗