An experimental study of 226Ra and 45Ca accumulation from the aquatic medium by freshwater turtles (fam. Chelidae) under varying Ca and Mg water concentrations

Snapping turtles Elseya dentata (Gray) from Magela Creek, Northern Territory, were exposed under laboratory conditions for up to 30 days to waters resembling the inorganic composition of Magela Creek water during the Wet season, with background and elevated Ca and Mg concentrations, that were labelled with ²²⁶Ra and ⁴⁵Ca. The resulting concentrations of ⁴⁵Ca in muscle, skin, gut, liver, shell bone and leg bone of E. dentata equilibrated or approached equilibrium by 12–18 days. Among the experiments, the concentrations of ⁴⁵Ca in all six tissues were inversely related to turtle mass. An increase in the Ca water concentration by a factor of 15 increased the ⁴⁵Ca concentration in all six tissues. The arithmetic factors of increase in the concentration in each tissue were proportional or more than proportional to the factor of increase in Ca water concentration; this factor was highest for muscle tissue (26.6). An increase in the Mg water concentration by a factor of 48 reduced the ⁴⁵Ca concentration in all tissues except skin where it increased. The concentration of ²²⁶Ra in each tissue (except the gut) was positively related to its ⁴⁵Ca concentration and inversely related to turtle mass in muscle, skin and liver. With the exception of the skin, the increased Ca water concentration did not reduce the ²²⁶Ra in the tissues but increased the ²²⁶Ra concentration in bone and muscle. The increased Mg water concentration had an inverse effect on the ²²⁶Ra concentrations in all tissues except shell. With the exception of the skin the effects of increased Ca and Mg water concentrations and turtle size on ²²⁶Ra concentrations in the tissues of E. dentata were similar to their effects on ⁴⁵Ca tissue concentrations, indicating the similar metabolic behaviour of ²²⁶Ra and ⁴⁵Ca in E. dentata.Exposures of the species Elseya latisternum (Gray), Emydura signata (Ahl) and Chelodina longicollis (Shaw), which are the same or closely related to species reported to occur in Magela Creek, to ⁴⁵Ca-labelled Sydney tap water for 7 days demonstrated their ability to also accumulate ⁴⁵Ca from their aquatic medium. The patterns of ⁴⁵Ca concentrations in the tissues of these species indicated that they were inversely related to turtle mass, as demonstrated in E. dentata. The concentrations of ⁴⁵Ca accumulated in the tissues were also comparable to those found in single specimens of E. dentata and E. victoriae (Gray) that were exposed for 7 days to simulated Magela Creek water. The data also indicated the larger long-necked C. longicollis accumulated less ⁴⁵Ca per gram of tissue than similar-sized, short-necked species E. signata and E. latisternum, suggesting that long-necked turtles from Magela Creek would accumulate less ²²⁶Ra from their aquatic medium than similar-sized short-necked species.The capacity of E. dentata to accumulate ²²⁶Ra from the aquatic medium is about two orders of magnitude less than that of the tissue of the freshwater mussel Velesunio angasi (Sowerby) exposed under similar experimental conditions.     

Sources and fluxes of submarine groundwater discharge delineated by radium isotopes

This paper reports the results derived fromradium isotopes of a submarine groundwaterdischarge (SGD) intercomparison in thenortheast Gulf of Mexico. Radium isotopesamples were collected from seepage meters,piezometers, and surface and deep ocean waters.Samples collected within the near-shore SGDexperimental area were highly enriched in allfour radium isotopes; offshore samples wereselectively enriched. Samples collected fromseepage meters were about a factor of 2–3higher in radium activity compared to theoverlying waters. Samples from piezometers,which sampled 1–4 meters below the sea bed were1–2 orders of magnitude higher in radiumisotopes than surface waters. The twolong-lived Ra isotopes, ²²⁸Ra and²²⁶Ra, provide convincing evidence thatthere are two sources of SGD to the study area:shallow seepage from the surficial aquifer andinput from a deeper aquifer. A three end-membermixing model can describe the Ra distributionin these samples. The short-lived radium isotopes, ²²³Ra and²²⁴Ra, were used to establish mixing ratesfor the near-shore study area. Mixing wasretarded within 3 km of shore due to a strongsalinity gradient. The product of the mixingrate and the offshore ²²⁶Ra gradientestablished the ²²⁶Ra flux. This flux mustbe balanced by Ra input from SGD. The flux ofSGD within 200 m of shore based on the²²⁶Ra budget was 1.5 m³ min^?1.This flux agreed well with other estimatesbased on seepage meters and ²²²Rn.     

Bone cancer risk of (239)pu in humans derived from animal models

Two-mutation model fits to bone cancer mortality data from mice, rats and beagle dogs injected with (239)Pu or (226)Ra show that (1) it is possible to fit the radiation-related parameters for animals from different strains of the same species together; (2) for every species the same significant parameters are found in the models for (239)Pu and in the models for (226)Ra, and the only difference is in the value of the linear mutation coefficient; and (3) the toxicity ratio, when defined as the ratio of the linear mutation coefficients for (239)Pu over (226)Ra, has a relatively uniform value of approximately 8 for the species considered. This relatively constant ratio enables the development of a (239)Pu model for humans that is based on the radium dial painters and the toxicity ratio for beagles. The model predictions agree well with published risk estimates based on other data and derived using alternative approaches. This has two important implications: (1) The two-mutation model appears to be a useful tool in translating from animal models to humans in a meaningful way; and (2) once a two-mutation model for humans has been derived, radiation risks can be calculated that depend on doses, dose rates and ages at exposure. Such a model therefore supplements published risk estimates that often lack such dependences.     

Activation and significance of vacuolar H+-ATPase in Saccharomyces cerevisiae adaptation and resistance to the herbicide 2,4-dichlorophenoxyacetic acid

The stimulation of the activity of the H(+)-ATPase present in the vacuolar membrane (V-ATPase) of Saccharomyces cerevisiae is here described in response to a moderate stress induced by 2,4-dichlorophenoxyacetic acid (2,4-D). This in vivo activation (up to 5-fold) took place essentially during the adaptation period, preceding cell division under herbicide stress, in coordination with a marked activation of plasma membrane H(+)-ATPase (PM-ATPase) (up to 30-fold) and the decrease of intracellular and vacuolar pH values, suggesting that activation may be triggered by acidification. Single deletion of VMA1 and genes encoding other V-ATPase subunits led to a more extended period of adaptation and to slower growth under 2,4-D stress. Results suggest that a functional V-ATPase is required to counteract, more rapidly and efficiently, the dissipation of the physiological H(+)-gradient across vacuolar membrane registered during 2,4-D adaptation.     

Comparative evaluation of 238U and 226Ra absorption by herbaceous and woody species under conditions of technogenic pollution

The results of studies on uranium and radium distribution in soil and vegetation at the territory contaminated by wastes of radium plant in 1930-1950 are presented. Specific activities of 226Ra and 238U in soil varied as 0.01-200 and 0.004-7.58 Bq/g per ash respectively. The radionuclides are mainly concentrated in the upper layer of soil profile. In the vegetation concentrations of 226Ra and 238U varied as 0.64-132 0.001-0.02 Bq/g per ash respectively. Among the woody species studied, Betula pubescens and Sorbus aucuparia are characterized by the highest absorption of radionuclides. For all plants studied a negative linear dependence is shown between logarithm of TF (TF = [226Ra, 238U in plant, Bq/g per ash]/[226Ra, 238U in soil, Bq/g per ash]) and logarithm of 226Ra and 238U concentrations in soil. The findings obtained can be used in modeling of biological absorption in radioecological investigations.     

Functional characterization of a NapA Na(+)/H(+) antiporter from Thermus thermophilus

Na(+)/H(+) antiporters are ubiquitous membrane proteins and play an important role in cell homeostasis. We amplified a gene encoding a member of the monovalent cation:proton antiporter-2 (CPA2) family (TC 2.A.37) from the Thermus thermophilus genome and expressed it in Escherichia coli. The gene product was identified as a member of the NapA subfamily and was found to be an active Na(+)(Li(+))/H(+) antiporter as it conferred resistance to the Na(+) and Li(+) sensitive strain E. coli EP432 (DeltanhaA, DeltanhaB) upon exposure to high concentration of these salts in the growth medium. Fluorescence measurements using the pH sensitive dye 9-amino-6-chloro-2-methoxyacridine in everted membrane vesicles of complemented E. coli EP432 showed high Li(+)/H(+) exchange activity at pH 6, but marginal Na(+)/H(+) antiport activity. Towards more alkaline conditions, Na(+)/H(+) exchange activity increased to a relative maximum at pH 8, where by contrast the Li(+)/H(+) exchange activity reached its relative minimum. Substitution of conserved residues D156 and D157 (located in the putative transmembrane helix 6) with Ala resulted in the complete loss of Na(+)/H(+) activity. Mutation of K305 (putative transmembrane helix 10) to Ala resulted in a compromised phenotype characterized by an increase in apparent K(m) for Na(+) (36 vs. 7.6 mM for the wildtype) and Li(+) (17 vs. 0.22 mM), In summary, the Na(+)/H(+) antiport activity profile of the NapA type transporter of T. thermophilus resembles that of NhaA from E. coli, whereas in contrast to NhaA the T. thermophilus NapA antiporter is characterized by high Li(+)/H(+) antiport activity at acidic pH.     

Activity regulation of the betaine transporter BetP of Corynebacterium glutamicum in response to osmotic compensation

As a response to hyperosmotic stress bacterial cells accumulate compatible solutes by synthesis or by uptake. Beside the instant activation of uptake systems after an osmotic upshift, transport systems show also a second, equally important type of regulation. In order to adapt the pool size of compatible solutes in the cytoplasm to the actual extent of osmotic stress, cells down-regulate solute uptake when the initial osmotic stress is compensated. Here we describe the role of the betaine transporter BetP, the major uptake carrier for compatible solutes in Corynebacterium glutamicum, in this adaptation process. For this purpose, betP was expressed in cells (C. glutamicum and Escherichia coli), which lack all known uptake systems for compatible solutes. Betaine uptake mediated by BetP as well as by a truncated form of BetP, which is deregulated in its response to hyperosmotic stress, was dissected into the individual substrate fluxes of unidirectional uptake, unidirectional efflux and net uptake. We determined a strong decrease of unidirectional betaine uptake by BetP in the adaptation phase. The observed decrease in net uptake was thus mainly due to a decrease of V_max of BetP and not a consequence of the presence of separate efflux system(s). These results indicate that adaptation of BetP to osmotic compensation is different from activation by osmotic stress and also different from previously described adaptation mechanisms in other organisms. Cytoplasmic K⁺, which was shown to be responsible for activation of BetP upon osmotic stress, as well as a number of other factors was ruled out as triggers for the adaptation process. Our results thus indicate the presence of a second type of signal input in the adaptive regulation of osmoregulated carrier proteins.     

Regulation of intracellular level of Na+, K+ and glycerol in Saccharomyces cerevisiae under osmotic stress

The intracellular level of Na⁺ and K⁺ of S. cerevisiae strain AB1375 revealed that under KCl as well as sorbitol stress, the cationic level was comparable to the level under no stress conditions. On the other hand, there was a sharp drop in the intracellular K⁺ content and increase in the Na⁺ content on addition of NaCl to the medium. However, the total cationic level was close to that under control conditions. In addition to changes in the cationic level, an enhanced production and accumulation of glycerol were also observed under osmotic stress. A regulatory mechanism co-ordinating the intracellular concentration of glycerol as well as Na⁺, K⁺ content under osmotic stress conditions has been proposed.     

The metabolism of radium-226 during pregnancy in the rat

Metabolism of radium including the transfer to the fetus through the placenta was studied during three successive pregnancies 92, 155, and 213 days after injection of 226Ra in young female rats. The cumulative fecal and urinary excretions of 226Ra in a 213-day period following injection were about 30 and 15% injected dose (%ID), respectively, most of them occurring during the first 42 days. The excretions were similar in both the pregnant and control (unmated) rats. The whole-body burden of radium (mostly in the skeleton) determined by actual analysis of the entire body was similar in the two groups and was about 53, 48, and 44 %ID at the first, second, and third pregnancy, respectively. Pregnancy alone, therefore, did not significantly affect metabolism of radium. At 20 days of gestation the mean placental content of radium was 0.005, 0.0045, and 0.0036 %ID in the first, second, and third litter, respectively; the corresponding mean fetal content was 0.01, 0.008, and 0.005 %ID. The radium burden of the full-term neonate (21-22 days) was 0.014 and 0.011 %ID for the first and second delivery, respectively. The total amount calculated of radium transferred from the mother to the 8-10 fetuses in a litter did not exceed about 0.3% of the maternal content per each pregnancy. Comparison of the ratio of radium and calcium in the fetus and maternal skeleton shows that there is a Ra-Ca discrimination during their passage from the mother to the fetus.     

Effect of inactivation of ccpA and aerobic growth in Lactobacillus plantarum: A proteomic perspective

Lactobacillus plantarum is a facultative heterofermentative lactic acid bacterium widely used in the production of most fermented food due to its ability to thrive in several environmental niches, including the human gut. In order to cope with different growth conditions, it has developed complex molecular response mechanisms, characterized by the induction of a large set of proteins mainly regulated by HrcA and CtsR repressors as well as by global regulators such as carbon catabolite control protein A (CcpA). In this study, the role of CcpA in the regulation of growth under anaerobiosis and aerobiosis, and the adaptation to aeration in L. plantarum WCFS1 were comprehensively investigated by differential proteomics. The inactivation of ccpA, in both growth conditions, significantly changed the expression level of 76 proteins, mainly associated with carbohydrate and energy metabolism, membrane transport, nucleotide metabolism, protein biosynthesis and folding. The role of CcpA as pleiotropic regulator was particularly evident at the shift from homolactic fermentation to mixed fermentation. Proteomic results also indicated that the mutant strain was more responsive to aerobic growth condition.     

A proteomic approach to study Cry1Ac binding proteins and their alterations in resistant Heliothis virescens larvae

Binding of the Bacillus thuringiensis Cry1Ac toxin to specific receptors in the midgut brush border membrane is required for toxicity. Alteration of these receptors is the most reported mechanism of resistance. We used a proteomic approach to identify Cry1Ac binding proteins from intestinal brush border membrane (BBM) prepared from Heliothis virescens larvae. Cry1Ac binding BBM proteins were detected in 2D blots and identified using peptide mass fingerprinting (PMF) or de novo sequencing. Among other proteins, the membrane bound alkaline phosphatase (HvALP), and a novel phosphatase, were identified as Cry1Ac binding proteins. Reduction of HvALP expression levels correlated directly with resistance to Cry1Ac in the YHD2-B strain of H. virescens. To study additional proteomic alterations in resistant H. virescens larvae, we used two-dimensional differential in-gel electrophoresis (2D-DIGE) to compare three independent resistant strains with a susceptible strain. Our results validate the use of proteomic approaches to identify toxin binding proteins and proteome alterations in resistant insects.     

Characteristics of erythrocyte lipids in blood of tundra voles (Microtus oeconomus Pall.) inhabiting areas with increased natural radioactivity

Interrelations between the lipid characteristics of the blood erythrocytes and 226Ra accumulation in the body of tundra voles (Microtus oeconomus Pall.) inhabiting areas with different levels of the radiation background were investigated. It is shown that the ratio of the phospholipid (PL) fractions which cause the blood erythrocyte structure depends on the phase of the population cycle, as well as on the sex and age of tundra voles. The statistically significant interrelation between lysoforms and the sphingomielin content has been revealed in the blood erythrocyte PL of tundra voles; its scale somewhat differs for the animals from the reference and Ra areas. The peroxide concentration in the blood erythrocyte lipids of tundra voles from the Ra area exceeded the control values in all mature groups of the animals trapped at the depression phase of the population density. The 226Ra content in the bodies of the tundra voles which were trapped in the Ra area at the increased phase of the population cycle is for certain higher than that for the animals from the reference area. Interrelations between the lipid peroxidation parameters in the blood erythrocytes and the body 226Ra content for separate sex-age groups of tundra voles have been revealed.     

No single way to understand singlet oxygen signalling in plants

When plant cells are under environmental stress, several chemically distinct reactive oxygen species (ROS) are generated simultaneously in various intracellular compartments and these can cause oxidative damage or act as signals. The conditional flu mutant of Arabidopsis, which generates singlet oxygen in plastids during a dark-to-light transition, has allowed the biological activity of singlet oxygen to be determined, and the criteria to distinguish between cytotoxicity and signalling of this particular ROS to be defined. The genetic basis of singlet-oxygen-mediated signalling has been revealed by the mutation of two nuclear genes encoding the plastid proteins EXECUTER (EX)1 and EX2, which are sufficient to abrogate singlet-oxygen-dependent stress responses. Conversely, responses due to higher cytotoxic levels of singlet oxygen are not suppressed in the ex1/ex2 background. Whether singlet oxygen levels lower than those that trigger genetically controlled cell death activate acclimation is now under investigation.