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Callosal neurones give rise to zinc-rich boutons in the rat visual cortex.

Sodium selenide was used as a retrograde tracer to assess the callosal origin of zinc-rich boutons in the neocortex of the rat. Selenide injections were placed in the lateral end of area Oc1 (area 17). Zinc present in synaptic boutons precipitated as zinc selenide and was transported retrogradely to the parent cell bodies. In addition to the ipsilateral labelling, contralateral retrogradely labelled somata were mainly observed along the border between areas Oc1 and Oc2L (area 18a). Labelled neurones were present in layer 2-3 and layer 6. In contrast to tracing studies with peroxidase, no labelled neurone was observed in layers 4 and 5 except at the inner border of layer 5. This study reveals the chemical heterogeneity of callosal projections, which may be divided into zinc-rich and zinc-poor systems.

Animals↗

Is the postganglionic sympathetic neuron zinc-enriched? A stop-flow nerve crush study on rat sciatic nerve.

Axonal transport of endogenous zinc ions in the rat sciatic nerve was studied by a stop-flow/nerve crush technique combined with zinc selenide autometallography (ZnSeAMG) at light and electron microscopic levels. Distinct accumulations of ZnSeAMG grains were detected, in particular proximal but also distal to the crushes, 1.5 h after the operation, and the amounts of zinc ions increased further in the following 3-8 h. Ultrastructurally, ZnSeAMG grains were located predominantly in unmyelinated axons. The data suggest that a subpopulation of sciatic nerve axons contains and transports zinc ions both antero- and retrogradely, indicating that the second neuron in the sympathetic nervous system is zinc enriched (ZEN).

Animals↗

Effects of sodium selenate treatment on altered responses of left and right atria from streptozotocin-induced diabetic rats.

Experiments were designed to determine whether experimental diabetes alters inotropic and chronotropic effects of adrenergic, adenosinergic, and cholinergic agonists and whether the observed changes are prevented by sodium selenate therapy. Thirty-two rats were divided into four groups of eight subjects each. Diabetes induced by streptozotocin caused significant decreases in isoproterenol-invoked contraction of the left atria with preservation of the right atrial chronotropic responses. The diminished response of the left atrial muscle to isoproterenol did not respond to treatment with sodium selenate. The left atria adenosine-induced direct- and indirect inotropic responses were diminished in the diabetic rats. After treatment with sodium selenate the direct response was completely normalized, but the indirect response was only partially corrected. Adenosine-induced negative chronotropic effects are accompanied by changed responses in diabetic right atria that are corrected after treatment. The carbachol-induced inotropic and chronotropic responses were not altered in tests of the acetylcholine system. We conclude that in diabetic rats, sodium selenate treatment reverses the deficits of adenosine-induced negative inotropic responses of left and right atria, but not those of isoproterenol-induced positive inotropic responses.

Animals↗

Effects of 5 HIV protease inhibitors on vasomotor function and superoxide anion production in porcine coronary arteries.

HIV protease inhibitors (PIs) have been implicated to cause cardiovascular complications. Previous studies demonstrated that the PI ritonavir (RTV) caused endothelial dysfunction in porcine arteries. This study investigated and compared the effects of 5 commonly used PIs on vasomotor function, endothelial nitric oxide synthase (eNOS) expression, and oxidative stress in porcine coronary arteries. Vessel rings were incubated with 15 microM of RTV, amprenavir (APV), saquinavir (SQV), indinavir (IDV), or nelfinavir (NFV) for 24 hours. Vasomotor function was studied using a myograph system. The contractility of the rings was significantly reduced for RTV and SQV. In response to bradykinin at 10(-5) M, the endothelium-dependent relaxation was significantly reduced for RTV, APV, and SQV. The eNOS mRNA levels were significantly reduced for RTV, APV, and SQV. Furthermore, the superoxide anion (O(2)(-)) levels of the vessels were significantly increased for RTV and APV. It was found that nitric oxide production was decreased, whereas the level of nitrotyrosine proteins was increased in RTV-treated vessels. Furthermore, antioxidant seleno-L-methionine (SeMet) reversed RTV-induced O(2)(-) production and vasomotor dysfunction. Thus, the HIV PIs RTV, APV, and SQV at 15 microM have more potent in vitro effects on vasomotor dysfunction, eNOS downregulation, and O(2)(-) production than IDV and NFV. The antioxidant SeMet can block these adverse effects of RTV. The results suggest that antioxidant therapy may have applications for controlling PI-associated cardiovascular complications.

Animals↗

Sulfurospirillum barnesii sp. nov. and Sulfurospirillum arsenophilum sp. nov., new members of the Sulfurospirillum clade of the epsilon Proteobacteria.

Two strains of dissimilatory arsenate-reducing vibrio-shaped bacteria are assigned to the genus Sulfurospirillum. These two new species, Sulfurospirillum barnesii strain SES-3T and Sulfurospirillum arsenophilum strain MIT-13T, in addition to Sulfurospirillum sp. SM-5, two strains of Sulfurospirillum deleyianum, and Sulfurospirillum arcachonense, form a distinct clade within the epsilon subclass of the Proteobacteria based on 16S rRNA analysis.

Arsenates↗

Bordetella petrii sp. nov., isolated from an anaerobic bioreactor, and emended description of the genus Bordetella.

A novel Bordetella species was isolated from an anaerobic, dechlorinating bioreactor culture enriched from river sediment. The only strain, Se-1111R(T) (= DSM 12804T = CCUG 43448T), for which the name Bordetella petrii is proposed, is designated the type strain of the novel species. Strain Se-1111R(T) was isolated from the dechlorinating mixed culture due to its ability to anaerobically reduce selenate to elemental selenium. Comparative 16S rDNA sequence analysis showed a close relationship between Se-1111R(T) and members of the genus Bordetella within the beta-Proteobacteria. This close phylogenetic relatedness was also reflected in several metabolic properties of Se-1111R(T), including its incapacity to utilize carbohydrates, by the high G+C content (63.8 mol%) of its DNA and by the presence of Q-8 as the major isoprenoid quinone. DNA-DNA hybridization experiments with type strains of all species of the genus Bordetella and closely related species Achromobacter xylosoxidans subsp. denitrificans provided further evidence for the assignment of strain Se-1111R(T) as a novel species of the genus Bordetella. This genus currently consists of seven aerobic species, all of which are known to occur in close pathogenic, opportunistic or possibly commensal relationships with various host organisms. B. petrii is the first member of this genus isolated from the environment and capable of anaerobic growth. The proposal of the novel species and an emended description of the genus Bordetella is presented.

Anaerobiosis↗

Salana multivorans gen. nov., sp. nov., a novel actinobacterium isolated from an anaerobic bioreactor and capable of selenate reduction.

Three facultatively anaerobic, Gram-positive bacteria, strains Se-3111T, Se-13111 and Se-1311A, were isolated from an anaerobic, dechlorinating bioreactor culture enriched from sediment of the River Saale in Germany. All strains were isolated from the dechlorinating mixed culture through their ability to reduce selenate anaerobically to elemental selenium. All three strains shared identical 16S rDNA sequences and phylogenetic analysis revealed that strain Se-3111T forms a novel taxon within the suborder Micrococcineae of the class Actinobacteria, related most closely to Beutenbergia cavernae. On the basis of genotypic, chemotaxonomic and physiological characteristics, it is proposed that the novel strains Se-3111T, Se-13111 and Se-1311A be classified in a new genus as Salana multivorans gen. nov., sp. nov. The type strain of the novel species is Se-3111T (= DSM 13521T = NRRL B-24118T).

Actinobacteria↗

Inventory and functional characterization of the HAK potassium transporters of rice.

Plants take up large amounts of K(+) from the soil solution and distribute it to the cells of all organs, where it fulfills important physiological functions. Transport of K(+) from the soil solution to its final destination is mediated by channels and transporters. To better understand K(+) movements in plants, we intended to characterize the function of the large KT-HAK-KUP family of transporters in rice (Oryza sativa cv Nipponbare). By searching in databases and cDNA cloning, we have identified 17 genes (OsHAK1-17) encoding transporters of this family and obtained evidence of the existence of other two genes. Phylogenetic analysis of the encoded transporters reveals a great diversity among them, and three distant transporters, OsHAK1, OsHAK7, and OsHAK10, were expressed in yeast (Saccharomyces cerevisiae) and bacterial mutants to determine their functions. The three transporters mediate K(+) influxes or effluxes, depending on the conditions of the experiment. A comparative kinetic analysis of HAK-mediated K(+) influx in yeast and in roots of K(+)-starved rice seedlings demonstrated the involvement of HAK transporters in root K(+) uptake. We discuss that all HAK transporters may mediate K(+) transport, but probably not only in the plasma membrane. Transient expression of the OsHAK10-green fluorescent protein fusion protein in living onion epidermal cells targeted this protein to the tonoplast.

Amino Acid Sequence↗

Selenium assimilation and volatilization from selenocyanate-treated Indian mustard and muskgrass.

Selenocyanate (SeCN(-)) is a major contaminant in the effluents from some oil refineries, power plants, and in mine drainage water. In this study, we determined the potential of Indian mustard (Brassica juncea) and muskgrass (a macroalga, Chara canescens) for SeCN(-) phytoremediation in upland and wetland situations, respectively. The tolerance of Indian mustard to toxic levels of SeCN(-) was similar to or higher than other toxic forms of Se. Indian mustard treated with 20 microM SeCN(-) removed 30% (w/v) of the Se supplied in 5 d, accumulating 554 and 86 microg of Se g(-1) dry weight in roots and shoots, respectively. Under similar conditions, muskgrass removed approximately 9% (w/v) of the Se supplied as SeCN(-) and accumulated 27 microg of Se g(-1) dry weight. A biochemical pathway for SeCN(-) degradation was proposed for Indian mustard. Indian mustard and muskgrass efficiently degraded SeCN(-) as none of the Se accumulated by either organism remained in this form. Indian mustard accumulated predominantly organic Se, whereas muskgrass contained Se mainly as selenite and organic Se forms. Indian mustard produced volatile Se from SeCN(-) in the form of less toxic dimethylselenide. Se volatilization by Indian mustard accounted for only 0.7% (w/v) of the SeCN(-) removed, likely because the biochemical steps in the production of dimethylselenide from organic Se were rate limiting. Indian mustard is promising for the phytoremediation of SeCN(-) -contaminated soil and water because of its remarkable abilities to phytoextract SeCN(-) and degrade all the accumulated SeCN(-) to other Se forms.

Biodegradation, Environmental↗

Characterization of a selenate-resistant Arabidopsis mutant. Root growth as a potential target for selenate toxicity.

Screening an Arabidopsis (Arabidopsis thaliana) T-DNA mutant library for selenate resistance enabled us to isolate a selenate-resistant mutant line (sel1-11). Molecular and genetic characterization showed that the mutant contained a lesion in the SULTR1;2 gene that encodes a high affinity root sulfate transporter. We showed that SULTR1;2 is the only gene among 13 mutated genes of the Arabidopsis sulfate transporter family whose mutation conferred selenate resistance to Arabidopsis. The selenate resistance phenotype of the sel1-11 mutant was mirrored by an 8-fold increase of root growth in the presence of selenate as shown by the calculated lethal concentration values. The impairment of SULTR1;2 activity in sel1-11 resulted in a reduced (35)S-sulfate uptake capacity by both roots and calli and a reduced sulfate and selenate content in root, shoot, and calli. Comparing sulfate-to-selenate ratios instead of absolute sulfate and selenate contents in roots and shoots enabled us to gain better insight into the mechanism of selenate toxicity in Arabidopsis. Roots of the sel1-11 mutant line showed a higher sulfate to selenate ratio than that of wild-type roots, while there were no significant differences in sulfate to selenate ratios in shoots of wild-type and mutant lines. These results indicated that the mechanism that confers the selenate resistance phenotype to the sel1-11 line takes place rather in the roots. It might be in part the result of a lower selenate uptake and of a protective effect of sulfate against the toxic effects of selenate on root growth. These results revealed in plants a central and specific role of the transporter SULTR1;2 in selenate sensitivity; they further suggested that root growth and potentially the root tip activity might be a specific target of selenate toxicity in Arabidopsis.

Anion Transport Proteins↗

Effect of ATP sulfurylase overexpression in bright yellow 2 tobacco cells. Regulation Of atp sulfurylase and SO4(2-) transport activities.

To determine if the ATP sulfurylase reaction is a regulatory step for the SO4(2-)-assimilation pathway in plants, an Arabidopsis thaliana ATP sulfurylase cDNA, APS2, was fused to the 35S promoter of the cauliflower mosaic virus and introduced by Agrobacterium tumefaciens-mediated transformation into isolated Bright Yellow 2 tobacco (Nicotiana tabacum) cells. The ATP sulfurylase activity in transgenic cells was 8-fold that in control cells, and was correlated with the expression of a specific polypeptide revealed by western analysis using an anti-ATP sulfurylase antibody. The molecular mass of this polypeptide agreed with that for the overexpressed mature protein. ATP sulfurylase overexpression had no effect on [35S]SO4(2-) influx or ATP sulfurylase activity regulation by S availability, except that ATP sulfurylase activity variations in response to S starvation in transgenic cells were 8 times higher than in the wild type. There were also no differences in cell growth or sensitivity to SeO4(2-) (a toxic SO4(2-) analog) between transgenic and wild-type cells. We propose that in Bright Yellow 2 tobacco cells, the ATP sulfurylase derepression by S deficiency may involve a posttranscriptional mechanism, and that the ATP sulfurylase abundance is not limiting for cell metabolism.

Arabidopsis↗

X-ray multiple-wave coherent interaction in a quasi-two-dimensional material NbSe2-2H.

The first observation of the X-ray multiple-wave interaction in an incommensurate charge-density-wave (CDW) modulated structure at low temperatures is reported for an example of a quasi-two-dimensional material, NbSe(2)-2H. Via the coherent interaction between the X-ray waves propagating in the CDW-modulated structure and the host structure, the phase-dependent intensity variations of a CDW reflection were detected. In accord with a centrosymmetric structure, the phases of the structure-factor triplets of two CDW reflections and a Bragg reflection of the host structure were determined to be either 0 or 180 degrees, and not to vary with temperature. Relative phase differences of the two CDW reflections are also deduced.

Crystallography, X-Ray↗

Crystal chemistry and binding of NO2, SCN and SeCN to Co in cobalamins.

Results of the accurate crystal structure determination of NO(2)Cbl.2LiCl (1), NO(2)Cbl.NaCl (2), NCSCbl (3) and NCSeCbl (4), based on synchrotron diffraction data collected at 100 K, are described. The nitro group in (1) was found to be disordered with two orientations that differ by a rotation of approximately 60 degrees about the Co-NO(2) bond, whereas in (2) the nitro group has only one orientation. The first X-ray structural determination of a cobalamin with a Co-Se bond is reported. Comparison of the axial distances indicates that SeCN has a bond length of 2.384 (3) A and that the trans influence on the Co-N bond is only slightly greater than that of SCN. The crystals of the thiocyanate cobalamin contain both the S- and N-bonded coordination isomers in a 3:2 ratio. The structural features of the Co-S bond in cobalamins are discussed. The crystal chemistry of cobalamins is discussed in terms of packing of roughly spherical molecules. The unit-cell parameters can be used to group the cobalamins' crystal structures in different arrays intermediate between distorted hexagonal close packing and primitive hexagonal arrangements. The structural features of cobalamins, and of cobaloximes that have the same axial fragment as the cobalamins, are reviewed and discussed in terms of the cis influence of the equatorial ligand.

Crystallization↗

Structural characterization of selenium and selenium-diiodine analogues of the antithyroid drug 6-n-propyl-2-thiouracil and its alkyl derivatives.

The structures of four selenium analogues of the antithyroid drug 6-n-propyl-2-thiouracil [systematic name: 2,3-dihydro-6-n-propyl-2-thioxopyrimidin-4(1H)-one], namely 6-methyl-2-selenouracil, C(5)H(6)N(2)OSe (1), 6-ethyl-2-selenouracil, C(6)H(8)N(2)OSe (2), 6-n-propyl-2-selenouracil, C(7)H(10)N(2)OSe (3), and 6-isopropyl-2-selenouracil, C(7)H(10)N(2)OSe (4), are described, along with that of the dichloromethane monosolvate of 6-isopropyl-2-selenouracil, C(7)H(10)N(2)OSe.CH(2)Cl(2) (4.CH(2)Cl(2)). The extended structure of (1) is a two-dimensional sheet of topology 6(3) with a brick-wall architecture. The extended structures of (2) and (4) are analogous, being based on a chain of eight-membered R(8)(6)(32) hydrogen-bonded rings. In (3) and (4.CH(2)Cl(2)), R(2)(2)(8) hydrogen bonding links molecules into chains. 6-n-Propyl-2-selenouracil.I(2), C(7)H(10)N(2)OSe.I(2) (7), is a charge-transfer complex with a ;spoke' structure, the extended structure of which is based on a linear chain formed principally by intermolecular N-H...O hydrogen bonds. Re-crystallization of 6-ethyl-2-selenouracil or (7) from acetone gave crystals of the diselenides [N-(6'-ethyl-4'-pyrimidone)(6-ethyl-2-selenouracil)(2)(Se-Se)].2H(2)O (9.2H(2)O) or [N-(6'-n-propyl-4'-pyrimidone)(6-n-propyl-2-selenouracil)(2)(Se-Se)] (10), respectively: these have similar extended chain structures formed via N-H...O and C-H...O hydrogen bonds, stacked to give two-dimensional sheets. Re-crystallization of (7) from methanol/acetonitrile led via deselenation to the formation of crystals of 6-n-propyl-2-uracil (11), in which six symmetry-related molecules combine to form a six-membered R(6)(6)(24) hydrogen-bonded ring, with each pair of molecules linked by an R(2)(2)(8) motif.

Antithyroid Agents↗

Solving the phase problem for carbohydrate-binding proteins using selenium derivatives of their ligands: a case study involving the bacterial F17-G adhesin.

The Escherichia coli adhesin F17-G is a carbohydrate-binding protein that allows the bacterium to attach to the intestinal epithelium of young ruminants. The structure of the 17 kDa lectin domain of F17-G was determined using the anomalous dispersion signal of a selenium-containing analogue of the monosaccharide ligand N-acetyl-d-glucosamine in which the anomeric oxygen was replaced by an Se atom. A three-wavelength MAD data set yielded good experimental phases to 2.6 A resolution. The structure was refined to 1.75 A resolution and was used to solve the structures of the ligand-free protein and the F17-G-N-acetyl-d-glucosamine complex. This selenium-carbohydrate phasing method could be of general use for determining the structures of carbohydrate-binding proteins.

Acetylglucosamine↗