Anisotropy studies around the galactic centre at EeV energies with the Auger Observatory

Data from the Pierre Auger Observatory are analyzed to search for anisotropies near the direction of the Galactic Centre at EeV energies. The exposure of the surface array in this part of the sky is already significantly larger than that of the fore-runner experiments. Our results do not support previous findings of localized excesses in the AGASA and SUGAR data. We set an upper bound on a point-like flux of cosmic rays arriving from the Galactic Centre which excludes several scenarios predicting sources of EeV neutrons from Sagittarius $A$. Also the events detected simultaneously by the surface and fluorescence detectors (the `hybrid' data set), which have better pointing accuracy but are less numerous than those of the surface array alone, do not show any significant localized excess from this direction.Comment: Matches published versio

Enzymological studies of one-carbon reactions in the pathway of acetate utilization by methanogenic bacteria

Several enzymes in the pathway of acetate conversion to methane and carbon dioxide have been purified from Methanosarcina thermophila. The mechanisms of these enzymes are under investigation utilizing biochemical, biophysical and molecular genetic approaches. Acetate kinase and phosphotransacetylase catalyzes the activation of acetate to acetyl-CoA. The primary structure of these enzymes will be determined through cloning and sequencing of the genes. Two protein components of the CO dehydrogenase complex are under investigations. The metal centers of each component have been characterized using EPR. Cloning and sequencing of the genes for the two subunits of each component is in progress. Results indicate that the Ni/Fe-S component cleaves the C-C and C-S bonds of acetyl-CoA followed by oxidation of the carbonyl group to carbon dioxide and transfer of the methyl group to the Co/Fe-S component. The enzymes and cofactors involved in transfer of the methyl group from the Co/Fe-S component to coenzyme M will be purified and characterized. Ferredoxin is an electron acceptor for the Ni/Fe-S component and also serves to reductively reactivate methylreductase which catalyzes the demethylation of methyl coenzyme M to methane. This ferredoxin is being characterized utilizing EPR and RR spectroscopic methods to determine the properties of the Fe-S centers. Genes encoding this and other ferredoxins have been cloned and sequenced to determine the primary structures. Carbonic anhydrase is being purified and characterized to determine the function of this enzyme in the pathway

Digesta flows in sheep fed poor-quality hay supplemented with urea and carbohydrates

Two metabolism trials were conducted with 12 yearling crossbred wethers per trial (34 and 38 kg for trials 1 and 2, respectively). The wethers, equipped with ruminal, abomasal and ileal cannulae, were randomly allotted for each trial to the following treatments: 1) hay alone or hay supplemented with 2) .9% urea, 3) 1% urea and 6.5% molasses or 4) 1% urea and 5.2% corn. Two digestive flow markers were used: Cr2O3 powder and Co-ethylenediaminetetraacetic acid (Co-EDTA). Urea and Co-EDTA were infused continuously into the rumen via cannula. Daily dry matter (DM) intake averaged 517 g. Urea supplementation improved N retention (P less than .01). Apparent digestibility of DM, acid detergent fiber (ADF) and energy was not affected by treatment. Urea and carbohydrate supplementation increased ruminal propionic acid molar proportions (P less than .05). Apparent ruminal DM digestion accounted for 41% of the total DM degraded, whereas 77.4% of the digestible ADF was degraded in the rumen. Urea supplementation increased ADF digestion in the large intestine (P less than .01). Urea and carbohydrate supplementation resulted in a stepwise increase in N flowing with the liquid phase at the abomasum. Mean retention times of the solid and liquid phases of digestive contents were similar across treatments. Overall, benefits of supplementation of poor-quality fescue hay diets by small amounts of urea and readily available carbohydrates remain questionable for sheep fed at a fixed level of intake below maintenance

Methanogenic Conversion of CO2 Into CH4

This SBIR project evaluated the potential to remediate geologic CO2 sequestration sites into useful methane gas fields by application of methanogenic bacteria. Such methanogens are present in a wide variety of natural environments, converting CO2 into CH4 under natural conditions. We conclude that the process is generally feasible to apply within many of the proposed CO2 storage reservoir settings. However, extensive further basic R&D still is needed to define the precise species, environments, nutrient growth accelerants, and economics of the methanogenic process. Consequently, the study team does not recommend Phase III commercial application of the technology at this early phase

Methanogenic Conversion of CO2 Into CH4

This SBIR project evaluated the potential to remediate geologic CO2 sequestration sites into useful methane gas fields by application of methanogenic bacteria. Such methanogens are present in a wide variety of natural environments, converting CO2 into CH4 under natural conditions. We conclude that the process is generally feasible to apply within many of the proposed CO2 storage reservoir settings. However, extensive further basic R&D still is needed to define the precise species, environments, nutrient growth accelerants, and economics of the methanogenic process. Consequently, the study team does not recommend Phase III commercial application of the technology at this early phase

Carbonic anhydrase inhibitors. Inhibition of the prokariotic beta and gamma-class enzymes from Archaea with sulfonamides

A detailed inhibition study of carbonic anhydrases (CAs, EC 4.2.1.1) belonging to the β- and γ-families from Archaea with sulfonamides has been performed. Compounds included in this study were the clinically used sulfonamide CA inhibitors, such as acetazolamide, methazolamide, ethoxzolamide, topiramate, valdecoxib, celecoxib, dorzolamide, sulfanilamide, dichlorophanamide, as well as sulfanilamide analogs, halogenated sulfanilamides, and some 1,3-benzenedisulfonamide derivatives. The two γ-CAs from Methanosarcina thermophila (Zn-Cam and Co-Cam) showed very different inhibitory properties with these compounds, as compared to the α-CA isozymes hCA I, II, and IX, and the β-CA from Methanobacterium thermoautotrophicum (Cab). The best Zn-Cam inhibitors were sulfamic acid and acetazolamide, with inhibition constants in the range of 63–96 nM, whereas other investigated aromatic/heterocylic sulfonamides showed a rather levelled behavior, with KIs in the range of 0.12–1.70 μM. The best Co-Cam inhibitors were topiramate and p-aminoethyl-benzenesulfonamide, with KIs in the range of 0.12–0.13 μM, whereas the worst one was homosulfanilamide (KI of 8.50 μM). In the case of Cab, the inhibitory power of these compounds varied to a much larger extent, with sulfamic acid and sulfamide showing millimolar affinities (KIs in the range of 44–103 mM), whereas the best inhibitor was ethoxzolamide, with a KI of 5.35 μM. Most of these sulfonamides showed inhibition constants in the range of 12–100 μM against Cab. Thus, the three CA families investigated up to now possess a very diverse affinity for sulfonamides, the inhibitors with important medicinal, and environmental applications

1G99 : AN ANCIENT ENZYME: ACETATE KINASE FROM METHANOSARCINA THERMOPHILA

Experimental Technique/Method:X-RAY DIFFRACTION Resolution:2.5 Classification:TRANSFERASE Release Date:2000-12-27 Deposition Date:2000-11-22 Revision Date:2008-04-27#2011-07-13#2018-01-31 Molecular Weight:89834.51 Macromolecule Type:Protein Residue Count:816 Atom Site Count:6120 DOI:10.2210/pdb1g99/pdb Abstract: Acetate kinase, an enzyme widely distributed in the Bacteria and Archaea domains, catalyzes the phosphorylation of acetate. We have determined the three-dimensional structure of Methanosarcina thermophila acetate kinase bound to ADP through crystallography. As we previously predicted, acetate kinase contains a core fold that is topologically identical to that of the ADP-binding domains of glycerol kinase, hexokinase, the 70-kDa heat shock cognate (Hsc70), and actin. Numerous charged active-site residues are conserved within acetate kinases, but few are conserved within the phosphotransferase superfamily. The identity of the points of insertion of polypeptide segments into the core fold of the superfamily members indicates that the insertions existed in the common ancestor of the phosphotransferases. Another remarkable shared feature is the unusual, epsilon conformation of the residue that directly precedes a conserved glycine residue (Gly-331 in acetate kinase) that binds the alpha-phosphate of ADP. Structural, biochemical, and geochemical considerations indicate that an acetate kinase may be the ancestral enzyme of the ASKHA (acetate and sugar kinases/Hsc70/actin) superfamily of phosphotransferases