Purification and Characterization of an L-Amino Amidase from Mycobacterium neoaurum ATCC 25795

An L-amino amidase from Mycobacterium neoaurum ATCC 25795 responsible for the enantioselective resolution of DL-α-methyl valine amide was purified and characterized. The purification procedure included ammonium sulfate fractionation, gel filtration, and anion-exchange chromatography, which resulted in a homogeneous preparation of the enzyme with a native molecular mass of 136 kDa and a subunit molecular mass of 40 kDa. The purified enzyme displayed the highest activity at 50°C and at pH 8.0 and 9.5. The enzyme was strongly inhibited by the metal-chelating agent 1,10-phenanthroline, the disulfide-reducing agent dithiothreitol, and the cysteine proteinase inhibitor iodoacetamide. The purified amino amidase showed a unique L-enantioselective activity towards a broad range of both α-H- and α-alkyl-substituted amino acid amides, with the highest activity towards the cyclic amino acid amide DL-proline amide. No activity was measured with DL-mandelic acid amide nor with the dipeptide L-phenylalanine-L-leucine. The highest catalytic efficiency (kcat/Km ratio) was measured with DL-α-allyl alanine amide, DL-α-methyl phenylalanine amide, and DL-α-methyl leucine amide

Mercury emissions and stable isotopic compositions at Vulcano Island (Italy)

Sampling and analyses methods for determining the stable isotopic compositions of Hg in an active volcanic system were tested and optimized at the volcanic complex of Vulcano (Aeolian Islands, Italy). Condensed gaseous fumarole Hg(fum) T , plume gaseous elemental Hg(g) 0 and plume particulate Hg(p) II were obtained at fumaroles F0, F5, F11, and FA. The average total Hg emissions, based on HgT/SO2 in condensed fumarolic gases and plumes, range from 2.5 to 10.1 kg y−1, in agreement with published values [Ferrara, R., Mazzolai, B., Lanzillotta, E., Nucaro, E., Pirrone, N., 2000. Volcanoes as emission sources of atmospheric mercury in the Mediterranean Basin. Sci. Total Environ. 259(1–3), 115–121; Aiuppa, A., Bagnato, E., Witt, M.L.I., Mather, T.A., Parello, F., Pyle, D.M., Martin, R.S., 2007. Real-time simultaneous detection of volcanic Hg and SO2 at La Fossa Crater, Vulcano (Aeolian Islands, Sicily). Geophys. Res. Lett. 34(L21307).]. Plume Hg(p) II increases with distance from the fumarole vent, at the expense of Hg(g) 0 and indicates significant in-plume oxidation and condensation of fumarole Hg(fum) T . Relative to the NIST SRM3133 Hg standard, the stable isotopic compositions of Hg are δ202Hg(fum) T =−0.74‰±0.18 (2SD, n=4) for condensed gaseous fumarole Hg(fum) T , δ202Hg(g) 0 =−1.74‰±0.36 (2SD, n=1) for plume gaseous elemental Hg(g) 0 at the F0 fumarole, and δ202Hg(p) II =−0.11‰±0.18 (2SD, n=4) for plume particulate Hg(p) II . The enrichment of Hg(p) II in the heavy isotopes and Hg(g) 0 in the light isotopes relative to the total condensed fumarolic Hg(fum) T gas complements the speciation data and demonstrates a gas-particle fractionation occurring after the gas expulsion inambient T° atmosphere. A first order Rayleigh equilibriumcondensation isotope fractionation model yields a fractionation factor αcond-gas of 1.00135±0.00058

The giant eyes of giant squid are indeed unexpectedly large, but not if used for spotting sperm whales

© The Author(s), 2013. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in BMC Evolutionary Biology 13 (2013): 187, doi:10.1186/1471-2148-13-187.We recently reported (Curr Biol 22:683–688, 2012) that the eyes of giant and colossal squid can grow to three times the diameter of the eyes of any other animal, including large fishes and whales. As an explanation to this extreme absolute eye size, we developed a theory for visual performance in aquatic habitats, leading to the conclusion that the huge eyes of giant and colossal squid are uniquely suited for detection of sperm whales, which are important squid-predators in the depths where these squid live. A paper in this journal by Schmitz et al. (BMC Evol Biol 13:45, 2013) refutes our conclusions on the basis of two claims: (1) using allometric data they argue that the eyes of giant and colossal squid are not unexpectedly large for the size of the squid, and (2) a revision of the values used for modelling indicates that large eyes are not better for detection of approaching sperm whales than they are for any other task. We agree with Schmitz et al. that their revised values for intensity and abundance of planktonic bioluminescence may be more realistic, or at least more appropriately conservative, but argue that their conclusions are incorrect because they have not considered some of the main arguments put forward in our paper. We also present new modelling to demonstrate that our conclusions remain robust, even with the revised input values suggested by Schmitz et al

A multi-resolution strategy for a multi-objective deformable image registration framework that accommodates large anatomical differences

Currently, two major challenges dominate the field of deformable image registration. The first challenge is related to the tuning of the developed methods to specific problems (i.e. how to best combine different objectives such as similarity measure and transformation effort). This is one of the reasons why, despite significant progress, clinical implementation of such techniques has proven to be difficult. The second challenge is to account for large anatomical differences (e.g. large deformations, (dis)appearing structures) that occurred between image acquisitions. In this paper, we study a framework based on multi-objective optimization to improve registration robustness and to simplify tuning for specific applications. Within this framework we specifically consider the use of an advanced model-based evolutionary algorithm for optimization and a dual-dynamic transformation model (i.e. two "non-fixed" grids: one for the source- and one for the target image) to accommodate for large anatomical differences. The framework computes and presents multiple outcomes that represent efficient trade-offs between the different objectives (a so-called Pareto front). In image processing it is common practice, for reasons of robustness and accuracy, to use a multi-resolution strategy. This is, however, only well-established for single-objective registration methods. Here we describe how such a strategy can be realized for our multi-objective approach and compare its results with a single-resolution strategy. For this study we selected the case of prone-supine breast MRI registration. Results show that the well-known advantages of a multi-resolution strategy are successfully transferred to our multi-objective approach, resulting in superior (i.e. Pareto-dominating) outcomes

Purification and characterization of an L-aminopeptidase from Pseudomonas putida ATCC 12633

An L-aminopeptidase of Pseudomonas putida, used in an industrial process for the hydrolysis of D,L-amino acid amide racemates, was purified to homogeneity. The highly L-enantioselective enzyme resembled thiol reagent-sensitive alk. serine proteinases was strongly activated by divalent cations. It possessed a high substrate specificity for dipeptides and a-H amino acid amides, e.g., L-phenylglycine amide. [on SciFinder (R)

Purification and characterization of an L-aminopeptidase from Pseudomonas putida ATCC 12633

An L-aminopeptidase of Pseudomonas putida, used in an industrial process for the hydrolysis of D,L-amino acid amide racemates, was purified to homogeneity. The highly L-enantioselective enzyme resembled thiol reagent-sensitive alk. serine proteinases was strongly activated by divalent cations. It possessed a high substrate specificity for dipeptides and a-H amino acid amides, e.g., L-phenylglycine amide. [on SciFinder (R)