Esterase screening using whole cells of Brazilian soil microorganisms

A miniaturized enzymatic assay using fluorescent probes to reveal esterase producing microorganisms was optimized and applied to screen 64 soil bacterial strains. The best results were validated using traditional non-fluorogenic assays with acetyl and propanoyl phenylethanol to confirm the miniaturized results. The most active microorganisms belong to the genus Bacillus showing esterase activity and good enantiomeric ratios for the resolution of phenylethanol derivatives (E > 30). Part of the microorganisms are kept in our laboratory in glycerol or freeze-dried and the best microorganisms will be deposited in the CBMAI/CPQBA/Unicamp culture collection.Um ensaio enzimático miniaturizado com sondas fluorescentes para triagem rápida de microrganismos produtores de esterases foi implementado e aplicado a 64 linhagens de bactérias de solo. Os melhores resultados foram validados por métodos convencionais e substratos não fluorogênicos (feniletanol acetilado e propanoilado) e confirmaram os resultados obtidos nas triagens rápidas. Os microrganismos que apresentaram atividades enzimáticas mais relevantes (razão enantiomérica E > 30) na hidrólise de ésteres e na obtenção de boas razões enantioméricas foram identificados como pertencentes ao gênero Bacillus. Parte dos microrganismos se encontra preservada em glicerol e/ou liofilizada no IQ/Unicamp e três linhagens serão depositadas na CBMAI do CPQBA/Unicamp como depósitos abertos.14841489Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Esterase screening using whole cells of Brazilian soil microorganisms

A miniaturized enzymatic assay using fluorescent probes to reveal esterase producing microorganisms was optimized and applied to screen 64 soil bacterial strains. The best results were validated using traditional non-fluorogenic assays with acetyl and propanoyl phenylethanol to confirm the miniaturized results. The most active microorganisms belong to the genus Bacillus showing esterase activity and good enantiomeric ratios for the resolution of phenylethanol derivatives (E > 30). Part of the microorganisms are kept in our laboratory in glycerol or freeze-dried and the best microorganisms will be deposited in the CBMAI/CPQBA/Unicamp culture collection

New investigations on the 32S(3He,d)33Cl reaction at 9.6 MeV bombarding energy

The 32S(3He,d)33Cl one-proton transfer reaction is a powerful tool to investigate the spectroscopy of low-lying states in the proton-rich 33Cl nucleus. However, the extraction of firm differential cross-section data at various angles to benchmark and constrain theoretical models is made challenging by the presence of competitive reactions on target contaminants. In this paper we report on arecent measurement using a new generation hodoscope of silicon detectors, capable to detect and identify emitted deuterons down to energies of the order of 2 MeV. The high angular segmentation of our hodoscope combined with a suitable target to control possible contaminants, allowed to unambiguously disentangle the contribution of various states in 33Cl, in particular the 2.352 MeV state lying just few tens of keV above the proton separation energy

Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450_(BM3) for Selective Hydroxylation of Large Substrates

Made for each other: Combinatorial alanine substitution of active site residues in a thermostable cytochrome P450_(BM3) variant was used to generate an enzyme that is active with large substrates. Selective hydroxylation of methoxymethylated monosaccharides, alkaloids, and steroids was thus made possible (see Scheme). This approach could be useful for improving the activity of enzymes that show only limited activity with larger substrates

Charged particle effects: Experimental and theoretical studies on the mechanisms underlying the induction of molecular and cellular damage and the modulation of intercellular signalling

In this paper we present the main outcomes of a wide collaborative effort (carried out within the INFN project “EPICA” and in part within the European projects “RISC-RAD” and “NOTE” and the ASI project MoMa-COUNT), both experimental and theoretical, devoted to the characterization and quantification of the induction of DNA-targeted and non-DNA-targeted molecular and cellular biological endpoints, following irradiation of human cells with different charged particles. The work was mainly aimed at reaching a better understanding of the mechanisms governing the physical and biophysical pathways leading from the initial energy deposition by radiation in matter to the induction of observable radiobiological damage, with particular focus on the role played by radiation quality. More specifically, we characterized the induction of DNA DSB within different fragment-size ranges outlining the effectiveness of high-LET radiation at inducing small fragments and thus clustered DNA breaks, which can evolve in terms of endpoints like chromosome aberrations (CAs). This was confirmed by the development and application of a model of CA induction based on the assumption that only clustered DNA breaks can lead to aberrations. Concerning non-DNA-targeted damage, we quantified the time-dependent induction of medium-mediated DNA damage in bystander cells and we characterized the time and dose dependence of cytokine concentration in the culture medium of sham-irradiated and irradiated cells, since medium-mediated bystander damage is thought to arise from molecular signalling between irradiated and unirradiated cells. The mechanisms governing such signalling were investigated developing a model and a MC code simulating cytokine release, diffusion and internalization, showing good agreement with experimental data. Non-DNA-targeted effects were further characterized by MRS investigation of the radiation effects on lipids and oxidative metabolism, which are particularly relevant also considering that they may be differently expressed in different tumors and in normal tissues

A Search for Jet Handedness in Hadronic Z0Z^0 Decays

We have searched for signatures of polarization in hadronic jets from $Z^0 \to q \bar{q}$ decays using the ``jet handedness'' method. The polar angle asymmetry induced by the high SLC electron-beam polarization was used to separate quark jets from antiquark jets, expected to be left- and right-polarized, respectively. We find no evidence for jet handedness in our global sample or in a sample of light quark jets and we set upper limits at the 95% C.L. of 0.063 and 0.099 respectively on the magnitude of the analyzing power of the method proposed by Efremov {\it et al.}Comment: Revtex, 8 pages, 2 figure

Enabling FAIR research in Earth Science through research objects

Data-intensive science communities are progressively adopting FAIR practices that enhance the visibility of scientific breakthroughs and enable reuse. At the core of this movement, research objects contain and describe scientific information and resources in a way compliant with the FAIR principles and sustain the development of key infrastructure and tools. This paper provides an account of the challenges, experiences and solutions involved in the adoption of FAIR around research objects over several Earth Science disciplines. During this journey, our work has been comprehensive, with outcomes including: an extended research object model adapted to the needs of earth scientists; the provisioning of digital object identifiers (DOI) to enable persistent identification and to give due credit to authors; the generation of content-based, semantically rich, research object metadata through natural language processing, enhancing visibility and reuse through recommendation systems and third-party search engines; and various types of checklists that provide a compact representation of research object quality as a key enabler of scientific reuse. All these results have been integrated in ROHub, a platform that provides research object management functionality to a wealth of applications and interfaces across different scientific communities. To monitor and quantify the community uptake of research objects, we have defined indicators and obtained measures via ROHub that are also discussed herein.Published550-5645IT. Osservazioni satellitariJCR Journa