Isozyme-Specific Ligands for O-acetylserine sulfhydrylase, a Novel Antibiotic Target

Conceived and designed the experiments: FS PC BC ES AM. Performed the experiments: FS RS ES PF SR. Analyzed the data: FS BC ES PF GEK PFC AM. Contributed reagents/materials/analysis tools: PC PB GC. Wrote the paper: FS GEK BC AM.The last step of cysteine biosynthesis in bacteria and plants is catalyzed by O-acetylserine sulfhydrylase. In bacteria, two isozymes, O-acetylserine sulfhydrylase-A and O-acetylserine sulfhydrylase-B, have been identified that share similar binding sites, although the respective specific functions are still debated. O-acetylserine sulfhydrylase plays a key role in the adaptation of bacteria to the host environment, in the defense mechanisms to oxidative stress and in antibiotic resistance. Because mammals synthesize cysteine from methionine and lack O-acetylserine sulfhydrylase, the enzyme is a potential target for antimicrobials. With this aim, we first identified potential inhibitors of the two isozymes via a ligand- and structure-based in silico screening of a subset of the ZINC library using FLAP. The binding affinities of the most promising candidates were measured in vitro on purified O-acetylserine sulfhydrylase-A and O-acetylserine sulfhydrylase-B from Salmonella typhimurium by a direct method that exploits the change in the cofactor fluorescence. Two molecules were identified with dissociation constants of 3.7 and 33 µM for O-acetylserine sulfhydrylase-A and O-acetylserine sulfhydrylase-B, respectively. Because GRID analysis of the two isoenzymes indicates the presence of a few common pharmacophoric features, cross binding titrations were carried out. It was found that the best binder for O-acetylserine sulfhydrylase-B exhibits a dissociation constant of 29 µM for O-acetylserine sulfhydrylase-A, thus displaying a limited selectivity, whereas the best binder for O-acetylserine sulfhydrylase-A exhibits a dissociation constant of 50 µM for O-acetylserine sulfhydrylase-B and is thus 8-fold selective towards the former isozyme. Therefore, isoform-specific and isoform-independent ligands allow to either selectively target the isozyme that predominantly supports bacteria during infection and long-term survival or to completely block bacterial cysteine biosynthesis.Yeshttp://www.plosone.org/static/editorial#pee

Catalytic cracking of n-hexadecane on mixed Al-MCM-41/MFI catalyst systems with regard to selectivity for C-3 and C-3 products

Recently, the large pore MCM-41 material has gained a considerable amount of interest, as a solution to the diffusion limitation provided by the conventional microporous cracking catalysts for the cracking of long-chain or bulky hydrocarbon feeds. The present work deals with the synthesis and characterization of novel mixed Al-MCM-41/MFI catalyst systems. Furthermore, the catalytic behavior of the as-synthesized Al-MCM-41/MFI mixed systems for cracking hydrocarbons by MAT (Micro Activity Test), standardized by ASTM D-3907, have been investigated in the present work. In situ formation of the meso- and microporous structures Al-MCM-41 and ZSM-5 have been obtained using a two-template [C16H33(CH3)(3)NBr and (CH3CH2CH2)(4)NBr] synthesis gel system. The mixed phases were obtained by optimizing template ratios and reaction temperature. The comparison with conventional zeolite Y and pure MCM-41 with regard to activity and selectivity showed a better activity and selectivity towards C-3 and C-4 fractions. The obtained results reveal a potential of mixed Al-MCM-41/ZSM-5 catalyst systems as cracking catalysts and, in turn, demand further optimization of physicochemical properties in the future in order to be exploited as commercial cracking catalysts

The conundrum in smart city governance: Interoperability and compatibility in an ever-growing ecosystem of digital twins

Today, technological developments are ever-growing yet fragmented. Alongside inconsistent digital approaches and attitudes across city administrations, such developments have made it difficult to reap the benefits of city digital twins. Bringing together experiences from five research projects, this paper discusses these digital twins based on two digital integration methodologies - systems and semantic integration. We revisit the nature of the underlying technologies, and their implications for interoperability and compatibility in the context of planning processes and smart urbanism. Semantic approaches present a new opportunity for bidirectional data flows that can inform both governance processes and technological systems to co-create, cross-pollinate, and support optimal outcomes. Building on this opportunity, we suggest that considering the technological dimension as a new addition to the trifecta of economic, environmental, and social sustainability goals that guide planning processes, can aid governments to address this conundrum of fragmentation, interoperability, and compatibility

Multi-technique physico-chemical characterization of particles generated by a gasoline engine: Towards measuring tailpipe emissions below 23 nm

Particulate emissions from on-road motor vehicles are the focus of intensive current research due to the impact of the ambient particulate matter (PM) levels on climate and human health. Constant improvement in engine technology has led to significant decrease in the number and mass of emitted PM, but particular concern is raised nowadays by the ultrafine particles. In this context, there is a critical lack of certification procedures for the measurement of the smallest-size (<23 nm) particulate matter emissions. To support the engine development process as well as future certification procedures, a measurement technology for sub-23 nm particles must be designed. The development of a reliable measurement procedure entails understanding the formation and evolution of particles from the engine to the tailpipe via multiple analytical techniques and theoretical simulations. We present here extensive experimental characterization of ultrafine particles emitted by a gasoline direct injection single-cylinder engine as particle generator. The particles were sampled using a cascade impactor which allows size-separation into 13 different size bins. Chemical characterization of the collected size-selected particles was performed using mass spectrometry, which gives access to detailed molecular information on chemical classes of critical interest such as organosulphates, oxygenated hydrocarbons, nitrogenated hydrocarbons, metals, or polycyclic aromatic hydrocarbons. Additionally, the morphology of the emitted particles was probed with atomic force (AFM) and scanning electron microscopy (SEM). Tip-Enhanced Raman Spectroscopy (TERS) was applied for the first time to sub-10 nm combustion-generated particles to gather information on their nanostructure. The extensive database built from these multiple experimental characterizations has been used as input of a theoretical approach to simulate and validate engine out-emissions. These studies were performed in the framework of the H2020 PEMS4Nano project which aims to the development of a robust, reliable and reproducible measurement technology for particles down to 10 nm for both chassis dyno and real driving emissions (RDE)