Dynamic Reductive Kinetic Resolution of Benzyl Ketones using Alcohol Dehydrogenases and Anion Exchange Resins

Dynamic reductive kinetic resolutions of racemic 3‐arylalkanones have been performed by the proper combination of an alcohol dehydrogenase and a basic anionic resin. The best results were found for the bioreduction with the alcohol dehydrogenase type A from Rhodococcus ruber DSM 44541 overexpressed in Escherichia coli (E. coli/ADH‐A) and the commercially available evo‐1.1.200, while the Amberlite IRA‐440 C and the DOWEX‐MWA‐1 resins allowed efficient in situ racemizations. Reaction conditions were optimized in terms of enzyme source and loading, type and amount of resin, pH, temperature and reaction times, obtaining a series of (R,R)‐substituted propan‐2‐ols with good conversions and both diastereoselectivity and stereoselectivity. As a proof of concept, the subsequent intramolecular cyclization of a selected propan‐2‐ol substrate afforded a valuable isochroman heterocycle without any loss of the optical purity.

     

But‐2‐ene‐1,4‐diamine and But‐2‐ene‐1,4‐diol as Donors for Thermodynamically Favored Transaminase‐ and Alcohol Dehydrogenase‐Catalyzed Processes

Both cis‐ and trans‐but‐2‐ene‐1,4‐diamines have been prepared and efficiently applied as sacrificial cosubstrates in enzymatic transamination reactions. The best results were obtained with the cis‐diamine. The thermodynamic equilibrium of the stereoselective transamination process is shifted to the amine formation due to tautomerization of 5H‐pyrrole into 1H‐pyrrole, achieving high conversions (78–99%) and enantiomeric excess (up to >99%) by using a small excess of the amine donor. Furthermore, when the reaction proceeded, a strong coloration was observed due to polymerization of 1H‐pyrrole. A structurally related compound, cis‐but‐2‐ene‐1,4‐diol, has been utilized as cosubstrate in different alcohol dehydrogenase (ADH)‐mediated bioreductions. In this case, high conversions (91–99%) were observed due to a lactonization process. Both strategies are convenient from both synthetic and atom economy points of view in the production of valuable optically active products.

     

Respirometric calibration and validation of a biological nitrite oxidation model including biomass growth and substrate inhibition

The modelling of the nitrification process of high-strength ammonium wastewater must be designed to consider it as a two-step reaction with substrate inhibition. Consequently, kinetic and stoichiometric parameters of both steps are required. In this work, the second step in the nitrification process was studied: a biological nitrite oxidation model was formulated, calibrated and validated using only oxygen uptake rate (OUR) measurements. The model included biomass growth and substrate inhibition. First, the biomass yield coefficient for nitrite-oxidising biomass was determined. Then, a respirometric experiment with one nitrite pulse of 500 mg N-NO₂⁻ L⁻¹ was performed to estimate the rest of the model parameters. The practical identifiability study showed that the parameters were strongly correlated. Hence, a new experimental design consisting of two consecutive pulses and a delayed third one was designed to improve the parameter identifiability. Both experimental designs were compared using contour plots of the objective function and optimal experimental design criteria for parameter estimation. It was concluded that the parameter identifiability was improved with the new experimental design. Finally, the estimated parameters were validated and the pH effect on the inhibition coefficient was evaluated.     

Inorganic carbon limitations on nitrification: experimental assessment and modelling

Nitrification is a two-step process that involves two different biomass populations: ammonia oxidising biomass (AOB) and nitrite oxidising biomass (NOB). Both populations are autotrophic (i.e. their carbon source is inorganic). Therefore, a deficit of this substrate should result in a decrease of the process rate. Recent technology advances such as the SHARON process have brought new scenarios in biological nitrogen removal where these limitations should be considered. Hence, this work examines the inorganic carbon limitation using respirometric and titrimetric techniques. For this aim, the nitrification rate was measured at different total inorganic carbon (TIC) concentrations. The experimental results obtained show that AOB was limited at TIC concentrations lower than 3mmol CL(-1). At the same time, no carbon source limitation for NOB was observed in spite of the low TIC concentrations attained (lower than 0.1mmol CL(-1)). The AOB limitation could be successfully modelled using Monod, Tessier and sigmoidal kinetics. The best fit was obtained with sigmoidal kinetics. However, unexpected biomass activity (oxygen consumption) was observed despite a very low TIC concentration (lower than 0.1mmol CL(-1)).     

Cellular Antisense Activity of PNA–Oligo(bicycloguanidinium) Conjugates Forming Self‐Assembled Nanoaggregates

A series of peptide nucleic acid–oligo(bicycloguanidinium) (PNA–BG_n) conjugates were synthesized and characterized in terms of cellular antisense activity by using the pLuc750HeLa cell splice correction assay. PNA–BG₄ conjugates exhibited low micromolar antisense activity, and their cellular activity required the presence of a hydrophobic silyl terminal protecting group on the oligo(BG) ligand and a minimum of four guanidinium units. Surprisingly, a nonlinear dose–response with an activity threshold around 3–4 μM , indicative of large cooperativity, was observed. Supported by light scattering and electron microscopy analyses, we propose that the activity, and thus cellular delivery, of these lipo‐PNA–BG₄ conjugates is dependent on self‐assembled nanoaggregates. Finally, cellular activity was enhanced by the presence of serum. Therefore we conclude that the lipo‐BG‐PNA conjugates exhibit an unexpected mechanism for cell delivery and are of interest for further in vivo studies.     

Identifying behaviour patterns of construction safety using system archetypes

Construction safety management involves complex issues (e.g., different trades, multi-organizational project structure, constantly changing work environment, and transient workforce). Systems thinking is widely considered as an effective approach to understanding and managing the complexity. This paper aims to better understand dynamic complexity of construction safety management by exploring archetypes of construction safety. To achieve this, this paper adopted the ground theory method (GTM) and 22 interviews were conducted with participants in various positions (government safety inspector, client, health and safety manager, safety consultant, safety auditor, and safety researcher). Eight archetypes were emerged from the collected data: (1) safety regulations, (2) incentive programs, (3) procurement and safety, (4) safety management in small businesses (5) production and safety, (6) workers’ conflicting goals, (7) blame on workers, and (8) reactive and proactive learning. These archetypes capture the interactions between a wide range of factors within various hierarchical levels and subsystems. As a free-standing tool, they advance the understanding of dynamic complexity of construction safety management and provide systemic insights into dealing with the complexity. They also can facilitate system dynamics modelling of construction safety process.     

PbS/CdS/ZnS Quantum Dots: A Multifunctional Platform for In Vivo Near‐Infrared Low‐Dose Fluorescence Imaging

Over the past decade, near‐infrared (NIR)‐emitting nanoparticles have increasingly been investigated in biomedical research for use as fluorescent imaging probes. Here, high‐quality water‐dispersible core/shell/shell PbS/CdS/ZnS quantum dots (hereafter QDs) as NIR imaging probes fabricated through a rapid, cost‐effective microwave‐assisted cation exchange procedure are reported. These QDs have proven to be water dispersible, stable, and are expected to be nontoxic, resulting from the growth of an outer ZnS shell and the simultaneous surface functionalization with mercaptopropionic acid ligands. Care is taken to design the emission wavelength of the QDs probe lying within the second biological window (1000–1350 nm), which leads to higher penetration depths because of the low extinction coefficient of biological tissues in this spectral range. Furthermore, their intense fluorescence emission enables to follow the real‐time evolution of QD biodistribution among different organs of living mice, after low‐dose intravenous administration. In this paper, QD platform has proven to be capable (ex vivo and in vitro) of high‐resolution thermal sensing in the physiological temperature range. The investigation, together with the lack of noticeable toxicity from these PbS/CdS/ZnS QDs after preliminary studies, paves the way for their use as outstanding multifunctional probes both for in vitro and in vivo applications in biomedicine.     

中国街区改革的关键 ——空间尺度和层次体系

我国街区制与西班牙巴塞罗那超级街区是近年被热议的两种街区改革方向.首先,从概念辨析、历史梳理、规划方式三个方面进行对比,发现:两种改革都以可持续的城市生活为终极目标,并且在策略上并非相反而是互补.进一步比较两种改革的评价体系,具体通过路网密度、道路健康、交通效率三个指标诠释如何"以人为本"地重塑街道的生活功能.最后,针对我国街区改革中易被忽视的差异性和层次性,提出两个关键点:(1)街区应依差异需求确定空间尺度,弱化绝对的大小、开闭和疏密概念;(2)街区应按功能结构建立层次体系,避免形成均质化、碎片化的城市结构.结合技术发展趋势预测未来规划方向,为我国街区改革和"未来社区"建设提供新的思路.     

Regional growth and spatial spillovers: Evidence from an SpVAR for the Spanish regions

This paper provides a spatial vector autoregressive (SpVAR) analysis of growth spillovers for the Spanish regions over the period 1965–2003. First, a spatial Granger causality analysis is performed that indicates the relevant impact of spatial spillover effects across regions in Spain. Second, the empirical research offers a contribution in the context of the SpVAR modelling estimating the push‐in (from the neighbours to the region) and push‐out (from the region to its neighbours) effects of growth spillovers within a regional economic system. Finally, the proposed methodology reveals empirical evidence about both the short‐run and long‐term regional growth adjustment processes in space and time. The results for the Spanish regional panel data suggest the existence of strong spatiotemporal regional spillovers of growth output. This has important implications for the choice of regional policy goals and regional policy instruments.