Intrinsic Fluorescence of the Active and the Inactive Functional Forms of Human Thymidylate Synthase

The observables associated with protein intrinsic fluorescence – spectra, time decays, anisotropies – offer opportunities to monitor in real time and non-invasively a protein‘s functional form and its interchange with other forms with different functions. We employed these observables to sketch the fluorometric profiles of two functional forms of human thymidylate synthase (hTS), a homodimeric enzyme crucial for cell proliferation and thus targeted by anticancer drugs. The protein takes an active and an inactive form. Stabilization of the latter by peptides that, unlike classical hTS inhibitors, bind it at the monomer/monomer interface offers an alternative inhibition mechanism that promises to avoid the onset of drug resistance in anticancer therapy. The fluorescence features depicted herein can be used as tools to identify and quantify each of the two protein forms in solution, thus making it possible to investigate the kinetic and thermodynamic aspects of the active/inactive conformational interchange. Two examples of fluorometrically monitored interconversion kinetics are provided.     

Comprehensive characterization of the functional activities of pressurized liquid and ultrasound-assisted extracts from Chlorella vulgaris

Chlorella vulgaris has been referred as a potential source of bioactive compounds (carotenoids and fatty acids). In this work, the ability of an environmentally friendly extraction technique such as Pressurized Liquid Extraction (PLE) and a traditional extraction technique such as Ultrasound-Assisted Extraction (UAE) to obtain functional compounds from C. vulgaris, at analytical scale, has been demonstrated. Seeked bioactivities were antioxidant and antimicrobial, for their interest in the food industry. Therefore, a methodology including analytical extraction, in-vitro assays and chemical characterization via HPLC-DAD and GC–MS has been used to determine the interest of Chlorella as a source of functional food ingredients. Results demonstrated that PLE provide higher yields than UAE while similar bioactivities were obtained. Important concentration of carotenoids (lutein, neoxanthin, β-carotene, etc.), chlorophylls, sterols, phytols, and fatty acids (among others) have been found in Chlorella extracts that could be correlated to the observed biological activity.     

Recommendations for dispatching emergency vehicles under multitiered response via simulation

Emergency medical service (EMS) systems provide medical care and transportation. While many real‐world systems use multiple vehicle types to attend different call priorities, few guidelines exist about which vehicles to allocate in multitiered responses where more than one vehicle is sent per call. This paper makes recommendations for multiple‐unit dispatch to multiple call priorities based on simulation optimization and heuristics. The objective is to maximize the overall expected survival probability of patients classified as “life‐threatening”. We assume two types of medical units and three call priorities; and that information may be updated when the medical unit arrives on‐scene. First, we study the optimal dispatching policies through several examples. Numerical results show that dispatching while considering call priorities, rather than dispatching the closest units, improves EMS system effectiveness. A heuristic algorithm is developed for large‐scale problems. A comparison between the heuristic and closest policy is demonstrated using real‐world data.     

Supply chain configurations: a model to evaluate performance in customised productions

This paper describes an approach used to evaluate the performance of different supply chain configurations in customised contexts. Based on historical data collected from the supply chain of a shoe producer, different configurations are evaluated based on a discrete-event simulation by highlighting the performance of the supply chain (in terms of supply chain order lead-time and inventory volume) when the production switched from standard production (characterised by batches of large quantities of the same product) to customised production (characterised by a small of series batches with high product variability). The simulation approach relies on experimentation through executable configurations, which enables the creation of different scenarios, and is then applied to the case of an actual firm in the footwear industry. The managerial implications of these findings are discussed.     

Effects of worker size on the dynamics of fire ant tunnel construction

Social insects work together to complete tasks. However, different individuals within a colony may vary in task proficiency. We investigated if fire ant (Solenopsis invicta) worker body size influenced the ability to construct tunnels—a key component of subterranean nests. We monitored excavation by worker groups in a substrate of small wetted glass particles in quasi-two-dimensional arenas. Morphological and network features of the tunnel system were measured. Total tunnel area did not differ significantly between groups of large and small workers, although the tunnel area of control sized workers was significantly larger than that of large workers. Moreover, large workers created wider but shorter tunnels, with slower growth rate of tunnel number. However, edge–vertex scaling and degree distribution of the tunnel network were similar across all treatments. In all cases, the amount of excavated material was correlated with the number of active workers. Our study reveals that morphological features of excavated tunnels show modest variation when constructed by workers of varying sizes, but topological features associated with the tunnel network are conserved. These results suggest that important behavioural aspects of tunnel construction—and thus nest building—are similar among morphologically distinct members of fire ant societies.     

DNA Break Mapping Reveals Topoisomerase II Activity Genome-Wide

Genomic DNA is under constant assault by endogenous and exogenous DNA damaging agents. DNA breakage can represent a major threat to genome integrity but can also be necessary for genome function. Here we present approaches to map DNA double-strand breaks (DSBs) and single-strand breaks (SSBs) at the genome-wide scale by two methods called DSB- and SSB-Seq, respectively. We tested these methods in human colon cancer cells and validated the results using the Topoisomerase II (Top2)-poisoning agent etoposide (ETO). Our results show that the combination of ETO treatment with break-mapping techniques is a powerful method to elaborate the pattern of Top2 enzymatic activity across the genome.     

Evaluation of 4-(4-Fluorobenzyl)piperazin-1-yl]-Based Compounds as Competitive Tyrosinase Inhibitors Endowed with Antimelanogenic Effects

There is a considerable attention for the development of inhibitors of tyrosinase (TYR) as therapeutic strategy for the treatment of hyperpigmentation disorders in humans. Continuing in our efforts to identify TYR inhibitors, we describe the design, synthesis and pharmacophore exploration of new small molecules structurally characterized by the presence of the 4-fluorobenzylpiperazine moiety as key pharmacophoric feature for the inhibition of TYR from Agaricus bisporus (AbTYR). Our investigations resulted in the discovery of the competitive inhibitor [4-(4-fluorobenzyl)piperazin-1-yl]-(3-chloro-2-nitro-phenyl)methanone 26 (IC₅₀=0.18 μM) that proved to be ∼100-fold more active than reference compound kojic acid (IC₅₀=17.76 μM). Notably, compound 26 exerted antimelanogenic effect on B16F10 cells in absence of cytotoxicity. Docking analysis suggested its binding mode into AbTYR and into modelled human TYR.