Kinetic modeling of tricarboxylic acid cycle and glyoxylate bypass in Mycobacterium tuberculosis, and its application to assessment of drug targets

BACKGROUND: Targeting persistent tubercule bacilli has become an important challenge in the development of anti-tuberculous drugs. As the glyoxylate bypass is essential for persistent bacilli, interference with it holds the potential for designing new antibacterial drugs. We have developed kinetic models of the tricarboxylic acid cycle and glyoxylate bypass in Escherichia coli and Mycobacterium tuberculosis, and studied the effects of inhibition of various enzymes in the M. tuberculosis model. RESULTS: We used E. coli to validate the pathway-modeling protocol and showed that changes in metabolic flux can be estimated from gene expression data. The M. tuberculosis model reproduced the observation that deletion of one of the two isocitrate lyase genes has little effect on bacterial growth in macrophages, but deletion of both genes leads to the elimination of the bacilli from the lungs. It also substantiated the inhibition of isocitrate lyases by 3-nitropropionate. On the basis of our simulation studies, we propose that: (i) fractional inactivation of both isocitrate dehydrogenase 1 and isocitrate dehydrogenase 2 is required for a flux through the glyoxylate bypass in persistent mycobacteria; and (ii) increasing the amount of active isocitrate dehydrogenases can stop the flux through the glyoxylate bypass, so the kinase that inactivates isocitrate dehydrogenase 1 and/or the proposed inactivator of isocitrate dehydrogenase 2 is a potential target for drugs against persistent mycobacteria. In addition, competitive inhibition of isocitrate lyases along with a reduction in the inactivation of isocitrate dehydrogenases appears to be a feasible strategy for targeting persistent mycobacteria. CONCLUSION: We used kinetic modeling of biochemical pathways to assess various potential anti-tuberculous drug targets that interfere with the glyoxylate bypass flux, and indicated the type of inhibition needed to eliminate the pathogen. The advantage of such an approach to the assessment of drug targets is that it facilitates the study of systemic effect(s) of the modulation of the target enzyme(s) in the cellular environment

In Situ Compatibilization of Biopolymer Ternary Blends by Reactive Extrusion with Low-Functionality Epoxy-Based Styrene Acrylic Oligomer

[EN] The present study reports on the use of low-functionality epoxy-based styrene¿acrylic oligomer (ESAO) to compatibilize immiscible ternary blends made of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polylactide (PLA), and poly(butylene adipate-co-terephthalate) (PBAT). The addition during melt processing of low-functionality ESAO at two parts per hundred resin (phr) of biopolymer successfully changed the soften inclusion phase in the blend system to a thinner morphology, yielding biopolymer ternary blends with higher mechanical ductility and also improved oxygen barrier performance. The compatibilization achieved was ascribed to the in situ formation of a newly block terpolymer, i.e. PHBVb- PLA-b-PBAT, which was produced at the blend interface by the reaction of the multiple epoxy groups present in ESAO with the functional terminal groups of the biopolymers. This chemical reaction was mainly linear due to the inherently low functionality of ESAO and the more favorable reactivity of the epoxy groups with the carboxyl groups of the biopolymers, which avoided the formation of highly branched and/or cross-linked structures and thus facilitated the films processability. Therefore, the reactive blending of biopolymers at different mixing ratios with low-functionality ESAO represents a straightforward methodology to prepare sustainable plastics at industrial scale with different physical properties that can be of interest in, for instance, food packaging applications.This research was funded by the EU H2020 project YPACK (Reference number 773872) and by the Spanish Ministry of Science, Innovation, and Universities (MICIU) with project numbers MAT2017-84909-C2-2-R and AGL2015-63855-C2-1-R. L. Quiles-Carrillo wants to thank the Spanish Ministry of Education, Culture, and Sports (MECD) for financial support through his FPU Grant Number FPU15/03812. Torres-Giner also acknowledges the MICIU for his Juan de la Cierva contract (IJCI-2016-29675).Quiles-Carrillo, L.; Montanes, N.; Lagaron, J.; Balart, R.; Torres-Giner, S. (2019). In Situ Compatibilization of Biopolymer Ternary Blends by Reactive Extrusion with Low-Functionality Epoxy-Based Styrene Acrylic Oligomer. 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Brood‐stock management and early hatchery rearing of Arctic charr (Salvelinus alpinus (Linnaeus))

Arctic charr (Salvelinus alpinus (Linnaeus)) is a stenothermic cold‐water fish, which has been cultured in Northern Europe and North America since the 1980s. The industry has remained relatively small with an annual production between 6000 and 10 000 tonnes, and is still challenged by an unreliable offspring production. This review focuses on offspring production in Arctic charr aquaculture including holding conditions for brood‐stock, fertilisation and egg rearing until hatch. Brood‐stock requires low temperatures during summer (<12°C) with the optimum still unknown. The temperature maximum for egg incubation lies between 6 and 8°C. The composition of an optimal brood‐stock diet is debated regarding fatty acids. A demand for a freshwater‐based diet rich in omega‐6 fatty acids is indicated, but results remain inconclusive. Extensive knowledge has been gained on the timing of spawning and its manipulation through photoperiod, temperature and hormone treatments; spawning can be induced by short‐day photoperiod; and temperature drops to 5°C. Eggs are fertilised dry in ovarian fluid. Egg quality is highly variable and positively related to egg size and energy density. Contrary, little information is available on sperm quality and its impact on egg survival. There may also be profound differences between Arctic charr of stationary or anadromous origin regarding requirements for holding conditions of brood‐stock and their diet. However, these differences have received little attention, and direct comparative studies are in demand