Structural Investigation and Molecular Modeling Studies of Strobilurin-Based Fungicides Active against the Rice Blast Pathogen Pyricularia oryzae

The increasing emergence of fungicide-resistant pathogens requires urgent solutions for crop disease management. Here, we describe a structural investigation of new fungicides obtained by combining strobilurin and succinate dehydrogenase inhibitor pharmacophores. We identified compounds endowed with very good activity against wild-type Pyricularia oryzae, combined in some cases with promising activity against strobilurin-resistant strains. The first three-dimensional model of P. oryzae cytochrome bc1 complex containing azoxystrobin as a ligand was developed. The model was validated with a set of commercially available strobilurins, and it well explains both the resistance mechanism to strobilurins mediated by the mutation G143A and the activity of metyltetraprole against strobilurin-resistant strains. The obtained results shed light on the key recognition determinants of strobilurin-like derivatives in the cytochrome bc1 active site and will guide the further rational design of new fungicides able to overcome resistance caused by G143A mutation in the rice blast pathogen.     

Critical Review of the Evolution of Extracellular Vesicles’ Knowledge: From 1946 to Today

Extracellular vesicles (EVs) are a family of particles/vesicles present in blood and body fluids, composed of phospholipid bilayers that carry a variety of molecules that can mediate cell communication, modulating crucial cell processes such as homeostasis, induction/dampening of inflammation, and promotion of repair. Their existence, initially suspected in 1946 and confirmed in 1967, spurred a sharp increase in the number of scientific publications. Paradoxically, the increasing interest for EV content and function progressively reduced the relevance for a precise nomenclature in classifying EVs, therefore leading to a confusing scientific production. The aim of this review was to analyze the evolution of the progress in the knowledge and definition of EVs over the years, with an overview of the methodologies used for the identification of the vesicles, their cell of origin, and the detection of their cargo. The MISEV 2018 guidelines for the proper recognition nomenclature and ways to study EVs are summarized. The review finishes with a “more questions than answers” chapter, in which some of the problems we still face to fully understand the EV function and potential as a diagnostic and therapeutic tool are analyzed.     

A High Throughput Approach Based on Dynamic High Pressure for the Encapsulation of Active Compounds in Exosomes for Precision Medicine

In recent decades, endogenous nanocarrier-exosomes have received considerable scientific interest as drug delivery systems. The unique proteo-lipid architecture allows the crossing of various natural barriers and protects exosomes cargo from degradation in the bloodstream. However, the presence of this bilayer membrane as well as their endogenous content make loading of exogenous molecules challenging. In the present work, we will investigate how to promote the manipulation of vesicles curvature by a high-pressure microfluidic system as a ground-breaking method for exosomes encapsulation. Exosomes isolated from Uppsala 87 Malignant Glioma (U87-MG) cell culture media were characterized before and after the treatment with high-pressure homogenization. Once their structural and biological stability were validated, we applied this novel method for the encapsulation in the lipidic exosomal bilayer of the chemotherapeutic Irinotecan HCl Trihydrate-CPT 11. Finally, we performed in vitro preliminary test to validate the nanobiointeraction of exosomes, uptake mechanisms, and cytotoxic effect in cell culture model.     

Evaluation of direct light processing for the fabrication of bioactive ceramic scaffolds: Effect of pore/strut size on manufacturability and mechanical performance

Bioactive ceramic scaffolds for bone regeneration consisting of a three-dimensional mesh of interpenetrating struts with square section were fabricated via Digital Light Processing (DLP). The ability of the technique to manufacture 3D porous structures from β-tricalcium phosphate (β-TCP) powders with different dimensions of struts and pores was evaluated, identifying the possibilities and limitations of the manufacturing process. Small pore sizes were found to seriously complicate the elimination of excess slurry from the scaffold’s innermost pores. The effect of the strut/pore size on the mechanical performance of the scaffolds under compressive stresses was also evaluated, but no significant influence was found. Under compressive stresses, the structures resulted weaker when tested perpendicularly to the printing plane due to interlayer shear failure. Interlayer superficial grooves are proposed as potential failure-controlling defects, which could also explain the lack of a Weibull size effect on the mechanical strength of the fabricated DLP scaffolds.     

Effect of thymol on microbial diversity in the porcine jejunum

Thirty two weaned pigs (24 d-old) were fed a diet without or with 1% (w/w) thymol. Pigs from each dietary treatment remained unchallenged or were challenged with Salmonella enterica serovar typhimurium. Jejunal content was collected and molecular microbial diversity was investigated using 16S rRNA gene polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Shannon index analysis showed no differences in diversity but Pearson coefficient analysis showed clear clustering of profiles, which delineated thymol fed and control groups irrespective of Salmonella challenge. Moreover, absence of bands corresponding to Actinobacillus minor (98-99% identity) was observed in thymol groups. A band corresponding to Citrobacter freundii (98% identity) was observed in almost all pigs from thymol groups, and only in 4 samples from pigs fed no thymol in the diet. Supplementation of pig diet with thymol caused clear changes in small intestine microbacterial community. Surprisingly, Salmonella infection caused no major perturbations to the community structure.     

Relation Between the North-Atlantic Oscillation and Hydroclimatic Conditions in Mediterranean Areas

Mediterranean basins are known for their susceptibility to water stress conditions resulting from a reduction in winter-season precipitation. This region is also prone to the effects of the North Atlantic Oscillation (NAO), a large-scale climatic pattern associated with a displacement of air mass between the arctic and the subtropical Atlantic. Even though previous studies have demonstrated the impact of the NAO on winter climate conditions in Europe and Northern Africa, it is still unclear to what extent the NAO is related to hydroclimatic patterns in Mediterranean areas and whether it can explain its recent drought history. To this end, this study investigates the interdependence between the NAO and winter precipitation, river flow and temperature in two Mediterranean regions: Southern Italy and Nile Delta (Egypt). The outcomes show the presence of significant connections between the NAO, winter rainfall and river discharge.     

Iron reduction potentiates hydroxyl radical formation only in flavonols

Flavonoids, substantial components of the human diet, are generally considered to be beneficial. However, they may possess possible pro-oxidative effects, which could be based on their reducing potential. The aims of this study were to evaluate the ability of 26 flavonoids to reduce ferric ions at relevant pH conditions and to find a possible relationship with potentiation of hydroxyl radical production. A substantial ferric ions reduction was achieved under acidic conditions, particularly by flavonols and flavanols with the catecholic ring B. Apparently corresponding bell-shaped curves displaying the pro-oxidant effect of flavonols quercetin and kaempferol on iron-based Fenton reaction were documented. Several flavonoids were efficient antioxidants at very low concentrations but rather inefficient or pro-oxidative at higher concentrations. Flavonols, morin and rutin were progressively pro-oxidant, while 7-hydroxyflavone and hesperetin were the only flavonoids with dose-dependent inhibition of hydroxyl radical production. Conclusively, administration of flavonoids may lead to unpredictable consequences with few exceptions.     

Prospects of fuel cell auxiliary power units in the civil markets

Auxiliary power units (APUs), i.e. devices designed to provide additional onboard power in vehicles, are believed to be an important entry point for fuel cell (FC) technology into commercial markets. Three technologies are under consideration for this market: solid oxide fuel cells (SOFCs), proton exchange membrane fuel cells (PEMFCs) and direct methanol fuel cells (DMFCs). By using the concept of total addressable, potential and capturable markets, this paper discusses the opportunities and challenges of fuel cell auxiliary power units (FC APUs). A number of conclusions can be drawn. As FC APUs do not offer increased fuel efficiency when meeting electrical demand while the main engine is used for propulsion (as opposed to idling), applications on board transit vehicles such as buses seem limited. Potential markets begin to open up in vehicles that either have a very large electricity demand due to many high-energy onboard functions, such as luxury limousines, or that require electrical power whilst stationary. Examples in the latter category include law enforcement vehicles, recreational vehicles, and most importantly heavy duty trucks. Volume and start-up time seem to be the major technical challenges hindering the market penetration of FC APUs. However, whilst the functional benefits of FC APUs over existing technologies are limited, the former must also be able to compete on a cost basis. The intense activities of APU manufacturers suggest a confidence either in the potential for cost reductions or in the consumers’ willingness to pay. Similarly, the involvement of a number of big truck manufacturers casts doubt on the extent to which incumbents are taken by surprise by competence-destroying, disruptive and radical technologies like FC APUs.     

High efficiency process for the production of pure oxygen based on solid oxide fuel cell–solid oxide electrolyzer technology

This paper presents a novel system for production of pure oxygen based on the integration of a solid oxide fuel cell (SOFC) and a solid oxide electrolyzer (SOEC). In the proposed arrangement, the SOFC provides electricity, heat and H₂O in vapour phase to the SOEC which carries out the inverse reactions of the SOFC, that is the separation of H₂O into H₂ (used as a fuel for the SOFC) and O₂ (representing the yield of the system). Simulations carried out in different operating conditions show that when the integrated SOFC–SOEC device runs at low current densities (less than 1000 A m⁻²), pure oxygen can be generated with an electric consumption comparable to mid-size cryogenic air separation units, and significantly lower than small scale systems based on the PSA technology.