Engineering phosphatidylinositol-4,5-bisphosphate model membranes enriched in endocytic cargo: a neutron reflectometry, AFM and QCM-D structural study

The combination of in vitro models of biological membranes based on solid-supported lipid bilayers (SLBs) and of surface sensitive techniques, such as neutron reflectometry (NR), atomic force microscopy (AFM) and quartz crystal microbalance with dissipation monitoring (QCM-D), is well suited to provide quantitative information about molecular level interactions and lipid spatial distributions. In this work, cellular plasma membranes have been mimicked by designing complex SLB, containing phosphatidylinositol 4,5-bisphosphate (PtdIns4,5P2) lipids as well as incorporating synthetic lipo-peptides that simulate the cytoplasmic tails of transmembrane proteins. The QCM-D results revealed that the adsorption and fusion kinetics of PtdIns4,5P2 are highly dependent of Mg2+. Additionally, it was shown that increasing concentrations of PtdIns4,5P2 leads to the formation of SLBs with higher homogeneity. The presence of PtdIns4,5P2 clusters was visualized by AFM. NR provided important insights about the structural organization of the various components within the SLB, highlighting that the leaflet symmetry of these SLBs is broken by the presence of CD4-derived cargo peptides. Finally, we foresee our study to be a starting point for more sophisticated in vitro models of biological membranes with the incorporation of inositol phospholipids and synthetic endocytic motifs.publishe

Fed-batch fermentation of olive mill wastewaters for lipase production

In the Mediterranean basin countries, huge amounts of olive mill wastewaters (OMW) are produced by the olive oil industry. It constitutes a serious environmental problem, nevertheless its composition turns OMW into a potential growth medium to lipolytic microorganisms. The aim of this work was to study lipase production as well as OMW degradation in fed-batch cultures of Candida cylindracea CBS 7869, Candida rugosa CBS 2275 and Yarrowia lipolytica W29 (ATCC 20460). Besides the improvement of lipase production, the fed-batch approach enhanced the effluent degradation, since it led to good COD and lipids reduction, both higher than 50%. C. rugosa achieved the highest value of lipase productivity (130 U L−1 h−1), in parallel with highest lipids reduction (77%). This study demonstrates thatOMWare becoming a competitive and valuable growth medium in fermentation processes with lipolytic microorganisms. The fed-batch strategy used proved to be an efficient approach to enhance lipase production from OMW and to reduce significantly the final organic load of the medium.The authors acknowledge the financial support provided by 'Fundacao para a Ciencia e Tecnologia' (Project PTDC/AMB/69379/2006; Grant SFRH/BD/27915/2006)

Flipped spinfoam vertex and loop gravity

We introduce a vertex amplitude for 4d loop quantum gravity. We derive it from a conventional quantization of a Regge discretization of euclidean general relativity. This yields a spinfoam sum that corrects some difficulties of the Barrett-Crane theory. The second class simplicity constraints are imposed weakly, and not strongly as in Barrett-Crane theory. Thanks to a flip in the quantum algebra, the boundary states turn out to match those of SO(3) loop quantum gravity -- the two can be identified as eigenstates of the same physical quantities -- providing a solution to the problem of connecting the covariant SO(4) spinfoam formalism with the canonical SO(3) spin-network one. The vertex amplitude is SO(3) and SO(4)-covariant. It rectifies the triviality of the intertwiner dependence of the Barrett-Crane vertex, which is responsible for its failure to yield the correct propagator tensorial structure. The construction provides also an independent derivation of the kinematics of loop quantum gravity and of the result that geometry is quantized.Comment: 37 pages, 4 figure

Evaluation of hydrogen sulfide supply to biostimulate the nutritive and phytochemical quality and the antioxidant capacity of Cabbage (<i>Brassica oleracea</i> L. ‘Bronco’)

The potential effects of the hydrogen sulfide on shoot biomass, nutritional quality and antioxidant capacity of Brassica oleracea, were investigated through the application of increasing doses of NaHS (H2S donor NaHS; 0.5, 1, 2.5, and 5 mM). The results showed that the 0.5 and 1 mM NaHS treatments increased biomass and the quality composition of ‘Bronco’ cabbage (i.e. chlorophylls, carotenoids, anthocyanins, flavonols, total phenolics and sinigrin). On the other hand, there was an increase in lipid peroxidation and hydrogen peroxide content with the application of doses higher than 2.5 mM NaHS. Therefore, we selected the 0.5 and 1 mM NaHS dosages as optimal for cabbage. The 2.5 and 5 mM NaHS produced an excessive lipid peroxidation, decreases in plant biomass and losses of chlorophylls, being all considered negative effects, and clear evidences of stressful situation for the plants. For practical purposes, this study suggested that exogenous application of H2S donor NaHS at 0.5 and 1 mM may be useful as bio-stimulant to boost the yield and the health-promoting composition of ‘Bronco’ cabbage (Brassica oleracea L.).   The online version of theis article (doi: 10.5073/JABFQ.2016.089.038) contains a supplementary file

A Immirzi-like parameter for 3d quantum gravity

We study an Immirzi-like ambiguity in three-dimensional quantum gravity. It shares some features with the Immirzi parameter of four-dimensional loop quantum gravity: it does not affect the equations of motion, but modifies the Poisson brackets and the constraint algebra at the canonical level. We focus on the length operator and show how to define it through non-commuting fluxes. We compute its spectrum and show the effect of this Immirzi-like ambiguity. Finally, we extend these considerations to 4d gravity and show how the different topological modifications of the action affect the canonical structure of loop quantum gravity.Comment: 14 pages, v2: one reference added, more comments on the 3d/4d compariso

Influence of ohmic heating in the composition of extracts from Gracilaria vermiculophylla

Electric field-based technologies for extraction processes have been gaining importance due to sustainability concerns. This work aims to assess the potential of ohmic heating as an efficient and feasible tool for the extraction of different biocompounds from Gracilaria vermiculophylla and its effect on the extracts' composition. Different ratios of water/ethanol (0 to 75% ethanol, v/v) were used to target different families of biocompounds. The ohmic heating-based extraction was performed at 82 °C under electric field and frequency of 28 V/cm and 25 kHz, respectively. Conventional extractions without the presence of electric field were made keeping a temperature profile identical to the ohmic heating treatments, thus addressing the potential occurrence of electrical (non-thermal) effects. Extraction yields and extracts composition (content in polysaccharides, proteins, phenolic compounds and pigments) were evaluated. Further, as agar is the major commertially exploited compound from Gracilaria spp., the effect of ohmic heating on the extracted agar in terms of yield, carbohydrates' composition, monosaccharides profile, and gelling ability was also envisaged. Overall, significant differences in the extraction of each family of compounds between ohmic and conventional extractions were observed, being more pronounced at the best solvent for each compound (100% water for carbohydrates, 75:25 water/ethanol for proteins, 75:25 and 50:50 water/ethanol for phenolic compounds and 25:75 water/ethanol for pigments). Higher extraction yields were achieved for ohmic heating at 1 h, except for 100% water, probably indicating accelerated extraction kinetics promoted by the presence of electric field effects. Furthermore, the gelling ability of agar and the antioxidant activity were not impaired by the use of moderate electric fields. Therefore, ohmic heating is an interesting alternative, with reduced energy consumption and improved extraction performances, to recover functional ingredients or additives from seaweeds for the food industry.This work was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of project OH2O – PTDC/EQU/029145/2017 and the strategic funding of UID/BIO/04469/2019 unit, and by the European Fund for Regional Development (FEDER) and COMPETE 2020 – Competitiveness and Internationalization Operational Program under the scope the projects OH2O (POCI-01-0145-FEDER 029145) and VALORMAR – Full valorization of marine resources: po tential, technological innovation and new applications (call 10/SI/2016; reference: 24517). This work was also developed within the scope of the stratigic funding of CICECO-Aveiro Institute of Materials (UIDB/50011/2020 & UIDP/50011/2020), QOPNA (UID/QUI/00062/2019) and LAQV-REQUIMTE (UIDB/50006/2020), financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement. This work was also funded by national funds (OE), through FCT, I.P., within the scope of the frame work contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19.info:eu-repo/semantics/publishedVersio

In Vitro Antifungal Activity of Ibrexafungerp (SCY-078) Against Contemporary Blood Isolates From Medically Relevant Species of Candida: A European Study

BackgroundIbrexafungerp (SCY-078) is the newest oral and intravenous antifungal drug with broad activity, currently undergoing clinical trials for invasive candidiasis. ObjectiveThe aim of this study was to assess the in vitro activity of ibrexafungerp and comparators against a collection of 434 European blood isolates of Candida. MethodsIbrexafungerp, caspofungin, fluconazole, and micafungin minimum inhibitory concentrations (MICs) were collected from 12 European laboratories for 434 blood isolates, including 163 Candida albicans, 108 Candida parapsilosis, 60 Candida glabrata, 40 Candida tropicalis, 29 Candida krusei, 20 Candida orthopsilosis, 6 Candida guilliermondii, 2 Candida famata, 2 Candida lusitaniae, and 1 isolate each of Candida bracarensis, Candida catenulata, Candida dubliniensis, and Candida kefyr. MICs were determined by the EUCAST broth microdilution method, and isolates were classified according to recommended clinical breakpoints and epidemiological cutoffs. Additionally, 22 Candida auris from different clinical specimens were evaluated. ResultsIbrexafungerp MICs ranged from 0.016 to >= 8 mg/L. The lowest ibrexafungerp MICs were observed for C. albicans (geometric MIC 0.062 mg/L, MIC range 0.016-0.5 mg/L) and the highest ibrexafungerp MICs were observed for C. tropicalis (geometric MIC 0.517 mg/L, MIC range 0.06->= 8 mg/L). Modal MICs/MIC(50)s (mg/L) against Candida spp. were 0.125/0.06 for C. albicans, 0.5/0.5 for C. parapsilosis, 0.25/0.25 for C. glabrata, 0.5/0.5 for C. tropicalis, 1/1 for C. krusei, 4/2 for C. orthopsilosis, and 0.5/0.5 for C. auris. Ibrexafungerp showed activity against fluconazole- and echinocandin-resistant isolates. If adopting wild-type upper limits, a non-wild-type phenotype for ibrexafungerp was only observed for 16/434 (3.7%) isolates: 11 (4.6%) C. parapsilosis, 4 (5%) C. glabrata, and 1 (2.5%) C. tropicalis. ConclusionIbrexafungerp showed a potent in vitro activity against Candida.This study received funding from SCYNEXIS. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of the article, or the decision to submit it for publication. CM-A is a recipient of a grant from Fundació n ONCE (Oportunidad al Talento). EE, AG, NJ, CM-A, and GQ have received grant support from Consejerı́a de Educación, Universidades e Investigación del Gobierno Vasco (GIC15 IT-990-16), Fondo de Investigación Sanitaria del Gobierno de España (FIS PI11/00203), and UPV/EHU (UFI 11/25). All authors declare no other competing interests

Phase space descriptions for simplicial 4d geometries

Starting from the canonical phase space for discretised (4d) BF-theory, we implement a canonical version of the simplicity constraints and construct phase spaces for simplicial geometries. Our construction allows us to study the connection between different versions of Regge calculus and approaches using connection variables, such as loop quantum gravity. We find that on a fixed triangulation the (gauge invariant) phase space associated to loop quantum gravity is genuinely larger than the one for length and even area Regge calculus. Rather, it corresponds to the phase space of area-angle Regge calculus, as defined by Dittrich and Speziale in [arXiv:0802.0864] (prior to the imposition of gluing constraints, that ensure the metricity of the triangulation). We argue that this is due to the fact that the simplicity constraints are not fully implemented in canonical loop quantum gravity. Finally, we show that for a subclass of triangulations one can construct first class Hamiltonian and Diffeomorphism constraints leading to flat 4d space-times.Comment: corrected structure constants, several references ad