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© 2021 The Authors., [Background and Purpose]: Local anaesthetics block sodium and a variety of potassium channels. Although previous studies identified a residue in the pore signature sequence together with three residues in the S6 segment as a putative binding site, the precise molecular basis of inhibition of Kv channels by local anaesthetics remained unknown. Crystal structures of Kv channels predict that some of these residues point away from the central cavity and face into a drug binding site called side pockets. Thus, the question arises whether the binding site of local anaesthetics is exclusively located in the central cavity or also involves the side pockets. [Experimental Approach]: A systematic functional alanine mutagenesis approach, scanning 58 mutants, together with in silico docking experiments and molecular dynamics simulations was utilized to elucidate the binding site of bupivacaine and ropivacaine. [Key Results]: Inhibition of Kv1.5 channels by local anaesthetics requires binding to the central cavity and the side pockets, and the latter requires interactions with residues of the S5 and the back of the S6 segments. Mutations in the side pockets remove stereoselectivity of inhibition of Kv1.5 channels by bupivacaine. Although binding to the side pockets is conserved for different local anaesthetics, the binding mode in the central cavity and the side pockets shows considerable variations. [Conclusion and Implications]: Local anaesthetics bind to the central cavity and the side pockets, which provide a crucial key to the molecular understanding of their Kv channel affinity and stereoselectivity, as well as their spectrum of side effects., The study was supported by the Ministerio de Ciencia e Innovación (MICINN, Spain) Grants SAF2016-75021-R and PID2019-104366RB-C21 (to C.V. and T.G.); the European Regional Development Fund (Fondo Europeo de Desarrollo Regional [FEDER]) and the Instituto de Salud Carlos III CIBERCV programme CB/11/00222 (to C.V. and T.G.); the Consejo Superior de Investigaciones Científicas (CSIC) Grants PIE201820E104 and 2019AEP148 (to C.V.); the Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT) 1191133 and the Fondo de Equipamiento Científico y Tecnológico (FONDEQUIP) 160063 grants from ANID (to W.G.); and the Deutsche Forschungsgemeinschaft (DFG) Grant DE1482-4/1 to N.D.

The inclusion of a medicinal plant leaf extract (MPLE) from sage (Salvia officinalis) and lemon verbena (Lippia citriodora), rich in verbascoside and triterpenic compounds like ursolic acid, was evaluated in gilthead seabream (Sparus aurata) fed a low fishmeal-based diet (48% crude protein, 17% crude fat, 21.7 MJ kg-1, 7% fishmeal, 15% fish oil) for 92 days. In particular, the study focused on the effect of these phytogenic compounds on the gut condition by analyzing the transcriptomic profiling (microarray analysis) and histological structure of the intestinal mucosa, as well as the histochemical properties of mucins stored in goblet cells. A total number of 506 differentially expressed genes (285 up- and 221 down-regulated) were found when comparing the transcriptomic profiling of the intestine from fish fed the control and MPLE diets. The gut transcripteractome revealed an expression profile that favored biological mechanisms associated to the 1) immune system, particularly involving T cell activation and differentiation, 2) gut integrity (i.e., adherens and tight junctions) and cellular proliferation, and 3) cellular proteolytic pathways. The histological analysis showed that the MPLE dietary supplementation promoted an increase in the number of intestinal goblet cells and modified the composition of mucins’ glycoproteins stored in goblet cells, with an increase in the staining intensity of neutral mucins, as well as in mucins rich in carboxylated and weakly sulfated glycoconjugates, particularly those rich in sialic acid residues. The integration of transcriptomic and histological results showed that the evaluated MPLE from sage and lemon verbena is responsible for the maintenance of intestinal health, supporting gut homeostasis and increasing the integrity of the intestinal epithelium, which suggests that this phytogenic may be considered as a promising sustainable functional additive for aquafeeds., This work has been financially supported by the project “Nutritional strategies for the improvement of productive performance: the use of functional feeds and health diets in aquaculture (DIETAplus)”, funded by JACUMAR (Ministry of Agriculture, Fisheries and Environment of Spain, MAPAMA) and FEMP (EU), as well as by the ERC (European Research Council) in MedAID project (Mediterranean Aquaculture Integrated Development; Grant Agreement Nb. 727315). FR-L thanks the support of Fondecyt Regular grant (Nb. 1211841; ANID; Government of Chile). RS is supported by a PhD grant from the government of Paraguay (BECAL). JF have been subsidized by the Industrial PhD program of Generalitat de Catalunya and TECNOVIT-FARMFAES S.L. (Nb. 2017 DI 017). EV-V was financially granted with DICYT-USACH Postdoctoral fellowship (Nb. 022043IB). Collaboration between Ibero-American researchers has been done under the framework of the network LARVAplus “Strategies for the development and improvement of fish larvae production in Ibero-America” (117RT0521) funded by the Ibero-American Program of Science and Technology for Development (CYTED, Spain).

Oral mouthwashes decrease the infectivity of several respiratory viruses including SARS-CoV-2. However, the precise agents with antiviral activity in these oral rinses and their exact mechanism of action remain unknown. Here we show that cetylpyridinium chloride (CPC), a quaternary ammonium compound in many oral mouthwashes, reduces SARS-CoV-2 infectivity by inhibiting the viral fusion step with target cells after disrupting the integrity of the viral envelope. We also found that CPC-containing mouth rinses decreased more than a thousand times the infectivity of SARS-CoV-2 in vitro, while the corresponding vehicles had no effect. This activity was effective for different SARS-CoV-2 variants, including the B.1.1.7 or Alpha variant originally identified in United Kingdom, and in the presence of sterilized saliva. CPC-containing mouth rinses could therefore represent a cost-effective measure to reduce SARS-CoV-2 infectivity in saliva, aiding to reduce viral transmission from infected individuals regardless of the variants they are infected with., This research was funded by Dentaid SL. The authors also acknowledge the crowdfunding initiative #Yomecorono. N. I-U. is supported by grant PID2020-117145RB-I00 from the Spanish Ministry of Science and Innovation. E. Pradenas was supported by a doctoral grant from the National Agency for Research and Development of Chile (ANID 72180406).

This study aimed to examine how interactions at both plant genotype and arbuscular mycorrhizal fungus species levels affected the expression of root traits and the subsequent effect on plant nutrition and growth. We used two wheat cultivars with contrasting phosphorus (P) acquisition efficiencies (Tukan and Crac) and two arbuscular mycorrhizal (AM) fungi (Rhizophagus intraradices and Claroideoglomus claroideum). Plant growth, as well as morphological and architectural root traits, were highly dependent on the myco-symbiotic partner in the case of the less P-acquisition efficient cultivar Tukan, with mycorrhizal responses ranging from -45 to 54 % with respect to non-mycorrhizal plants. Meanwhile, these responses were between only -7 and 5 % in the P-acquisition efficient cultivar Crac. The AM fungal species produced contrasting mechanisms in the improvement of plant nutrition and root trait responses. Colonization by R. intraradices increased Ca accumulation, regardless of the cultivar, but reduced root growth on Tukan plants. On the other hand, C. claroideum increased P content in both cultivars, with a concomitant increase in root growth and diffusion-based nutrient acquisition by Tukan. Moreover, plants in symbiosis with R. intraradices showed greater organic acid concentration in their rhizosphere compared to C. claroideum-colonized plants, especially Tukan (24 and 35 % more citrate and oxalate, respectively). Our results suggest that the responses in plant-AM fungal interactions related to nutrient dynamics are highly influenced at the fungus level and also by intra-specific variations in root traits at the genotype level, while growth responses related to improved nutrition depend on plant intrinsic acquisition efficiency., This work was supported by the Agencia Nacional de Investigacion y Desarrollo (ANID) Chile, grants ANID/FONDECYT/1170264, ANID/ FONDECYT/11160385, ANID/FONDAP/15130015, ANID/DOCTORADO NACIONAL/21161474, and ANID/PAI/T7817120011); and by Ministerio de Ciencia, Innovacion y Universidades [grant number RTI2018-094350-B-C31], Ministerio de Economia y Empresa [grant number AGL2015-64990-C2-1R], Plan Nacional de I+D+I [grant number CGL2015-69118-C2-2-P-COEXMED-II], Plan 6-UJA postdoctoral fellowship from University of Jaén (Spain); and the European Regional Development Fund (ERDF).

The aim was to identify the effects of AM symbiosis on the expression patterns of genes associated with K and Na compartmentalization and translocation and on K/Na homeostasis in some lettuce (Lactuca sativa) cultivars as well as the effects of the relative abundance of plant AQPs on plant water status. Two AM fungi species (Funneliformis mosseae and Claroideoglomus lamellosum) isolated from the hyper-arid Atacama Desert (northern Chile) were inoculated to two lettuce cultivars (Grand Rapids and Lollo Bionda), and watered with 0 and 60 mM NaCl. At 60 days of plant growth, the AM symbiotic development, biomass production, nutrient content (Pi, Na, K), physiological parameters, gene expressions of ion channels and transporters (NHX and HKT1), and aquaporins proteins abundance (phosphorylated and non-phosphorylated) were evaluated. Salinity increased the AM root colonization by both inocula. AM lettuce plants showed an improved growth, increased relative water content and improved of K/Na ratio in root. In Grand Rapids cultivar, the high efficiency of photosystem II was higher than Lollo Bionda cultivar; on the contrary, stomatal conductance was higher in Lollo Bionda. Nevertheless, both parameters were increased by AM colonization. In the same way, LsaHKT1;1, LsaHKT1;6, LsaNHX2, LsaNHX4, LsaNHX6 and LsaNHX8 genes and aquaporins PIP2 were up-regulated differentially by both AM fungi. The improved plant growth was closely related to a higher water status due to increased PIP2 abundance, as well as to the upregulation of LsaNHX gene expression, which concomitantly improved plant nutrition and K/Na homeostasis maintenance., This work was funded by the National Agency for Research and Development (ANID, Chile) through the grants ANID/FONDAP/15130015 and ANID/FONDECYT/1170264.

The interaction of the alternative oxidase (AOX) pathway with nutrient metabolism is important for understanding how respiration modulates ATP synthesis and carbon economy in plants under nutrient deficiency. Although AOX activity reduces the energy yield of respiration, this enzymatic activity is upregulated under stress conditions to maintain the functioning of primary metabolism. The in vivo metabolic regulation of AOX activity by phosphorus (P) and nitrogen (N) and during plant symbioses with Arbuscular mycorrhizal fungi (AMF) and Rhizobium bacteria is still not fully understood. We highlight several findings and open questions concerning the in vivo regulation of AOX activity and its impact on plant metabolism during P deficiency and symbiosis with AMF. We also highlight the need for the identification of which metabolic regulatory factors of AOX activity are related to N availability and nitrogen-fixing legume-rhizobia symbiosis in order to improve our understanding of N assimilation and biological nitrogen fixation., FONDECYT from National Agency for Research and Development (ANID) 1191118, Chilean Scholarship Program/Becas de doctorado nacional/2017 21180329, European Union (EU) 753301

© The Author(s) 2020., The phytohormone jasmonoyl-isoleucine (JA-Ile) regulates fundamental plant processes as developmental and defense responses. JA-Ile mediates the interaction between the F-box protein COI1 (part of the SCF E3 ubiquitin ligase) and a JAZ repressor leading to early jasmonate responses. The Arabidopsis JAZ1 protein contains the canonical LPIARR degron sequence, which is responsible for the stabilization of the AtCOI1-JA-Ile-AtJAZ1 complex. In strawberry (Fragaria × ananassa) JAZ family was described at the transcriptional level during fruit development but the information about the interaction mode of this complex is still scarce at the molecular level. To gain insight into the strawberry JA-Ile receptor complex, we evaluated the interaction at the structural level, and protein models were built and analyzed for FaCOI1 and FaJAZ1, FaJAZ8.1, and FaJAZ10. The interaction between FaCOI1 and FaJAZ1, FaJAZ8.1 and FaJAZ10 were explored using several ligands, through molecular docking and molecular dynamics (MD) simulations, finding the strongest interaction with (+)-7-iso-JA-Ile than other ligands. Additionally, we tested interactions between FaCOI1 and FaJAZs by yeast two-hybrid assays in the presence of coronatine (COR, a JA-Ile mimic). We detected strong COR-dependent interactions between FaCOI1 and FaJAZ1. Interestingly, FaJAZ1 contains a new non-canonical (IPMQRK) functional degron sequence, in which Arg and Lys are the key residues for maintaining the interaction of the FaCOI1–COR–FaJAZ1 complex as we observed in mutated versions of the FaJAZ1 degron. Phylogenetic analysis showed that the IPMQRK degron is only present in orthologs belonging to the Rosoideae but not in other Rosaceae subfamilies. Together, this study uncovers a new degron sequence in plants, which could be required to make an alternative and functional JA-Ile perception complex in strawberry., This research was funded by the National Research and Development Agency (ANID, Chile), grant FONDECYT/Regular 1181310 to Carlos R. Figueroa. Adrián Garrido-Bigotes acknowledges ANID (grant FONDECYT/Postdoctorado 3190894). F.V-R acknowledges Universidad de Talca for a doctoral scholarship. Work in R. Solano’s laboratory was funded by the Spanish Ministry of Economy and Competitivity grant BIO2016-77216-R (MINECO/FEDER).