The insertion/deletion in the DNA-binding region allows the discrimination and subsequent identification of the glucocorticoid receptor 1 (gr1) and gr2 nucleotide sequences in gilthead sea bream (Sparus aurata): Standardizing the gr nomenclature for a better understanding of the stress response in teleost fish species

Cortisol carries out its physiological mechanism of action through the recognition by the mineralocorticoid receptor (MR) and the glucocorticoid receptor (GR) 1 (GR1) and GR2. Previous studies reported that the main difference between gr1 and gr2 nucleotide sequences resides in a 27-nucleotide insertion/deletion in the DNA-binding region, respectively. However, in gilthead sea bream (Sparus aurata) the annotation for gr1 and gr2 seems contradictory. The gr2 sequence possesses the characteristic 27-nucleotide insertion that, in fact, is associated with the gr1 nucleotide sequence. Thus, this study aimed to elucidate the nucleotide sequences for the gr1 and gr2 in gilthead sea bream. The Clustal Omega alignment for different fish species corroborated the presence of such 27-nucleotide insertion/deletion in the DNA-binding region for gr1 and gr2, respectively. Then, we design specific primers set for the amplification of the gilthead sea bream gr1 by polymerase chain reaction (PCR). Importantly, the gr1 nucleotide partial sequence has a high similarity with other gr1 sequences already published for other fish species, being present in all of them the 27-nucleotide insertion in the DNA-binding region. We also detected that in European sea bass the gr1 and gr2 sequences had not been named according to the 27-nucleotide insertion/deletion criteria in the DNA-binding region. Thus, our study makes an urgent call to the scientific community to discuss the establishment of an updated agreement that allows homogenizing the criteria for the nomenclature defining the gr1 and gr2 nucleotide sequences for a better understanding of the stress response in teleost fish species.This study thanks to the AGL2016-76069-C2-2- R, PID2020-117557RB-C21, PID2020-117557RB-C22 grants (AEI-MINECO; Spain). EV-V thanks the support of Fondecyt iniciacion grant (project number 11221308; Agencia Nacional de Investigacion y Desarrollo de Chile, Government of Chile). AK was the recipient of a Ministry of Science, Research, and Technology (Iran) fellowship. MT thanks for the support of the post-doctoral fellowship "Ramon y Cajal" (ref. RYC2019-026841-I) (Ministerio de Ciencia e Innovacion, Spanish Government). FER-L thanks the support of Fondecyt regular grant (project number: 1211841; Agencia Nacional de Investigacion y Desarrollo de Chile, Government of Chile)

Fluorescent Transgenic Zebrafish Tg(nkx2.2a:mEGFP) Provides a Highly Sensitive Monitoring Tool for Neurotoxins

10.1371/journal.pone.0055474PLoS ONE82

WldS Reduces Paraquat-Induced Cytotoxicity via SIRT1 in Non-Neuronal Cells by Attenuating the Depletion of NAD

WldS is a fusion protein with NAD synthesis activity, and has been reported to protect axonal and synaptic compartments of neurons from various mechanical, genetic and chemical insults. However, whether WldS can protect non-neuronal cells against toxic chemicals is largely unknown. Here we found that WldS significantly reduced the cytotoxicity of bipyridylium herbicides paraquat and diquat in mouse embryonic fibroblasts, but had no effect on the cytotoxicity induced by chromium (VI), hydrogen peroxide, etoposide, tunicamycin or brefeldin A. WldS also slowed down the death of mice induced by intraperitoneal injection of paraquat. Further studies demonstrated that WldS markedly attenuated mitochondrial injury including disruption of mitochondrial membrane potential, structural damage and decline of ATP induced by paraquat. Disruption of the NAD synthesis activity of WldS by an H112A or F116S point mutation resulted in loss of its protective function against paraquat-induced cell death. Furthermore, WldS delayed the decrease of intracellular NAD levels induced by paraquat. Similarly, treatment with NAD or its precursor nicotinamide mononucleotide attenuated paraquat-induced cytotoxicity and decline of ATP and NAD levels. In addition, we showed that SIRT1 was required for both exogenous NAD and WldS-mediated cellular protection against paraquat. These findings suggest that NAD and SIRT1 mediate the protective function of WldS against the cytotoxicity induced by paraquat, which provides new clues for the mechanisms underlying the protective function of WldS in both neuronal and non-neuronal cells, and implies that attenuation of NAD depletion may be effective to alleviate paraquat poisoning

Review of the techniques used in motor‐cognitive human‐robot skill transfer

Abstract A conventional robot programming method extensively limits the reusability of skills in the developmental aspect. Engineers programme a robot in a targeted manner for the realisation of predefined skills. The low reusability of general‐purpose robot skills is mainly reflected in inability in novel and complex scenarios. Skill transfer aims to transfer human skills to general‐purpose manipulators or mobile robots to replicate human‐like behaviours. Skill transfer methods that are commonly used at present, such as learning from demonstrated (LfD) or imitation learning, endow the robot with the expert's low‐level motor and high‐level decision‐making ability, so that skills can be reproduced and generalised according to perceived context. The improvement of robot cognition usually relates to an improvement in the autonomous high‐level decision‐making ability. Based on the idea of establishing a generic or specialised robot skill library, robots are expected to autonomously reason about the needs for using skills and plan compound movements according to sensory input. In recent years, in this area, many successful studies have demonstrated their effectiveness. Herein, a detailed review is provided on the transferring techniques of skills, applications, advancements, and limitations, especially in the LfD. Future research directions are also suggested

Vaccination and immune response of the pituitary in fish

Article from the Special Issue on ISFSI 2019; Edited by Daniel Montero.-- Trabajo presentado en el 3rd International Conference of Fish & Shellfish Immunology (ISFSI 2019), celebrado en Las Palmas de Gran Canaria del 16 al 20 de junio de 2019.Although it is known that pituitary hormones can have a direct or indirect influence stimulating or suppressing the immune responses, whether there is a local immune response in this tissue or what is the effect of the immune stimulus on the pituitary function in fish has not received specific attention. In order to understand the immuno-endocrine interaction at the pituitary level, particularly the Hypothalamus-Pituitary-Interrenal axis, different experiments were carried out in rainbow trout and gilthead seabream using both in vitro and an in vivo approaches. Pituitaries of rainbow trout Oncorhynchus mykiss were cultured in vitro, incubated for 3h with Vibrio anguillarum bacterin, bacterin plus CRH, cortisol, human recombinant IL1β, or spleen medium, and then genes involved in pro-inflammation (il1β, il8, tnfα1, ifnγ), anti-inflammation (tgfβ1b, il10), or innate immune modulation (mhcIIa, c3, mif) were tested. Data showed that, incubation with bacterin alone and bacterin plus recombinant IL1β or CRH, as well as medium from bacterin-treated spleen caused significant up-regulation of pro-inflammatory genes, while down-regulated the anti-inflammatory gene tgfβ1b. Besides, recombinant IL1β plus bacterin or alone caused raise of mhcIIa and tnfa, respectively. A second experiment assessed the response of seabream vaccinated by means of an injection with Lactococcus garveiae and immune gene modulation was determined after 1h, 6h and 24 hours looking at the expression of the proinflammatory genes il1β, tnfa and cox2, the anti-inflammatory genes tgfβ1b, il10, and the innate genes lys and c3 corresponding to lysozyme and complement proteins. The immune genes il1β, cox2 and lys, showed a strong expression in the pituitary tissue after injection vaccination, notably il1β which showed more than 10 fold raise, thus indicating both a high sensitivity to the vaccine and the onset of a robust immune response in the pituitary at in vivo level. The overall results indicate that pituitary shows a relevant local immune gene equipment, and also the potential of fish pituitary to develop both innate and adaptive immune responses.Peer reviewe

Anion Complexation Studies of 3‑Nitrophenyl-Substituted Tripodal Thiourea Receptor: A Naked-Eye Detection of Sulfate via Fluoride Displacement Assay

A thiourea-based tripodal receptor <b>L</b> substituted with 3-nitrophenyl groups has been synthesized, and the binding affinity for a variety of anions has been studied by ¹H NMR titrations and nuclear Overhauser enhancement spectroscopy experiments in dimethyl sulfoxide-<i>d</i>₆. As investigated by ¹H NMR titrations, the receptor binds an anion in a 1:1 binding mode, showing the highest binding and strong selectivity for sulfate anion. A competitive colorimetric assay in the presence of fluoride suggests that the sulfate is capable of displacing the bound fluoride, showing a sharp visible color change. The strong affinity of <b>L</b> for sulfate was further supported by UV–vis titrations and density functional theory (DFT) calculations. Time-dependent DFT calculations indicate that the fluoride complex possesses a different optical absorption spectrum (due to charge transfer between the fluoride and the surrounding ligand) than the sulfate complex, reflecting the observed colorimetric change in these two complexes. The receptor was further tested for its biocompatibility on primary human foreskin fibroblasts and HeLa cells, exhibiting an excellent cell viability up to 100 μM concentration