Browsing by Author "Power, Deborah Mary"
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- Activation profile of the Atlantic salmon (Salmo salar) calcium-sensing receptor (Casr) by selected L-amino acidsPublication . Gomes, Ana; Gélébart, Virginie; Félix, Rute; Cardoso, João; Zimmermann, Fabian; Lai, Floriana; Power, Deborah Mary; Ronnestad, IvarIn mammals, the calcium-sensing receptor (CaSR) is involved in nutrient sensing and modulated by several amino acids. In teleosts, sequence homologues of the mammalian CaSR have been described but their function in sensing amino acids remains elusive, including in Atlantic salmon (Salmo salar), an important aquaculture species. This study investigated the activation of Atlantic salmon Casr (asCasr)-mediated signaling pathways-Gq, Gi, and ERK1/2-by six selected L-amino acids (histidine, tryptophan, phenylalanine, isoleucine, leucine and valine) and by Ca2+. Using a Flp-In-HEK293 cell line stably expressing asCasr, we confirmed activation of all three pathways. L-histidine, L-phenylalanine, and L-tryptophan triggered Gi signaling independent of Ca-2(+). Notably, no Ca-2(+) concentrations induced Gi activation, but IP1 production increased in a concentration-dependent manner. L-histidine was the only amino acid to activate the Gq pathway without Ca-2(+), and this response was amplified by the presence of Ca-2(+). In the presence of 2.5 mM Ca-2(+), L-phenylalanine and L-tryptophan also activated Gq signaling in a concentration-dependent manner. Additionally, in the presence of 10 mM Ca-2(+), L-histidine, L-phenylalanine, and L-tryptophan triggered ERK phosphorylation. These findings establish asCasr as a functional homologue of mammalian CaSR, activated in a concentration-dependent manner by L-amino acids with an aromatic ring.
- Bacterial 16S ribosomal gene fingerprints as a tool to diagnose and mitigate fish larvae gut dysbiosisPublication . NAJAFPOUR, BABAK; Canario, Adelino; Power, Deborah MaryDysbiosis is associated with shifts in the diversity or relative abundance of beneficial versus harmful bacteria, leading to health issues in organisms. This study investigated gut bacterial dysbiosis associated with larval quality using 16S rRNA gene sequencing. The gut microbiome of gilthead sea bream and European sea bass, key commercial species and vertebrate models, was examined in high- and low-quality larvae batches from several European hatcheries. Larval quality, hatchery site and species influenced bacterial diversity in the gut. Individuals from larval batches that performed well had higher microbial diversity in the gut and individuals from batches that performed poorly had a gut microbiota dominated by pathogenic Vibrio (e.g., V. aestuarianus and V. cortegadensis). The bacterial dysbiosis index revealed a notable predominance of Fusobacteriota and Firmicutes phyla, Thermoanaerobacteria class and Lactobacillaceae, Moritellaceae, Clostridiaceae, Thiotrichaceae and Shewanellaceae families in good-quality larvae batches, and a prevalence of the Proteobacteria phylum, Gammaproteobacteria class, Sphingomonadaceae and Vibrionaceae families in the gut of individuals from poor-quality larvae batches. A positive dysbiosis index (cutoff >0.4) was associated with a high risk of decreased larval performance and quality. Additionally, the abundance of Clostridium_sensu_ stricto_15, Shewanellaceae_unclassified, Cetobacterium, Psychrilyobacter, Moritella and Latilactobacillus genera in the gut of good production batches, and the Vibrio genus in the gut of poor production batches, identified these genus as potential markers for diagnosing and mitigating bacterial dysbiosis in fish and potentially other vertebrates.
- Cartilage acidic protein a novel therapeutic factor to improve skin damage repair?Publication . Félix, Rute; Anjos, Liliana; Costa, Rita; Letsiou, Sophia; Power, Deborah MaryFish skin has been gaining attention due to its efficacy as a human-wound-treatment product and to identify factors promoting its enhanced action. Skin fibroblasts have a central role in maintaining skin integrity and secrete extra cellular matrix (ECM) proteins, growth factors and cytokines to rapidly repair lesions and prevent further damage or infection. The effects on scratch repair of the ubiquitous but poorly characterized ECM protein, cartilage acidic protein 1 (CRTAC1), from piscine and human sources were compared using a zebrafish SJD.1 primary fibroblast cell line. A classic in vitro cell scratch assay, immunofluorescence, biosensor and gene expression analysis were used. Our results demonstrated that the duplicate sea bass Crtac1a and Crtac1b proteins and human CRTAC-1A all promoted SJD.1 primary fibroblast migration in a classic scratch assay and in an electric cell impedance sensing assay. The immunofluorescence analysis revealed that CRTAC1 enhanced cell migration was most likely caused by actin-driven cytoskeletal changes and the cellular transcriptional response was most affected in the early stage (6 h) of scratch repair. In summary, our results suggest that CRTAC1 may be an important factor in fish skin promoting damage repair.
- Core genes of biomineralization and cis-regulatory long non-coding RNA regulate shell growth in bivalvesPublication . Peng, Maoxiao; Cardoso, João; Pearson, Gareth Anthony; Canario, Adelino; Power, Deborah MaryBivalve molluscs are abundant in marine and freshwater systems and contribute essential ecosystem services. They are characterized by an exuberant diversity of biomineralized shells and typically have two symmetric valves (a.k.a shells), but oysters (Ostreidae), some clams (Anomiidae and Chamidae) and scallops (Pectinida) have two asymmetrical valves. Predicting and modelling the likely consequences of ocean acidification on bivalve survival, biodiversity and aquaculture makes understanding shell biomineralization and its regulation a priority. Objectives: This study aimed to a) exploit the atypical asymmetric shell growth of some bivalves and through comparative analysis of the genome and transcriptome pinpoint candidate biomineralization-related genes and regulatory long non-coding RNAs (LncRNAs) and b) demonstrate their roles in regulating shell biomineralization/growth. Methods: Meta-analysis of genomes, de novo generated mantle transcriptomes or transcriptomes and proteomes from public databases for six asymmetric to symmetric bivalve species was used to identify biomineralization-related genes. Bioinformatics filtering uncovered genes and regulatory modules characteristic of bivalves with asymmetric shells and identified candidate biomineralization-related genes and lncRNAs with a biased expression in asymmetric valves. A shell regrowth model in oyster and gene silencing experiments, were used to characterize candidate gene function. Results: Shell matrix genes with asymmetric expression in the mantle of the two valves were identified and unique cis-regulatory lncRNA modules characterized in Ostreidae. LncRNAs that regulate the expression of the tissue inhibitor of metalloproteinases gene family (TIMPDR) and of the shell matrix protein domain family (SMPDR) were identified. In vitro and in vivo silencing experiments revealed the candidate genes and lncRNA were associated with divergent shell growth rates and modified the microstructure of calcium carbonate (CaCO3) crystals. Conclusion: LncRNAs are putative regulatory factors of the bivalve biomineralization toolbox. In the Ostreidae family of bivalves biomineralization-related genes are cis-regulated by lncRNA and modify the planar growth rate and spatial orientation of crystals in the shell.
- Corrigendum: Evidence for a genetic contribution to the ossification of spinal ligaments in ossification of posterior longitudinal ligament and diffuse idiopathic skeletal hyperostosis: a narrative reviewPublication . Couto, Ana Rita; Parreira, Bruna; Power, Deborah Mary; Pinheiro, Luís; Dias, João Madruga; Novofastovski, Irina; Eshed, Iris; Sarzi-Puttini, Piercarlo; Pappone, Nicola; Atzeni, Fabiola; Verlaan, Jorrit-Jan; Kuperus, Jonneke; Bieber, Amir; Ambrosino, Pasquale; Kiefer, David; Khan, Muhammad Asim; Mader, Reuven; Baraliakos, Xenofon; Bruges-Armas, Jácome
- Dual-compartment-gate organic transistors for monitoring biogenic amines from foodPublication . Sergi, Ilenia; Sensi, Matteo; Zanotti, Rian; Tsironi, Theofania; Flemetakis, Emmanouil; Power, Deborah Mary; Bortolotti, Carlo Augusto; Biscarini, FabioAccording to the Food and Agriculture Organization of the United Nations (FAO) more than 14% of the world's food production is lost every year before reaching retail, and another 17% is lost during the retail stage. The use of the expiration date as the main estimator of the life-end of food products creates unjustified food waste. Sensors capable of quantifying the effective food freshness and quality could substantially reduce food waste and enable more effective management of food chain. We propose an electrolyte-gated organic transistor (EGOT) that responds to the release of biogenic amines, like diamines and tyramine, generated by degradation of protein-rich food. The EGOT sensor features a polymeric poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) gate electrode fabricated in the shape of a miniaturized beaker containing an aqueous solution in the inner side (to be exposed to food) and capacitively coupled through a hydrogel to the transistor channel on the outside (not in contact with food). The hydrogen bonds formed by the water-dissolved amines with PEDOT:PSS modulate the EGOT channel across a wide range of amine concentrations. We demonstrate that our sensor can detect different amines by the combinatorial analysis of the response from different channel materials, PEDOT:PSS and the other DPP-DTT, with a limit of detection as low as 100 pM.
- Evolution of chitin-synthase in molluscs and their response to ocean acidificationPublication . Cardoso, João; Power, Deborah Mary; Peng, MaoxiaoChitin-synthase (CHS) is found in most eukaryotes and has a complex evolutionary history. Research into CHS has mainly been in the context of biomineralization of mollusc shells an area of high interest due to the consequences of ocean acidification. Exploration of CHS at the genomic level in molluscs, the evolution of isoforms, their tissue distribution, and response to environmental challenges are largely unknown. Exploiting the extensive molecular resources for mollusc species it is revealed that bivalves possess the largest number of CHS genes (12-22) reported to date in eukaryotes. The evolutionary tree constructed at the class level of molluscs indicates four CHS Type II isoforms (A-D) probably existed in the most recent common ancestor, and Type II-A (Type II-A1/Type II-A-2) and Type II-C (Type II-C-1/Type II-C-2) underwent further differentiation. Non-specific loss of CHS isoforms occurred at the class level, and in some Type II (B-D groups) isoforms the myosin head domain, which is associated with shell formation, was not preserved and highly species-specific tissue expression of CHS isoforms occurred. These observations strongly support the idea of CHS functional diversification with shell biomineralization being one of several important functions. Analysis of transcriptome data uncovered the species-specific potential of CHS isoforms in shell formation and a species-specific response to ocean acidification (OA). The impact of OA was not CHS isoform-dependent although in Mytilus, Type I-B and Type II-D gene expression was down-regulated in both M. galloprovincialis and M. coruscus. In summary, during CHS evolution the gene family expanded in bivalves generating a large diversity of isoforms with different structures and with a ubiquitous tissue distribution suggesting that chitin is involved in many biological functions. These findings provide insight into CHS evolution in molluscs and lay the foundation for research into their function and response to environmental changes.
- Examination of the effects of excess microalgae availability on the disruption of mussel byssus secretionPublication . Ji-Yue Ni; Yan Zhou; Yu-Qing Wang; Shi-Hui Huang; Qian-Wen Cui; Wen-Yi Wang; Xiao-Ying Yang; Power, Deborah Mary; Yi-Feng LiThe present study evaluated the effects of food availability on byssus secretion in the hard-shelled mussel Mytilus coruscus. Byssus production was evaluated, and foot tissue transcriptomes, the site of byssus production, were generated. The results showed that byssus secretion was disrupted in mussels fed high levels of microalgae (HFL, 88.8 mg/L day−1), compared to the control group (p < 0.05), and that more byssus production occurred in the HFL group during the recovery period. Byssus secretion was not affected in mussels fed low levels of microalgae (8.88 × 10−3 mg/L day−1), but the shedding of byssus filaments was significantly increased compared to the control mussels (p < 0.05). Transcriptome data of the foot tissue revealed that lysosome, FoxO signaling, and autophagy pathways involved in autophagic cell death and apoptosis were significantly affected (p < 0.05) and may explain differences in byssus growth under modified food availability in M. coruscus. High feed levels modified metabolic pathways, such as sphingolipid, tyrosine, and phenylalanine, and downregulated genes coding for mussel foot proteins, which may explain reduced byssus production. The study found that contrary to expectation increased food availability reduced mussel byssus production by altering foot metabolism and promoting autophagy of foot tissue.
- Extracellular bioelectrical lexicon: detecting rhythmic patterns within dermal fibroblast populationsPublication . Félix, Rute; Medeiros, Maria do Carmo; Elamine, Youssef; Power, Deborah Mary; Gomes, Henrique LeonelThis study uses a bioelectronic-based method to establish how non-electrogenic cells, like dermal fibroblast, employ bioelectrical signals to convey information. Electrophysiology using large-area Multielectrode Arrays (MEAs) devices revealed how populations of non-electrogenic cells in vitro generate patterns of bioelectrical signals. The period of the bioelectrical patterns depends on cell population activity. In a fully formed, healthy monolayer, bioelectrical activity is minimal. But during the formation of a monolayer, signals appear randomly, with a dominant period of 4.2 min. Occasionally, quasi-periodic bursts occur with a period between 1.6 and 2 min. When a mechanical wound is inflicted and during subsequent monolayer repair, quasi-periodic signal bursts occur, with an average period ranging from 60 to 110 min. The study uncovers a short-range non humoral communication system and a lexicon of bioelectrical signals linked to cell states.
- Extracting protein from microalgae (Tetraselmis chuii) for proteome analysisPublication . L, Anjos; Estêvão, J.; Infante, Carlos; Mantecón, Lalia; Power, Deborah MaryMicroalgae have high potential as a resource for sustainable and green protein for food or bioactive molecules. Nonetheless, despite the high protein content of microalgae (40 - 70% dry weight) progress in the characterization of their protein composition remains challenging. This is due to the highly variable chemical composition of microalgae strains and factors such as their rigid thick cell wall, polysaccharide content, protein stability, pH. The method described herein was developed to optimize protein extraction for proteome analysis of microalgae (Tetraselmis chuii) biomass. The effects on protein solubility of solvent type (organic, denaturing, and non-denaturing) combined with three customized microalgae disruption methods were investigated. The proteome targeted high quality protein extracts were for hydro-soluble proteins recovered by cell disruption using bead milling coupled to centrifugation (protein yield approximate to 13%). The developed method is inexpensive, efficient (yielding high-quality protein extracts with a low content of interfering compounds) and from an industrial perspective easily scalable and compatible with other applications. To add value to the end product we additionally propose the use of stabilizing agents to maintain protein solubility during refrigerated storage and a method targeting the fractionation of low molecular weight proteins. An inexpensive easy-to-do 5 step protocol for microalgae protein extracts. A protein extraction method free from dangerous or highly polluting chemicals. Production of high yield aqueous protein extracts suitable for proteomics. (C) 2022 The Authors. Published by Elsevier B.V.
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