Complement component C4-like protein in Atlantic cod (<i>Gadus morhua</i> L.) - Detection in ontogeny and identification of post-translational deimination in serum and extracellular vesicles

The complement system is a critical part of teleost immune defences, with complement component C4 forming part of the classical and lectin pathways. Cod C4-like protein was isolated from plasma, specific antibodies generated and C4-like protein was assessed in cod sera, mucus and in extracellular vesicles (EVs) from serum and mucus. Higher levels of C4-like protein were detected in serum- than mucus-derived EVs. Post-translational deimination, caused by conversion of arginine into citrulline, can affect protein structure and function. Here we detected deiminated forms of C4-like protein in cod serum and at lower levels in mucus. C4-like protein was also found in deiminated form at low levels in EVs from both serum and mucus. C4-like protein was assessed by immunohistochemistry in cod larvae and detected in a range of organs including in liver, kidney, gut, muscle, skin and mucus, as well as in neuronal tissues of the brain, spinal cord and eye. This abundance of C4-like protein during early development may indicate roles in tissue remodelling, in addition to immune defences. The presence of deiminated C4-like protein in serum and mucosa, as well as in EVs, may suggest C4 protein moonlighting via post-translational deimination

Physiological responses of reared sea bream (Sparus aurata Linnaeus, 1758) to an Amyloodinium ocellatum outbreak

Amyloodiniosis represents a major bottleneck for semi-intensive aquaculture production in Southern Europe, causing extremely high mortalities. Amyloodinium ocellatum is a parasitic dinoflagellate that can infest almost all fish, crustacean and bivalves that live within its ecological range. Fish mortalities are usually attributed to anoxia, associated with serious gill hyperplasia, inflammation, haemorrhage and necrosis in heavy infestations; or with osmoregulatory impairment and secondary microbial infections due to severe epithelial damage in mild infestation. However, physiological information about the host responses to A.ocellatum infestation is scarce. In this work, we analysed the proteome of gilthead sea bream (Sparus aurata) plasma and relate it with haematological and immunological indicators, in order to enlighten the different physiological responses when exposed to an A.ocellatum outbreak. Using 2D-DIGE, immunological and haematological analysis and in response to the A.ocellatum contamination we have identified several proteins associated with acute-phase response, inflammation, lipid transport, homoeostasis, and osmoregulation, wound healing, neoplasia and iron transport. Overall, this preliminary study revealed that amyloodiniosis affects some fish functional pathways as revealed by the changes in the plasma proteome of S. aurata, and that the innate immunological system is not activated in the presence of the parasite.DIVERSIAQUA, Portugal [MAR2020]Fundacao para a Ciencia e Tecnologia [SFRH/BD/118601/2016]info:eu-repo/semantics/publishedVersio

Dietary b-glucan (MacroGard®) enhances survival of first feeding turbot (Scophthalmus maximus) larvae by altering immunity, metabolism and microbiota

Reflecting the natural biology of mass spawning fish aquaculture production of fish larvae is often hampered by high and unpredictable mortality rates. The present study aimed to enhance larval performance and immunity via the oral administration of an immunomodulator, β-glucan (MacroGard®) in turbot (Scophthalmus maximus). Rotifers (Brachionus plicatilis) were incubated with or without yeast β-1,3/1,6-glucan in form of MacroGard® at a concentration of 0.5 g/L. Rotifers were fed to first feeding turbot larvae once a day. From day 13 dph onwards all tanks were additionally fed untreated Artemia sp. nauplii (1 nauplius ml/L). Daily mortality was monitored and larvae were sampled at 11 and 24 dph for expression of 30 genes, microbiota analysis, trypsin activity and size measurements. Along with the feeding of β-glucan daily mortality was significantly reduced by ca. 15% and an alteration of the larval microbiota was observed. At 11 dph gene expression of trypsin and chymotrypsin was elevated in the MacroGard® fed fish, which resulted in heightened tryptic enzyme activity. No effect on genes encoding antioxidative proteins was observed, whilst the immune response was clearly modulated by β-glucan. At 11 dph complement component c3 was elevated whilst cytokines, antimicrobial peptides, toll like receptor 3 and heat shock protein 70 were not affected. At the later time point (24 dph) an anti-inflammatory effect in form of a down-regulation of hsp 70, tnf-α and il-1β was observed. We conclude that the administration of MacroGard® induced an immunomodulatory response and could be used as an effective measure to increase survival in rearing of turbot

Symptomer på spiseforstyrrelser og selvfølelse blant unge utholdenhetsutøvere og utøvere i estetiske idretter: En tverrsnittstudie ved norske toppidrettsgymnas, gymnas med idrettsfag og ordinære videregående skoler

Masteroppgave - Norges idrettshøgskole, 2021Bakgrunn: Risikoen for å utvikle spiseforstyrrelser er økt for både kvinnelige- og mannlige idrettsutøvere sammenlignet med ikke-idrettsutøvere. Idrettsutøvere som konkurrerer i vektsensitive idretter er særlig utsatt. Hensikten med denne studien var å undersøke om unge idrettsutøvere som konkurrerer i utholdenhets- og estetiske idretter har høyere forekomst av symptomer på spiseforstyrrelser sammenlignet med jevn aldrende kontroller. Kjønnsforskjeller når det gjelder forekomst av symptomer på spiseforstyrrelser ble også undersøkt. Selvfølelse og spiseforstyrrelser har tidligere vist å ha en sammenheng, og det var derfor ønskelig å se om det var gjeldende for dette utvalget også. Metode: Tverrsnittstudie med baselinedata fra et pågående doktorgrads-prosjekt om seksuell trakassering ved Norges idrettshøgskole. Elever fra andre året ved norske videregående skoler ble inkludert (292 kvinner og 208 menn), og delt inn i to grupper: Idrettselever (elever ved toppidrettsgymnas og idrettslinjer) (n=241) og kontrollelever (elever ved studiespesialiserende linjer ved ordinære videregående skoler) (n=259). Kun idrettselever som representerte utholdenhetsidretter (n=192) og estetiske idretter (n=49) ble inkludert. Eating Disorder Examination - Questionnaire (EDE-Q) ble brukt for å undersøke symptomer på spiseforstyrrelser. Rosenberg Selvfølelsesskala (RSES) ble brukt for å undersøke selvfølelse. Resultater: Kontrollelevene hadde signifikant høyere EDE-Q global skår blant kvinner (p=0.004) og menn (p=0.013). For kvinner var det signifikant flere kontrollelever over cut-off (21.6%) sammenlignet med idrettselevene (10.7%) (p=0.015). I den mannlige gruppen ble det derimot ikke funnet signifikant forskjell i andel over cut-off mellom idrettselevene (6.7%) og kontrollelevene (12.5%) (p=0.150). Det var signifikant forskjell i andel over cut-off mellom utholdenhetsidrett (20.2%) og estetiske idretter (33.9%) for kvinner (p=0.023). Det var en sammenheng mellom RSES og EDE-Q hos menn (r=-.343) og kvinner (r=-.519). Konklusjon: Flere kontrollelever hadde symptomer på spiseforstyrrelser sammenlignet med idrettselever. Kvinner hadde en høyere forekomst av symptomer på spiseforstyrrelser sammenlignet med menn. Idrettselever i estetiske idretter hadde høyere forekomst av symptomer på spiseforstyrrelser sammenlignet med elever i utholdenhetsidretter for kvinner. RSES og EDE-Q hadde en negativ korrelasjon for menn (svak) og for kvinner (moderat) i det totale utvalget.publishedVersionInstitutt for idrettsmedisinske fag / Department of Sports Medicin

Poly-β-hydroxybutyrate administration during early life: effects on performance, immunity and microbial community of European sea bass yolk-sac larvae

The reliable production of marine fish larvae is one of the major bottlenecks in aquaculture due to high mortalities mainly caused by infectious diseases. To evaluate if the compound poly-β-hydroxybutyrate (PHB) might be a suitable immunoprophylactic measure in fish larviculture, its capacity to improve immunity and performance in European sea bass (Dicentrarchus labrax) yolk-sac larvae was explored. PHB was applied from mouth opening onwards to stimulate the developing larval immune system at the earliest possible point in time. Larval survival, growth, microbiota composition, gene expression profiles and disease resistance were assessed. PHB administration improved larval survival and, furthermore, altered the larva-associated microbiota composition. The bacterial challenge test using pathogenic Vibrio anguillarum revealed that the larval disease resistance was not influenced by PHB. The expression profiles of 26 genes involved e.g. in the immune response showed that PHB affected the expression of the antimicrobial peptides ferritin (fer) and dicentracin (dic), however, the response to PHB was inconsistent and weaker than previously demonstrated for sea bass post-larvae. Hence, the present study highlights the need for more research focusing on the immunostimulation of different early developmental stages for gaining a more comprehensive picture and advancing a sustainable production of high quality fry

Immunostimulatory effects of dietary poly-β-hydroxybutyrate in European sea bass post-larvae

The stable production of high quality fry in marine aquaculture is still hampered by unpredictable mortality caused by infectious diseases during larval rearing. Consequently, the development of new biocontrol agents is crucial for a viable aquaculture industry. The bacterial energy storage compound poly-β-hydroxybutyrate (PHB) has been shown to exhibit beneficial properties on aquatic organisms such as enhanced survival, growth, disease resistance and a controlling effect on the gastrointestinal microbiota. However, the effect of PHB on the developing immune system of fish larvae has so far not been investigated. In the present study, the effect of feeding PHB-enriched Artemia nauplii on survival, growth and immune response in European sea bass (Dicentrarchus labrax) post-larvae was examined. Amorphous PHB was administered to 28 days old sea bass larvae over a period of 10 days. The survival and growth performance were monitored and the expression of 29 genes involved in immunity, growth, metabolism and stress-response was measured. While the expression of the insulin-like growth factor 1 (igf1), an indicator of relative growth, was upregulated in response to feeding PHB, the larval survival and growth performance remained unaffected. After 10 days of PHB treatment, the expression of the antimicrobial peptides dicentracin (dic) and hepcidin (hep) as well as mhc class IIa and mhc class IIb was elevated in the PHB fed larvae. This indicates that PHB is capable of stimulating the immune system of fish early life stages, which may be the cause of the increased resistance to diseases and robustness observed in previous studies

A novel ladder-like lectin relates to sites of mucosal immunity in Atlantic Halibut (Hippoglossus hippoglossus L.)

A novel 27 kDa ladder-lectin-like protein, showing a multimeric structure under non-reducing conditions, was isolated from halibut serum by binding to N-acetyl glucosamine. Mass-spectrometry analysis did not show significant homology with known proteins. Specific antibodies were produced and used in immunohistochemistry on tissue sections of early halibut ontogeny from 119 until 1050 °d post hatching. A strong positive response was detected in the mucosal cells of the skin, gills and gut, indicating a role in the mucosal immune defence at these sites. Further immunopositivity was detected in liver, myeloma of kidney and the brain at different developmental stages but predominant expression was found in mucosal surfaces at later stages of development tested (1050 °d). It is still uncertain whether this ladder-like lectin forms part of the complement pathway, as a lectin or ficolin, or if it belongs to galectins. A strong detection in mucosal surfaces on skin, gills and gut, show similar patterns of expression as both mucosal lectins and galectins in other fish. Detection in neuronal tissue may indicate putative roles in tissue remodelling of brain and in ongoing neurogenesis in the fish eye

Extracellular vesicles, deiminated protein cargo and microRNAs are novel serum biomarkers for environmental rearing temperature in Atlantic cod (<i>Gadus morhua</i> L.)

Extracellular vesicles (EVs) are released from cells and carry protein and genetic cargo involved in cell communication. EVs isolated from bodyfluids, including plasma and serum, can also be used as markers of pathophysiological changes. Peptidylarginine deiminases (PADs) are phylogenetically conserved enzymes with physiological and pathophysiological roles and cause post-translational protein deimination. This can affect function of target proteins and deimination is also linked to EV release. Atlantic cod (Gadus morhua L.) reared at 4 °C and 9 °C respectively for 18 months, were here assessed for changes in serum derived EVs, including analysis of deiminated protein and micro-RNA cargo markers related to stress and growth. We found that cod reared at 9 °C showed significantly reduced numbers of EVs in serum, compared to cod reared at 4 °C. Some deiminated protein targets, including complement component C3, were found to be considerably higher in EVs of cod reared at 4 °C. Proteomic analysis revealed further differences in deiminated protein targets in EVs isolated from sera of the two temperature groups. Whole cod sera from the two temperature groups furthermore showed differences in deiminated protein targets, including C3, CRP and histone H3, which is a marker of neutrophil extracellular trap formation. MicroRNAs related to inflammation (miRNA-21) and stress (miRNA-155) were elevated in both total cod serum and serum-derived EVs from the 9 °C group, while the growth-related miRNA-206 was higher in the 4 °C group. Our findings highlight EVs as novel biomarkers to assess fish health in response to environmental rearing temperature

Serum bactericidal activity as indicator of innate immunity in pacu Piaractus mesopotamicus (Holmberg, 1887)

The immune system of teleost fish has mechanisms responsible for the defense against bacteria through protective proteins in several tissues. The protein action can be evaluated by serum bactericidal activity and this is an important tool to analyze the immune system. Pacu, Piaractus mesopotamicus, is one of the most important fish in national aquaculture. However there is a lack of studies on its immune responses. In order to standardize and assess the accuracy of the serum bactericidal activity assay, fish were briefly challenged with Aeromonas hydrophila and sampled one week after the challenge. The bacterial infection increased the concentration of protective proteins, resulting in a decrease of colony-forming unit values expressed as well as an enhanced serum bactericidal activity. The protocol showed a reliable assay, appropriate to determine the serum bactericidal activity of pacu in the present experimental conditions.O sistema imune de peixes teleósteos tem mecanismos responsáveis pela defesa contra bactérias e atua através de proteínas presentes em diversos tecidos. A ação destas proteínas pode ser avaliada pela atividade bactericida do soro, sendo esta uma importante ferramenta para analisar o sistema imune. O pacu, Piaractus mesopotamicus, é um peixe nativo muito importante para aquicultura nacional, entretanto há pouco conhecimento sobre o funcionamento de seu sistema imune. Assim foi realizado experimento para padronizar e avaliar a eficiência do ensaio de atividade bactericida. Resumidamente, peixes foram desafiados por Aeromonas hydrophila e amostradas uma semana após o desafio. A infecção bacteriana promoveu um aumento na concentração de proteínas protetoras, resultando em diminuição dos valores de unidades formadoras de colônias ou expressos também como aumento da atividade bactericida do soro. O protocolo se mostrou confiável, sendo apropriado para determinar a atividade bactericida do soro de pacu nas condições experimentais.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Estadual PaulistaUniversidade Estadual Paulista Faculdade de ZootecniaUniversidade Estadual PaulistaUniversidade Estadual Paulista Faculdade de Zootecni

Gender differences in work-life balance of European neurosurgeons

Neurosurgery is one of the most demanding medical specialties. For neurosurgeons, balancing professional activity with personal life can be challenging. To evaluate gender differences in contribution of neurosurgeons in the household and child-rearing, as well as their impact on personal life and career. An anonymous, electronic, 59-item web-based survey was administered to National Neurosurgical Societies of Europe, and European Member Societies of the European Association of Neurosurgical Societies (June-October 2021). A total of 205 European neurosurgeons (87 females and 118 males, mean age 40.7) are included in our survey. In neurosurgery, females are significantly more likely to be alone (37.9%), while males are significantly more likely to have children (66.9%). In terms of household efforts, females spend more time than males on the same tasks. Most participants (71.2%) view gender issues as a disadvantage in career pursuing. Women feel less accepted (54.3%) and having fewer opportunities (58.6%), while men believe that pregnancy/child-rearing (65.8%) and having many roles (51.3%) are the main obstacles. Both genders (77.6%) favor more convenient working conditions for young parents. Discussion and Conclusion. In our study we found that, women neurosurgeons take more responsibilities at home, especially in the child-rearing years. Female neurosurgeons are more likely to live alone or stay childless more often compared to their male colleagues. Supportive facilities, flexible programs, universal life policies and presumably curbing of the social stereotypes are of importance to overcome gender inequities that women are still facing in neurosurgery