Heat shock protein (HSP) expression and mitogen-activated protein kinase (MAPK) phosphorylation during early embryonic developmental stages of the Gilthead sea bream (Sparus aurata)

Both heat shock proteins (HSPs), which have key roles in vital cell functions, as well as members of the mitogen-activated protein kinases (MAPKs), which adjust gene expression by transducing cellular signals to the nucleus, are necessary for normal embryonic development in vertebrates. Therefore, protein expression levels of HSP70 and HSP90 and the activation of members of the MAPK protein family, such as p38 MAPK, ERKs, and JNKs were studied in the early developmental stages of the Gilthead sea bream, Sparus aurata Linnaeus, 1758. The protein expression of HSP70 and the phosphorylation ratio of JNKs remained at equal levels at all examined developmental stages, while the other examined proteins exhibited a differential profile. HSP90 levels were mostly increased at the 16-cell stage and towards the morula stages, and the lowest values were observed at the two- to four-cell and one-half epiboly stages. While p38 MAPK phosphorylation ratio exhibited increased values mostly in the early developmental stages, the opposite was observed concerning ERK phosphorylation ratio, where increased values were observed in the later embryonic stages (high blastula to one-half epiboly stages). These differential profiles of the examined protein expression levels highlight the importance of these proteins during embryogenesis and pave the way for further research to unveil their distinct role in early development

Heat hardening enhances metabolite-driven thermoprotection in the Mediterranean mussel Mytilus galloprovincialis

Introduction: Temperature affects organisms’ metabolism and ecological performance. Owing to climate change, sea warming constituting a severe source of environmental stress for marine organisms, since it increases at alarming rates. Rapid warming can exceed resilience of marine organisms leading to fitness loss and mortality. However, organisms can improve their thermal tolerance when briefly exposed to sublethal thermal stress (heat hardening), thus generating heat tolerant phenotypes.Methods: We investigated the “stress memory” effect caused by heat hardening on M. galloprovincialis metabolite profile of in order to identify the underlying biochemical mechanisms, which enhance mussels’ thermal tolerance.Results: The heat hardening led to accumulation of amino acids (e.g., leucine, isoleucine and valine), including osmolytes and cytoprotective agents with antioxidant and anti-inflammatory properties that can contribute to thermal protection of the mussels. Moreover, proteolysis was inhibited and protein turnover regulated by the heat hardening. Heat stress alters the metabolic profile of heat stressed mussels, benefiting the heat-hardened individuals in increasing their heat tolerance compared to the non-heat-hardened ones.Discussion: These findings provide new insights in the metabolic mechanisms that may reinforce mussels’ tolerance against thermal stress providing both natural protection and potential manipulative tools (e.g., in aquaculture) against the devastating climate change effects on marine organisms

Epididymal Adenomatoid Tumor: A Very Rare Paratesticular Tumor of Childhood

Adenomatoid tumor is an uncommon benign mesothelial neoplasm, usually localized in the epididymis. It is the most common paratesticular tumor of middle-aged patients (average age of clinical presentation: 36 years). However, these tumors in pediatric and pubertal patients are extremely rare. Due to their rarity, we present a case of adenomatoid tumor of the tail of the epididymis in a 16-year-old patient. After systematic research of the current literature, we did not find another case report of epididymal adenomatoid tumor in a male patient aged 16 years old or less. This notice and our concern, as well, about the patient’s surveillance protocol during the postoperative period were the motive for this case study

Dietary Tenebrio molitor Larvae Meal Inclusion Exerts Tissue-Specific Effects on Cellular, Metabolic, and Antioxidant Status in European Sea Bass (Dicentrarchus labrax) and Gilthead Seabream (Sparus aurata)

The present study addresses the effects of dietary Tenebrio molitor (TM) larvae meal inclusion on cytoprotective, cell death pathways, antioxidant defence, and intermediate metabolism in the heart, muscle, and digestive tract of gilthead seabream (Sparus aurata) and European sea bass (Dicentrarchus labrax). Three experimental diets were formulated to contain 0%, 25%, or 50% inclusion TM levels. Heat Shock Proteins (HSPs) induction was apparent in both species’ muscle at 50% inclusion. Conversely, p44/42 Mitogen-Activated Protein Kinase (MAPK) activation was increased () in both species’ muscle and digestive tract at 25% inclusion. Regarding the apoptotic machinery, TM inclusion exerted no influence on gilthead seabream, while suppression through autophagy may have occurred in the muscle. However, significant apoptosis () was evident in European sea bass muscle and digestive tract. Both fish species’ heart seemed to additionally rely on lipids compared to muscle and digestive tract. In contrast to gilthead seabream, European sea bass exhibited increased () antioxidant activity at 50% TM inclusion. The present findings highlight the dietary derived induction of cellular responses in a species- and tissue-specific manner, whereas European sea bass appears to be more susceptible to TM inclusion

Heat hardening enhances mitochondrial potential for respiration and oxidative defence capacity in the mantle of thermally stressed Mytilus galloprovincialis

Ectotherms are exposed to a range of environmental temperatures and may face extremes beyond their upper thermal limits. Such temperature extremes can stimulate aerobic metabolism toward its maximum, a decline in aerobic substrate oxidation, and a parallel increase of anaerobic metabolism, combined with ROS generation and oxidative stress. Under these stressful conditions, marine organisms recruit several defensive strategies for their maintenance and survival. However, thermal tolerance of ectothermic organisms may be increased after a brief exposure to sub- lethal temperatures, a process known as "hardening". In our study, we examined the ability of M. galloprovincialis to increase its thermal tolerance under the effect of elevated temperatures (24, 26 and 28 °C) through the "hardening" process. Our results demonstrate that this process can increase the heat tolerance and antioxidant defense of heat hardened mussels through more efficient ETS activity when exposed to temperatures beyond 24 °C, compared to non-hardened individuals. Enhanced cell protection is reflected in better adaptive strategies of heat hardened mussels, and thus decreased mortality. Although hardening seems a promising process for the maintenance of aquacultured populations under increased seasonal temperatures, further investigation of the molecular and cellular mechanisms regulating mussels’ heat resistance is required

Tenebrio molitor larvae meal inclusion affects hepatic proteome and apoptosis and/or autophagy of three farmed fish species

Acknowledgements Financial support for the trial on European sea bass was provided by the AQUAEXEL Project PROINSECTLIFE (Ref. No. 0013/03/05/15B), the AQUAEXEL Project INDIFISH (Ref. No. 0125/08/05/15/TNA), and by the University of Turin (ex 60%) Grant (Es. fn. 2014). NP (Scholarship Code: 1752) has been fnancially supported by the General Secretariat for Research and Technology (GSRT) of Greece and the Hellenic Foundation for Research and Innovation (HFRI) and MM by the Operational Programme “Human Resources Development, Education and Lifelong Learning” in the context of the project “Strengthening Human Resources Research Potential via Doctorate Research” (MIS-5000432) as implemented by the State Scholarships Foundation (ΙΚΥ). Tanks to Evelyn Argo and Craig Pattinson (University of Aberdeen) for providing help with 2DE. EM was fnancially supported by Marine Alliance for Science and Technology Scotland (MASTS) visiting Fellowship.Peer reviewedPublisher PD

Case Report Multiple Gastric Erosion Early after a 3 V Lithium Battery (CR2025) Ingestion in an 18-Month-Old Male Patient: Consideration about the Proper Time of Intervention

Introduction. Button battery ingestion is considered an emergency situation in pediatric patients that needs to be managed promptly; otherwise, it may lead to serious and potentially lethal complications, especially when it is impacted in the esophagus. Less attention has been given in cases where the battery passes into the stomach, with guidelines for emergency intervention being based on the presence of symptoms. Case Report. We present a case of an 18-month-old male patient who presented to our emergency department after button battery ingestion. He did not have any symptoms and no pathological findings were encountered during clinical examination. X-ray investigation revealed the presence of the battery in the stomach. The patient was admitted for observation and two hours later he had two episodes of vomiting. He underwent urgent endoscopic removal of the battery where multiple acute gastric mucosal erosion in place of direct contact of the battery's negative pole with the mucosa of the gastric antrum was found. Conclusion. In specific cases the urgent endoscopic intervention for removal of an ingested button battery that is located in the stomach even in asymptomatic patients should be suggested

Effects of dietary substitution of fishmeal by black soldier fly (Hermetia illucens) meal on growth performance, whole-body chemical composition, and fatty acid profile of Pontastacus leptodactylus juveniles

Freshwater crayfish are considered as aquatic products of high quality and high nutritional value. The increasing demand has led to populations reduction in several locations throughout their range. Thus, the development of appropriate rearing conditions is considered necessary, among which, optimization of their diet is a basic part. Towards this direction, in the present study, a 98-day feeding trial was carried out to evaluate the impact of dietary fishmeal substitution by Hermetia illucens meal on Pontastacus leptodactylus juveniles kept under laboratory conditions. Insect meals represent an environmentally friendly alternative solution, considered as a high-value feed source, rich in nutrients such as protein and fat. Three dietary regimens were utilized with a fishmeal-based without Hermetia meal (HM) defined as the control diet (HM0), and two diets, the first with 50% (HM50) and the second with 100% (HM100) of fishmeal substitution by HM, respectively. Growth performance, whole-body composition, and fatty acid profiles of individuals were studied in the different treatments. At the end of the feeding trial, statistically significant differences were observed in the mean survival rate (SR), specific growth rate (SGR), feed conversion ratio (FCR) and weight gain (WG) values. More specifically, animals fed with HM-based diets had higher mean SR, while the control group performed better regarding FCR and SGR. The HM inclusion in the diet significantly altered the whole-body chemical composition of the crayfish signifying a different metabolic utilization compared to fishmeal (FM). The fatty acid analysis revealed that 16:0 (palmitic acid) was the predominant saturated fatty acid (SFA), 18:1ω9 (oleic acid) was found to be the main monounsaturated fatty acid (MUFA), while 18:2ω6 (linoleic acid) represented the major polyunsaturated fatty acid (PUFA) followed by C20:3 cis ω3 (cis-11-14-17-eicosatrienoate) and C22:6 cis ω3 (cis-4,7,10,13,16,19-Docosahexaenoic) fatty acids. The inclusion of dietary HM significantly reduced the contents of ∑SFAs, ∑PUFAs and ∑ω6 fatty acids, as well as those of C22:6 cis ω3 and increased the ω6/ω3 and hypocholesterolemic to hypercholesterolemic ratios in the body. In parallel with improvements in balanced diets and in culture conditions that need to be optimised for rearing of freshwater crayfish, our study provides new data that enlighten the suitability of insect meals in the nutrition of P. leptodactylus

Seasonal biochemical and physiological responses of the gilthead sea bream ( Sparus aurata ): correlation to the sea temperature and the climate change

Increased atmospheric levels of CO2 due to human activities and the subsequent Climate Change with effects summarizing except of the world temperature increase, to the ocean acidification, the reduction of the dissolved oxygen in marine ecosystems and the increase of the sea level, trigger a series of physical and chemical alterations that affect all levels of biological organization. Therefore, temperature is an environmental factor of great importance, since it significantly affects organisms’ activity as well as growth and developmental rates. Fish is the main group of aquatic organisms, and as a diet it is one of the main protein sources, in many countries, including Greece. The aim of the present study was to determine the biochemical and metabolic responses of one well studied and commercial species, the gilthead sea bream (Sparus aurata), in different water temperatures with or without the presence of CO2, and the seasonal pattern of these responses. Specifically, in several tissues of the particular species, we determined the induced levels of heat shock proteins Hsp70 and Hsp90, and the phosphorylated levels p38 MAPK, p54/46 JNK, p44/42 MAPK (ERKs), as well as expression levels of the transcriptional factor Hydroxy - Hifα and Ubiquitin. Cell metabolism adjustments were assessed by determining the activity of enzymes such as lactate dehydrogenase (L-LDH), citrate synthetase (CS) and β-hydroxyacyl CoA dehydrogenase (HOAD). Additionally we studied the kinetics of the above mentioned enzymes and antioxidant enzymes such as xanthine oxidase (XO), catalase (CAT), glutathione peroxidise (GPx) and superoxide dismoutase (SOD), as well as the determination of thiobarbituric acid reactive substances (TBARs). Levels of glucose, triglycerides and lactate were determined in blood plasma. A few reports have shown a direct relationship between phosphorylation of p38 MAPK and induction of Hsp70. This evidence is derived only from studies on mammalian tissues. A particular experimental procedure species has also shown the existence of this molecular mechanism in the particular species. Water temperature in the Mediterranean and the northeast Aegean, where gilthead sea bream is usually cultivated, varies from 10oC in the winter to 26oC in the summer. Seasonal biochemical and metabolic variations in Hsps’ induction, MAPKs’ phosphorylation, as well as in the activity of key enzymes and the activation of antioxidant mechanisms, reflect a seasonal temperature impact on the gilthead sea bream’s metabolic and molecular profile. In order to determine whether physiological processes such as reproduction are involved in the specific seasonal pattern, we conducted a study in reproduction parameters, showing a possible involvement in the metabolic mechanisms, as the activity of certain enzymes is reduced, a phenomena known as “energy allocation”. Laboratory studies showed that elevated temperature with/without the presence of CO2 activated cellular response to stress, especially under the simultaneous effect of temperature and hypercapnia. From 17οC to 22οC, gilthead sea bream demonstrates a normal behaviour. Above and below these temperatures, stress responses are activated. The above results are of importance if they could possibly serve in management programmes of this highly commercial speciesΤο αυξημένο, εξαιτίας των ανθρωπογενών δραστηριοτήτων, CO2 στην ατμόσφαιρα και η επακόλουθη Κλιματική Αλλαγή με κύριες επιπτώσεις εκτός από την αύξηση της μέσης παγκόσμιας θερμοκρασίας, την οξείδωση των ωκεανών και την άνοδο της στάθμης της θάλασσας, ενεργοποιεί μία σειρά φυσικών και χημικών αλλαγών που επηρεάζουν αρκετά επίπεδα βιολογικής οργάνωσης. Η θερμοκρασία είναι ένας κρίσιμος περιβαλλοντικός παράγοντας, θέτοντας τα όρια για επιβίωση, καθώς επηρεάζει σημαντικά τη δραστηριότητα των οργανισμών και τους ρυθμούς αύξησης και ανάπτυξής τους. Τα ψάρια, είναι η κυρίαρχη ζωική ομάδα των θαλάσσιων οικοσυστημάτων και συνάμα αποτελούν μία από τις κυριότερες πηγές πρωτεϊνών. Για το λόγο αυτό, τα αποτελέσματα της θαλάσσιας υπερθέρμανσης στα ψάρια χρήζουν προσεκτικής ανάλυσης. Σκοπός της παρούσας διατριβής ήταν η μελέτη των βιοχημικών και μεταβολικών αποκρίσεων της τσιπούρας (Sparus aurata), ενός από τα καλύτερα μελετημένα και μεγάλης εμπορικής αξίας είδη, σε διαφορετικές θερμοκρασίες θαλασσινού νερού με και χωρίς την παρουσία σε αυτό διοξειδίου του άνθρακα και το εποχικό πρότυπο αυτών. Συγκεκριμένα, σε ιστούς του συγκεκριμένου ψαριού, προσδιορίστηκαν οι πρωτεΐνες θερμικού πλήγματος Hsp70 και Hsp90, η φωσφορυλιωμένη p38 MAPK, οι φωσφορυλιωμένες JNKs, οι φωσφορυλιωμένες μορφές τις p44/42 MAPK, ο μεταγραφικός παράγοντας Hydroxy - Hifα και η ουβικουϊτίνη. Η μελέτη του κυτταρικού μεταβολισμού έγινε με τον προσδιορισμό της δραστικότητας ενζύμων όπως η αφυδρογονάση του γαλακτικού οξέος (L-LDH), η συνθετάση του κιτρικού οξέος (CS) και η αφυδρογονάση του β-υδροξυακυλο-CoA (HOAD). Μελετήθηκε και η κινητική των παραπάνω ενζύμων και η δραστικότητα αντιοξειδωτικών ενζύμων όπως η οξειδάση της ξανθίνης (XO), η καταλάση (CAT), η υπεροξειδάση της γλουταθειόνης (GPx) και η δισμουτάση του υπεροξειδίου (SOD), αλλά και η υπεροξείδωση των λιπιδίων με τη μέθοδο TBARs. Στο πλάσμα του αίματος προσδιορίστηκαν η γλυκόζη, τα τριγλυκερίδια και το γαλακτικό οξύ. Προηγούμενες έρευνες έχουν δείξει μια στενή σχέση μεταξύ της φωσφορυλίωσης της p38 MAPK και της επαγωγής της Hsp70, αλλά μόνο σε ιστούς θηλαστικών. Για τον λόγο αυτό διεξήχθη πειραματική διαδικασία η οποία απέδειξε τη σχέση αυτή στο μελετώμενο είδος. Η θερμοκρασία στη Μεσόγειο Θάλασσα και ιδιαίτερα στο Βορειοανατολικό Αιγαίο, όπου και η τσιπούρα εκτρέφεται, κυμαίνεται εποχικά από τους 10oC τον χειμώνα έως τους 26oC το καλοκαίρι. Οι εποχικές βιοχημικές και μεταβολικές αλλαγές, τόσο στις Hsps και στις MAPKs, όσο και στη δραστικότητα ενζύμων, αντικατοπτρίζουν μια εποχική επίδραση της θερμοκρασίας στο μεταβολικό και μοριακό πρότυπο της τσιπούρας. Για να διαπιστωθεί η συμμετοχή ή όχι βασικών φυσιολογικών διαδικασιών όπως είναι η αναπαραγωγή στο συγκεκριμένο εποχικό πρότυπο, διεξήχθη μελέτη των αναπαραγωγικών παραμέτρων, αποδεικνύοντας πιθανή εμπλοκή αυτών στη δραστικότητα βασικών ενζύμων του μεταβολισμού και τελικά μείωση των μεταβολικών μηχανισμών στο είδος αυτό, ενός φαινομένου που είναι γνωστό και ως «μετατόπιση ενέργειας». Σε εργαστηριακές συνθήκες, η επίδραση της θερμοκρασίας με/χωρίς CO2, ενεργοποιεί επίσης τους μηχανισμούς κυτταρικής απόκρισης στο στρες, με εντονότερες μεταβολές κάτω από την ταυτόχρονη επίδραση της θερμοκρασίας και της υπερκαπνίας. Συγκεκριμένα, η τσιπούρα παρουσιάζει φυσιολογική συμπεριφορά από τους 17οC μέχρι και τους 22οC, κάτω και πάνω από τους οποίους αντίστοιχα, αρχίζουν και παρατηρούνται αντιδράσεις στρες. Τα αποτελέσματα αυτά είναι σημαντικά εφόσον βοηθήσουν σε προγράμματα διαχείρισης αυτού του εμπορικού εκτρεφόμενου είδου

Synergistic effects of acute warming and low pH on cellular stress responses of the gilthead seabream Sparus aurata

The present study assesses the resilience of the Mediterranean gilthead seabream (Sparus aurata) to acute warming and water acidification, using cellular indicators of systemic to molecular responses to various temperatures and CO2 concentrations. Tissue metabolic capacity derived from enzyme measurements, citrate synthase, 3-hydroxyacyl CoA dehydrogenase (HOAD), as well as lactate dehydrogenase. Cellular stress and signaling responses were identified from expression patterns of Hsp70 and Hsp90, the phosphorylation of p38 MAPK, JNKs and ERKs, from protein ubiquitylation and finally from the levels of transcription factor Hif-1α as an indicator of systemic hypoxemia. Exposure to elevated CO2 levels at temperatures higher than 24 °C generally caused an increase in fish mortality above the rate caused by warming alone, indicating effects of the two factors and a failure of acclimation and thus the limits of phenotypic plasticity to be reached. As a potential reason, tissue-dependent induction and stabilization of Hif-1α indicate hypoxemic conditions. Their exacerbation by enhanced CO2 levels is linked to the persistent expression of Hsp70 and Hsp90, oxidative stress and activation of MAPK and ubiquitin pathways. Antioxidant defence is enhanced by expression of catalase and glutathione reductase, however, leaving superoxide dismutase suppressed by elevated CO2 levels. On longer timescales in specimens surviving warming and CO2 exposures, various metabolic adjustments initiate a preference to oxidize lipid via HOAD for energy supply. These processes indicate significant acclimation up to a limit and a time-limited capacity to survive extreme conditions passively by exploiting mechanisms of cellular resilience