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Gilthead seabream (Sparus aurata Linnaeus, 1758 (Perciformes, Sparidae)) is an important aquaculture species in the Mediterranean Sea basin. Yet, quantitative data on its dentition under standard farming conditions are currently lacking. Furthermore, it is unknown if the dentition can adapt to food of different sizes. Here, we describe the lower jaw dentition of juvenile S. aurata fed a standard pellet size (4 mm) and present a detailed analysis of 11 representative teeth. Overall, the number of teeth showed large individual variation, but it was not significantly related to fish length. Considerable left-right differences were observed, without clear side dominance. We also assessed the influence of feeding S. aurata a smaller (2 mm) or larger (6 mm) pellet size. Four months of feeding with different pellet sizes did not cause detectable differences in total tooth number on the dentaries at the time of harvest, nor in size of the teeth assumed to be most relevant in food processing. If and how different pellet sizes may nevertheless affect digestion, and eventually fish health, is subject for further studies. Key words: gilthead seabream, Sparus aurata, lower jaw, dentary, dentition, teeth, pellet size. Bien que la dorade royale (Sparus aurata Linnaeus, 1758 (perciformes, sparides)) soit une importante espece aquacole dans le bassin de la mer Mediterranee, des donnees quantitatives sur sa dentition dans des conditions d'elevage normales ne sont actuellement pas disponibles. En outre, la capacite d'adaptation de cette dentition a des aliments de differentes tailles n'est pas etablie. Nous decrivons la dentition de la machoire inferieure de S. aurata juveniles nourries de granules de taille standard (4 mm) et presentons une analyse detaillee de 11 dents representatives. Globalement, il y a de grandes variations individuelles du nombre de dents, mais elles ne presentent pas une relation significative avec la longueur des poissons. Des differences considerables entre les cotes gauche et droit sont observees, sans predominance claire d'un cote ou de l'autre. Nous evaluons egalement l'effet de donner aux S. aurata des granules de plus petite (2 mm) ou de plus grande (6 mm) taille. Apres 4 mois d'alimentation avec des granules de differentes tailles, aucune difference detectable n'a ete relevee du nombre de dents sur les dentaires au moment de la recolte ni de la taille des dents presumees etre les plus importantes pour le traitement de la nourriture. L'incidence eventuelle de la taille des granules sur la digestion et, consequemment, la sante des poissons devra faire l'objet d'autres etudes. [Traduit par la Redaction] Mots-cles: dorade royale, Sparus aurata, machoire inferieure, os dentaire, dentition, dents, taille des granules., Introduction Gilthead seabream (Sparus aurata Linnaeus, 1758 (Perciformes, Sparidae)) is a warm-temperate coastal teleost fish species in the Eastern Atlantic and is a species intensely produced in aquaculture in the [...]

Background: Plaque angiogenesis is associated with atherosclerotic lesion growth, plaque instability and negative clinical outcome. Plaque angiogenesis is a natural occurring process to fulfil the increasing demand of oxygen and nourishment of the vessel wall. However, inadequate formed, immature plaque neovessels are leaky and cause intraplaque haemorrhage.Objective: Blockade of VEGFR2 normalizes the unbridled process of plaque neovessel formation and induces maturation of nascent vessels resulting in prevention of intraplaque haemorrhage and influx of inflammatory cells into the plaque and subsequently increases plaque stability.Methods and Results: In human carotid and vein graft atherosclerotic lesions, leaky plaque neovessels and intraplaque haemorrhage co-localize with VEGF/VEGFR2 and angiopoietins. Using hypercholesterolaemic ApoE3*Leiden mice that received a donor caval vein interposition in the carotid artery, we demonstrate that atherosclerotic vein graft lesions at t28 are associated with hypoxia, Hif1α and Sdf1 up-regulation. Local VEGF administration results in increased plaque angiogenesis. VEGFR2 blockade in this model results in a significant 44% decrease in intraplaque haemorrhage and 80% less extravasated erythrocytes compared to controls. VEGFR2 blockade in vivo results in a 32% of reduction in vein graft size and more stable lesions with significantly reduced macrophage content (30%), and increased collagen (54%) and smooth muscle cell content (123%). Significant decreased VEGF, angiopoietin-2 and increased Connexin 40 expression levels demonstrate increased plaque neovessel maturation in the vein grafts. VEGFR2 blockade in an aortic ring assay showed increased pericyte coverage of the capillary sprouts.Conclusion: Inhibition of intraplaque haemorrhage by controlling neovessels maturation holds promise to improve plaque stability. [ABSTRACT FROM AUTHOR]

The present study was carried out to test different mussel meal (MM) dietary levels in combination with fishmeal (FM) on the growth performance, fatty acid composition and liver histology of common sole, Solea solea juveniles to highlight the growth potential of this species. Four isoproteic (53%) and isolipidic (11%) pelletized diets were formulated to contain graded levels of mussel meal, MM0 (0%), MM25 (25%), MM50 (50%) and MM75 (75%), up to 75%. Sole juveniles (initial individual mean body weight 13.1 ± 2.3 g, n = 840) were fed to satiation for 91 days. Seventy fish per tank (500-L, 0.64 m² bottom surface) were reared in 12 tanks (3 tanks per treatment) at 20 ± 1°C. Diets containing MM (MM25, MM50 and MM75) gave a significantly higher specific growth rate (SGR, 1.27 ± 0.01, 1.38 ± 0.06 and 1.40 ± 0.05, respectively), higher feed intake and lower feed conversion rate (FCR, 1.09 ± 0.01, 1.00 ± 0.04 and 0.98 ± 0.02, respectively) when compared to the FM-based diet (MM0, SGR, 0.98 ± 0.11, FCR, 1.52 ± 0.13). Carcass proximate composition was not influenced by dietary treatments, with the exception of the significantly lower lipid content in the MM75 group. Protein efficiency ratio (PER) and gross protein efficiency (GPE) were significantly improved by the mussel meal inclusion (PER, 1.29 ± 0.12, 1.76 ± 0.01, 1.89 ± 0.06, 1.95 ± 0.08; GPE, 25.29 ± 1.85, 33.38 ± 0.89, 35.96 ± 1.36, 36.59 ± 1.05 in MM0, MM25, MM50 and MM75, respectively). A significant decrease in the viscerosomatic index was observed in fish fed with MM50 and MM75 in comparison to MM0. The hepatosomatic index of fish fed with MM0 and MM25 was higher than that of fish fed with MM75, although the histological examination of liver parenchyma in all experimental groups showed a uniformly abundant accumulation of lipid droplets. Carcass fatty acid composition was significantly affected by dietary treatments, reflecting the dietary fatty acid profile. According to these results, the inclusion of MM in experimental FM-based diets improved the performance and feed utilization of common sole juveniles. The inclusion of MM in the present trial allowed a higher SGR than that registered in previous growth trials on common sole. This study could provide useful information to detect effective ingredients for practical diets in Solea solea. It also seems advisable to consider an inclusion of at least 25% MM in the experimental reference diet to be used for further application towards the development of specific diets for this species. [ABSTRACT FROM AUTHOR]

A 9-week feeding trial was performed to evaluate the effects of fishmeal ( FM) replacement by a mixture of plant proteins ( PP), consisted of wheat gluten, soybean meal and soy protein concentrate, on the growth performance and welfare of turbot juveniles (initial weight 9.7 ± 0.2 g). Four isonitrogenous and isolipidic diets contained FM at 500 g kg−1 ( FM50), 350 g kg−1 ( FM35), 200 g kg−1 ( FM20) and 50 g kg−1 ( FM5). A decreased feed intake and an increased feed conversion rate was observed in FM5 group. Specific growth rate was significantly reduced in FM20 and FM5 groups, whereas protein and lipid utilization and proximate whole body composition were significantly different in FM5 group. Serum cortisol significantly increased in FM20 and FM5 groups whereas cholesterol, triglycerides, NEFA, total protein and urea concentrations significantly decreased. Serum lysozyme and blood phagocytes increased in FM20 and FM5 groups. FM35 ensured growth close to FM50, without significant effects on health and welfare of animals. FM20 and FM5 groups displayed reduced growth, metabolic stress and an immune response with effects on health and welfare. Results highlighted the consistency between growth performance and welfare status, suggesting the usefulness of their combined assessment for evaluating the suitability of PP and to improve dietary formulation for turbot. [ABSTRACT FROM AUTHOR]

Contents The study of lipids and fatty acids ( FAs) has been used in the assessment of egg quality because their composition can influence the fertilization rate, hatching, survival and growth of marine fish larvae. For these reasons, the lipid content ( TL) and fatty acid composition of common sole ( Solea solea) eggs were measured and correlated to egg and larval viability parameters throughout an entire reproductive season. Seventeen batches of fertile eggs obtained from natural spawning of captive breeders were characterized for the TL, FA profile, hatching rate ( HR) and survival rate of larvae ( SR) at 0-6 days post-hatching (dph). The egg FA composition reflected the composition of the feed supplied to the broodstock during summer and autumn (before and during vitellogenesis) rather than that supplied during the spawning season. In general, the egg FA profile showed minimal differences among the early-, mid- and late-spawning periods (possibly due to the change of the diet and/or water temperature) indicating that it is possible to obtain a similar egg quality in terms of egg FA profile over 2 months of spawning. Saturated FAs and monounsaturated FAs ( MUFA) were positively correlated with HR, while TL, 22 : 6n-3 ( DHA), 20 : 4n-6 ( ARA), polyunsaturated FAs of the (n-3) series (n-3 PUFA) and polyunsaturated FAs of the (n-6) series were negatively correlated (p ≤ 0.05). MUFA, 20 : 5n-3 ( EPA), n-6/n-3 were positively correlated with SR, while DHA, n-3 PUFA, DHA/ EPA were negatively correlated (p ≤ 0.05). In conclusion, the feed supplied before and during vitellogenesis has a major role in determining the egg FA profile in common sole. The relationships found between TL and FAs with egg and larval viability parameters differ from many other farmed marine fish species, which may suggest the need for a specific broodstock feed for this species. [ABSTRACT FROM AUTHOR]

Histological and morphometric techniques were employed in juvenile Senegalese sole ( Solea senegalensis) livers to evaluate diet-related adaptive changes and to establish how an integrated morphofunctional approach could improve the understanding of rearing strategies. Four isoenergetic diets with variable protein/lipid content (44/22, 49/20, 54/18 and 59/16) were formulated. Data compared statistically showed that an increase in protein and decrease of lipid in the diet was associated with a significantly higher specific growth rate (SGR) and reduced feed conversion rate (FCR). Histologic evaluation (visual scoring of lipid storage) and morphometry (area and perimeter) of hepatocytes showed that a high lipid diet resulted in increased lipid storage. The data confirmed that a concomitant decrease of protein with an increase in lipids in the diet led to a massive storage of unused lipid within hepatocytes associated with a reduced growth performance. [ABSTRACT FROM AUTHOR]

The marine sponge Chondrosia reniformis Nardo consists largely of a collagenous tissue, the mesohyl, which confers a cartilaginous consistency on the whole animal. This investigation was prompted by the incidental observation that, despite a paucity of potentially contractile elements in the mesohyl, intact C. reniformis stiffen noticeably when touched. By measuring the deflection under gravity of beam-shaped tissue samples, it was demonstrated that the flexural stiffness of the mesohyl is altered by treatments that influence cellular activities, including [Ca2+] manipulation, inorganic and organic calcium channel-blockers and cell membrane disrupters, and that it is also sensitive to extracts of C. reniformis tissue that have been repeatedly frozen then thawed. Since the membrane disrupters and tissue extracts cause marked stiffening of mesohyl samples, it is hypothesised that cells in the mesohyl store a stiffening factor and that the physiologically controlled release of this factor is responsible for the touch-induced stiffening of intact animals. [ABSTRACT FROM AUTHOR]

The results of study of Pb influence on high temperature magnetic and thermal properties of the chalcogenides (PbySn1-y)2P2S6 are presented. The increasing Pb content shifts phase transition to the ferroelectric state at about 337 K towards lower temperatures while magnetic field till 3 T has no influence on this transition. The measured susceptibility and magnetisation data are discussed. [ABSTRACT FROM AUTHOR]

The striped venus clam (Chamelea gallina) is a relevant economic resource in the Adriatic Sea. This study explored the physiological status of C. gallina at four sites selected along a gradient from high to low incidence of recorded historical mortality events and low to high productivity in the Northwestern Adriatic Sea. Investigations were performed during the marine heatwave in 2022 (from July to November). The optimal temperature range for C. gallina was exceeded in July and September, exacerbating stress conditions and a poor nutritional status, particularly at the low productivity sites. Transcriptional profiles assessed in digestive glands showed that clams from the low productivity sites up-regulated transcripts related to feeding/digestive functions as a possible compensatory mechanism to withstand adverse environmental conditions. Clams from the high productivity sites, that in a previous study showed enrichment of health-promoting microbiome components, displayed a healthier metabolic makeup (IDH up-regulation) and induction of protective antioxidant and immune responses. These features are hallmarks of putative enhanced resilience of the species towards environmental stress. Despite the well-known high sensitivity of C. gallina to environmental variations and its narrow window of acclimatory potential, results highlight that local conditions may influence physiological plasticity of this clam species and shape either positively or negatively its response capabilities to environmental changes. The identification of health-promoting endogenous mechanisms both from the animal (this study) and from its associated microbiome may provide the foundation for developing novel tools and strategies to improve clam health and production in low productivity areas or under adverse environmental conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

CCL17 is produced by conventional dendritic cells (cDCs), signals through CCR4 on regulatory T cells (Tregs), and drives atherosclerosis by suppressing Treg functions through yet undefined mechanisms. Here we show that cDCs from CCL17-deficient mice display a pro-tolerogenic phenotype and transcriptome that is not phenocopied in mice lacking its cognate receptor CCR4. In the plasma of CCL17-deficient mice, CCL3 was the only decreased cytokine/chemokine. We found that CCL17 signaled through CCR8 as an alternate high-affinity receptor, which induced CCL3 expression and suppressed Treg functions in the absence of CCR4. Genetic ablation of CCL3 and CCR8 in CD4 + T cells reduced CCL3 secretion, boosted FoxP3 + Treg numbers, and limited atherosclerosis. Conversely, CCL3 administration exacerbated atherosclerosis and restrained Treg differentiation. In symptomatic versus asymptomatic human carotid atheroma, CCL3 expression was increased, while FoxP3 expression was reduced. Together, we identified a non-canonical chemokine pathway whereby CCL17 interacts with CCR8 to yield a CCL3-dependent suppression of atheroprotective Tregs., Competing Interests: Competing Financial Interests The authors declare no competing financial interests.

Background: The clam Chamelea gallina is an ecologically and economically important marine species in the Northwestern Adriatic Sea, which currently suffers from occasional, and still unexplained, widespread mortality events. In order to provide some glimpses in this direction, this study explores the connections between microbiome variations at the clam-sediment interface and the nutritional status of clams collected at four Italian production sites along the Emilia Romagna coast, with different mortality incidence, higher in the Northern sites and lower in the Southern sites., Results: According to our findings, each production site showed a peculiar microbiome arrangement at the clam-sediment interface, with features that clearly differentiate the Northern and Southern sites, with the latter also being associated with a better nutritional status of the animal. Interestingly, the C. gallina digestive gland microbiome from the Southern sites was enriched in some health-promoting microbiome components, capable of supplying the host with essential nutrients and defensive molecules. Furthermore, in experiments conducted under controlled conditions in aquaria, we provided preliminary evidence of the prebiotic action of sediments from the Southern sites, allowing to boost the acquisition of previously identified health-promoting components of the digestive gland microbiome by clams from the Northern sites., Conclusions: Taken together, our findings may help define innovative microbiome-based management strategies for the preservation of the productivity of C. gallina clams in the Adriatic Sea, through the identification and maintenance of a probiotic niche at the animal-sediment interface., (© 2023. The Author(s).)

The present study was designed to evaluate the effects of dietary levels of bioactive peptides (BPs) derived from salmon processing by-products on the presence and distribution of peptic cells (oxyntopeptic cells, OPs) and enteric endocrine cells (EECs) that contain GHR, NPY and SOM in the gastric mucosa of European seabass and gilthead seabream. In this study, 27 seabass and 27 seabreams were divided into three experimental groups: a control group (CTR) fed a control diet and two groups fed different levels of BP to replace fishmeal: 5% BP (BP5%) and 10% BP (BP10%). The stomach of each fish was sampled and processed for immunohistochemistry. Some SOM, NPY and GHR-IR cells exhibited alternating "open type" and "closed type" EECs morphologies. The BP10% group (16.8 ± 7.5) showed an increase in the number of NPY-IR cells compared to CTR (CTR 8.5 ± 4.8) and BP5% (BP10% vs. CTR p ≤ 0.01; BP10% vs. BP5% p ≤ 0.05) in the seabream gastric mucosa. In addition, in seabream gastric tissue, SOM-IR cells in the BP 10% diet (16.8 ± 3.5) were different from those in CTR (12.5 ± 5) (CTR vs. BP 10% p ≤ 0.05) and BP 5% (12.9 ± 2.5) (BP 5% vs. BP 10% p ≤ 0.01). EEC SOM-IR cells increased at 10% BP (5.3 ± 0.7) compared to 5% BP (4.4 ± 0.8) (5% BP vs. 10% BP p ≤ 0.05) in seabass. The results obtained may provide a good basis for a better understanding of the potential of salmon BPs as feed ingredients for seabass and seabream.

The entire shellfish farming sector is negatively affected by heat waves. Predictive models show that while heat waves are not predicted to exceed 28 °C in the northern Adriatic Sea over the coming decades, their duration will increase to periods of up to 30 days. Knowledge regarding the effects of heat waves on bivalves at physiological and molecular level is still limited. This study attempted to simulate what will happen in the future in Pacific oysters exposed to prolonged heat waves, assessing morphometric and physiological indices, and investigating the expression level of a number of genes, including the chaperone heat shock proteins HSP70, HSP72 and HSP90, and the factor P53. A state of stress in the heat wave-exposed animals was found, with loss of body weight and energy resources: despite showing a higher clearance rate, these animals were unable to absorb the nutrients required to maintain homeostasis, as well as demonstrating an alteration in hemolymphatic AST activity, total calcium and magnesium concentration. mRNA levels of all examined genes increased in response to thermal stress, with long-term overexpression, activating cell stress defense mechanisms and modulating the cycle cell. The results of this study indicate that heat waves affect oyster welfare, with consequences for the productivity of the sector due to the lack of salable products., Competing Interests: Declaration of competing interest The authors confirm they have no known conflicts of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Practicing hot yoga may bring significant psychological benefits, but it is largely unstudied. We examined the effects of hot yoga on multifaceted well-being indicators with 290 healthy yoga-naïve volunteers partaking in a six-week randomized controlled trial. Participants completed questionnaires pre- and post-intervention, and reported their emotional experiences four times per day throughout an experience-sampling study. Results revealed that the hot yoga group ( n = 137) improved their well-being from pre- to post-treatment, comparing to the wait-list control group ( n = 153). These improvements included life satisfaction, general health, mindfulness, peace of mind, and eudaimonic well-being (Δ R 2 ranging from .01 to .08)-but not flourishing, which describes major aspects of social-psychological functioning. Multilevel analyses demonstrated that momentary positive emotional experiences increased significantly throughout the trial in the yoga group only (conditional R 2 = .68), particularly when attending a yoga class (conditional R 2 = .50). Interestingly, this increase in momentary positive emotion explained the improvement in post-intervention mindfulness, peace of mind, and general health by 21%, 31%, and 11%, respectively. Finally, the benefits of hot yoga were more notable in individuals with lower levels of baseline eudaimonic well-being (conditional R 2 = .45), flourishing (conditional R 2 = .61), and mental well-being (conditional R 2 = .65), even after ruling out any possible ceiling effects. To sum up, this study demonstrated multiple psychological benefits of hot yoga and its potential to be an effective positive psychology intervention. Future research-especially considering an active control group-is warranted., Competing Interests: Conflict of Interest: The authors of this article declare no conflict of interest., (Copyright © 2022, Colegio Oficial de la Psicología de Madrid.)

The current work was designed to assess the effect of feed supplemented with essential oils (EOs) on the histological features in sea bass's gastric mucosa. Fish were fed three diets: control diet (CTR), HERBAL MIX ® made with natural EOs (N-EOs), or HERBAL MIX ® made with artificial EOs obtained by synthesis (S-EOs) during a 117-day feeding trial. Thereafter, the oxyntopeptic cells (OPs) and the ghrelin (GHR) and somatostatin (SOM) enteroendocrine cells (EECs) in the gastric mucosa were evaluated. The Na + K + -ATPase antibody was used to label OPs, while, for the EECs, anti-SOM and anti-GHR antibody were used. The highest density of OP immunoreactive (IR) area was in the CTR group (0.66 mm 2 ± 0.1). The OP-IR area was reduced in the N-EO diet group (0.22 mm 2 ± 1; CTR vs. N-EOs, p < 0.005), while in the S-EO diet group (0.39 mm 2 ± 1) a trend was observed. We observed an increase of the number of SOM-IR cells in the N-EO diet (15.6 ± 4.2) compared to that in the CTR (11.8 ± 3.7) (N-EOs vs. CTR; p < 0.05), but not in the S-EOs diet. These observations will provide a basis to advance current knowledge on the anatomy and digestive physiology of this species in relation to pro-heath feeds.

A 71-day study was conducted to explore the effect of increasing dietary levels (0, 250, 500, 1000 mg kg feed -1 ; D0, D250, D500 and D1000, respectively) of a blend of microencapsulated organic acids (OA, specifically citric and sorbic acid) and nature identical compounds (NIC, specifically thymol and vanillin), on growth, intestinal immune parameters and gut microbiota (GM) of European sea bass juveniles reared under normal and subsequently suboptimal environmental conditions (high temperature, 30.0 ± 0.4 °C and low oxygen, 4.6 ± 0.6 mg L -1 ). OA and NIC did not promote growth, feed utilisation and feed intake at the inclusion tested but induced a significantly upregulation of IL-8, IL-10 and TGFβ. GM analyzed by next-generation sequencing showed that OA and NIC were able to exert prebiotic properties stimulating the development of beneficial bacteria taxa such as Lactobacillus, Leuconostoc, and Bacillus sp. Picrust analyses displayed a significant potential functional reconfiguration of GM promoting a decrease in inflammation-promoting and homeostatic functions at increasing OA and NIC administration. For the first time on this species the exposure to suboptimal rearing conditions was able to modify GM structure reducing LAB and increasing Proteobacteria, findings which were consistent with the inflammatory process observed at mRNA level.

Organic acids (OA) and nature-identical compounds (NIC) such as monoterpenes and aldehydes are well-known growth and health promoters in terrestrial livestock while their application for fish production is recent and their mechanisms of action require further study. Hence, this study tested the increasing dietary level (D0, D250, D500, D1000; 0, 250, 500 and 1000 mg kg feed -1 respectively) of a microencapsulated blend containing citric and sorbic acid, thymol and vanillin over 82 days on rainbow trout to assess the effects on growth, feed utilization, intestine cytokine gene expression and gut microbiota (GM). Furthermore, the effects on intestinal cytokine gene expression and GM were also explored after one week at high water temperature (23 °C). OA and NIC improved specific growth rate (SGR) and feed conversion rate (FCR) during the second half (day 40-82) of the feeding trial, while at the end of the trial protein (PER) and lipid efficiency (LER) increased with increasing dietary level. GM diversity and composition and cytokine gene expression analysis showed no significant differences in fish fed with increasing doses of OA and NIC (82 days) demonstrating the absence of inflammatory activity in the intestinal mucosa. Although there were no statistical differences, GM structure showed a tendency in clustering D0 group separately from the other dietary groups and a trend towards reduction of Streptococcus spp. was observed in the D250 and D1000 groups. After exposure to high water temperature, lower GM diversity and increased gene expression of inflammatory intestinal cytokines were observed for both inclusions (D0 vs. D1000) compared to groups in standard condition. However, the gene up-regulation involved a limited number of cytokines showing the absence of a substantial inflammation process able to compromise the functional activity of the intestine. Despite further study should be conducted to fully clarify this mechanism, cytokines up-regulation seems to be concomitant to the reduction of the GM diversity and, particularly, to the reduction of specific lactic acid bacteria such as Leuconostoc. The application of the microencapsulate blend tested can be a useful strategy to improve growth and feed utilization in rainbow trout under normal temperature conditions. According to the results organic acids and nature-identical compounds did not revert the effects triggered by the increased temperature of water., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Intraplaque angiogenesis increases the chance of unstable atherosclerotic plaque rupture and thrombus formation leading to myocardial infarction. Basic Fibroblast Growth Factor (bFGF) plays a key role in angiogenesis and inflammation and is involved in the pathogenesis of atherosclerosis. Therefore, we aim to test K5, a small molecule bFGF-inhibitor, on remodelling of accelerated atherosclerotic vein grafts lesions in ApoE3*Leiden mice. K5-mediated bFGF-signalling blockade strongly decreased intraplaque angiogenesis and intraplaque hemorrhage. Moreover, it reduced macrophage infiltration in the lesions by modulating CCL2 and VCAM1 expression. Therefore, K5 increases plaque stability. To study the isolated effect of K5 on angiogenesis and SMCs-mediated intimal hyperplasia formation, we used an in vivo Matrigel-plug mouse model that reveals the effects on in vivo angiogenesis and femoral artery cuff model to exclusively looks at SMCs. K5 drastically reduced in vivo angiogenesis in the matrigel plug model while no effect on SMCs migration nor proliferation could be seen in the femoral artery cuff model. Moreover, in vitro K5 impaired endothelial cells functions, decreasing migration, proliferation and tube formation. Our data show that K5-mediated bFGF signalling blockade in hypercholesterolemic ApoE3*Leiden mice reduces intraplaque angiogenesis, haemorrhage and inflammation. Therefore, K5 is a promising candidate to stabilize advanced atherosclerotic plaques.

The reproductive performance of Octopus vulgaris broodstocks fed two different diets (mixed fish [F group, BW 1,048.14 g] or mixed crustaceans [C group, BW 998.44 g]) was analyzed using an experimental recirculating aquaculture system consisting of a tank equipped with spawning and incubation chambers. A total of 8 females (F1-4; C1-4), and 8 males (M1-M8) were selected. DI of the C group females was significantly (p < 0.05) higher (3.0 ± 0.29%) than the F group (2.16 ± 0.67%). SGR in C group was significantly higher (1.43 ± 0.12%) than the F group (1.18 ± 0.25%). Egg clusters, number of clusters, number of clusters/kg BW, and total length were more favorable in the C group than the F group. The number of clusters/kg BW of C females was 2.5 times higher than that of F females (78.1 ± 6.5 vs 31.1 ± 13.3). The total eggs number, number of eggs/cm, number of eggs/kg BW in the C group were significantly (p < 0.05) higher compared with the F group; the number of eggs/kg BW and paralarvae/kg BW were 5 times higher in the C group (115,928 ± 12,513 C vs 22,109 ± 7912 F and 114,953 ± 12,591 vs 20,729 ± 7104, respectively). Hatching rate of the C group was significantly (p < 0.05) higher compared to the F group.

Adenosine-to-inosine (A-to-I) editing in the seed sequence of microRNAs can shift the microRNAs' targetomes and thus their function. Using public RNA-sequencing data, we identified 35 vasoactive microRNAs that are A-to-I edited. We quantified A-to-I editing of the primary (pri-)microRNAs in vascular fibroblasts and endothelial cells. Nine pri-microRNAs were indeed edited, and editing consistently increased under ischemia. We determined mature microRNA editing for the highest expressed microRNAs, i.e., miR-376a-3p, miR-376c-3p, miR-381-3p, and miR-411-5p. All four mature microRNAs were edited in their seed sequence. We show that both ADAR1 and ADAR2 (adenosine deaminase acting on RNA 1 and RNA 2) can edit pri-microRNAs in a microRNA-specific manner. MicroRNA editing also increased under ischemia in vivo in a murine hindlimb ischemia model and ex vivo in human veins. For each edited microRNA, we confirmed a shift in targetome. Expression of the edited microRNA targetomes, not the wild-type targetomes, was downregulated under ischemia in vivo. Furthermore, microRNA editing enhanced angiogenesis in vitro and ex vivo. In conclusion, we show that microRNA A-to-I editing is a widespread phenomenon, induced by ischemia. Each editing event results in a novel microRNA with a unique targetome, leading to increased angiogenesis., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

VEGFR2 and VEGF-A play a pivotal role in the process of angiogenesis. VEGFR2 activation is regulated by protein tyrosine phosphatases (PTPs), enzymes that dephosphorylate the receptor and reduce angiogenesis. We aim to study the effect of PTPs blockade using bis(maltolato)oxovanadium(IV) (BMOV) on in vivo wound healing and in vitro angiogenesis. BMOV significantly improves in vivo wound closure by 45% in C57BL/6JRj mice. We found that upon VEGFR2 phosphorylation induced by endogenously produced VEGF-A, the addition of BMOV results in increased cell migration (45%), proliferation (40%) and tube formation (27%) in HUVECs compared to control. In a mouse ex vivo, aortic ring assay BMOV increased the number of sprouts by 3 folds when compared to control. However, BMOV coadministered with exogenous VEGF-A increased ECs migration, proliferation and tube formation by only 41%, 18% and 12% respectively and aortic ring sprouting by only 1-fold. We also found that BMOV enhances VEGFR2 Y951 and p38MAPK phosphorylation, but not ERK1/2. The level of phosphorylation of these residues was the same in the groups treated with BMOV supplemented with exogenous VEGF-A and exogenous VEGF-A only. Our study demonstrates that BMOV is able to enhance wound closure in vivo. Moreover, in the presence of endogenous VEGF-A, BMOV is able to stimulate in vitro angiogenesis by increasing the phosphorylation of VEGFR2 and its downstream proangiogenic enzymes. Importantly, BMOV had a stronger proangiogenic effect compared to its effect in coadministration with exogenous VEGF-A.

Objective: Neutrophil extracellular traps (NET) expose modified antigens for autoantibodies in vasculitis. Little is known about levels and removal pathways of NET in systemic lupus erythematosus (SLE), especially in lupus nephritis (LN). We determined circulating levels and defined NET removal in large subsets of patients with incident SLE (iSLE), some of whom had new-onset nephritis., Methods: Serum levels of NET (ELISA), DNase1/DNase1L3 (ELISA), and DNase activity (functional assay) were determined in 216 patients with iSLE [103 had incident LN (iLN)], in 50 patients with other primary glomerulonephritis, and in healthy controls. Ex vivo NET production by neutrophils purified from a random selection of patients was quantified as elastase/DNA release and by immunofluorescence techniques., Results: Serum NET levels were very high in iSLE/iLN compared to all groups of controls and correlated with anti-dsDNA, C3-C4, and proteinuria; iLN had the highest levels. DNase activity was decreased in iLN compared to SLE (20% had one-half DNase activity) despite similar serum levels of DNase1/DNase1L3. In these cases, pretreatment of serum with protein A restored DNase efficiency; 1 patient was homozygous for a c.289&#95;290delAC variant of DNASE1L3 . Ex vivo NET production by neutrophils purified from LN, SLE, and normal controls was similar in all cases., Conclusion: Patients with iLN have increased circulating NET and reduced DNase activity, the latter being explained by the presence of inhibitory substances in circulation and/or by rare DNase1L3 mutations. Accumulation of NET derives from a multifactorial mechanism, and is associated and may contribute to disease severity in SLE, in particular to renal lesions. (Clinical trial registration: The Zeus study was registered at ClinicalTrials.gov, study number NCT02403115).

Unstable atherosclerotic plaques frequently show plaque angiogenesis which increases the chance of rupture and thrombus formation leading to infarctions. Hypoxia plays a role in angiogenesis and inflammation, two processes involved in the pathogenesis of atherosclerosis. We aim to study the effect of resolution of hypoxia using carbogen gas (95% O 2 , 5% CO 2 ) on the remodeling of vein graft accelerated atherosclerotic lesions in ApoE3*Leiden mice which harbor plaque angiogenesis. Single treatment resulted in a drastic decrease of intraplaque hypoxia, without affecting plaque composition. Daily treatment for three weeks resulted in 34.5% increase in vein graft patency and increased lumen size. However, after three weeks intraplaque hypoxia was comparable to the controls, as were the number of neovessels and the degree of intraplaque hemorrhage. To our surprise we found that three weeks of treatment triggered ROS accumulation and subsequent Hif1a induction, paralleled with a reduction in the macrophage content, pointing to an increase in lesion stability. Similar to what we observed in vivo, in vitro induction of ROS in bone marrow derived macrophages lead to increased Hif1a expression and extensive DNA damage and apoptosis. Our study demonstrates that carbogen treatment did improve vein graft patency and plaque stability and reduced intraplaque macrophage accumulation via ROS mediated DNA damage and apoptosis but failed to have long term effects on hypoxia and intraplaque angiogenesis.

We introduce an improved semiclassical dynamics approach to quantum vibrational spectroscopy. In this method, a harmonic-based phase space sampling is preliminarily driven toward non-harmonic quantization by slowly switching on the actual potential. The new coordinates and momenta serve as initial conditions for the semiclassical dynamics calculation, leading to a substantial decrease in the number of chaotic trajectories to deal with. Applications are presented for model and molecular systems of increasing dimensionality characterized by moderate or high chaoticity. They include a bidimensional Henon-Heiles potential, water, formaldehyde, and methane. The method improves accuracy and precision of semiclassical results and it can be easily interfaced with all pre-existing semiclassical theories.

The authenticity of fish products has become an imperative issue for authorities involved in the protection of consumers against fraudulent practices and market stabilization. The present study aimed to provide a method for authentication of European sea bass ( Dicentrarchus labrax) according to the requirements for seafood labels (Regulation 1379/2013/EU). Data on biometric traits, fatty acid profile, elemental composition, and isotopic abundance of wild and reared (intensively, semi-intensively, and extensively) specimens from 18 southern European sources ( n = 160) were collected, clustered in six sets of parameters, and then subjected to multivariate analysis. Correct allocations of subjects according to their production method, origin, and stocking density were demonstrated with good approximation rates (94, 92, and 92%, respectively) using fatty acid profiles. Less satisfying results were obtained using isotopic abundance, biometric traits, and elemental composition. The multivariate analysis also revealed that extensively reared subjects cannot be analytically discriminated from wild subjects.

Rationale: Adenosine-to-inosine editing of microRNAs has the potential to cause a shift in target site selection. 2'-O-ribose-methylation of adenosine residues, however, has been shown to inhibit adenosine-to-inosine editing., Objective: To investigate whether angiomiR miR487b is subject to adenosine-to-inosine editing or 2'-O-ribose-methylation during neovascularization., Methods and Results: Complementary DNA was prepared from C57BL/6-mice subjected to hindlimb ischemia. Using Sanger sequencing and endonuclease digestion, we identified and validated adenosine-to-inosine editing of the miR487b seed sequence. In the gastrocnemius muscle, pri-miR487b editing increased from 6.7±0.4% before to 11.7±1.6% ( P =0.02) 1 day after ischemia. Edited pri-miR487b is processed into a novel microRNA, edited miR487b, which is also upregulated after ischemia. We confirmed editing of miR487b in multiple human primary vascular cell types. Short interfering RNA-mediated knockdown demonstrated that editing is adenosine deaminase acting on RNA 1 and 2 dependent. Using reverse-transcription at low dNTP concentrations followed by quantitative-PCR, we found that the same adenosine residue is methylated in mice and human primary cells. In the murine gastrocnemius, the estimated methylation fraction increased from 32.8±14% before to 53.6±12% 1 day after ischemia. Short interfering RNA knockdown confirmed that methylation is fibrillarin dependent. Although we could not confirm that methylation directly inhibits editing, we do show that adenosine deaminase acting on RNA 1 and 2 and fibrillarin negatively influence each other's expression. Using multiple luciferase reporter gene assays, we could demonstrate that editing results in a complete switch of target site selection. In human primary cells, we confirmed the shift in miR487b targeting after editing, resulting in a edited miR487b targetome that is enriched for multiple proangiogenic pathways. Furthermore, overexpression of edited miR487b, but not wild-type miR487b, stimulates angiogenesis in both in vitro and ex vivo assays., Conclusions: MiR487b is edited in the seed sequence in mice and humans, resulting in a novel, proangiogenic microRNA with a unique targetome. The rate of miR487b editing, as well as 2'-O-ribose-methylation, is increased in murine muscle tissue during postischemic neovascularization. Our findings suggest miR487b editing plays an intricate role in postischemic neovascularization., (© 2017 American Heart Association, Inc.)

Acute cardiovascular events, due to rupture or erosion of an atherosclerotic plaque, represent the major cause of morbidity and mortality in patients. Growing evidence suggests that plaque neovascularization is an important contributor to plaque growth and instability. The vessels' immaturity, with profound structural and functional abnormalities, leads to recurrent intraplaque hemorrhage. This review discusses new insights of atherosclerotic neovascularization, including the effects of leaky neovessels on intraplaque hemorrhage, both in experimental models and humans. Furthermore, modalities for in vivo imaging and therapeutic interventions to target plaque angiogenesis will be discussed., (Copyright © 2017. Published by Elsevier B.V.)

Background and Aims: We aimed at investigating the role of 14q32 microRNAs in intimal hyperplasia and accelerated atherosclerosis; two major contributors to restenosis. Restenosis occurs regularly in patients treated for coronary artery disease and peripheral arterial disease. We have previously shown that inhibition of 14q32 microRNAs leads to increased post-ischemic neovascularization, and microRNA miR-494 also decreased atherosclerosis, while increasing plaque stability. We hypothesized that 14q32 microRNA inhibition has beneficial effects on intimal hyperplasia, as well as accelerated atherosclerosis., Methods: Non-constrictive cuffs were placed around both femoral arteries of C57BL/6J mice to induce intimal hyperplasia. Accelerated atherosclerotic plaque formation was induced in hypercholesterolemic ApoE -/- mice by placing semi-constrictive collars around both carotid arteries. 14q32 microRNAs miR-329, miR-494 and miR-495 were inhibited in vivo using Gene Silencing Oligonucleotides (GSOs)., Results: GSO-495 administration led to a 32% reduction of intimal hyperplasia. Moreover, the number of macrophages in the arterial wall of mice treated with GSO-495 was reduced by 55%. Inhibition of miR-329 and miR-494 had less profound effects on intimal hyperplasia. GSO-495 administration also decreased atherosclerotic plaque formation by 52% and plaques of GSO-495 treated animals showed a more stable phenotype. Finally, cholesterol levels were also decreased in GSO-495 treated animals, via reduction of the VLDL-fraction., Conclusions: GSO-495 administration decreased our primary outcomes, namely intimal hyperplasia, and accelerated atherosclerosis. GSO-495 administration also favourably affected multiple secondary outcomes, including macrophage influx, plaque stability and total plasma cholesterol levels. We conclude that 14q32 microRNA miR-495 is a promising target for prevention of restenosis., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

Fish are free-living organisms since initial stages of development and are exposed to numerous pathogens before their lymphoid organs have matured and adaptive immunity has developed. Susceptibility to diseases and juvenile mortality represent key critical factors for aquaculture. In this context, the characterization of the appearance kinetics of the immune system key members will be useful in understanding the ability of a particular species in generating immune protection against invading pathogens at different developmental stages. The present study characterized, for the first time, the transcriptional onset of un-explored relevant genes of both innate and adaptive immune system during the Solea solea ontogenesis. Gene expression profiles of immune relevant genes was investigated, by means of DNA microarray, in ten developmental stages, from hatching (1 day post-hatching, dph) to accomplishment of the juvenile form (33 dph). The obtained results revealed that transcripts encoding relevant members of innate immune repertoire, such as lysozyme, AMPs (hepcidin, β-defensin), PPRs and complement components are generally characterized by high expression levels at first stages (i.e. hatch and first feeding) indicating protection from environmental pathogens even at early development. Transcription of adaptive immune genes (i.e. Class I and class II MHC, TCRs) differs from that of the innate immune system. Their onset coincides with metamorphosis and larvae-to-juvenile transition, and likely overlaps with the appearance and maturation of the main lymphoid organs. Finally, data collected suggest that at the end of metamorphosis S. solea cell-mediated immune system hasn't still undergone full maturation., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This study evaluated the therapeutic efficacy of laser therapy in treating oral human papilloma virus (HPV) lesions. In particular, mode of action, healing, postoperative patient compliance, visual numeric scale (VNS) pain index, and recurrence were analyzed. During 2001-2012, in 170 patients (80 women and 90 men), 174 intraoral and lip HPV lesions were detected and excised by diode laser of different wavelengths (810-980 nm), with an average power of 2.1 W, in continuous wave mode, using 300 to 320 μm optical fibers. In most cases (95.4%), complete healing occurred in the first 30 days. There were no adverse effects and all patients were carefully followed up until complete healing occurred, documenting any complications. There was only one recurrence, which was later treated successfully; the mean VNS pain score was below one. In treating HPV lesions, the diode laser is not only a valuable tool for their eradication but especially it reduces relapses, thanks to the characteristics of the laser light.

Among echinoderms, crinoids are well known for their remarkable regenerative potential. Regeneration depends mainly on progenitor cells (undifferentiated or differentiated), which migrate and proliferate in the lesion site. The crucial role of the "progenitor" elements involved in the regenerative processes, in terms of cell recruitment, sources, and fate, is a central problem in view of its topical interest and biological implications. The spectacular regenerative potential of crinoids is used to replace lost internal and external organs. In particular, the process of arm regeneration in the feather star Antedon mediterranea is the regeneration model most extensively explored to date. We have addressed the morphological and functional characterization of the cell phenotypes responsible for the arm regenerative processes by using an in vitro approach. This represents the first successful attempt to culture cells involved in crinoid regeneration. A comparison of these results with others from previous in vivo investigations confirms the diverse cell types contributing to regeneration and underscores their involvement in migration, proliferation, and dedifferentiation processes.