Avocado Paste Phenolics Mitigate a High-Fat Diet-Induced Plasma HDL Decrease in Male Wistar Rats, by Altering the mRNA Expression of Hepatic SCARB1.

Avocado paste (AP) is the main industrial byproduct of its processing, and retains various phenolic compounds (PCs). PCs are known to normalize the plasma lipid profile, but those from avocado byproducts have been minimally studied. We report the normalizing effects of an AP-derived phenolic extract (PE) on the plasma lipid profile of male Wistar rats. A standard (SD) and high-fat diet (HFD) were formulated, and the same diets were supplemented with 1 g/kg of diet of PE (SD + PE and HFD + PE). Rats were fed these diets during an 8-week period. The HFD induced signs of dyslipidemia, but PE treatment countered the decrease in HDL. Relative mRNA expression (real-time PCR) of the hepatic HDL receptor (SCARB1) increased in both groups (SD + PE and HFD + PE), while the LDR receptor (LDLR) increased in SD + PE group. The mRNA expression of apolipoproteins APOA1 and APOB was unaffected. We conclude that PCs from AP can counter a diet-induced decrease in plasma HDL by acting on the mRNA expression of its hepatic receptor.

Daytime-Restricted Feeding Ameliorates Oxidative Stress by Increasing NRF2 Transcriptional Factor in the Rat Hippocampus in the Pilocarpine-Induced Acute Seizure Model.

Seizure-mediated oxidative stress is a crucial mechanism in the pathophysiology of epilepsy. This study evaluated the antioxidant effects of daytime-restricted feeding (DRF) and the role of the Nrf2 signaling pathway in a lithium-pilocarpine model seizure model that induces status epilepticus (SE). We performed a lipoperoxidation assay and dihydroethidium fluorescence to measure oxidative stress markers in the hippocampus (malondialdehyde and reactive oxygen species). The protein content of Nrf2 and its downstream protein SOD2 was evaluated using Western blotting. The cellular distribution of the Nrf2 and SOD2 proteins in the pyramidal cell layer of both the CA1 and CA3 hippocampal subfields and astrocytes (GFAP marker) were quantified using immunofluorescence and immunohistochemistry, respectively. Our results indicate that DRF reduced the malondialdehyde levels and the production of reactive oxygen species. Furthermore, a significant increase in Nrf2 and SOD2 protein content was observed in animals subjected to restrictive diet. In addition, DRF increased the relative intensity of the Nrf2 fluorescence in the perinuclear and nuclear compartments of pyramidal neurons in the CA1 subfield. Nrf2 immunoreactivity and the astrocyte marker GFAP also increased their colocalization under DRF conditions. Additionally, SOD2 immunoreactivity was increased in CA1 pyramidal neurons but not in the CA3 region. Our findings suggest that DRF partially prevents oxidative stress by increasing the Nrf2 transcriptional factor and the SOD2 enzyme during the development of SE.

N-(levodopa) chitosan derivative based on click chemistry shows biological functionality in brain cells.

OBJECTIVE: To develop N-(levodopa) chitosan derivatives through click chemistry to study their effect in brain cells.Significance: This study presents a proof-of-concept that macromolecules such as N-(Levodopa) chitosan derivatives traverse brain cell membranes and induce biomedical functionalities. METHODS: Through click chemistry, we developed N-(levodopa) chitosan derivatives. They were physically and chemically characterized by FT-IR, 1H-NMR, TGA and Dynamic Light Scattering analyses. Solution and nanoparticles of N-(levodopa) chitosan derivatives were tested in primary cell cultures from the postnatal rat olfactory bulb, substantia nigra and corpus callosum. Ca2+ imaging and UPLC experiments were used to investigate if the biomaterial modulated the brain cell physiology. RESULTS: N-(levodopa) chitosan derivatives induced intracellular Ca2+ responses in primary cell cultures of the rat brain. UPLC experiments indicated that levodopa attached to chitosan was converted into dopamine by brain cells. CONCLUSION: The present study shows that N-(levodopa) chitosan may be useful to develop new treatment strategies, which could serve as molecular reservoirs of biomedical drugs to treat degenerative disorders of the nervous system.

Phenolic compounds can induce systemic and central immunomodulation, which result in a neuroprotective effect.

Inflammation may negatively impact health, particularly that of the central nervous system. Phenolic compounds are bioactive molecules present in fruits and vegetables with potential anti-inflammatory effects. The purpose of the present work is to review the immunomodulatory bioactivities of phenolic compounds in the periphery and in the central nervous system. Results show that various types of phenolics are able to counter diet- or pathogen-induced systemic inflammation (among others) in various models. In vitro data show significant effects of flavonoids and phenolic acids in particular; similar bioactivities were reported in vivo, when administering them as pure compounds or from fruit and vegetable extracts that contain them. In the central nervous system, phenolics counter chronic inflammation and aggressive acute inflammatory processes, such as ischemic events, when administered preemptively and even therapeutically. We therefore conclude that the immunomodulatory potential of phenolic compounds can maintain an adequate immune response; their regular consumption should therefore be prioritized in order to maintain health. PRACTICAL APPLICATIONS: The immune response must be carefully regulated in order to avoid its deleterious effects. The present work highlights how phenolic compounds, dietary components ubiquitous in everyday diet, are able to maintain it within an adequate range. As humans are exposed to more proinflammatory stimuli (inadequate dietary pattern, mental stress, environmental pollution, chronic diseases, etc.), it becomes necessary to counter them, and consuming adequate amounts of foods that contain compounds with this ability is a rather simple strategy. Thus, the present work highlights how fruits and vegetables can help to maintain an adequate immune response that can preserve systemic health and that of the central nervous system. Furthermore, specific compounds contained in them can also be ideal candidates for additional in-depth studies, which can potentially lead to the development of potent, targeted, and safe anti-inflammatory molecules.

Mango "Ataulfo" Peel Extract Improves Metabolic Dysregulation in Prediabetic Wistar Rats.

The hypoglycemic effect of functional phytochemicals has been evaluated in diabetic rodents but scarcely in its premorbid condition (prediabetes; PD). This study aimed to evaluate a mango (cv. Ataulfo) peel hydroethanolic (20:80) extract (MPE) for in vivo glycemic/lipidemic-normalizing effect and in vitro enzyme inhibitory (α-amylase/α-glucosidase) activity. The polyphenolic MPE (138 mg EAG.g−1, mainly gallic acid and mangiferin) with antioxidant capacity (DPPH• 34 mgTE.g−1) was fed to PD rats (induction: high-fat diet (60% energy) + single dose streptozotocin (35 mg·kg−1), 4 weeks). At the 8th week, fasting glycemia (FG), oral glucose tolerance test, and insulin sensitivity indexes (HOMA-IR, HOMA-ß) > blood lipid-normalizing effect were documented as healthy controls > MPE > disease (PD) controls, which was possibly related to the extract's concentration−response in vitro enzyme inhibitory activity (IC50 ≈ 0.085 mg·mL−1). MPE is a rich source of glucose-lowering phytochemicals for the primary prevention of type 2 diabetes.

Sub-chronic consumption of a phenolic-rich avocado paste extract induces GLP-1-, leptin-, and adiponectin-mediated satiety in Wistar rats.

Avocado paste (AP) is a phenolic-rich byproduct of avocado oil extraction. The effects of sub-chronic consumption of diets supplemented with an AP phenolic extract (PE) were analyzed. A standard diet (SD), high-fat diet (HFD), and these supplemented with PE (SD + PE and HFD + PE) were used. Significantly increased satiety was observed in PE-supplemented groups, according to less food consumption (-15% in SD + PE vs. SD, and -11% in HFD + PE vs. HFD), without changes in weight gain or percentage of adipose tissue. PE-supplemented groups had an increased plasma concentration ( + 16% in SD + PE vs. SD, and +26% in HFD + PE vs. HFD) and relative mRNA expression (+74% in SD + PE vs. SD, and +46% in HFD + PE vs. HFD) of GLP-1; an increase in plasma leptin and adiponectin was independent of their mRNA expression. Our results suggest that AP-derived PE exerts a satiety effect in vivo, possibly mediated by GLP-1, leptin, and adiponectin. PRACTICAL APPLICATIONS: Minimizing food waste is a top priority in most of the world, thus, researchers seek methods to reintroduce industrial fruit and vegetable byproducts into the food processing chain. The present work highlights the potential of avocado byproducts as sources of bioactive phenolic compounds, whose sub-chronic consumption (8 weeks) exerts a satiety action in vivo. Avocado farming is resource-intensive, making it of relevance to producers and processing industries to avoid discarding its byproducts as much as possible.

Phenolic compounds that cross the blood-brain barrier exert positive health effects as central nervous system antioxidants.

The blood-brain barrier (BBB) is a physical structure whose main function is to strictly regulate access to circulating compounds into the central nervous system (CNS). Vegetable-derived phenolic compounds have been widely studied, with numerous epidemiologic and interventional studies confirming their health-related bioactivities across multiple cells, organs and models. Phenolics are non-essential xenobiotics, and should theoretically be unable to cross the BBB. The present work summarizes current experimental evidence that reveals that not only are phenolic compounds able to cross the BBB and bioaccumulate in the brain, but there is some stereoselectivity, which suggests the presence of specific transporters that allow them to reach the brain. Some molecules cross the BBB intact, while others do so only after being biotransformed or metabolized elsewhere. Once inside the CNS, they prevent or counter oxidative stress, which maintains the molecular, cellular, structural and functional integrity of the brain, and subsequently, overall human health.

Influence of Dietary Lipids and Environmental Salinity on the n-3 Long-Chain Polyunsaturated Fatty Acids Biosynthesis Capacity of the Marine Teleost Solea senegalensis.

Fish vary in their ability to biosynthesise long-chain polyunsaturated fatty acids (LC-PUFA) depending upon the complement and function of key enzymes commonly known as fatty acyl desaturases and elongases. It has been reported in Solea senegalensis the existence of a Δ4 desaturase, enabling the biosynthesis of docosahexaenoic acid (DHA) from eicosapentaenoic acid (EPA), which can be modulated by the diet. The present study aims to evaluate the combined effects of the partial replacement of fish oil (FO) with vegetable oils and reduced environmental salinity in the fatty acid composition of relevant body compartments (muscle, hepatocytes and enterocytes), the enzymatic activity over α-linolenic acid (ALA) to form n-3 LC-PUFA through the incubation of isolated hepatocytes and enterocytes with [1-14C] 18:3 n-3, and the regulation of the S. senegalensis fads2 and elovl5 in the liver and intestine. The presence of radiolabelled products, including 18:4n-3, 20:4n-3 and EPA, provided compelling evidence that a complete pathway enabling the biosynthesis of EPA from ALA, establishing S. senegalensis, has at least one Fads2 with ∆6 activity. Dietary composition prevailed over salinity in regulating the expression of fads2, while salinity did so over dietary composition for elovl5. FO replacement enhanced the proportion of DHA in S. senegalensis muscle and the combination with 20 ppt salinity increased the amount of n-3 LC-PUFA in hepatocytes.

Angiotensin II, ATP and high extracellular potassium induced intracellular calcium responses in primary rat brain endothelial cell cultures.

The meninges shield the nervous system from diverse, rather harmful stimuli and pathogens from the periphery. This tissue is composed of brain endothelial cells (BECs) that express diverse ion channels and chemical-transmitter receptors also expressed by neurons and glial cells to communicate with each other. However, information about the effects of ATP and angiotensin II on BECs is scarce, despite their essential roles in blood physiology. This work investigated in vitro if BECs from the meninges from rat forebrain respond to ATP, angiotensin II and high extracellular potassium, with intracellular calcium mobilizations and its second messenger-associated pathways. We found that in primary BEC cultures, both ATP and angiotensin II produced intracellular calcium responses linked to the activation of inositol trisphosphate receptors and ryanodine receptors, which led to calcium release from intracellular stores. We also used RT-PCR to explore what potassium channel subunits are expressed by primary BEC cultures and freshly isolated meningeal tissue, and which might be linked to the observed effects. We found that BECs mainly expressed the inward rectifier potassium channel subunits Kir1.1, Kir3.3, Kir 4.1 and Kir6.2. This study contributes to the understanding of the functions elicited by ATP and angiotensin II in BECs from rat meninges. SIGNIFICANCE OF THE STUDY: Brain endothelial cells (BECs) express diverse ion channels and membrane receptors, which they might use to communicate with neurons and glia. This work investigated in vitro, if BECs from the rat forebrain respond to angiotensin II and ATP with intracellular calcium mobilizations. We found that these cells did respond to said substances with intracellular calcium mobilizations linked to inositol trisphosphate and ryanodine receptor activation, which led to calcium release from intracellular stores. These findings are important because they might uncover routes of active communication between brain cells and endothelial cells.

Effects of Dietary Phenylalanine and Tyrosine Supplements on the Chronic Stress Response in the Seabream (Sparus aurata).

The increase of aquaculture production is associated with a growing interest in improving physiological status and welfare in fish. For this reason, the search for strategies for mitigating stress has been intensified, with one of these strategies being food supplementation with different amino acids (AA). The objective of this study was to evaluate the effects of dietary phenylalanine (Phe) and tyrosine (Tyr) supplements on the endocrine and physiological state of seabreams (Sparus aurata) subjected to chronic stress. The fish were stocked at 30 fish/tank in a recirculation aquatic system, fed one control diet and two diets supplemented with 5% Phe or Tyr for 90 days. Blood was drawn from 10 fish per tank every 30 days, and the weight and length were measured every 15 days. At the end of the experiment, length/weight of the fish were measured, and they were sacrificed for the extraction of blood, head kidney, liver, and brain. Classic plasma stress markers (glucose, lactate, proteins, and cortisol), as well as hormones derived from Phe and Tyr (adrenaline, norepinephrine, and dopamine) and the accumulation of AA were analyzed. Fish fed with diets supplemented with Phe or Tyr showed a reduction in various stress markers and physiological parameters. In addition, the stress condition favored a mobilization of AA toward the tissues, especially in supplemented diets, so this excess of AA could be used as an energy substrate to cope with stress.

Neuroprotective effects of mango cv. 'Ataulfo' peel and pulp against oxidative stress in streptozotocin-induced diabetic rats.

BACKGROUND: Oxidative stress has been implicated in the pathogenesis and progression of diabetes mellitus. Both can damage the brain. Mango and its by-products are sources of bioactive compounds with antioxidant properties. We hypothesized that mango cv. 'Ataulfo' peel and pulp mitigate oxidative stress in the brain of streptozotocin-induced diabetic rats. RESULTS: Twenty-four male Wistar rats were divided into four groups: control, untreated diabetic (UD), diabetic treated with a mango-supplemented diet (MTD), and diabetic pretreated with a mango-supplemented diet (MPD). The rats were fed the different diets for 4 weeks after diabetes induction (MTD), or 2 weeks before and 4 weeks after induction (MPD). After the intervention, serum and brain (cerebellum and cortex) were collected to evaluate gene expression, enzyme activity, and redox biomarkers. Superoxide dismutase 2 (SOD2) expression increased in the cortex of the MTD group, whereas glutathione-S-transferase p1 (GSTp1) expression was higher in the cortex of the MTD group, and cortex and cerebellum of the MPD group. SOD1 activity was higher in the cerebellum and cortex of all diabetic groups, whereas GST activity increased in the cerebellum and cortex of the MPD group. Lipid peroxidation increased in the cerebellum and cortex of the UD group; however, a mango-supplemented diet prevented this increase in both regions, while also mitigating polyphagia and weight loss, and maintaining stable glycemia in diabetic rats. CONCLUSION: We propose that mango exerts potent neuroprotective properties against diabetes-induced oxidative stress. It can be an alternative to prevent and treat biochemical alterations caused by diabetes. © 2020 Society of Chemical Industry.

Phenolic Compounds Promote Diversity of Gut Microbiota and Maintain Colonic Health.

The role of non-energy-yielding nutrients on health has been meticulously studied, and the evidence shows that a compound can exert significant effects on health even if not strictly required by the organism. Phenolic compounds are among the most widely studied molecules that fit this description; they are found in plants as secondary metabolites and are not required by humans for growth or development, but they can influence a wide array of processes that modulate health across multiple organs and systems. The lower gastrointestinal tract is a prime site of action of phenolic compounds, namely, by their effects on gut microbiota and colonic health. As with humans, phenolic compounds are not required by most bacteria but can be substrates of others; in fact, some phenolic compounds exert antibacterial actions. A diet rich in phenolic compounds can lead to qualitative and quantitative effects on gut microbiota, thereby inducing indirect health effects in mammals through the action of these microorganisms. Moreover, phenolic compounds may be fermented by the gut microbiota, thereby modulating the compounds bioactivity. In the colon, phenolic compounds promote anti-inflammatory, anti-oxidant and antiproliferative actions. The aim of the present review is to highlight the role of phenolic compounds on maintaining or restoring a healthy microbiota and overall colonic health. Mechanisms of action that substantiate the reported evidence will also be discussed.

Phenylalanine and Tyrosine as Feed Additives for Reducing Stress and Enhancing Welfare in Gilthead Seabream and Meagre.

Increased aquaculture production is associated with a growing interest in improving fish welfare. For this reason, the search for strategies to mitigate stress has intensified, one of these strategies being food supplementation with amino acids. The objective of this study was to evaluate the effects of dietary phenylalanine (Phe) and Tyrosine (Tyr) on the stress response and metabolism of juvenile gilthead seabreams (Sparus aurata) and meagres (Argyrosomus regius). Fish batches were fed a control diet and two diets supplemented with 5% Phe or Tyr for seven days. At the end of the experiment fish were stressed by air exposure for 3 min and then sacrificed for the extraction of blood and brain. Classical plasma stress markers were analyzed (glucose, lactate, proteins, cortisol), as well as hormones derived from those amino acids (adrenaline, noradrenaline and dopamine). Despite interspecific differences, fish fed the diets supplemented with Phe or Tyr showed a reduction on several stress markers. However, interspecific differences were detected for many indicators. Concretely, hormonal stress markers were significantly attenuated in meagres fed the enriched diets. Moreover, the stress condition favored a mobilization of amino acids towards the brain, especially in supplemented diets, hence this amino acid excess could be used as an energy substrate to cope with stress.

The Use of Dietary Additives in Fish Stress Mitigation: Comparative Endocrine and Physiological Responses.

In the last years, studies on stress attenuation in fish have progressively grown. This is mainly due to the interest of institutions, producers, aquarists and consumers in improving the welfare of farmed fish. In addition to the development of new technologies to improve environmental conditions of cultured fish, the inclusion of beneficial additives in the daily meal in order to mitigate the stress response to typical stressors (netting, overcrowding, handling, etc.) has been an important research topic. Fish are a highly diverse paraphyletic group (over 27,000 species) though teleost infraclass include around 96% of fish species. Since those species are distributed world-wide, a high number of different habitats and vital requirements exist, including a wide range of environmental conditions determining specifically the stress response. Although the generalized endocrine response to stress (based on the release of catecholamines and corticosteroids) is detectable and therefore provides essential information, a high diversity of physiological effects have been described depending on species. Moreover, recent omics techniques have provided a powerful tool for detecting specific differences regarding the stress response. For instance, for transcriptomic approaches, the gene expression of neuropeptides and other proteins acting as hormonal precursors during stress has been assessed in some fish species. The use of different additives in fish diets to mitigate stress responses has been deeply studied. Besides the species factor, the additive type also plays a pivotal role in the differentiation of the stress response. In the literature, several types of feed supplements in different species have been assayed, deriving in a series of physiological responses which have not focused exclusively on the stress system. Immunological, nutritional and metabolic changes have been reported in these experiments, always associated to endocrine processes. The biochemical nature and physiological functionality of those feed additives strongly affect the stress response and, in fact, these can act as neurotransmitters or hormone precursors, energy substrates, cofactors and other essential elements, implying multi-systematic and multi-organic responses. In this review, the different physiological responses among fish species fed stress-attenuating diets based on biomolecules and minerals have been assessed, focusing on the endocrine regulation and its physiological effects.

Metabolic and Stress Responses in Senegalese Soles (Solea senegalensis Kaup) Fed Tryptophan Supplements: Effects of Concentration and Feeding Period.

: The objective of this study was to assess the impact of different dietary Trp concentrations on the stress and metabolism response of juvenile Senegalese soles (Solea senegalensis). Fish (38.1 ± 1.9 g) were fed different Trp-enriched feeds (0%, 1% and 2% Trp added) for two and eight days, and later exposed to air stress for three min. Samples were taken pre- and 1 h post-stress (condition). Plasma cortisol, lactate, glucose and proteins were significantly affected by the sampling time, showing higher values at 1 h post-stress. Trp concentration in food also had significant effects on lactate and glucose levels. However, the feeding period did not affect these parameters. Post-stress values were higher than in the pre-stress condition for every plasma parameter, except for lactate in two days and 1% Trp treatment. Nevertheless, cortisol, glucose and lactate did not vary significantly between pre- and post-stress samplings in fish fed the 1% Trp-enriched diet for two days. The lack of variability in cortisol response was also due to the high pre-stress value, significantly superior to pre-stress control. The exposure time to Trp feeding did not significantly affect any enzyme activity; however, Trp added and condition influenced protein-related enzyme activities. In spite of decreasing stress markers, Trp-enriched diets altered the protein metabolism.

Stressors Due to Handling Impair Gut Immunity in Meagre (Argyrosomus regius): The Compensatory Role of Dietary L-Tryptophan.

In the context of intensive aquaculture, meagre (Argyrosomus regius) is one of the most important new aquaculture species in Southern Europe and several studies are focused on the optimization of its culture. Nevertheless, stressors such as handling during transport or culture maintenance may affect the immune system, thereby impairing some immune responses or provoking cellular damage. One strategy that has been used to avert this type of negative stress response is the supplementation of amino acids to improve resistance to stress. In this experiment, meagre (105.0 ± 2.6 g, mean ± standard deviation) juveniles were fed two diets for a period of 7 days, the first a commercial diet supplemented with 1% tryptophan (Trp) and second, the same commercial diet without tryptophan supplementation (control group). The effects of two types of handling stressors (air exposure and confinement/netting) on fish fed both diets was evaluated in terms of gene expression of the selected gut immunity markers, such as (1) innate immune response processes: c3 complement (c3), lysozyme (lys), and cyclooxygenase (cox2); (2) humoral immune response processes: interferon type 1 (ifn1), mx protein (mxp), interleukin 1b (il-1b), tumor necrosis factor 1a (tnf1a), and interleukin 10 (il-10); (3) antimicrobial peptides: defensin (def), hepcidin (hep), piscidin (pis), and a marker for mitochondrial respiration: glyceraldehyde 3-phosphate dehydrogenase (gapdh). Samples of the anterior intestine were collected at 1 and 6 h post-stress (hps). Results showed that in fish fed 1% Trp, the air exposure resulted in an upregulation of gene expression at 6 hps for c3, lys, cox2, ifn1, mxp, il-10 and gapdh, and il-1b and pis. The confinement/netting test for fish fed 1% Trp resulted in an upregulation of c3 and mxp and a downregulation of cox2, ifn1, il-1b, tnf1a, il-10, def, hep, and gapdh at both post-stress times (1 and 6 hps). According to the present study, dietary supplementation with 1% Trp may be considered as a proper nutritional strategy for improving tolerance and/or alleviating acute response to handling stressors.

Stress effects of amyloodiniosis in gilthead sea bream Sparus aurata.

Amyloodiniosis is a parasitological disease caused by one of the most common and important parasitic dinoflagellates in fish, Amyloodinium ocellatum (Brown), that represents a major bottleneck for gilthead seabream semi-intensive aquaculture in Southern Europe. In this experiment, we analyzed some metabolic, osmoregulatory and stress indicators to elucidate some of the physiological responses of gilthead sea bream when exposed to an A. ocellatum outbreak. We observed significant differences between Control and Infection groups in the cortisol, lactate and gill Na+/K+-ATPase (NKA) activity levels but that glucose, osmolarity, pH and total protein did not present such differences. This could indicate that the presence of the parasite induced a stress response, possibly enhancing the metabolization of glucose and subsequently lactate to cope with the higher energy requirements of the organism. There was also a decrease in gill NKA activity possibly due to severe epithelial damage and increased mucus production caused by the parasite A. ocellatum, which could induce anoxia and osmoregulatory impairment in the organism. However, further works must be performed to fully understand the physiological reactions of fish for A. ocellatum outbreaks.

Chitosan Derivatives: Introducing New Functionalities with a Controlled Molecular Architecture for Innovative Materials.

The functionalization of polymeric substances is of great interest for the development of innovative materials for advanced applications. For many decades, the functionalization of chitosan has been a convenient way to improve its properties with the aim of preparing new materials with specialized characteristics. In the present review, we summarize the latest methods for the modification and derivatization of chitin and chitosan under experimental conditions, which allow a control over the macromolecular architecture. This is because an understanding of the interdependence between chemical structure and properties is an important condition for proposing innovative materials. New advances in methods and strategies of functionalization such as the click chemistry approach, grafting onto copolymerization, coupling with cyclodextrins, and reactions in ionic liquids are discussed.

Skin mucus metabolites in response to physiological challenges: A valuable non-invasive method to study teleost marine species.

Knowledge concerning the health and welfare of fish is important to conserve species diversity. Fish mucosal surfaces, and particularly the skin, are of utmost importance to protect the integrity and homeostasis of the body and to prevent skin infections by pathogens. We performed three trials simulating different environmental and anthropogenic challenges: fish capture (air exposure), bacterial infection and fasting, with the aim of evaluating epidermal mucus as a non-invasive target of studies in fish. In this initial approach, we selected three well-known marine species: meagre (Argyrosomus regius), European sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) for our study. Mucus viscosity was measured in order to determine its rheological properties, and mucus metabolite (glucose, lactate, protein and cortisol) levels were analysed to establish their suitability as potential biomarkers. Skin mucus appeared as a viscous fluid exhibiting clearly non-Newtonian behaviour, with its viscosity being dependent on shear rate. The highest viscosity (p < 0.05) was observed in sea bream. Mucus metabolites composition responded to the different challenges. In particular, glucose increased significantly due to the air exposure challenge in meagre; and it decreased during food deprivation in sea bream by a half (p < 0.05). In contrast, mucus protein only decreased significantly after pathogenic bacterial infection in sea bass. In addition, mucus lactate immediately reflected changes closely related to an anaerobic condition; whereas cortisol was only modified by air exposure, doubling its mucus concentration (p < 0.05). The data provided herein demonstrate that mucus metabolites can be considered as good non-invasive biomarkers for evaluating fish physiological responses; with the glucose/protein ratio being the most valuable and reliable parameter. Determining these skin mucus metabolites and ratios will be very useful when studying the condition of critically threatened species whose conservation status prohibits the killing of specimens.

Effects of amino acid supplementations on metabolic and physiological parameters in Atlantic cod (Gadus morhua) under stress.

The effects of tryptophan (Trp) and phenylalanine (Phe) diet supplementation on the stress and metabolism of the Atlantic cod have been studied. Fish were fed diet supplemented with Trp or Phe or control diet for 1 week. At the end of the feeding trial, fish were subjected to air exposure or heat shock. Following samples of blood, liver and muscle were taken from the fish and were analyzed for stress and metabolic indicators. After an air exposure, plasma cortisol levels in fish fed with Trp and Phe diets were lower compared to the fish fed the control diet. Diets containing both amino acids increased significantly the liver transaminase activities in juvenile cod. During thermal stress, high Trp contents had significant effects on fructose biphosphatase activity though Phe did not. Overall, activities of glucose 6-phosphate dehydrogenase, pyruvate kinase, and phosphofructokinase increased significantly for both amino acid diets. For the thermal stress, fish had the highest values of those activities for the 3Trp diet. Trp content in the diet had significant effects on the transaminase activity in muscle during air stress compared to fish fed control and Phe diets. Muscle alanine transaminase activity for thermal stress in fish fed any diet was not significantly different from the control. Both Trp and Phe supplementations reduced the stress markers in the cod; hence, they could be used as additives for the stress attenuation. However, they also raised the activity of key enzymes in glycolysis and gluconeogenesis, mainly the Trp diets.