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Simple Summary: The salmonid industry is susceptible to opportunistic pathogens particularly affecting fish in early developmental stages. Understanding the immunological capacity during these stages is crucial for effective disease control. The receptor for Interleukin 8 (CXCR1), plays a key role in various inflammatory conditions. This study analyzed the expression of IL-8R and IL8 in rainbow trout larvae in response to bacterial lipopolysaccharide (LPS). The larvae 19 day post-hatching (dph) exhibited strong immune responses after 8 h of LPS stimulation, with IL-8 and omCXCR1 protein expression increasing. The use of specific antisera allowed localization of both proteins in mucosal tissue cells. This study highlights the effectiveness of LPS immersion in activating the innate immune system of trout larvae using IL-8/CXCR1 as immunological markers. The salmonid industry faces challenges due to the susceptibility of fish to opportunistic pathogens, particularly in early developmental stages. Understanding the immunological capacity during these stages is crucial for developing effective disease control strategies. IL-8R, a member of the G-protein-coupled receptor family, acts as a receptor for Interleukin 8 (IL-8). The binding of IL-8 to IL-8R plays a major role in the pathophysiology of a wide spectrum of inflammatory conditions. This study focused on the immune response capacity of rainbow trout (Oncorhynchus mykiss) larvae by analyzing IL-8/CXCR1 response to lipopolysaccharide (LPS) from Pseudomonas aeruginosa. Previous research demonstrated that LPS from P. aeruginosa acts as a potent immunostimulant in teleost, enhancing pro-inflammatory cytokines. The methodology included in silico analysis and the synthesis and characterization of an omCXCR1-derived epitope peptide, which was used to produce omCXCR1-specific anti98 serum in mice. The research revealed that rainbow trout larvae 19 days post-hatching (dph) exhibited pronounced immune responses post-stimulation with 1 µg/mL of LPS. This was evidenced by the upregulated protein expression of IL-8 and omCXCR1 in trout larvae 2 and 8 h after LPS challenge, as analyzed by ELISA and immunohistochemistry. Furthermore, fluorescence microscopy successfully revealed the colocalization of IL-8 and its receptor in cells from mucosal tissues after LPS challenge in larvae 19 dph. These findings underscore the efficacy of LPS immersion as a method to activate the innate immune system in trout larvae. Furthermore, we propose IL-8 and its receptor as molecular markers for evaluating immunostimulation in the early developmental stages of salmonids. [ABSTRACT FROM AUTHOR]

The skin of fish is a physicochemical barrier that is characterized by being formed by cells that secrete molecules responsible for the first defense against pathogenic organisms. In this study, the biological activity of peptides from mucus of Seriola lalandi and Seriolella violacea were identified and characterized. To this purpose, peptide extraction was carried out from epidermal mucus samples of juveniles of both species, using chromatographic strategies for purification. Then, the peptide extracts were characterized to obtain the amino acid sequence by mass spectrometry. Using bioinformatics tools for predicting antimicrobial and antioxidant activity, 12 peptides were selected that were chemically produced by simultaneous synthesis using the Fmoc-Tbu strategy. The results revealed that the synthetic peptides presented a random coil or extended secondary structure. The analysis of antimicrobial activity allowed it to be discriminated that four peptides, named by their synthesis code 5065, 5069, 5070, and 5076, had the ability to inhibit the growth of Vibrio anguillarum and affected the copepodite stage of C. rogercresseyi. On the other hand, peptides 5066, 5067, 5070, and 5077 had the highest antioxidant capacity. Finally, peptides 5067, 5069, 5070, and 5076 were the most effective for inducing respiratory burst in fish leukocytes. The analysis of association between composition and biological function revealed that the antimicrobial activity depended on the presence of basic and aromatic amino acids, while the presence of cysteine residues increased the antioxidant activity of the peptides. Additionally, it was observed that those peptides that presented the highest antimicrobial capacity were those that also stimulated respiratory burst in leukocytes. This is the first work that demonstrates the presence of functional peptides in the epidermal mucus of Chilean marine fish, which provide different biological properties when the fish face opportunistic pathogens. [ABSTRACT FROM AUTHOR]

Currently, one of the primary challenges in salmon farming is caligidosis, caused by the copepod ectoparasites Caligus spp. The infection process is determined by the copepod's ability to adhere to the fish skin through the insertion of its chitin-composed filament. In this study, we examined several antimicrobial peptides previously identified in salmonid mucosal secretions, with a primary focus on their potential to bind to chitin as an initial step. The binding capacity to chitin was tested, with hepcidin and piscidin showing positive results. Further assessments involving cytotoxicity in salmonid cells RTgill-W1, SHK-1, RTS-11, and RT-gut indicated that the peptides did not adversely affect cell viability. However, hemolysis assays unveiled the hemolytic capacity of piscidin at lower concentrations, leading to the selection of hepcidin for antiparasitic assays. The results demonstrated that the nauplius II stage of C. rogercresseyi exhibited higher susceptibility to hepcidin treatments, achieving a 50% reduction in parasitic involvement at 50 µM. Utilizing fluorescence and scanning electron microscopy, we observed the localization of hepcidin on the surface of the parasite, inducing significant spherical protuberances along the exoskeleton of C. rogercresseyi. These findings suggest that cysteine-rich AMPs derived from fish mucosa possess the capability to alter the development of the chitin exoskeleton in copepod ectoparasites, making them therapeutic targets to combat recurrent parasitic diseases in salmon farming. [ABSTRACT FROM AUTHOR]

The SARS-CoV-2 worldwide outbreak prompted the development of several tools to detect and treat the disease. Among the new detection proposals, the use of peptides mimetics has surged as an alternative to avoid the use of antibodies, of which there has been a shortage during the COVID-19 pandemic. However, the use of peptides in detection systems still presents some questions to be answered, mainly referring to their stability under different environmental conditions. In this work, we synthesized an ACE2 peptide mimic and evaluated its stability in different pH, salinity, polarity, and temperature conditions. Further, the same conditions were assessed when using the ability of the peptide mimic to detect the recombinant SARS-CoV-2 spike protein in a biotin-streptavidin-enzyme-linked assay. Finally, we also tested the capacity of the peptide to detect SARS-CoV-2 from patients' samples. The results indicate that the peptide is structurally sensitive to the medium conditions, with relevance to the pH, where basic pH favored its performance when used as a SARS-CoV-2 detector. Further, the proposed peptide mimic was able to detect SARS-CoV-2 comparably to RT-qPCR results. Therefore, the present study promotes knowledge advancement, particularly in terms of stability considerations, in the application of peptide mimics as a replacement for antibodies in detection systems. [ABSTRACT FROM AUTHOR]

Simple Summary: In mammals, autophagy plays a fundamental role in the defense against intracellular pathogens; however, in fish, this noncanonical function has not been described. In this context, it was proposed to study whether autophagy was modulated/activated in muscle cells challenged with a bacterial pathogen. Muscle cell cultures were performed and challenged with Piscirickettsia salmonis, the main threat to the salmon industry. Genes associated with immune response and autophagy were evaluated. In addition, the protein content of the LC3-II-specific marker of the autophagic process was evaluated via Western blot. Additionally, genes associated with vesicular traffic and endocytosis were evaluated, finding that P. salmonis promotes these processes. The results show a concomitant modulation of the genes associated with the immune response, vesicular trafficking, and autophagy, suggesting an early intracellular response by the muscle cell against this bacterium. Due to the necessity of seeking and discovering new alternatives and strategies to fight intracellular pathogens in the salmon industry, a better understanding of how autophagy participates in immune system responses may lead to the development of new technologies that allow for the effective control of intracellular pathogens, improving animal welfare and contributing to the sustainability of the global fish industry. Autophagy is a fundamental cellular process implicated in the health of the cell, acting as a cytoplasmatic quality control machinery by self-eating unfunctional organelles and protein aggregates. In mammals, autophagy can participate in the clearance of intracellular pathogens from the cell, and the activity of the toll-like receptors mediates its activation. However, in fish, the modulation of autophagy by these receptors in the muscle is unknown. This study describes and characterizes autophagic modulation during the immune response of fish muscle cells after a challenge with intracellular pathogen Piscirickettsia salmonis. For this, primary cultures of muscle cells were challenged with P. salmonis, and the expressions of immune markers il-1β, tnfα, il-8, hepcidin, tlr3, tlr9, mhc-I and mhc-II were analyzed through RT-qPCR. The expressions of several genes involved in autophagy (becn1, atg9, atg5, atg12, lc3, gabarap and atg4) were also evaluated with RT-qPCR to understand the autophagic modulation during an immune response. In addition, LC3-II protein content was measured via Western blot. The challenge of trout muscle cells with P. salmonis triggered a concomitant immune response to the activation of the autophagic process, suggesting a close relationship between these two processes. [ABSTRACT FROM AUTHOR]

Given the short span of instrumental precipitation records in the South American Altiplano, longer-term hydroclimatic records are needed to understand the nature of climate variability and to improve the predictability of precipitation, a key natural resource for the socioeconomic development in the Altiplano and adjacent arid lowlands. In this region grows Polylepis tarapacana, a long-lived tree species that is very sensitive to hydroclimatic changes and has been widely used for tree-ring studies in the central and southern Altiplano. However, in the northern sector of the Peruvian and Chilean Altiplano (16–19 ∘ S) still exists a gap of high-resolution hydroclimatic data based on tree-ring records. Our study provides an overview of the temporal evolution of the late-spring–mid-summer precipitation for the period 1625–2013 CE at the northern South American Altiplano, allowing for the identification of wet or dry periods based on a regional reconstruction from three P. tarapacana chronologies. An increase in the occurrence of extreme dry events, together with a decreasing trend in the reconstructed precipitation, has been recorded since the 1970s in the northern Altiplano within the context of the last ∼4 centuries. The average precipitation over the last 17 years stands out as the driest in our 389-year reconstruction. We reveal a temporal and spatial synchrony across the Altiplano region of dry conditions since the mid-1970s. Independent tree-ring-based hydroclimate reconstructions and several paleoclimatic records based on other proxies available for the tropical Andes record this synchrony. The influence of El Niño–Southern Oscillation (ENSO) on the northern Altiplano precipitation was detected by our rainfall reconstruction that showed past drier conditions in our study region associated with ENSO warm events. The spectral properties of the rainfall reconstruction showed strong imprints of ENSO variability at decadal, sub-decadal, and inter-annual timescales, in particular from the Pacific NIÑO 3 sector. Overall, the recent reduction in precipitation in comparison with previous centuries, the increase in extreme dry events and the coupling between precipitation and ENSO variability reported by this work is essential information in the context of the growing demand for water resources in the Altiplano. This study will contribute to a better understanding of the vulnerability and resilience of the region to the projected evapotranspiration increase for the 21st century associated with global warming. [ABSTRACT FROM AUTHOR]

The discovery and improvements of antimicrobial peptides (AMPs) have become an alternative to conventional antibiotics. They are usually small and heat-stable peptides, exhibiting inhibitory activity against Gram-negative and Gram-positive bacteria. In this way, studies on broad-spectrum AMPs found in amphibians with the remarkable capability to regenerate a wide array of tissues are of particular interest in the search for new strategies to treat multidrug-resistant bacterial strains. In this work, the use of bioinformatic approaches such as sequence alignment with Fasta36 and prediction of antimicrobial activity allowed the identification of the Ramosin peptide from the de novo assembled transcriptome of the plethodontid salamander Bolitoglossa ramosi obtained from post-amputation of the upper limb tissue, heart, and intestine samples. BLAST analysis revealed that the Ramosin peptide sequence is unique in Bolitoglossa ramosi. The peptide was chemically synthesized, and physicochemical properties were characterized. Furthermore, the in vitro antimicrobial activity against relevant Gram-positive and Gram-negative human pathogenic bacteria was demonstrated. Finally, no effect against eukaryotic cells or human red blood cells was evidenced. This is the first antibacterial peptide identified from a Colombian endemic salamander with interesting antimicrobial properties and no hemolytic activity. [ABSTRACT FROM AUTHOR]

Simple Summary: Antimicrobial peptides are part of the fish defense system, which can directly eliminate pathogenic microorganisms and, at the same time, regulate the immune response against them. This study evaluated the immunomodulatory effects of the antimicrobial peptide hepcidin in both juvenile fish and fish leukocyte cells. The results showed that hepcidin increased the expression of TNF-α, IL-1β, and IL-10 cytokines in leukocyte cells from trout. Moreover, the mRNA expressions of the same cytokines were up-regulated in different immune tissue of sea bass, confirming in vitro results. This study provides new insights into immunomodulatory function complementary to hepcidin's previously established antimicrobial activity modulating the pro- and anti-inflammatory responses in teleost fish. Antimicrobial peptides (AMP) play an essential role in the innate immune system, modulating the defense response. In a previous study, we demonstrated the antimicrobial activity of synthetic hepcidin (hep20) from rainbow trout (Oncorhynchus mykiss), and its protective effect in European sea bass (Dicentrarchus labrax) challenged with Vibrio anguillarum. Additionally, we described the uptake and distribution of hep20 in different tissues and leukocyte cells. Interestingly, various AMPs characterized in high vertebrates, called host defense peptides (HDPs), also possess immunomodulation activity. For that reason, the present study explores the immunomodulatory abilities of hep20 through in vitro and in vivo studies. First, a monocyte/macrophage RTS-11 cell line from rainbow trout was used to evaluate hep20 effects on pro- and anti-inflammatory cytokines in fish leukocyte cells. Next, the European sea bass juveniles were used to determine if hep20 can regulate the expression of cytokines in fish immune tissues. The results show that hep20 was uptake inner to RTS-11 cells and was able to induce the expression of IL-10, IL-1β, and TNFα at transcriptional and protein levels. Then, the European sea bass juveniles were given intraperitoneal injections of the peptide. At 1, 3, 7, 14, and 21 days post-injection (dpi), IL-10, IL -1β, and TNFα mRNA were quantified in the anterior gut, spleen, and head kidney. The hep20 was able to up-regulate cytokine gene expression in these tissues, mainly in the head kidney. Furthermore, the evaluated cytokines showed a cyclical tendency of higher to lesser expression. Finally, a bioinformatics analysis showed that the structure adopted by hep20 is similar to the γ-core domain described for cysteine-stabilized AMP, defined as immunomodulatory and antimicrobial, which could explain the ability of hep20 to regulate the cytokine expression. This study provides new insights into immunomodulatory function complementary to the previously established antimicrobial activity of hep20, suggesting a role as an HDP in teleost fish. These facts are likely to be associated with molecular functions underpinning the protective effect of fish hepcidin against pathogens. [ABSTRACT FROM AUTHOR]

Simple Summary: Various long-term stress conditions may exist in fish cultivation, damaging the physiological responses that regulate the fish growth and feed. Different signalers connect the brain with the gastrointestinal tract, including the perception of stress factors for the regulation of physiological responses. Here, we evaluated the effect of varying culture densities of Salmo salar post-smolt on the gene expression of some brain and gastrointestinal hormone signalers. We found that high stock densities could promote the levels of molecules associated with feed inhibition, which could be related to the stress pathway regulated by corticoids. Thus, the expression of these peptide hormones could be used as biological markers to improve production practices in fish aquaculture. A variety of long-term stress conditions may exist in fish cultivation, some of which are so severe that fish can no longer reestablish homeostasis. In teleost fish, the brain and gastrointestinal tract integrate signals that include the perception of stress factors regulating physiological responses, such as social stress by fish population density, where peripheral and central signals, such as peptide hormones, are the main regulators. Therefore, we proposed in this study to analyze the effect of different stock densities (SD) in the gene expression of brain neuropeptide Y (NPY) and calcitonin gene-related peptide (CGRP), together with the gastrointestinal peptide hormones leptin (Lep), vasointestinal peptide (VIP), and protachykinin-1 (Prk-1) in Salmo salar post-smolt. The coding sequence of S. salar VIP and Prk-1 precursors were firstly cloned and characterized. Then, the mRNA expression of these genes, together with the NPY, Lep, and CGRP genes, were evaluated in post-smolts kept at 11 Kg/m3, 20 Kg/m3, and 40 Kg/m3. At 14 days of culture, the brain CGRP and liver leptin mRNA levels increased three and tenfold in the post-smolt salmons kept at the highest SD, respectively. The high levels of leptin were kept during all the fish culture experiments. In addition, the highest expression of intestine VIP mRNA was obtained on Day 21 in the group of 40 Kg/m3 returning to baseline on Day 40. In terms of stress biochemical parameters, cortisol levels were increased in the 20 Kg/m3 and 40 Kg/m3 groups on Day 40 and were the highest in the 20 Kg/m3 group on Day 14. This study provides new insight into the gastrointestinal signals that could be affected by chronic stress induced by high stock density in fish farming. Thus, the expression of these peptide hormones could be used as molecular markers to improve production practices in fish aquaculture. [ABSTRACT FROM AUTHOR]

Given the short span of instrumental precipitation records in the South American Altiplano, long-term 25 hydroclimatic records are needed to understand the nature of climate variability and to improve the predictability of precipitation, a key natural resource for the socio-economic development in the Altiplano and adjacent arid lowlands. In this region grows Polylepis tarapacana, a long-lived tree species that is very sensitive to hydroclimatic changes and have been widely used for tree-ring studies in the central and southern Altiplano. However, in the northern sector of the Peruvian and Chilean Altiplano (16°-19° S) still exist a gap of hydroclimatic tree-ring records. Our study provides an overview of the 30 temporal evolution of annual precipitation for the period 1625-2013 CE at the northern South American Altiplano, allowing for the identification of wet or dry periods based on a regional reconstruction composed by three P. tarapacana chronologies. An increase in the occurrence rate of extreme dry events, together with a decreasing trend in the reconstructed precipitation, have been recorded since the 1970s decade in the northern Altiplano within the context of the last ~four centuries. The average precipitation of the last 17-year stands out as the driest in our 389-years reconstruction. We revealed a 35 temporal and spatial synchrony across the Altiplano region of wet conditions during the first half of the 19th century and the drought conditions since mid 1970s recorded by independent tree-ring based hydroclimate reconstructions and several paleoclimatic records based on other proxies available for the tropical Andes. The rainfall reconstruction provides also valuable information about the ENSO influences in the northern Altiplano precipitation. The spectral properties of the rainfall reconstruction showed strong imprints of ENSO variability at decadal, sub-decadal and inter-annual time-scale, in 40 particular from the Pacific N3 sector. Overall, the remarkable recent reduction in precipitation in comparison with previous centuries, the increase in extreme dry events and the coupling between precipitation and ENSO variability reported by this work is essential information in the context of the growing demand for water resources in the Altiplano that will contribute to a better understanding of the vulnerability/resilience of the region to the projected evapotranspiration increase for the 21st century associated to global warming. [ABSTRACT FROM AUTHOR]

The analysis of the food environment is used to identify areas with gaps in the availability of healthy foods and can be used as a public policy assessment tool. In recent decades, Chile has implemented several strategies and regulations to improve food environments, with encouraging results. Little is known about the scope of these measures in socially vulnerable environments. This study is part of a project that seeks to build an integrated intervention model for healthy school environments in a vulnerable area of Santiago, Chile. The objective of this study was to evaluate the availability of healthy and unhealthy foods around schools and the relationship between it and socioeconomic determinants of the school community in the Chilean context. A cross-sectional study to measure the food environment of informal markets (street food), formal markets (stores), and institutions (schools) was conducted in and around 12 schools (100 m surrounding schools) in a vulnerable urban area of Santiago, Chile. A lack of healthy foods was observed, which was related to some socio-economic determinants and the multidimensional poverty was the most relevant. The diagnosis of food environments around schools can represent an important target for governments to implement policies focused at improving the availability of healthy foods. [ABSTRACT FROM AUTHOR]

Prumnopitys andina is a South American podocarp with a restricted distribution in southern Andean temperate forests, and is severely threatened due to habitat loss. Despite much dendrochronological work on other temperate South American conifers, little is known about the dendrochronological potential of P. andina. Here, we examine the main features of P. andina tree rings and evaluate the potential of this species for yielding tree-ring chronologies sensitive to climatic variability. In a population at the southern range limit of the species, we collected tree cores and cross-section samples of P. andina for analyses following standard dendrochronological techniques. 76 tree-ring series from 45 trees were examined, 32 (42%) of which were successfully cross-dated and included in a chronology. We found that the species forms well-defined tree rings with distinct boundaries delimiting annual growth. The chronology began in 1778, with notable periods of suppression and release occurring in the late nineteenth and early twentieth centuries, respectively. The chronology showed a high inter-series correlation value of 0.51, suggesting a strong common signal among trees. December to January precipitation showed a positive relationship with radial growth. Conversely, maximum and mean temperature showed negative relationships with radial growth during the austral summer and autumn. These significant relationships of climate variables with radial growth of P. andina indicate that tree-ring series of this species are suitable for dendroclimatological studies. [ABSTRACT FROM AUTHOR]

Several factors have influenced the increasing presence of peptides as an important class of Active Pharmaceutical Ingredients. One is the continued development of synthetic methodologies for peptide synthesis. Herein, we investigated the Fmoc removal step, using the tea-bag strategy. In this regard, three different secondary amines: piperidine, 4-methylpiperidine, and piperazine, were evaluated. As a result of this study, 4-methyl piperidine showed to be an excellent alternative to the usually used piperidine in terms of purity and compliance with green chemistry principles as well. [ABSTRACT FROM AUTHOR]

Polycystic ovary syndrome (PCOS) is the most common endocrine disorder among reproductive-aged women and the main cause of infertility due to anovulation. However, this syndrome spans the lives of women affecting them from in-utero life until death, leading to several health risks that can impair quality of life and increase morbidity and mortality rates. Fetal programming may represent the beginning of the condition characterized by hyperandrogenism and insulin resistance which leads to a series of medical consequences in adolescence, adulthood, and old age. Menstrual and fertility problems evolve into metabolic complications as age advances. An early and precise diagnosis is important for an adequate management of PCOS, especially at the extreme ends of the reproductive lifespan. However, many different phenotypes are included under the same condition, being important to look at these different phenotypes separately, as they may require different treatments and have different consequences. In this way, PCOS exhibits a great metabolic complexity and its diagnosis needs to be revised once again and adapted to recent data obtained by new technologies. According to the current medical literature, lifestyle therapy constitutes the first step in the management, especially when excess body weight is associated. Pharmacotherapy is frequently used to treat the most predominant manifestations in each age group, such as irregular menses and hirsutism in adolescence, fertility problems in adulthood, and metabolic problems and risk of cancer in old age. Close surveillance is mandatory in each stage of life to avoid health risks which may also affect the offspring, since fetal and post-natal complications seem to be increased in PCOS women. [ABSTRACT FROM AUTHOR]

Nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) are efficient soluble intracellular sensors that activate defense mechanisms against pathogens. In teleost fish, the involvement of NLRs in the immune response is not well understood. However, recent work has evidenced the expression of different NLRs in response to some pathogen associated molecular patterns (PAMPs). In the present work, the cDNA sequence encoding three new NOD-like receptors were identified in Oncorhynchus mykiss , namely OmNLRC3, OmNLRC5 and OmNLRX1. Results showed that their sequences coded for proteins of 1135, 836 and 1010 amino acids, respectively. The deduced protein sequences of all receptors showed characteristic domains of this receptor family, such as leucine rich repeats and NACHT domain. Phylogenetic analysis revealed a high degree of identity with other NOD-like receptors and they are clustered into different families. Transcript expression analysis indicated that OmNLRs are constitutively expressed in liver, spleen, intestine, gill, skin and brain. OmNLR expression was upregulated in kidney and gills from rainbow trout in response to LPS. In order to give new insights into the function of these new NLR members, an in vitro model of immune stimulation was established using the rainbow trout cell line RTgill-W1. Expression analysis revealed that RTgill-W1 overexpressed proinflammatory cytokines in response to LPS and poly I:C alongside with a differential overexpression of OmNLRC3, OmNLRC5 and OmNLRX1. The expression of OmNLRC5 was further verified at the protein level by immunofluorescence. Finally, the effect of the overexpressed cytokines on the OmNLR expression by RTgill-W1 cells was assessed, suggesting a regulatory mechanism on OmNLRC3 expression. Overall, results suggest that O. mykiss NOD-like receptors could play a key role in the defense mechanisms of teleost through PAMP recognition. Future studies will focus on gills which could be related with a key sensor mucosal system in one of the most environmentally fish exposed tissues. [ABSTRACT FROM AUTHOR]