Environmental influence on the Atlantic salmon transcriptome and methylome during sea lice infestations.

The fish's immune response is affected by different factors, including a wide range of environmental conditions that can also disrupt or promote changes in the host-pathogen interactions. How environmental conditions modulate the salmon genome during parasitism is poorly understood here. This study aimed to explore the environmental influence on the Salmo salar transcriptome and methylome infected with the sea louse Caligus rogercresseyi. Atlantic salmon were experimentally infected with lice at two temperatures (8 and 16 °C) and salinity conditions (32 and 26PSU). Fish tissues were collected from the infected Atlantic salmon for reduced representation bisulfite sequencing (RRBS) and whole transcriptome sequencing (RNA-seq) analysis. The parasitic load was highly divergent in the evaluated environmental conditions, where the lowest lice abundance was observed in fish infected at 8 °C/26PSU. Notably, transcriptome profile differences were statistically associated with the number of alternative splicing events in fish exposed to low temperature/salinity conditions. Furthermore, the temperature significantly affected the methylation level, where high values of differential methylation regions were observed at 16 °C. Also, the association between expression levels of spliced transcripts and their methylation levels was determined, revealing significant correlations with Ferroptosis and TLR KEEG pathways. This study supports the relevance of the environmental conditions during host-parasite interactions in marine ecosystems. The discovery of alternative splicing transcripts associated with DMRs is also discussed as a novel player in fish biology.

Liquid-Liquid Dispersion Performance Prediction and Uncertainty Quantification Using Recurrent Neural Networks.

We demonstrate the application of a recurrent neural network (RNN) to perform multistep and multivariate time-series performance predictions for stirred and static mixers as exemplars of complex multiphase systems. We employ two network architectures in this study, fitted with either long short-term memory and gated recurrent unit cells, which are trained on high-fidelity, three-dimensional, computational fluid dynamics simulations of the mixer performance, in the presence and absence of surfactants, in terms of drop size distributions and interfacial areas as a function of system parameters; these include physicochemical properties, mixer geometry, and operating conditions. Our results demonstrate that while it is possible to train RNNs with a single fully connected layer more efficiently than with an encoder-decoder structure, the latter is shown to be more capable of learning long-term dynamics underlying dispersion metrics. Details of the methodology are presented, which include data preprocessing, RNN model exploration, and methods for model performance visualization; an ensemble-based procedure is also introduced to provide a measure of the model uncertainty. The workflow is designed to be generic and can be deployed to make predictions in other industrial applications with similar time-series data.

Grape Pomace Polyphenols Reduce Acute Inflammatory Response Induced by Carrageenan in a Murine Model.

Grape pomace (GP), a by-product of wine production, contains bioactive polyphenols with potential health benefits. This study investigates the anti-inflammatory properties of a polyphenolic fraction derived from GP, obtained by ultrasound-microwave hybrid extraction and purified using ion-exchange chromatography. In the inflammation model, mice were divided into six groups: intact, carrageenan, indomethacin, and three GP polyphenols treatment groups. Paw edema was induced by subplantar injection of carrageenan, and the GP polyphenols were administered intraperitoneally at doses of 10, 20, and 40 mg/kg. The anti-inflammatory effect was evaluated by measuring paw volume, and expression of inflammatory markers: cyclooxygenase-2 (COX-2), myeloperoxidase (MPO), and cytokines (IL-1ß and IL-6), along with lipid peroxidation levels. The GP polyphenols significantly reduced paw edema and expression levels of COX-2, MPO, and cytokines in a dose-dependent manner effect, with the highest dose showing the greatest reduction. Additionally, lipid peroxidation levels were also decreased by GP polyphenols treatment at doses of 10 and 20 mg/kg. These findings suggest that ultrasound-microwave extraction combined with amberlite purification proved to be effective in obtaining a polyphenolic-rich fraction from GP. Thus, GP polyphenols may serve as a natural anti-inflammatory and antioxidant agent for treating inflammation and oxidative stress-related diseases.

Identification and expression analysis of two steamer-like retrotransposons in the Chilean blue mussel (Mytilus chilensis).

BACKGROUND: Disseminated neoplasia (DN) is a proliferative cell disorder of the circulatory system of bivalve mollusks. The disease is transmitted between individuals and can also be induced by external chemical agents such as bromodeoxyuridine. In Mya arenaria, we have cloned and characterized an LTR-retrotransposon named Steamer. Steamer mRNA levels and gene copy number correlates with DN and can be used as a marker of the disease. So far, the only mollusk where a retrotransposon expression relates to DN is Mya arenaria. On the other hand, it has been reported that the Chilean blue mussel Mytilus chilensis can also suffers DN. Our aim was to identify retrotransposons in Mytilus chilensis and to study their expression levels in the context of disseminated neoplasia. RESULTS: Here we show that 7.1% of individuals collected in August 2018, from two farming areas, presents morphological characteristics described in DN. Using Steamer sequence to interrogate the transcriptome of M. chilensis we found two putative retrotransposons, named Steamer-like elements (MchSLEs). MchSLEs are present in the genome of M. chilensis and MchSLE1 is indeed an LTR-retrotransposon. Neither expression, nor copy number of the reported MchSLEs correlate with DN status but both are expressed at different levels among individual animals. We also report that in cultured M. chilensis haemocytes MchSLEs1 expression can be induced by bromodeoxyuridine. CONCLUSIONS: We conclude that SLEs present in Mytilus chilensis are differentially expressed among individuals and do not correlate with disseminated neoplasia. Treatment of haemocytes with a stressor like bromodeoxyuridine induces expression of MchSLE1 suggesting that in Mytilus chilensis environmental stressors can induce activation of LTR-retrotransposon.

Gills de novo assembly reveals oxidative stress, unfolded protein, and immune response on red cusk-eel (Genypterus chilensis) under thermal stress.

The heat waves on the South Pacific coast could lead to thermal stress in native fish. The red cusk-eel (Genypterus chilensis) is relevant for Chilean artisanal fisheries and aquaculture diversification. This study examined the effect of high-temperature stress in the gills of G. chilensis in control (14 °C) and high-temperature stress (19 °C) conditions. High-temperature stress induces a significant increase in gills cortisol levels. Additionally, oxidative damage was observed in gills (protein carbonylation and lipoperoxidation). RNA-seq data was used to build the first transcriptome assembly of gills in this species (23,656 annotated transcripts). A total of 1138 down-regulated and 1531 up-regulated transcripts were observed in response to high-temperature stress in gills. The enrichment analysis showed immune response and replication enriched processes (on down-regulated transcripts), and processes related to the folding of proteins, endoplasmic reticulum, and transporter activity (on up-regulated transcripts). The present study showed how gills could be affected by high-temperature stress.

Antiproliferative and cytotoxic effects of polyphenols from pomegranate peel and coffee pulp on cancer cells.

Pomegranate peel (PP) and coffee pulp (CP) are by-products of the food industry that can cause environmental pollution if not handled adequately. These by-products contain a significant amount of polyphenolic compounds which have antioxidant and possibly anticancer properties. We investigated the antiproliferative and cytotoxic activities of polyphenols from PP, CP and a 50-50% mixture of both against HeLa, A549, MDA-MB and Hek-293 cells. The total phenolic content of the PP and CP extracts was determined by high performance liquid chromatography/electrospray ionisation/mass spectrometry, and the antiproliferative and cytotoxic potentials were evaluated by MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and Lactate Dehydogenase assays, respectively. Results showed antiproliferative and cytotoxic effects of polyphenols from PP and CP when administered at different concentrations or mixtures on HeLa, A549 and MDA-MB cells. No significant antiproliferative effects were observed on Hek-293 cells treated under similar conditions. These results suggest the potential of PP and CP polyphenols, individually or in combination, to modulate biological mechanisms involved in cervical, breast and lung cancer.

Development of Groundwater Levels Dataset for Chile since 1970.

Access to accurate spatio-temporal groundwater level data is crucial for sustainable water management in Chile. Despite this importance, a lack of unified, quality-controlled datasets have hindered large-scale groundwater studies. Our objective was to establish a comprehensive, reliable nationwide groundwater dataset. We curated over 120,000 records from 640 wells, spanning 1970-2021, provided by the General Water Resources Directorate. One notable enhancement to our dataset is the incorporation of elevation data. This addition allows for a more comprehensive estimation of groundwater elevation. Rigorous data quality analysis was executed through a classification scheme applied to raw groundwater level records. This resource is invaluable for researchers, decision-makers, and stakeholders, offering insights into groundwater trends to support informed, sustainable water management. Our study bridges a crucial gap by providing a dependable dataset for expansive studies, aiding water management strategies in Chile.

11-Deoxycorticosterone (DOC)'s Action on the Gill Osmoregulation of Juvenile Rainbow Trout (Oncorhynchus mykiss).

In aquaculture, stress can negatively affect fish growth. For years, the cortisol hormone has been thought to play both glucocorticoid and mineralocorticoid functions. Nevertheless, recent research has suggested that 11-deoxycorticosterone (DOC) released during stress could contribute to cortisol actions, though this process is still misunderstood. Here, we evaluated the DOC effects on physiological and early transcriptional responses by RNA-seq. Juvenile rainbow trout were treated with DOC and/or glucocorticoids (mifepristone) or mineralocorticoid (eplerenone) receptor antagonists. Subsequently, plasma was collected, and cDNA libraries were generated from the gills of vehicle (control), DOC, mifepristone, mifepristone with DOC, eplerenone, and eplerenone with DOC groups. Calcium and phosphate levels in plasma were changed. Results revealed 914 differentially expressed transcripts (DETs) induced by DOC compared with control, mainly associated with sodium ion transmembrane transport, gluconeogenesis, negative regulation of transmembrane transport, and activation of innate immune response. DOC versus eplerenone with DOC comparison displayed 444 DETs related to cell-cell junction organization, canonical glycolysis, positive regulation of immune response, and potassium ion transport. Conversely, no DETs were detected in DOC versus mifepristone with DOC comparison. These data suggest that DOC has a relevant role in gill stress response and ion transport, which is differentially regulated by mineralocorticoid receptors.

Weighted Hermite-Hadamard integral inequalities for general convex functions.

In this article, starting with an equation for weighted integrals, we obtained several extensions of the well-known Hermite-Hadamard inequality. We used generalized weighted integral operators, which contain the Riemann-Liouville and the $ k $-Riemann-Liouville fractional integral operators. The functions for which the operators were considered satisfy various conditions such as the $ h $-convexity, modified $ h $-convexity and $ s $-convexity.

Sprouts as probiotic carriers: A new trend to improve consumer nutrition.

Over the past few decades, efforts to eradicate hunger in the world have led to the generation of sustainable development goals to reduce poverty and inequality. It is estimated that the current coronavirus pandemic could add between 83 and 132 million to the total number of undernourished people in the world by 2021. Food insecurity is a contributing factor to the increase in malnutrition, overweight and obesity due to the quality of diets to which people have access. It is therefore necessary to develop functional foods that meet the needs of the population, such as the incorporation of sprouts in their formulation to enhance nutritional quality. Germination of grains and seeds can be used as a low-cost bioprocessing technique that provides higher nutritional value and better bioavailability of nutrients. Consequently, the manuscript describes relevant information about the germination process in different seeds, the changes caused in their nutritional value and the use of techniques within the imbibition phase to modify the metabolic profiles within the sprouts such as inoculation with lactic acid bacteria and yeasts, to generate a functional symbiotic food.

10­Gingerol, a novel ginger compound, exhibits antiadipogenic effects without compromising cell viability in 3T3­L1 cells.

Obesity is defined as excessive fat accumulation that can be detrimental to health and currently affects a large part of the global population. Obesity arises from excessive energy intake along with a sedentary lifestyle and leads to adipocytes with aggravated hypertrophy. Strategies have been designed to prevent and treat obesity. Nutrigenomics may serve a role in prevention of obesity using bioactive compounds present in certain foods with anti-obesogenic effects. Ginger (Zingiber officinale Roscoe) contains gingerols, key bioactive compounds that inhibit hypertrophy and hyperplasia of adipocytes. The present study aimed to evaluate the antiadipogenic activity of 10-gingerol (10-G) in the 3T3-L1 cell line. Three study groups were formed: Negative (3T3-L1 preadipocytes) and positive control (mature 3T3-L1 adipocytes) and 10-G (3T3-L1 preadipocytes stimulated with 10-G during adipogenic differentiation). Cell viability and lipid content were evaluated by MTT assay and Oil Red O staining, respectively. mRNA expression of CCAAT enhancer-binding protein α (C/ebpα), peroxisome proliferator-activated receptor γ (Pparγ), mechanistic target of rapamycin complex (Mtor), sterol regulatory element binding transcription factor 1 (Srebf1), acetyl-coenzyme A carboxylase (Acaca), fatty acid binding protein 4 (Fabp4), and 18S rRNA (Rn18s), was quantified by quantitative PCR. The protein expression of C/EPBα was analyzed by western blot. In the 10-G group, lipid content was decreased by 28.83% (P<0.0001) compared with the positive control; notably, cell viability was not affected (P=0.336). The mRNA expression in the 10-G group was higher for C/ebpα (P<0.001) and lower for Acaca (P<0.001), Fabp4 (P<0.001), Mtor (P<0.0001) and Srebf1 (P<0.0001) compared with the positive control group, while gene expression of Pparγ did not present significant changes. The presence of 10-G notably decreased C/EBPα protein levels in 3T3-L1 adipocytes. In summary, the antiadipogenic effect of 10-G during the differentiation of 3T3-L1 cells into adipocytes may be explained by mRNA downregulation of adipogenic transcriptional factors and lipid metabolism-associated genes.

Polyphenolic characterisation and antiprotozoal effect of extracts obtained by maceration, ultrasound, microwave and ultrasound/microwave of Porophyllum ruderale (Jacq.) Cass.

Porophyllum ruderale (Jacq.) Cass. (Asteraceae) has antiprotozoal properties and contains extractable phenolic compounds by the maceration method (M). However, new extraction proposals such as ultrasound (U), microwaves (MW), and ultrasound/microwaves (U/MW) have emerged to optimise yields, but it is unknown if these methods modify effectiveness. Therefore, the study consisted of extracting the aerial part of P. ruderale with ethanol using the M, U, MW and U/MW methods to study its composition by RP-HPLC-ESI-MS, its total polyphenol content and its effect against Entamoeba histolytica. The study showed that U, MW and U/MW did not modify the extraction yield compared to M, but they did change the composition and the total polyphenol content. All extracts contain phloretin, caffeic acid 4-O-glucoside, todolactol A, quercetin 3-O-glucoside, quercetin 3-O-rhamnoside, luteolin and 3,7-dimethylquercetin, and affected the growth of E. histolytica. However, M and U extracts were the most effective at 5 mg/mL.

Sugarcane Bagasse Ash as an Alternative Source of Silicon Dioxide in Sodium Silicate Synthesis.

To reduce the environmental impacts from sodium silicate synthesis, a ceramic method was suggested, with sugarcane bagasse ash (SCBA) as the source of silicon dioxide and sodium carbonate. Although the production of sodium silicate is carried out on a large scale, it should be noted that its process requires temperatures above 1000 °C; it also requires the use of highly corrosive agents such as sodium hydroxide and chlorine gas to neutralize the remaining sodium hydroxide. In the present study, the synthesis temperatures were reduced to 800 °C with a reaction time of 3 h by pressing equimolar mixtures of previously purified SCBA and sodium carbonate; then, heat treatment was carried out under the indicated conditions. The resulting materials were analyzed with Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). Among the crystalline phases, calcium disodium silicate was identified, in addition to sodium silicate; thus, it was inferred that the other components of the ash can interfere with the synthesis of silicate. Therefore, in order to obtain the highest composition of sodium silicate, a leaching treatment of the SCBA is required.

Jean Irvine and Lesley Yellowlees, first female presidents of the Royal Pharmaceutical Society and of the Royal Society of Chemistry, respectively / Jean Irvine y Lesley Yellowlees, primeras mujeres presidentas de la Royal Pharmaceutical Society y de la Royal Society of Chemistry, respectivamente

Despite the indisputable progress made in the fight against gender inequalities since the last years of the last century, there are still many differences between the roles played by women and men when it comes to holding the top positions on boards of directors of relevant companies or in the presidencies of scientific societies, in which the presence of women does not reach at all the percentage of 40% that is estimated as necessary to achieve that equality. In this article, we show the biographies of Jean Irvine and Lesley Yellowlees, the first female presidents of the Royal Pharmaceutical Society and the Royal Society of Chemistry, respectively, although their appointments occurred many years after the aforementioned societies had been founded. The objective is to make their figures known to society and place them as references and models to be followed by the rest of women who wish to practice a profession. Some brief biographical data of other women who played a prominent role in scientific and academic societies in the early years of the 20th century are also shown.(AU)

A pesar de los indiscutibles avances habidos en la lucha contra las desigualdades de género desde los últimos años del siglo pasado, aún existen muchas diferencias entre los roles que desempeñan las mujeres y los hombres a la hora de ocupar los primeros puestos en los consejos de administración de empresas relevantes o en las presidencias de sociedades científicas, en las que la presencia de mujeres no alcanza en absoluto el porcentaje del 40% que se estima necesario para alcanzar esa igualdad. En este artículo mostramos las biografías de Jean Irvine y Lesley Yellowlees, las primeras mujeres presidentas de la Royal Pharmaceutical Society y de la Royal Society of Chemistry, respectivamente, aunque sus nombramientos se produjeron muchos años después de la fundación de dichas sociedades. El objetivo es dar a conocer sus figuras ante la sociedad y situarlas como referentes y modelos a seguir por el resto de las mujeres que deseen ejercer una profesión. También se muestran algunos breves datos biográficos de otras mujeres que desempeñaron un papel destacado en sociedades científicas y académicas en los primeros años del siglo XX.(AU)

Intravenous Administration of Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSC) for Acute Respiratory Distress Syndrome Due to COVID-19 Infection.

The COVID-19 pandemic has posed significant therapeutic challenges in addressing acute respiratory distress syndrome (ARDS). This serious illness has caused numerous fatalities worldwide and has had profound health and economic impacts. Previous studies have shown that mesenchymal stem cells (MSCs) can suppress ARDS. In this case series, we report on the treatment of nine patients with a single intravenous dose of 100 million hypoxic cultured umbilical cord-derived MSCs (UC-MSCs). Following the intravenous administration of UC-MSCs, obtained from the lining of the umbilical cord, longitudinal laboratory analysis revealed a sustained decrease in inflammatory markers and stabilized pulmonary function in eight out of nine patients. UC-MSCs possess immunomodulatory and anti-inflammatory properties, enabling them to attenuate the cytokine storm and potentially aid in lung repair. Importantly, no adverse events associated with the treatment were observed. These findings collectively suggest that a cell-based approach significantly enhances the survival rate of ARDS induced by SARS-CoV-2 and offers a promising treatment option in both preclinical and clinical settings.

A Balanced Dietary Ratio of n-6:n-3 Polyunsaturated Fatty Acids Exerts an Effect on Total Fatty Acid Profile in RBCs and Inflammatory Markers in Subjects with Obesity.

The n-3 polyunsaturated fatty acids (PUFAs) can reduce inflammatory markers and may therefore be useful in obesity management. The aim of this study was to analyze the effect of supplementation with n-3 PUFAs on total fatty acid profile in red blood cells (RBCs), as well as biochemical and inflammatory markers, in subjects with obesity. The study consisted in a randomized placebo-controlled, double-blind clinical trial involving 41 subjects with obesity during a 4-month follow-up. Individuals were randomly assigned to two groups: n-3 PUFA supplementation (1.5 g fish oil) and placebo (1.5 g sunflower oil). Anthropometric, biochemical, dietetic, cytokine and total fatty acid profiles in RBCs were measured. Both groups increased their PUFA intake and DHA incorporation in RBCs. However, the placebo group showed a reduction in serum IL-8 and MCP-1 at the end of the study. A multiple linear regression model adjusted by body fat mass and sex showed that an increase in DHA in RBCs decreased the serum IL-8 levels in both study groups at the end of the study. Our results highlight the role of dietary DHA and n-3 supplementation usefulness in exerting beneficial anti-inflammatory effects.

A mix of ginger phenols exhibits anti­adipogenic and lipolytic effects in mature adipocytes derived from 3T3­L1 cells.

The prevalence of obesity has increased rapidly worldwide. Obesity is characterized by excessive adipose tissue in the body, which is related to hyperplasia and hypertrophy in adipocytes. Ginger (Zingiber officinale Roscoe) is a medicinal plant that possesses an anti-obesogenic effect mostly attributed to gingerols, the most abundant bioactive compounds in ginger. The anti-adipogenic and lipolytic effects of these phenols have been shown when investigated individually. Therefore, the present study aimed to evaluate the lipolytic and anti-adipogenic activity of a mix of the main ginger phenols 6-gingerol, 8-gingerol, 10-gingerol, 6-shogaol, 8-shogaol and 10-shogaol on the 3T3-L1 cell line. A total of four study groups were designed: Negative control (3T3-L1 preadipocytes); positive control (mature 3T3-L1 adipocytes); phenols-pre (3T3-L1 cells stimulated with the phenols mix during adipogenic differentiation); and phenols-post (mature 3T3-L1 adipocytes stimulated with the phenols mix). MTT viability cell assay and Oil Red O staining were performed. Glycerol concentration supernatants were determined using the VITROS 350 Chemistry System. Expression of mRNA was measured using qPCR. The treatment with a 2 µg/ml ginger phenol dose reduced the lipid content by 45.52±7.8 and 35.95±0.76% in the phenols-pre and -post group, respectively, compared with that in the positive control group. The phenols-post group presented a higher glycerol concentration in the supernatant compared with that in the positive control and the phenols-pre groups. The mRNA expression levels of CCAAT/enhancer-binding protein alpha, peroxisome proliferator activated receptor-γ, fatty acid-binding protein 4 and fatty acid synthase were higher in the phenols-pre and lower in the phenols-post groups, compared with those in the positive control group. To the best of our knowledge, the current study demonstrated for the first time the anti-adipogenic and lipolytic effects of a mix of the main bioactive compounds found in ginger, and it also established the basis to use this mix of phenols in in vivo studies and clinical trials.

Photovoltaic Glass Waste Recycling in the Development of Glass Substrates for Photovoltaic Applications.

Because of the increasing demand for photovoltaic energy and the generation of end-of-life photovoltaic waste forecast, the feasibility to produce glass substrates for photovoltaic application by recycling photovoltaic glass waste (PVWG) material was analyzed. PVWG was recovered from photovoltaic house roof panels for developing windows glass substrates; PVWG was used as the main material mixed with other industrial waste materials (wSG). The glass was casted by air quenching, annealed, and polished to obtain transparent substrates samples. Fluorine-doped tin oxide (FTO) was deposited as back contact on the glass substrates by spray pyrolysis. The chemical composition of the glass materials was evaluated by X-ray fluorescence (XRF), the thermal stability was measured by differential thermal analysis (DTA) and the transmittance was determined by UV-VIS spectroscopy. The surface of the glass substrates and the deposited FTO were observed by scanning electron microscopy (SEM), the amorphous or crystalline state of the specimens were determined by X-ray diffraction (XRD) and the sheet resistance was evaluated by the four-point probe method. The sheet resistance of the deposited FTO on the wSG substrate was 7.84 ± 3.11 Ω/□, lower than that deposited on commercial soda-lime glass (8.48 ± 3.67 Ω/□), meaning that this material could present improved conduction of the produced electrons by the photovoltaic effect. This process may represent an alternative to produce glass substrates from waste materials that could be destined for photovoltaic applications, especially the production of ecological photovoltaic windows.

Early regulation of corticosteroid receptor expression in rainbow trout (Oncorhynchus mykiss) gills is mediated by membrane-initiated cortisol signaling.

Cortisol is a key stress-related hormone involved in the physiological adjustments of fish. In gills, cortisol contributes to acclimatization to changes in environmental salinity, promoting both ion uptake or salt excretion. Cortisol exerts its biological effects through its interaction with specific intracellular glucocorticoid (GR) and mineralocorticoid (MR) receptors. Additionally, the further identification of GR and MR on the surface of different tissues, together with the existence of cortisol-mediated effects observed using membrane-impermeable analogs (e.g., cortisol-BSA), supports the existence of membrane-initiated cortisol actions in fish. Nevertheless, the impact of this alternative cortisol mechanism in relevant tissues for fish salinity acclimation, such as gill, is unknown. In this work, we sought to explore the contribution of rapid membrane-initiated cortisol on GR and MR regulation in rainbow trout (Oncorhynchus mykiss) gills using in vivo and in vitro approaches. Juvenile rainbow trout intraperitoneally injected with cortisol or cortisol-BSA showed increased gr2 but no gr1 or mr mRNA levels in gills after one hour of treatment. This result was further confirmed using RT-gills-W1 cell lines stimulated with both versions of cortisol. Interestingly, after three and six hours of cortisol or cortisol-BSA treatment, there were no changes in the mRNA levels of any corticosteroid receptor in RT-gills-W1 cells. Finally, using immunofluorescence analysis, we identified GR and MR in rainbow trout gill cells localized on the cell surface. Considering the in vivo and in vitro results of this work, we suggest that membrane-initiated cortisol action contributes to the early expression of gr2 in rainbow trout gills during salinity acclimation.

Production of a Fungal Punicalagin-Degrading Enzyme by Solid-State Fermentation: Studies of Purification and Characterization.

The present work describes the purification of an enzyme capable of degrading punicalagin. The enzyme was produced by Aspergillus niger GH1 by solid-state fermentation, and the enzyme production was induced by using ellagitannins as the sole carbon source. The purification steps included the concentration by lyophilization, desalting, anionic exchange, and gel filtration chromatography. The enzyme kinetic constants were calculated by using punicalagin, methyl gallate, and sugar beet arabinans. The molecular mass of the protein was estimated by SDS-PAGE. The identified bands were excised and digested using trypsin, and the peptides were submitted to HPLC-MS/MS analysis. The docking analysis was conducted, and a 3D model was created. The purification fold increases 75 times compared with the cell-free extract. The obtained Km values were 0.053 mM, 0.53% and 6.66 mM for punicalagin, sugar beet arabinans and methyl gallate, respectively. The optimal pH and temperature for the reaction were 5 and 40 °C, respectively. The SDS-PAGE and native PAGE analysis revealed the presence of two bands identified as α-l-arabinofuranosidase. Both enzymes were capable of degrading punicalagin and releasing ellagic acid.