Supercritical CO2 extraction increases the recovery levels of omega-3 fatty acids in Tetraselmis chuii extracts.

Tetraselmis chuii is a microalgae marketed as ingredient meeting the acceptance criteria for novel foods established by the European Union and can be an important source of healthy fatty acids (FA). The aim of this research was to characterize the FA profile of T. chuii fractions obtained by supercritical carbon dioxide (SCCO2) extraction operating with two sequential co-solvents and to evaluate the effect of biomass pretreatment (freeze/thaw cycles followed by ultrasounds). T. chuii biomass was confirmed to be an important source of omega-3 FA, mainly due to the abundance of α-linolenic acid, and pre-treatment significantly improved the lipid yield. Other omega-3 FA, such as 16:3, 16:4, 18:4, 18:5, 20:3 and 20:5, were also detected in different proportions. When SCCO2 extraction of pretreated and un-pretreated T. chuii was compared with conventional solvent extraction, the nutritional quality indices of the extracts were improved by the use of SCCO2.

Impact of the biomass pretreatment and simulated gastrointestinal digestion on the digestibility and antioxidant activity of microalgae Chlorella vulgaris and Tetraselmis chuii.

Chlorella vulgaris and Tetraselmis chuii are two microalgae species already marketed because of their richness in high-value and health-beneficial compounds. Previous studies have demonstrated the biological properties of compounds isolated from both microalgae, although data are not yet available on the impact that pre-treatment and gastrointestinal digestion could exert on these properties. The aim of the present study was to analyze the impact of the biomass pre-treatment (freeze/thaw cycles plus ultrasounds) and simulated gastrointestinal digestion in the bioaccessibility and in vitro antioxidant activity (ABTS, ORAC, Q-FRAP, Q-DPPH) of the released digests. The cell wall from microalgae were susceptible to the pre-treatment and the action of saliva and gastric enzymes, releasing bioactive peptides and phenolic compounds that contributed to the potent antioxidant activity of digests through their radical scavenging and iron reduction capacities. Our findings suggest the potential of these microalgae against oxidative stress-associated diseases at both, intestinal and systemic level.

Correction: Transcription factor induced conversion of human fibroblasts towards the hair cell lineage.

[This corrects the article DOI: 10.1371/journal.pone.0200210.].

The mitochondrial signature of cultured endothelial cells in sepsis: Identifying potential targets for treatment.

Sepsis is the most common cause of death from infection in the world. Unfortunately, there is no specific treatment for patients with sepsis, and management relies on infection control and support of organ function. A better understanding of the underlying pathophysiology of this syndrome will help to develop innovative therapies. In this regard, it has been widely reported that endothelial cell activation and dysfunction are major contributors to the development of sepsis. This review aims to provide a comprehensive overview of emerging findings highlighting the prominent role of mitochondria in the endothelial response in in vitro experimental models of sepsis. Additionally, we discuss potential mitochondrial targets that have demonstrated protective effects in preclinical investigations against sepsis. These promising findings hold the potential to pave the way for future clinical trials in the field.

Bioactivity and Digestibility of Microalgae Tetraselmis sp. and Nannochloropsis sp. as Basis of Their Potential as Novel Functional Foods.

It is estimated that by 2050, the world's population will exceed 10 billion people, which will lead to a deterioration in global food security. To avoid aggravating this problem, FAO and WHO have recommended dietary changes to reduce the intake of animal calories and increase the consumption of sustainable, nutrient-rich, and calorie-efficient products. Moreover, due to the worldwide rising incidence of non-communicable diseases and the demonstrated impact of diet on the risk of these disorders, the current established food pattern is focused on the consumption of foods that have functionality for health. Among promising sources of functional foods, microalgae are gaining worldwide attention because of their richness in high-value compounds with potential health benefits. However, despite the great opportunities to exploit microalgae in functional food industry, their use remains limited by challenges related to species diversity and variations in cultivation factors, changes in functional composition during extraction procedures, and limited evidence on the safety and bioavailability of microalgae bioactives. The aim of this review is to provide an updated and comprehensive discussion on the nutritional value, biological effects, and digestibility of two microalgae genera, Tetraselmis and Nannochloropsis, as basis of their potential as ingredients for the development of functional foods.

Influence of Dietary Algae Meal on Lipid Oxidation and Volatile Profile of Meat from Lambs with Competent Reticular Groove Reflex.

Dietary lipid sources influence intramuscular fatty acid composition, which in turn may affect the volatile profile of meat. The aim of this work was to investigate the effects of marine algae supplementation (Aurantiochytrium limacinum) on volatile compounds of cooked lamb meat. Forty-eight lambs with 42 days of age were divided into three groups: lambs fed a conventional diet without algae meal supplementation (NOALG), lambs with competent reticular groove reflex (RGR) fed the same diet supplemented with 2.5% marine algae meal mixed in the concentrate (ALGCON), and lambs with competent RGR, receiving the same diet and fed with 2.5% marine algae meal in a milk replacer to bypass the rumen (ALGMILK). Lipid and protein oxidation in raw meat was assessed and volatile compounds in grilled meat were determined. The highest and lowest lipid oxidations were observed in the ALGMILK and NOALG groups, respectively. Protein oxidation was unaffected. Out of 56 identified compounds, 12 volatiles significantly increased in both algae groups and 6 of them exclusively in the ALGCON treatment. Algae meal supplementation and its form of administration, either protected or not from rumen degradation, are important factors to consider in lipid oxidation and the aromatic profile of lamb meat.

Enrichment of Ewe's Milk with Dietary n-3 Fatty Acids from Palm, Linseed and Algae Oils in Isoenergetic Rations.

Increasing the levels of n-3 fatty acids (FA) in dairy products is an important goal in terms of enhancing the nutritional value of these foods for the consumer. The purpose of this research was to evaluate the effects of linseed and algae oil supplements in ovine isoenergetic diets on healthy milk fatty acid composition, mainly n-3. Seventy-two Churra dairy ewes were divided and randomly assigned to four experimental treatments for 6 weeks. The treatments consisted of a TMR (40:60 forage:concentrate ratio) that varied according to the inclusion of different types of fat (23 g/100 g TMR): hydrogenated palm oil (control), linseed oil (LO), calcium soap of linseed oil (CaS-LO) and marine algae oil (AO). The most effective lipid supplement to increase n-3 FA in milk was AO. 22:6 n-3 and total n-3 PUFA content increased from 0.02 and 0.60% (control) to 2.63 and 3.53% (AO), respectively. All diets supplemented with n-3 FA diminished the content of saturated FA in milk and its atherogenic index, while the levels of trans-11 18:1 and cis-9 trans-11 18:2 significantly increased. Overall, the enhancement of n-3 FA in ewe's milk would be advantageous for the manufacture of nutritionally improved cheeses.

Effects of Extending Milk Replacer Feeding during the Fattening Period on the Behaviour and Welfare of Lambs: A Preliminary Study.

There is a lack of information on the behavioural and welfare effects of sustaining artificial milk feeding in fattening lambs. Therefore, the present work aimed to study the effects of prolonged artificial milk feeding during fattening with a high concentrate diet on the behaviour of lambs. The behaviour of 16 non castrated male lambs of the Manchega sheep breed (eight lambs were in the group that were fed daily a bottle of milk, and the other eight were in the weaned group) was recorded with four fixed cameras just before bottle feeding (~8:30 a.m.) of the unweaned group till four hours later, every day for 7 weeks. The solid diet (pelleted concentrate plus cereal straw) and housing conditions were the same in both groups. Solid feeds were offered ad libitum. There were no differences between groups in time spent eating nor in drinking, playing, scratching and oral activity behaviours (p > 0.05), but resting episodes were longer in weaned lambs (p < 0.05). Weaned lambs presented a higher frequency of self-grooming behaviour (p < 0.05), while the unweaned group performed a higher frequency of interaction behaviour (p < 0.05). In conclusion, the behaviours of lambs that were fed daily a bottle of milk during the fattening period did not substantially differ from the weaned individuals.

Cell Therapy for Colorectal Cancer: The Promise of Chimeric Antigen Receptor (CAR)-T Cells.

Colorectal cancer (CRC) is a global public health problem as it is the third most prevalent and the second most lethal cancer worldwide. Major efforts are underway to understand its molecular pathways as well as to define the tumour-associated antigens (TAAs) and tumour-specific antigens (TSAs) or neoantigens, in order to develop an effective treatment. Cell therapies are currently gaining importance, and more specifically chimeric antigen receptor (CAR)-T cell therapy, in which genetically modified T cells are redirected against the tumour antigen of interest. This immunotherapy has emerged as one of the most promising advances in cancer treatment, having successfully demonstrated its efficacy in haematological malignancies. However, in solid tumours, such as colon cancer, it is proving difficult to achieve the same results due to the shortage of TSAs, on-target off-tumour effects, low CAR-T cell infiltration and the immunosuppressive microenvironment. To address these challenges in CRC, new approaches are proposed, including combined therapies, the regional administration of CAR-T cells and more complex CAR structures, among others. This review comprehensively summarises the current landscape of CAR-T cell therapy in CRC from the potential tumour targets to the preclinical studies and clinical trials, as well as the limitations and future perspectives of this novel antitumour strategy.

Intramuscular Fatty Acids in Meat Could Predict Enteric Methane Production by Fattening Lambs.

Methane (CH4) emissions pose a serious problem for the environmental sustainability of ruminant production. The aim of the present study was to explore the usefulness of the intramuscular fatty acid (FA) profile to estimate CH4 production of lambs fattened under intensive feeding systems. A statistical regression analysis of intramuscular FA derived from ruminal metabolism was carried out to assess the best predictive model of CH4 production (g/d) in lambs fed with different diets. CH4 was calculated with three distinct equations based on organic matter digestibility (OMD) at maintenance feeding levels. The OMD of the experimental diets was determined in an in vivo digestibility trial by means of the indicator method. Regression models were obtained by stepwise regression analysis. The three optimized models showed high adjusted coefficients of determination (R2adj = 0.74-0.93) and concordance correlation coefficients (CCC = 0.89-0.98), as well as small root mean square prediction errors (RMSPE = 0.29-0.40 g/d). The best single predictor was vaccenic acid (trans-11 C18:1), a bioactive FA that is formed in the rumen to a different extent depending on dietary composition. Based on our data and further published lamb research, we propose a novel regression model for CH4 production with excellent outcomes: CH4 (g/d) = -1.98 (±1.284)-0.87 (±0.231) × trans-11 C18:1 + 0.79 (±0.045) × BW (R2adj = 0.97; RMSPE = 0.76 g/d; CCC = 0.98). In conclusion, these results indicate that specific intramuscular FA and average BW during fattening could be useful to predict CH4 production of lambs fed high concentrate diets.

Cyclopropane Fatty Acids as Quality Biomarkers of Cheeses from Ewes Fed Hay- and Silage-Based Diets.

This study aimed to investigate cyclopropane fatty acids (CPFAs) as quality biomarkers of forage feedings in cheese fat obtained from ewe's milk, based on two different dietary treatments (hay and silage). The gas chromatography-mass spectrometry (GC-MS) analysis detected CPFAs in most cheese samples, both from hay and silage-based diets. CPFA levels in cheese fat from hay feeding were positively correlated to the total trans-monounsaturated fatty acids (MUFAs) and n-6 polyunsaturated fatty acids (PUFAs), whereas they were negatively correlated to cis-MUFAs, odd- and branched-chain fatty acids (i.e., C13:0 anteiso, C16:0 iso, and C17:1), and C22:5n-3, which are mainly associated with a low starch intake and grass pasture. Overall, the presence of CPFAs in ovine cheese fat suggests the use of silage, but it can also be an indicator of poor-quality hay forages. This approach confirmed the reliability of CPFAs as biomarkers of forage quality, especially in relation to the use of conserved forages and good livestock practices.

Effects of Algae Meal Supplementation in Feedlot Lambs with Competent Reticular Groove Reflex on Growth Performance, Carcass Traits and Meat Characteristics.

There is growing interest in increasing omega-3 fatty acid (FA) contents in ruminant meat by means of dietary manipulation, but the effects of such manipulation on productive results and meat quality need to be ascertained. The aim of the present study was to assess the effects of supplementing lambs with competent reticular groove reflex (RGR) with marine algae as a source of omega-3 fatty acids on growth performance, carcass traits, and meat quality characteristics. Forty-eight feedlot lambs were distributed into three equal groups: the control group neither consumed marine algae nor had competent RGR, the second group received daily 2.5% of algae meal mixed in the concentrate, and the last group consumed the same amount of algae meal, but emulsified in a milk replacer and bottle-fed. Lambs in the second and third groups had competent RGR. There were not any negative effects on performance, carcass or meat quality parameters with algae supplementation. However, the results of the oxidative stability parameters were not conclusive. Ageing for 6 days improved meat tenderness and color, and increased lipid oxidation. In conclusion, algae meal inclusion in the diet of fattening lambs with competent RGR has no detrimental effects on animal performance, carcass traits or meat quality characteristics.

Feeding Algae Meal to Feedlot Lambs with Competent Reticular Groove Reflex Increases Omega-3 Fatty Acids in Meat.

The aim of this study was to compare the effects of supplementing marine algae as a source of omega-3 fatty acids (FA) in the diet, mixed in the concentrate or bottle-fed, on intramuscular fat FA composition of lambs with competent reticular groove reflex (RGR). Forty-eight feedlot lambs were distributed in three equal groups: one group did not consume marine algae nor had competent RGR, the second group received a daily dose (2.5%) of algae meal in the concentrate and the last group consumed the same dose of algae meal emulsified in milk replacer and bottle-fed. Marine algae raised the contents of EPA, DPA, and mainly DHA in the intramuscular fat, but the increase was significantly higher when algae meal was administered with a bottle via RGR. This strategy could contribute to improvements in the marketing of lamb meat by optimizing its status as a healthier food.

Odd- and Branched-Chain Fatty Acids in Lamb Meat as Potential Indicators of Fattening Diet Characteristics.

There is a growing interest of researchers in meat authentication in terms of geographical and dietary background of animals, and several analytical methods have been proposed for the purpose of investigating this. We hypothesized that the odd- and branched-chain fatty acid (OBCFA) profile in intramuscular fat (IMF) might suffice to distinguish lamb meat entering the food chain supply on the basis of the type of diet fed to lambs during the fattening period. A total of 30 individual OBCFA profiles, quantified by gas chromatography, of IMF of Manchego lambs were used. During the fattening period (42 days), the lambs were fed three diets differing in concentrate composition: (i) Control, concentrate typical of commercial fattening rations, rich in starch and based on cereals and soybean meal; (ii) Camelina, similar to Control but replacing 50% of the soybean meal with camelina meal; and (iii) Fibrous, concentrate rich in neutral detergent fiber (NDF), based on fibrous by-products and not including cereals nor soybean meal. The OBCFA were grouped into three classes (linear odd, iso and anteiso fatty acids) and were then submitted to a linear discriminant analysis, using the feeding treatments as grouping variable and the OBCFA class contents in IMF as quantitative variables. The results suggested that a high NDF to starch ratio of the concentrate, being the lowest for Control (CON) treatment and the highest for Fibrous (FIB) treatment, would be negatively related to the odd/anteiso ratio and positively related to the iso/(anteiso+odd) FA ratio in IMF. Determination of OBCFA profile in lamb meat would be useful to monitor the feeding regime (starch- or NDF-rich) of lambs entering the food chain supply.

CRISPR/CasRx Proof-of-Concept for RNA Degradation: A Future Tool against RNA Viruses?

Influenza viruses provide a great threat for the human population, causing highly contagious respiratory infections that can lead to serious clinical complications. There are a limited variety of influenza antivirals, and these antivirals are subjected to the constant emergence of resistances. Therefore, the development of new antiviral strategies to combat influenza viruses and other RNA viruses must be promoted. In this work, we design a proof-of-concept of a recently described CRISPR/Cas tool that has been proposed as a possible future RNA virus antiviral, named CRISPR/CasRx. For this, we verified the efficiency of the CasRx endonuclease in the degradation of the eGFP mRNA reporter gene and we established the best conditions for, and the efficient performance of, the CRISPR/CasRx system. The results were measured by fluorescence microscopy, flow cytometry, and qRT-PCR. The analyses demonstrated a reduction in fluorescence, regardless of the amount of eGFP reporter plasmid transfected. The analyses showed an 86-90% reduction in fluorescence by flow cytometry and a 51-80% reduction in mRNA expression by qRT-PCR. Our results demonstrate that the CasRx endonuclease is an efficient tool for eGFP mRNA knockdown. Therefore, subsequent experiments could be useful for the development of a new antiviral tool.

Prediction of Fatty Acids in Chocolates with an Emphasis on C18:1 trans Fatty Acid Positional Isomers Using ATR-FTIR Associated with Multivariate Calibration.

This work aimed to predict C18:1 TFA isomers as well as other groups of fatty acids (saturated, monounsaturated, polyunsaturated, and total TFA) in chocolates by ATR-FTIR and partial least square regression. The quantification of fatty acids in representative samples (white, dark, and milk chocolates) was rapid (<30 s) and did not require derivatization. The optimized models showed satisfactory linear correlations compared to a reference gas chromatographic method. Coefficients of correlation for prediction considering C18:1 positional isomers were 0.973 (trans 6-8), 0.991 (trans 9), 0.991 (trans 10), 0.988 (trans 11), and 0.998 (trans 12). When considering fatty acids groups, these coefficients ranged from 0.965 to 0.999. The obtained results indicate that this straightforward procedure is suitable for chocolate analysis, for determining its general lipid composition and TFA isomeric profile, which would be of great interest for quality control programs in the face of the new TFA regulations.

Fast capillary electrophoresis method for determination of docosahexaenoic and eicosapentaenoic acids in marine oils omega-3 supplements.

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are essential omega-3 fatty acids (FA) in human nutrition. Dietary supplements containing these FA are available in the market and there is a need to assess their quality to prevent frauds. The purpose of this work was to optimize and validate a fast capillary electrophoresis (CE) method to determine EPA and DHA content in marine oils omega-3 supplements. Sample preparation comprised only a saponification step and analysis time was 8 min. Validation was performed according to limits of detection and quantification, linearity, accuracy and repeatability. Ten real samples of marine oil supplements were analyzed by CE and compared to conventional gas chromatography method. No significant differences between both methods were found within 95% confidence interval. Overall, CE method was successful to FA quantification and it could be used for fast FA monitoring during omega-3 supplement manufacture, final product quality assurance and labelling.

Application of a linear regression model to study the origin of C17 branched-chain fatty acids in caprine milk fat.

This research communications addresses the hypothesis that a part of iso 17:0 and anteiso 17:0 in milk fat could come from endogenous extraruminal tissue synthesis. In order to confirm this a linear regression model was applied to calculate the proportions of iso 17:0 and anteiso 17:0 in milk fat that could come from elongation of their putative precursors iso 15:0 and anteiso 15:0, respectively. Sixteen dairy goats were allocated to two simultaneous experiments, in a crossover design with four animals per treatment and two experimental periods of 25 d. In both experiments, alfalfa hay was the sole forage and the forage to concentrate ratio (33 : 67) remained constant. Experimental diets differed on the concentrate composition, either rich in starch or neutral detergent fibre, and they were administered alone or in combination with 30 g/d of linseed oil. Iso 15:0, anteiso 15:0, iso 17:0 and anteiso 17:0, the most abundant branched-chain fatty acids in milk fat, were determined by gas chromatography using two different capillary columns. The regression model resolved that 49% of iso 17:0 and 60% of anteiso 17:0 in milk fat was formed extraruminally from iso 15:0 and anteiso 15:0 elongation.

Lipid Composition of Brazilian Chocolates and Chocolate Products with Special Emphasis on Their Fat Origin and Trans C18:1 Isomeric Profile.

The aim of this work was to provide recent fatty acid (FA) profiling of chocolates and chocolate products, principally C18:1 trans FAs (TFAs). Thirty-two samples were analyzed by gas chromatography and FAs were quantified. The total TFA content declared in chocolate labeling and the real TFA content were compared. The TFA content ranged from 0.04 to 2.51 g/100 g of sample, and it was noticed that several manufacturers were underestimating the total TFA content in their labeling. The main TFA isomers quantified were C18:1 trans-9 (0.006-0.244%), C18:1 trans-10 (0.009-0.392%), and C18:1 trans-11 (0.013-0.464%), expressed in g/100 g of sample. Principal component analysis was used to discriminate industrial fats from natural trans fats based on the isomeric TFA profile and dairy fat (DF) biomarkers allowing to group samples in four clusters: high TFA content and high DF content, high TFA content and low DF content, low TFA content and high DF content, and low TFA content and low DF content.

[Trans fatty acids and conjugated linoleic acid in food: origin and biological properties]. / Ácidos grasos trans y ácido linoleico conjugado en alimentos: origen y propiedades biológicas.

INTRODUCTION: Trans fatty acids (TFA) are minor lipid components present in different foods, including ruminant derived products, which have received great attention due to their relationship with cardiovascular disease risk. The origin of TFA in food is mainly related to the industrial hydrogenation processes of unsaturated vegetable oils, but they can also occur naturally in the digestive tract of ruminants by enzymatic biohydrogenation reactions. Both mechanisms generate similar TFA compounds. However, TFA consumption may exert different biological effects depending on the isomeric distribution, which is strongly influenced by the dietary source (i.e., industrial or natural). Industrial hydrogenated vegetable fats are rich in elaidic (trans-9 18:1) and trans-10 18:1 fatty acids, among others. In contrast, vaccenic acid (trans-11 18:1) is the major TFA isomer detected in milk and other ruminant derived products. Vaccenic acid is the physiological precursor of conjugated linoleic acid, a bioactive lipid with beneficial effects on human health. This article provides updated information on the biological effects and potential bioactive properties of TFA considering both, their chemical structure and provenance.

INTRODUCCIÓN: Los ácidos grasos trans (AGT) son componentes lipídicos minoritarios que se encuentran en distintos alimentos, entre ellos, aquellos derivados de animales rumiantes, que han merecido atención por su relación con el riesgo de incidir en enfermedades cardiovasculares. El origen de los AGT en los alimentos se encuentra mayoritariamente en los procesos de hidrogenación industrial de aceites vegetales insaturados y en las reacciones enzimáticas de biohidrogenación que tienen lugar, de forma natural, en el tracto digestivo de los rumiantes. Aunque las moléculas que se generan por ambos mecanismos son similares, la distribución isomérica de los AGT es muy diferente, lo que puede generar diferencias a la hora de evaluar los efectos biológicos derivados de su consumo. Las grasas vegetales hidrogenadas son abundantes en ácido elaídico (trans-9 18:1) y trans-10 18:1 entre otros. En contraste, el ácido vacénico (trans-11 18:1) es el principal AGT presente en la leche y otros productos derivados de rumiantes, siendo además precursor fisiológico del ácido linoleico conjugado, un componente al que se atribuyen numerosos efectos beneficiosos para la salud. En este artículo se actualizan los efectos biológicos y las potenciales propiedades bioactivas de estos ácidos grasos.