Microencapsulation by spray drying of omega-3 lipids extracted from oilseeds and microalgae: Effect on polyunsaturated fatty acid composition

In this work, microencapsulation of omega-3 lipids from sustainable sources, including oilseeds such as Salvia hispanica L., Echium plantagineum L. and Camelina sativa L., microalgal lipids (from Nannochloropsis gaditana) and enzymatically produced fatty acid ethyl esters (FAEEs), was successfully done by spray drying and their outcomes studied. Previous to microencapsulation process, lipid extracts from new omega-3 sources (chia, camelina and echium oilseeds and wet microalgal biomass of Nannochloropsis) were produced using Pressurized Liquid Extraction. The best microencapsulation efficiency was achieved for chia FAEEs (76.9%), while the lowest was obtained for camelina oil (58.8%) and Nannochloropsis gaditana lipids (57.0%) microcapsules, which also showed low emulsion stability. Analysis by GC-MS showed that microencapsulation process did not negatively affect omega-3 fatty acid profile of microencapsulated extracts. The percentage of omega-3 in each used lipid was similar before and after spray drying (no significant differences at 5% level). Under the optimal conditions, produced microcapsules ranged from 1.5 to 30 μm and showed spherical shape and smooth surface without cracks. Thus, the proposed method provides an original approach to produce omega-3 microencapsulated lipid extracts from different nature, with high microencapsulation efficiency and payload, for potential applications in the development of foods enriched in omega-3 fatty acidsAuthors thank the Spanish Ministry of Education, Culture and Sport for the pre-doctoral contract (FPU 2013–01796) granted to Natalia Castejón. The authors gratefully acknowledge the financial support from Comunidad de Madrid provided through project ALGATEC-CM (P2018/BAA-4532), co-financed by the European Social Fund. Authors thank Camelina Company (Spain) for kindly provide camelina seeds, Technology Crops Int. (UK) for providing echium seeds and Novozymes (Denmark) for donating commercial lipases. The assistance of Javier Agundez (ICP–CSIC, Spain) for getting SEM images is also acknowledged. Authors also thank AlgaEnergy S.A. (Spain) and Dr. Federico Witt for kindly provide Nannochloropsis gaditana biomas

Building paths to success: a multilevel analysis of the effects of an emotional intelligence development program on the academic achievement of future teachers

Introduction: The situation generated by the recent pandemic has had several effects on education, one of them being the necessary but hasty efforts of teachers and students to adapt to the demands of a virtual classroom environment. Thus, it is essential to promote the development of digital competencies in future teachers, enabling them to create effective learning situations in the digital medium. Moreover, the European university curriculum includes a set of specific competencies (specific to each degree) and a series of generic competencies, many of which are related to emotional intelligence. Our work emerges from these specific premises and the more general need to develop emotional skills and learning abilities in virtual environments for future educators. Methods: The sample comprised 240 students in higher education, pursuing a bachelor’s degree in education at the University of Alicante, Spain (68.3% were female and average age of the participants was 19.43 years SD = 4.127). Using a quasi-experimental design with pretests, posttests, and a control group, we present the effects of an online program aimed at improving the emotional skills and academic achievement of future teachers in higher education. In this study, a 30-h emotional intelligence training program was implemented. Since the student sample was grouped into different classes, we analyzed, using a generalized linear mixed-effects model, whether students who took the program showed a significant improvement in their academic achievement at the end of their studies, compared to those who did not take it. Results: The findings show a significant improvement in the academic achievement at the end of the bachelor’s degree studies of the students in the experimental group. Discussion: These results open up a field of possibilities for the implementation of this type of training using virtual environments, enabling interventions to enhance the emotional development of the teaching staff (and, subsequently, in their students), develop their teaching careers adequately, and employ the ideal strategies to address educational programs. Looking ahead, the integration of emotional development programs that incorporate emerging technology into university curricula could enhance the preparation of educators profoundly and create adaptable learning environments for contemporary students.The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research has been funded by MCIN/AEI/10.13039/501100011033/FEDER, UE (Project Ref. PID2021-125279OB-I00

Small RNAs of Haloferax mediterranei: Identification and Potential Involvement in Nitrogen Metabolism

Small RNAs have been studied in detail in domains Bacteria and Eukarya but, in the case of the domain Archaea, the knowledge is scarce and the physiological function of these small RNAs (sRNAs) is still uncertain. To extend the knowledge of sRNAs in the domain Archaea and their possible role in the regulation of the nitrogen assimilation metabolism in haloarchaea, Haloferax mediterranei has been used as a model microorganism. The bioinformatic approach has allowed for the prediction of 295 putative sRNAs genes in the genome of H. mediterranei, 88 of which have been verified by means of RNA-Sequencing (RNA-Seq). The secondary structure of these sRNAs and their possible targets have been identified. Curiously, some of them present as possible target genes relating to nitrogen assimilation, such as glutamate dehydrogenase and the nitrogen regulatory PII protein. Analysis of RNA-Seq data has also revealed differences in the expression pattern of 16 sRNAs according to the nitrogen source. Consequently, RNomic and bioinformatic approaches used in this work have allowed for the identification of new sRNAs in H. mediterranei, some of which show different expression patterns depending on the nitrogen source. This suggests that these sRNAs could be involved in the regulation of nitrogen assimilation and can constitute an important gene regulatory network.This work was supported by grant BIO2013-42921-P from the Spanish Ministry of Economy and Competitiveness. Funds from “Programa Propio para el fomento de la I+D+i del Vicerrectorado de Investigación y Transferencia de Conocimiento” de la Universidad de Alicante are also acknowledged

Identification of clinical variants beyond the exome in inborn errors of metabolism

Inborn errors of metabolism (IEM) constitute a huge group of rare diseases affecting 1 in every 1000 newborns. Next-generation sequencing has transformed the diagnosis of IEM, leading to its proposed use as a second-tier technology for confirming cases detected by clinical/biochemical studies or newborn screening. The diagnosis rate is, however, still not 100%. This paper reports the use of a personalized multi-omics (metabolomic, genomic and transcriptomic) pipeline plus functional genomics to aid in the genetic diagnosis of six unsolved cases, with a clinical and/or biochemical diagnosis of galactosemia, mucopolysaccharidosis type I (MPS I), maple syrup urine disease (MSUD), hyperphenylalaninemia (HPA), citrullinemia, or urea cycle deficiency. Eight novel variants in six genes were identified: six (four of them deep intronic) located in GALE, IDUA, PTS, ASS1 and OTC, all affecting the splicing process, and two located in the promoters of IDUA and PTS, thus affecting these genes’ expression. All the new variants were subjected to functional analysis to verify their pathogenic effects. This work underscores how the combination of different omics technologies and functional analysis can solve elusive cases in clinical practic

Coordinación interuniversitaria para la realización virtual de prácticas sanitarias a través de videoconferencias

El Proyecto de Innovación presente tuvo como objetivo fundamental continuar con la experiencia de la I Jornada Interuniversitaria online de Práctica Logopédica adecuándola al sentido de un proyecto de innovación docente propiamente. Para ello, no se trataba únicamente de ofrecer conferencias a estudiantes con necesidades de formación práctica, sino dotar de una estructura y sentido formativo a los documentos audiovisuales. El objetivo general fue entonces el dotar de estructura tanto a nivel teórico como a nivel aplicado a un banco de vídeos destinados a la formación práctica de estudiantes del Grado de Logopedia

Herramientas bioinformáticas para el diagnóstico genético preimplantacional mediante secuenciación masiva

El diagnóstico genético preimplantacional de aneuploidías (DGP-A) se define como el procedimiento que analiza si los embriones están libres de anomalías cromosómicas para ser transferidos al útero materno. La técnica preferida empléa la secuenciación masiva (NGS). Durante la elaboración de la librería los fragmentos de ADN a secuenciar no se originan por fragmentación del material original sino por amplificación a través de cebadores aleatorios. Ésto produce sesgos que generan dos fuentes de artefactos: una por hibridación de cebadores aleatorios en ADN amplificado en lugar de ADN genómico; la segunda por amplificación de la librería tras la ligación del adaptador. Esto se denomina duplicados de PCR y puede enmascarar los resultados. La NGS puso de manifiesto el fenómeno del mosaicismo (dos o más líneas celulares distintas en un embrión). La bibliografía recoge que estas anomalías producen bajos ratios de implantación en procesos de reproducción asistida. Algunos estudios afirman que las tasas de éxito en transferencia de embriones con bajo porcentaje de mosaicismo son en realidad falsos positivos. Otras corrientes acusan que las técnicas presentan falsos negativos, describiendo casos donde embriones determinados como euploides engendraron bebés mosaico. En cualquier caso, todos coinciden en que la determinación del porcentaje exacto de aneuplodía es un elemento crítico para determinar la probabilidad de implantación de un embrión mosiaico. La presente tesis desarrolla un algoritmo destinado al filtrado de duplicados y artefactos de PCR, MiNFilterDups y un algoritmo que permite la detección de porcentajes bajos de aneuploidía y determinación del nivel de mosaicismo, el MiNmos. Para su validación se generaron varios conjuntos de archivos a partir de los datos de embriones de pacientes. El DGP también puede ser aplicado en determinación de embriones libres de enfermedades monogénicas (DGP-M). Las técnicas son variadas pero todas deben enfrentarse al fenómeno ADO (Allele Drop-Out) o amplificación preferencial de un alelo frente al otro. Esto genera incertidumbre respecto al resultado emitido ya que si un locus aparece como homocigoto y no se detecta la alteración en estudio, cabe la posibilidad de que el alelo mutado no haya sido amplificado, constituyendo un falso negativo desencadene la transferencia de un embrión enfermo. Para evitar esto, se analizan varios polimorfismos adjacentes al locus mutado y se determina si cosegregan con el alelo mutado o el silvestre. Aunque la gran densidad a lo largo del genoma constituye una ventaja, secuenciar todos los polimorfismos de una región es redundante y costoso; se puede reducir el número empleando estrategias de selección de tagSNPs (polimorfismos que representan a otros) basadas en el desequilibrio de ligamiento. Multitud de parámetros permiten calcular si un SNP puede ser o no útil en la predicción de otros, pero la selección de polimorfismos útiles para DGP-M requiere considerar más factores que la correlación, pues la mayoría de técnicas obtienen paneles que no son útiles para distinguir fenómenos de recombinación. MiNtagSNP es un algoritmo de selección de tagSNPs útiles en DGP-M, que permite, determinar los polimorfismos presentes en cada embrión y descartar aquellos que porten la alteración. La validación in silico se realizó empleando datos simulados a partir de las principales bases de datos de polimorfismos. Para la validación in vitro, se comprobó la concordancia de los resultados con respecto a los métodos tradicionales. Finalmente, se implementó en el laboratorio, y se realizó un seguimiento de los casos en un periodo de tiempo. Podemos concluir que el principal objetivo de los algoritmos desarrollados en el marco de esta tesis persigue el diseño de un método de análisis rápido y eficaz que permita aunar los procesos de análisis de DGP-A y DGP-M mediante secuenciación por NGS a partir de una biopsia única.Preimplantation Genetic Testing for Aneuploidy (PGT-A) is defined as the analysis of embryos for chromosomal abnormalities before being transferred to the mother's uterus. The preferred technique for PGT-A is Next Generation Sequencing (NGS). Library elaboration protocol is slightly different with respect to other genetic diagnostic techniques, since DNA fragments to be sequenced do not originate by fragmentation but by amplification through random primers that bind to the original DNA. Biases occur during the process that generate artifacts due to the hybridization of random primers in amplified DNA instead of genomic DNA or the amplification of the library after adapter ligation, named PCR duplicates, which can mask the results. NGS techniques reveal a phenomenon that had gone unnoticed, mosaicism (several cell lines in an embryo). Bibliography shows these anomalies produce low implantation rates. Furthermore, some studies affirm that the success rates in embryo transfers with a low percentage of mosaicism are actually false positives. On the other hand, other studies accuse that the techniques present false negatives, reporting cases where embryos determined as euploids generated mosaic newborns. Due to this fact, developing a precise method for determining mosaicism, controlling levels of sensitivity and specificity is an essential. We present MiNFilterDups, an algorithm for filtering duplicates and PCR artifacts and a second algorithm, MiNmos, to detect low percentages of aneuploidy and determine the level of mosaicism. For validation, several sets of files were generated from embryos of patients who had undergone an assisted reproduction process. Molecular diagnosis can also be applied in the determination of monogenic diseases named, PGT-M (Preimplantation Genetic Testing to Monogenic Diseases). All of the techniques must face the ADO (Allele Drop-Out) phenomenon (preferential amplification of one allele) which causes a heterozygous locus appears as homozygous because one of the two alleles is not amplified. This cause some uncertainty regarding the result issued since the alteration under study is not detected, it is possible that the mutated allele has not been amplified, constituting a false negative that can trigger the transfer of a diseased embryo. To avoid this problem, generally not only the mutation is studied, but several polymorphisms adjacent to the mutated locus are also analysed. Through a linkage study, it is determined whether these polymorphisms cosegregate with the mutated allele or with the wild one. The advantage of using SNPs is the high density that present throughout the entire genome, allowing embryos to be discarded if the polymorphisms associated with the affected chromosome are present. Sequencing all SNPs in a region is redundant and expensive; Fortunately, the number can be reduced by tagSNP strategies (polymorphisms that represent others). There are many parameters that allow us to calculate whether or not one SNP may be useful in predicting others, but in the specific case of selecting useful polymorphisms for PGT-M, consideration of more factors than the correlation between them is required to obtain panels that are useful when it comes to distinguishing recombination phenomena. We present MiNtagSNP, an algorithm for the selection of tagSNPs useful in PGT-M, which allows to discard embryos that carry the alteration. In silico validation was performed using simulated data from the main polymorphism databases. For the in vitro validation, results were checked for agreement with traditional methods. Finally, methodology was implemented in the laboratory, and the cases were followed up over a period of time. Thus, we can conclude that the main objective of the algorithms presented within the framework of this thesis pursues the design of a fast and efficient analysis method that allows combining the analysis processes of PGT-A and PGT-M by NGS techniques from of a single biopsy

Nuevas fuentes de aceites ricos en omega-3 para la producción de lípidos saludables mediante tecnologías limpias de extracción y modificación enzimática

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Química Física Aplicada. Fecha de lectura: 31-10-2018Esta tesis tiene embargado el acceso al texto completo hasta el 30-04-2020En esta memoria se presentan los resultados más relevantes obtenidos a lo largo del desarrollo de la presente Tesis doctoral, en la que se ha estudiado la producción de lípidos saludables a partir de nuevas fuentes de ácidos grasos omega-3 incluyendo semillas naturales, semillas modificadas por técnicas de ingeniería genética y microalgas. Los procesos desarrollados se caracterizaron por combinar técnicas avanzadas de extracción y disolventes medioambientalmente limpios como acetato de etilo, etanol, agua o sus mezclas. En conjunto, con la modificación enzimática de lípidos, se lograron extractos lipídicos enriquecidos en ácidos grasos omega-3 con propiedades beneficiosas para la salud. Así, se llevó a cabo el estudio de la extracción de aceite de semillas oleaginosas de chía, camelina o echium mediante la utilización de técnicas avanzadas de extracción: líquidos presurizados (PLE), extracción asistida por ultrasonidos (UAE) y extracción asistida por microondas (MAE). Además, se optimizó la influencia de los parámetros tiempo, temperatura y potencia en el rendimiento y composición del aceite extraído. Se obtuvieron rendimientos de extracción de aceite similares al método tradicional Soxhlet empleando PLE y disolventes verdes. UAE se caracterizó por lograr importantes rendimientos de aceite a baja temperatura (entre 30 y 55 °C) y, MAE en las condiciones óptimas alcanzó en solo 2 minutos el 60% del aceite que se obtiene en 8 horas con el método tradicional. El perfil de ácidos grasos de los aceites obtenidos no varió, independientemente del método y técnica de extracción empleado, con un contenido de α-linolénico (ALA) del 65% para el aceite de chía, 30% para el aceite de camelina y, el aceite de echium se caracterizó por un 35% de ALA y un 15% de ácido estearidónico (SDA), obteniendo una relación ω-6/ω-3 extraordinariamente baja en comparación con los aceites habituales de consumo. Al mismo tiempo que el aceite extraído con PLE y disolventes verdes a partir de semillas de camelina modificada genéticamente se caracterizó por un importante porcentaje en los ácidos grasos omega-3 eicosapentaenoico (EPA) y docosahexaenoico (DHA), 6 y 4,5%, respectivamente, que no están presentes en el aceite de camelina silvestre, ni en ninguna otra planta cultivada en la actualidad. Por otro lado, se estudió la aplicación de líquidos presurizados en la extracción y fraccionamiento simultáneo de acilgliceroles y glucolípidos enriquecidos en ácidos grasos omega-3 a partir de la pasta húmeda de la microalga Nannochloropsis gaditana. La combinación del pretratamiento enzimático de la muestra y la extracción con líquidos presurizados permitió obtener fracciones enriquecidas de hasta el 53% del omega-3 EPA. Asimismo, se llevó a cabo la producción de lípidos estructurados enriquecidos en ácidos grasos omega-3 a partir de los aceites obtenidos anteriormente y aplicando biocatalizadores inmovilizados producidos en el laboratorio y mejorados por técnicas fisicoquímicas, como la pegilación, para mejorar su actividad y estabilidad, lo que permitió desarrollar procesos viables, eficaces y selectivos. Por último, los extractos lipídicos se microencapsularon con altas eficiencias superiores al 60%, para posibles aplicaciones potenciales en el desarrollo de productos fortificados y nuevos alimentos con ácidos grasos omega-3 de gran interés para la industria alimentari

Fatty Acid Composition and Cytotoxic Activity of Lipid Extracts from Nannochloropsis gaditana Produced by Green Technologies

Microalgae are alternatives and sustainable sources of omega-3 long chain-polyunsaturated fatty acids (LC-PUFA). However, the eco-friendly extraction of these bioactives remains unexplored. In this work, the use of enzyme-based methods in combination with ultrasounds was evaluated as green approaches to extract the omega-3 lipids from Nannochloropsis gaditana. Three commercial enzymatic solutions (Viscozyme® L, Celluclast® 1.5 L, and Saczyme®) were investigated, and results were compared with the traditional Folch method. A promising extraction approach was developed by using Saczyme®, achieving a lipid yield of 25.7% ± 0.5, comparable to the traditional method (27.3% ± 0.7) (p > 0.05). Similar omega-3 content was found by GC–MS analysis for both lipid extracts (30.2% ± 2.4 and 29.3% ± 0.8 for the green and the traditional method, respectively), showing that the green approaches did not affect the fatty acid profile. Moreover, the cytotoxic activity of produced lipids was assessed by comparing human colon cancer cells (HCT-116) and epithelial nontumorigenic immortalized cells (HCEC-1CT). Results suggest that the lipid extracts have a selective effect, reducing the viability of the colon carcinoma cells but not the nontumorigenic cells. Thus, this study provides new eco-innovative approaches for extracting the omega-3 LC-PUFA from microalgae with promising biological properties

Advanced Extraction of Lipids with DHA from Isochrysis galbana with Enzymatic Pre-Treatment Combined with Pressurized Liquids and Ultrasound Assisted Extractions

Microalgal biomass is a sustainable and valuable source of lipids with omega-3 fatty acids. The efficient extraction of lipids from microalgae requires fast and alternative extraction methods, frequently combined with biomass pre-treatment by different procedures. In this work, Pressurized liquid extraction (PLE) was optimized and compared with traditional lipid extraction methods, Folch and Bligh and Dyer, and with a new Ultrasound Assisted Extraction (UAE) method for lipids from microalgae Isochrysis galbana. To further optimize PLE and UAE, enzymatic pre-treatment of microalga Isochrysis galbana was studied with commercial enzymes Viscozyme and Celluclast. No significant differences were found for lipid yields among different extraction techniques used. However, advanced extraction techniques with or without pre-treatment are a green, fast, and toxic solvent free alternative to traditional techniques. Lipid composition of Isochrysis was determined by HPLC-ELSD and included neutral and polar lipids, showing that each fraction comprised different contents in omega-3 polyunsaturated fatty acids (PUFA). The highest polar lipids content was achieved with UAE (50 °C and 15 min) and PLE (100 °C) techniques. Moreover, the highest omega-3 PUFA (33.2%), eicosapentaenoic acid (EPA) (3.3%) and docosahexaenoic acid (DHA) (12.0%) contents were achieved with the advanced technique UAE, showing the optimized method as a practical alternative to produce valuable lipids for food and nutraceutical applications

Exploring the potential of icelandic seaweeds extracts produced by aqueous pulsed electric fields-assisted extraction for cosmetic applications

Funding Information: The project was partly supported by the Science and technology assistance project of Xinjiang (No. 2014AB033), the science and technology plan project Technology Research Development plan project of Shaanxi Province, China (No. 2014K01-17-07). Science and Technology Major Project of Xianyang city (No. 2014k01-15), Shaanxi Province, China. Funding Information: The project was partly supported by the Science and technology assistance project of Xinjiang (No. 2014AB033), the science and technology plan project of Xi’an city (No. NC1317 (1)), the Science and Technology Research Development plan project of Shaanxi Province, China (No. 2014K01-17-07). Science and Technology Major Project of Xianyang city (No. 2014k01-15), Shaanxi Province, China. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.A growing concern for overall health is driving a global market of natural ingredients not only in the food industry but also in the cosmetic field. In this study, a screening on potential cosmetic applications of aqueous extracts from three Icelandic seaweeds produced by pulsed electric fields (PEF) was performed. Produced extracts by PEF from Ulva lactuca, Alaria esculenta and Palmaria palmata were compared with the traditional hot water extraction in terms of polyphenol, flavonoid and carbohydrate content. Moreover, antioxidant properties and enzymatic inhibitory activities were evaluated by using in vitro assays. PEF exhibited similar results to the traditional method, showing several advantages such as its non-thermal nature and shorter extraction time. Amongst the three Icelandic species, Alaria esculenta showed the highest content of phenolic (mean value 8869.7 µg GAE/g dw) and flavonoid (mean value 12,098.7 µg QE/g dw) compounds, also exhibiting the highest antioxidant capacities. Moreover, Alaria esculenta extracts exhibited excellent anti-enzymatic activities (76.9, 72.8, 93.0 and 100% for collagenase, elastase, tyrosinase and hyaluronidase, respectively) for their use in skin whitening and anti-aging products. Thus, our preliminary study suggests that Icelandic Alaria esculenta-based extracts produced by PEF could be used as potential ingredients for natural cosmetic and cosmeceutical formulations.Peer reviewe