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Introducción: Las úlceras de extremidades inferiores (UEEII) tienen una incidencia de 3 a 5 nuevos casos cada 1000 personas y año. Existen diferentes etiologías que pueden producir este tipo de lesiones, diferenciándolas entre lesiones típicas y atípicas en función de su prevalencia. Las úlceras de Martorell (UM) pertenecen al grupo de lesiones atípicas con una prevalencia del 0,5%-1%. Son lesiones producidas por una HTA mal controlada y de larga evolución, cuyo desconocimiento deriva en errores e infradiagnóstico.Objetivo: Identificar los signos y síntomas diferenciales de las UM respecto a otras (UEEII).Metodología: Se llevó a cabo una scoping review mediante la búsqueda de artículos en WOS, Pubmed, CUIDEN y Dialnet, utilizando términos DeCS/MESH, palabras clave y operadores booleanos, e incluyendo criterios de selección para acotar la búsqueda. También se obtuvo información del grupo GNEAUPP.Resultados: Se analizaron 12 artículos buscando información relevante para Enfermería: historia clínica, signos y síntomas, diagnóstico, tratamiento, intervenciones de enfermería y resolución de la UM. Estos artículos se contrastaron con 2 guías de práctica clínica.Discusión: Las UM son producidas por una HTA diastólica mal controlada y de larga evolución, más frecuentes en mujeres mayores de 50 años. Su aparición es lenta, evolucionando a lesiones con necrosis y eritema perilesional. El dolor intenso y su localización son características diferenciales de la UM. El diagnóstico se confirma con biopsia, y el tratamiento consiste en antihipertensivos, analgesia y curas locales donde destaca el desbridamiento. Las intervenciones de Enfermería incluyen control de constantes, curas según evolución, y actividades de prevención y educación sanitaria.Conclusiones y aportaciones para enfermería: Las UM son lesiones desconocidas por muchos profesionales sanitarios. Dar a conocer estas lesiones es fundamental para la implantación del diagnóstico y tratamiento tempranos y correctos, evitando afectar a la calidad de vida de los pacientes. No existe un protocolo estandarizado para el abordaje de las UM, por lo que ampliar su investigación es primordial para reducir su impacto sociosanitario.

Póster presentado en: EIFS2022, the 2nd Iberian Meeting on Supercritical Fluids (2º Encontro Ibérico de Fluidos Supercríticos / 2º Encuentro Ibérico de Fluidos Supercríticos), to be held on 28.February - 2.March 2022 in Coimbra, Portugal., Increasing wastes as consequence of the growing population and higher quality of life, is one of the biggest problems that society faces with nowadays. The concept of biorefinery, in which residues are reincorporated into industrial processes to obtain base molecules, emerges as a solution. In this work, a complete valorization of the red alga Gelidium sesquipedale solid byproduct generated after industrial agar extraction (GBP) is proposed. Despite being usually discarded, it still contains high content of interesting molecules such as carbohydrates, proteins, amino acids, etc., that can be recovered. Traditional extraction methods have several drawbacks. They are usually time-consuming and have low selectivity and extraction yields. In addition, large volumes of organic solvents, which are dangerous, too expensive and can be harmful for environment and human health, are used. As an alternative, subcritical water treatment (SWT) is proposed in this work to recover the compounds of interest in the above mentioned Gelidium sesquipedale byproduct (GBP). Also, enzymatic assisted extraction (EAE) has been studied in order to compare SWT with other technologies. Different hydrolytic enzymes (cellulase and protease) have been also used for this purpose. Finally, different pressure- driven membrane technologies have been evaluated for fractionation and concentration of the subcritical water extracts, with the purpose of obtaining isolated and high-purity concentrates of the extracted bioactive compounds. The results obtained allow us to conclude that the extraction/hydrolysis taking place during SWT favor the recovery of several bioactive compounds from GBP (see Fig. 1), providing high yields through a complete byproduct valorization. SWT showed to be highly influenced by temperature, heating rate and residence time [1, 2]. SWT led to an efficient extraction/hydrolysis of the protein fraction of GBP. The best experimental conditions in a semi-continuous fix- bed reactor were 200 °C and 6 mL/min with nearly 70% of the solubilized protein content. The highest content of individual amino acids was obtained for small amino acids such as valine, alanine and glycine as well as aspartic acid. Therefore, an increase in the non- polar selectivity was observed by working at high severity factors. Positive and strong correlation was obtained between the phenolics recovered and the reducing capacity of the BP extracts. EAE has also been proven to be an efficient technology to valorize the GBP [3]. Longer times were needed and lower yields of biocompounds extraction/hydrolysis were observed for EAE in comparison with SWT. Ultrafiltration with tubular inorganic membranes has been proven to be a suitable separation technology to fractionate subcritical water extracts from macroalgae residue. The most influential parameter in separation process was the MWCO of the membrane. Further research about the functional properties of the concentrated and isolated biocompounds is needed in order to study their possible applications., AEI [grant numbers PID2019-104950RB-I00 and PID2020-116716RJ-I00 / AEI / https:// doi.org/10.13039/501100011033]. JCyL and ERDF [grant number BU05P20]. JCyL for E. Trigueros and P. Alonso-Riaño predoctoral contracts ORDEN EDU/574/2018 and EDU/556/2019. BG20/00185 for R. Melgosa Beatriz Galindo Research Fellowship. JCyL for AE Illera postdoctoral contract [BU05P20]

Trabajo presentado en: EIFS2022, the 2nd Iberian Meeting on Supercritical Fluids (2º Encontro Ibérico de Fluidos Supercríticos / 2º Encuentro Ibérico de Fluidos Supercríticos), to be held on 28.February - 2.March 2022 in Coimbra, Portugal., Nowadays, the society is immersed in the transition from a linear to a circular economy, in which the value of products, materials and resources is maintained in the economy for as long as possible and the generation of waste is minimized. This means that the byproduct of a process becomes the input of a new one where it acquires new value. Among other agricultural by-products, onion (Allium cepa L.) skin wastes offer great potential for valorization. Onion is the second most important horticultural crop worldwide. More specifically, Spain, in the year 2018 produced 1.27 Mt [1]. The onion industry generates every year more than 0.5 Mt of Onion Skin Wastes (OSW) worldwide [2], including skins (the outermost layers), roots and bulbs unfit for consumption. The non-edible brown skin and external layers of onions are rich in phenolic compounds, mainly flavonoids such as quercetin (QC) [3] and its derivatives: quercetin 4’-O-β- glycoside (QC4’), quercetin 3,4’-O-β-diglycoside (QC3,4’) and quercetin 3-O-β- glycoside (QC3). All of them are high-added value natural antioxidants [2]. The main drawback of quercetin and quercetin derivatives is the limited solubility in water, which limits their oral bioavailability [4] and extractability, and forces the use of an organic solvent to successfully extract them. Alternatively, Subcritical Water (SubW) can be used to extract flavonoids from onion skins. SubW refers to water at temperatures ranging from 100 °C (boiling point) to 374 °C (critical point) which remains in a liquid state due to the application of pressure. Changes in the working conditions change the properties of the SubW (among them, viscosity, surface tension and dielectric constant, which can be similar to those of some organic solvents), enhancing mass transfer and the extractability of barely water-soluble bioactive compounds, as summarized by Benito-Roman et al. [5], as SubW favors the hydrolysis of the bonds between phenolic compounds and the vegetable matrix. In this work the extraction, identification and quantification of phenolic compounds from OSW has been studied using SubW in a semicontinuous extractor (flow rate constant and equal to 2.5 mL/min; temperatures up to 180 °C with working pressure of 5 MPa, to keep water in liquid state). The extraction of flavonoids resulted to be fast (, This work was supported by AEI [PID2019-104950RB-I00 and PID2020-116716RJ-I00] and JCyL and ERDF [BU050P20] [BU301P18]

Póster presentado en: EIFS2022, the 2nd Iberian Meeting on Supercritical Fluids (2º Encontro Ibérico de Fluidos Supercríticos / 2º Encuentro Ibérico de Fluidos Supercríticos), to be held on 28.February - 2.March 2022 in Coimbra, Portugal., Brewer’s spent grain (BSG) is one of the most important by-products in large and small- scale breweries. BSG is the solid residue generated after mashing and wort filtration process, it is generated at an average rate of 20 kg per 100 L of beer and accounts for approximately 85 % of the total residues of the brewing process [4]. Nowadays, it is mainly used for animal feed (70 %), biogas production (10 %), or landfilled [5]. However, BSG presents a valuable chemical composition with a high content of protein and carbohydrates, as well as important quantities of phenolic compounds with potential bioactive properties. BSG also contains non-negligible amount of lipids (5 %) with more than 50 % being linoleic acid (C18:2 ω-6) [6]. Due to the valuable chemical composition of BSG, different techniques have been proposed to valorize this lignocellulosic biomass, such as enzymatic and chemical hydrolysis, ultrasound assisted extraction or microwave assisted extraction [7]. High pressure processing of biomass has been also proposed since it offers unique opportunities in the extraction and valorization of the bioactive compounds of BSG. Among the different high-pressure processes, the use of supercritical CO2 (sc-CO2) presents a great attractive since it is considered a green solvent and it presents gas-like (high diffusivities) and liquid-like (good solvation power) properties at supercritical conditions (Tc = 31.1 oC pc= 7.39 MPa). Sc-CO2 has been extensively studied as a green extracting agent over traditional organic solvents to valorize the lipophilic fraction of biomass [8]. From a biorefinery perspective and aiming at the integral valorization of BSG, sc-CO2 extraction and enzymatic hydrolysis processes have been applied to this by-product. First, the extraction of the lipophilic fraction of BSG with sc-CO2 has been systematically studied. The most influential extraction conditions, namely pressure and temperature, were varied from 20 to 40 MPa and from 313 to 353 K, respectively. A maximum yield of 5.70 ± 0.07 g /100 gBSG was obtained at 353 K and 40 MPa (see Graphical abstract, Fig. a)). High pressures and temperatures resulted in higher content of total phenolic and flavonoids compounds, as well as higher antioxidant capacity. Enzymatic hydrolysis was performed on samples after sc-CO2 treatment and non-treated BSG samples. Enzymatic hydrolysis was carried out at 323 K in an acetate buffer at pH=5 with a cellulase, 1,4-(1,3:1,4)-β-D-Glucan 4-glucanohydrolase, EC 3.2.1.4, from Aspergillus niger provided by Sigma-Aldrich. The graphical abstract b) represents the glucose yield along enzymatic hydrolysis for sc- CO2 treated and non-treated BSG at different enzyme concentrations. An improvement of the enzymatic hydrolysis yield by cellulase was observed in the sc-CO2 treated BSG compared to the non-treated. This improvement could be partially attributed to the removal of the lipid fraction and to morphological changes of BSG after sc-CO2. Based on this double benefit, sc-CO2 can play an important role on biomass valorization [8]., AEI [PID2019-104950RB-I00 and PID2020-116716RJ-I00] and JCyL and ERDF [BU050P20]. P. AlonsoRiaño and E. Trigueros acknowledge funding from JCyL and ESF [Orden EDU/556/2019 and EDU/574/2018, respectively]. R. Melgosa is supported by a Beatriz Galindo Research Fellowship [BG20/00182]. Ó. Benito-Román post-doctoral contract was funded by AEI [PID2020-116716RJ-I00/AEI/10.13039/501100011033]. A.E. Illera post-doctoral contract was funded by JCyL and ERDF [BU050P20].

Póster presentado en: EIFS2022, the 2nd Iberian Meeting on Supercritical Fluids (2º Encontro Ibérico de Fluidos Supercríticos / 2º Encuentro Ibérico de Fluidos Supercríticos), to be held on 28.February - 2.March 2022 in Coimbra, Portugal., Gelidium sesquipedale solid residue after industrial agar extraction contains high amounts of proteins with all essential amino acids and carbohydrates such as glucans, galactans or arabinans [1]. Therefore, although it is generally discarded, its reincorporation in the industry as a value-added product brings an interesting solution. Traditional methods used for bioactive compounds extraction from different raw materials present numerous drawbacks, namely, time-consuming, costly to dispose of used products and harmful to environment and human health [2]. Among green technologies, subcritical water extraction (SWE) stands out as a great alternative to traditional extraction processes. SWE consists of using hot pressurized water above its boiling point, 100 °C, and below its critical point, 374 °C, which causes many of the properties of water to change, such as density or dielectric constant [3,4]. Water dielectric constant, which is related to its polarity, decreases with increasing temperature, similar to organic solvents, at 200 °C. As a result, through the dielectric constant modulation with temperature, SWE is able to selectively extract polar or non-polar compounds [5]. In order to assess the feasibility of industrial-scale subcritical solvent extraction, a pilot- scale process must first be tested. Generally, the design of the industrial SWE equipment is preceded by the study of laboratory- and pilot-scale systems. Hence, although in many cases the pilot-scale study stage is eliminated, the scaling-up process would be much more efficient by incorporating the pilot-scale study to obtain quality data and determination of scale-up factor [6]. Therefore, the main goal of this research was to prove the feasibility of industrial-scale subcritical water system through scaling-up from lab to pilot system., by the comparison of lab- and pilot-scale subcritical water performance. For this, many analytical methods were applied for the comparison of the extraction yield of the two systems; such as, polysaccharide fraction identification and quantification, total protein content and free amino acids determination, and total polyphenol content (TPC) and antioxidant activity. Galactose was mainly recovered as oligomer fraction with maximum yields of 71.4 (36 minutes) and 74.5 % (45 minutes) for pilot and lab-scale, respectively (Figure 1a). Lower yields were determined for glucans, with maximum yields of 9.5 % for both systems, in which more than 6 % was extracted in the first minutes. Similar extraction curves and yields were determined for protein fraction with final extraction yields close to 40 % (Figure 1b), while free amino acids content was higher in laboratory scale. The greatest extraction yield was accounted for the smallest amino acids, such as glycine, alanine and aspartic acid, whereas polar amino acids such as glutamic acid and lysine were reduced, although lysine was not detected in pilot system. Differences in total polyphenolic compounds (TPC) extraction were observed for both systems. Increasing TPC content with time was determined for lab-scale system, while in pilot system a plateau phase was observed after 36 minutes of extraction. SWE has been proven to be an efficient technology for bioactive compounds recovery such as carbohydrates, protein and amino acids from algae residue. Scaling up of subcritical water system from laboratory to pilot scale resulted in good and reproducible results. Therefore, feasibility of industrial-scale subcritical water system through scaling- up from lab to pilot system has been showed., This work was supported by AEI [PID2019-104950RB-I00 and PID2020-116716RJ-I00] and JCyL and ERDF [BU050P20] P. Alonso-Riaño and E. Trigueros acknowledge funding from JCyL and ESF [Orden EDU/556/2019 and EDU/574/2018, respectively]. A.E. Illera post-doctoral contract was funded by JCyL and ERDF [BU050P20]. R. Melgosa is supported by a Beatriz Galindo Research Fellowship [BG20/00182].

Póster presentado en: EIFS2022, the 2nd Iberian Meeting on Supercritical Fluids (2º Encontro Ibérico de Fluidos Supercríticos / 2º Encuentro Ibérico de Fluidos Supercríticos), to be held on 28.February - 2.March 2022 in Coimbra, Portugal., Brewer ́s spent grain (BSG) is the solid by-product generated in breweries after the mashing and wort filtration process. It comprises about 85 % of the total by-products, generating approximately 20 kg per 100 L [1]. BSG presents a valuable chemical composition with a high content of protein and carbohydrates, 5 % of lipids and an important source of phenolic compounds. Among the different techniques proposed to valorize BSG, supercritical CO2 (sc-CO2) has become a promising technology to process biomass. The main aim of this work was to assess the effect of sc-CO2, after BSG oil extraction, on the subsequent enzymatic hydrolysis to hydrolyze the polysaccharide fraction into monomeric sugars by comparing the sugar yields of sc-CO2 and non-scCO2 treated BSG. After sc-CO2 treatment at 40 MPa and 80 oC, the raffinate obtained was subjected to enzymatic hydrolysis by cellulase at different enzyme dose. At the sc-CO2 extraction the carbohydrate fraction remained in the raffinate phase after extraction. Glucose yield increased with enzyme concentration for non-treated and sc-CO2 treated BSG. Higher yields of glucose were obtained for sc-CO2 treated compared to non-treated sc-CO2 for all the enzymes concentrations. The percentage increase in glucose yield for sc-CO2 treated and non-treated BSG was 8, 14 and 18 % for the three cellulase concentrations essayed in this work, 0.25, 0.5 and 1 %, respectively. The hydrolysis kinetics for xylose and arabinose have been also determined for non-treated and sc-CO2 treated BSG at 1 % of cellulase dose. Although not big differences were observed in the final sugar yield in the hydrolysates, the initial hydrolysis rate were significant higher for the sc-CO2 treated BSG than for the untreated samples. The higher enzymatic hydrolysis rate and yield obtained in the raffinate-BSG after sc- CO2 treatment compared with non-treated BSG could be attributed partially to the removal of the lipid fraction. As it has been described in literature [2], fats and oils could influence the susceptibility of carbohydrates to enzymes. This improvement was also due to surface morphology modification. The structural and chemical changes of untreated and sc-CO2 treated BSG was assessed by scanning electron microscopy and X-ray powder diffraction (XRD). Although the XRD pattern after sc-CO2 treatment indicated that the pre-treatment was not strong enough to modify the BSG crystallinity, the raffinate exhibited an irregular porosity and lamellar structure. sc-CO2 broke partially some structural barriers allowing a better enzyme access. Furthermore, some phenolic compounds were determined in the enzymatic hydrolysates at the end of the hydrolysis by using 1 % of cellulase (Table 1). It was observed that for p-coumaric acid and ferulic acid, a concentration 31 and 24 % higher, respectively, was obtained in the BSG hydrolysates after sc-CO2 treatment, while for vanillin similar concentration was obtained in both hydrolysates. The concentration of p-coumaric acid and vanillin after cellulase hydrolysis was lower than the values previously reported for the same BSG by alkaline, xylanase (1%) and subcritical water hydrolysis [3], [4]. Nevertheless, for ferulic acid, only a higher value was reached for alkaline hydrolysis., JCyL and ERDF for financial support of project BU050P20 Agencia Estatal de Investigación for financial support of project PID2019-104950RB-I00 / AEI / 10.13039/501100011033 JCyL and ESF for E. Trigueros (ORDEN EDU/574/2018) and P. Alonso-Riaño predoctoral (EDU/556/2019) contracts R. Melgosa is supported by a Beatriz Galindo Research Fellowship [BG20/00182]

Trabajo presentado en: 18th European Meeting on Supercritical Fluids. 2021, 4-6 de mayo, Burdeos (online), Introduction Gelidium sesquipedale residue after industrial agar extraction is a very valuable by-product because of its carbohydrate, protein and bioactive compounds content. Subcritical water (SW) extraction was carried out by using different solvent flow rates (4, 6 and 8ml/min) to study the effect of residence time (RT = 6.0, 4.2 and 3.0 minutes, respectively) on sugars hydrolysis and fractionation at 185°C. Results and discussion Macroalga residue after industrial agar extraction presents an important fraction of carbohydrate composed by glucans (23.4%), galactans (10.9%) and arabinans (2.9%). Accumulated hydrolysis curves for arabinans, galactans and glucans by using SW are shown in Figure 1. No monomers were observed under work conditions in subcritical water extracts so that carbohydrate fraction was recovered as sugar oligomers. Sugars extraction curves show how the lower the residence time, the higher extraction yields for all components. Moreover, a preferential hydrolysis was observed for galactans, followed by arabinans and glucans that showed the lowest yield. As has been seen, increasing and faster extraction has been found at increasing flow rate due to promotion of mass transfer of the solutes from by-product matrix1, that is translated into a greater compounds extraction. Similar results were found in literature about residence time effect on bioactive compounds extraction2,3. Conclusions SWE constitutes an interesting technique for sugars recovery, promoting a better use of the alga residue generated in the food industry without using organic solvents in compare to conventional treatments. Low residence times led to higher sugars recovery by SW treatment., JCyL and ERDF for financial support of projects BU301P18, BU050P20 and O. Benito-Román’s post-doctoral contract. To Agencia Estatal de Investigación for project funding PID2019-104950RB-I00/AEI/10.13039/501100011033. To the company Hiperbaric through project BIOLIGNO (10/16/BU/0017), funded by ERDF and JCyL through ICE.

Published

Trabajo presentado en: 1st GREENERING International Conference. 2021, 15-17 de febrero, Costa da Caparica, Agencia Estatal de Investigación [grant number PID2019-104950RB-I00 / AEI / 10.13039/501100011033] and the Junta de Castilla y León (JCyL) and the European Regional Development Fund (ERDF) [grant numbers BU301P18 and BU050P20

Trabajo presentado en: 18th European Meeting on Supercritical Fluids. 2021, 4-6 de mayo, Burdeos (online), 1. Introduction Brewer’s spent grain (BSG) is the most abundant brewing industry by-products, comprising 85 % of the total by products generated. BSG is mainly composed of carbohydrates (52%), lignin (18%), proteins (18%) and lipids (6%). BSG is also a valuable source of phenolic compounds, with approximately 1.2 % of mono- and di-meric phenolic acids, being hydroxycinnamic acids, such as ferulic and p-coumaric acids, the primary class of phenolic compounds that have demonstrated antioxidant effect. In this work, we proposed the use of subcritical water (SW) at 50 bar and 4 mL/min in a semi-continuous reactor at different temperatures, from 125 to 185 C, to extract phenolic compounds from BSG. 2. Results and discussion Total phenolic compounds (TPC) were determined according to the Folin-Ciocalteu assay. It was found that an increase in the operating temperature led to an increase in TPC with a maximum of accumulative TPC of 33.0 ± 0.3 mg GAE/g dry-BSG at 185 °C. Figure 1 shows the accumulative profile of individual phenolic compounds at different temperatures. Aldehyde phenolic compounds such as vanillin, syringic and protocatechuic aldehyde; reached the maximum level at the highest temperature studied in this work; however, for hydroxycinnamic acids, such as ferulic acid and p-coumaric, the maximum was obtained at 160 °C. The different behavior with temperature allowed fractionation of bioactive compounds by SW. Reducing capacity of SW extracts, according to the FRAP assay, showed that increasing the temperature up to 185 C significantly increased the reducing capacity (p < 0.05). A positive correlation between reducing capacity and total phenolic compounds was stablished according to the Pearson product moment correlation (R2 = 0.9511). The high TPC value and antioxidant capacity of the extracts obtained at the highest temperatures could be due to newly formed compounds related to Maillard reactions that influence TPC and FRAP responses. 3. Conclusions This study shows that phenolic compounds were successfully extracted from BSG using subcritical water. SW extraction of phenolic compounds was maximized at the highest temperature studied in this work, 185 °C, although hydroxycinnamic acids were more sensitive to temperature than aldehyde phenolic compounds. SW extracts showed a high antioxidant capacity and allowed extraction and fractionation of phenolic compounds., JCyL and ERDF for financial support of projects BU301P18 and BU050P20 and. Benito-Román´s post-doctoral contract. To Agencia Estatal de Investigación for project funding PID2019-104950RB-I00/AEI/10.13039/501100011033. To the company Hiperbaric through project BIOLIGNO (10/16/BU/0017), funded by ERDF and JCyL through ICE. To JCyL and ESF for P. Alonso-Riaño and E. Trigueros predoctoral contracts.

Trabajo presentado en: VII Jornadas de Doctorandos de la Universidad de Burgos. 2021, 9-10 de marzo, Burgos (online), organizadas por la Escuela de Doctorado, Líneade investigaciónfinanciadapor la AgenciaEstatalde Investigación(PID2019-104950RB-I00 / AEI / 10.13039/501100011033)y laJunta de Castilla y León JCyLy la European Regional DevelopmentFund(ERDF)(BU301P18yBU050P20) Contratospredoctoralesde E. Triguerosy P. Alonso-Riaño financiadospor JCyL(ORDEN EDU/574/2018, de 28 de mayoy ORDEN EDU/556/2019, de 5 de junio,respectivamente)

Póster presentado en: 18th European Meeting on Supercritical Fluids. 2021, 4-6 de mayo, Burdeos (online), Olive leaves (OL) are one of the by-products generated during the pruning of olives tress and during separation process in olive processing. OL are lignocellulosic biomass with important amounts of extractives and lignin (Table 1). OL have a significant amount of phenolic compounds and other bioactive compounds such as mannitol. Mannitol is a sugar alcohol with reduced caloric value compared with other sugars with different applications in the food and pharmaceutical industries due to its functional properties. Mannitol production depends on the sorbitol market since is obtained as a by-product in the catalytic hydrogenation process of sorbitol production. Therefore, alternative process to obtain mannitol from natural sources avoiding the use of high temperatures would be advantageous to obtain this polyol. In this work, valorization of OL was performed by subcritical water extraction in a batch pilot plant with 20 L capacity where 3 kg of the dry OL were put in contact with subcritical water at a medium temperature of 181.1  0.6 ºC. Figures 1 and 2 show the extraction curves of mannitol and some of the main polyphenols compounds identified by HPLC/DAD. By SW treatment, mannitol extraction yield was nearly 100 % after 20 min of extraction (OL has 99  1 mg mannitol/ g dry OL). The main PC extracted by SW was oleuropein nearly 1.5 mg/g dry OL, but also verbascoside and luteolin-7-glucoside. TPC of SW extracts was 33 mg of gallic acid equivalent/g dry OL., JCyL and ERDF for financial support of projects BU301P18 and BU050P20 and to Agencia Estatal de Investigación for project funding PID2019-104950RB-I00/AEI/10.13039/501100011033. To the company Hiperbaric through project BIOLIGNO (10/16/BU/0017), funded by ERDF and JCyL through ICE. To JCyL and ESF for E. Trigueros and P. Alonso-Riaño predoctoral contracts and for the contracts of D. Benito-Bedoya and D.M. Aymara-Caiza trough the YEI program.

Trabajo presentado en: VII Jornadas de Doctorandos de la Universidad de Burgos. 2021, 9-10 de marzo, Burgos (online), organizadas por la Escuela de Doctorado, Desarrollado en el grupo BIOINDy financiado por: AEI(PID2019-104950RB-I00/AEI/10.13039/501100011033) y JCyLy FEDER(BU301P18 y BU050P20). La JCyLy el Fondo Social Europeo ha financiado los contratos predoctorales de E. Trigueros y P. A. Riaño (ORDEN EDU/574/2018, de 28 de mayo y ORDEN EDU/556/2019, de 5 de junio, respectivamente)

El cáncer mama es uno de los canceres cada vez más habituales en las mujeres. Se pronostica que 1 de cada 8 mujeres desarrollara un cáncer de mama a lo largo de su vida. El cáncer supone un factor muy estresante en la vida de las pacientes, por ello, desarrollandiferentes técnicas, estrategias de afrontamiento, que les ayudaran a sobrellevar la enfermedad así como mejorar su calidad de vida. El equipo deenfermería debe tener una especial formación para brindar al paciente la mejor atención posible.

Póster presentado en: 1st GREENERING International Conference. 2021, 15-17 de febrero, Costa da Caparica, JCyL and ERDF for financial support of projects BU301P18 and BU050P20 and Benito-Román´s post-doctoral contract. Agencia Estatal de Investigación for project funding PID2019-104950RB-I00/AEI/10.13039/501100011033.

The semi-continuous hydrolysis of onion skin wastes (OSW) using subcritical water (SubW; working conditions: 105-180 ºC; 5 MPa; 2.5 mL/min) has been studied in this work. Liquids after hydrolysis showed a sharp increase in total organic carbon (>30 mg/g OSW) at temperatures above 145 ºC, when SubW began to promote the partial hydrolysis of the structural components of OSW. Among them, pectin was one of main components recovered (extraction yield up to 9% at 145 ºC), whereas cellulose was barely hydrolyzed in the range of temperatures studied. The composition of pectin demonstrated that SubW promoted the recovery of the valuable RG-I (21.1±1.1 mol %) regions compared to acid water extraction processes (10.8±0.9 mol %). The control of the hydrolysis conditions was found to be critical, since high temperatures and long hydrolysis times led to the formation of organic acids (acetic, formic and levulinic) and degradation products such as furfural (up to 0.8 mg/g OSW) from the C5 sugars. Finally, the High Heating Value (HHV) calculated for the solid residue obtained at 180 ºC (16.4±0.2 MJ/kg) indicated the potential use of this residue as a fuel, once the valuable compounds (phenolics and pectins) have been recovered., Agencia Estatal de Investigación (AEI) though projects PID2020-116716RJ-I00/ AEI / 10.13039/501100011033 and PID2019-104950RB-I00 / AEI / 10.13039/501100011033 and by the Junta de Castilla y León (JCyL) and the European Regional Development Fund (ERDF) through projects BU301P18 and BU050P20. Benito-Román post-doctoral contract was funded by JCyL and EDRF through project BU050P20 and by AEI through project PID2020-116716RJ-I00. E. Díaz de Cerio post-doctoral contract was funded by JCyL through project BU050P20. P. Alonso-Riaño predoctoral contract was funded by JCyL through ORDEN EDU/556/2019, de 5 de junio.

Póster presentado en: 1er Encuentro Ibérico de Fluidos Supercríticos/1º Encontro Ibérico de Fluidos Supercríticos. 2020, 18-19 de febrero, Santiago de Compostela, Rice bran oil is a source of bioactive molecules such as sterols, tocols, -oryzanols and unsaturated fatty acids [1,2]. In this work, the encapsulation of rice bran oil extracted using supercritical CO2 under the conditions optimized by Benito-Román et al. [3] has been studied. Microencapsulation processes are sequential and involve the emulsion formation and then, the emulsion drying. In a first stage, the emulsification process by high pressure homogenization was studied and optimized. High pressure homogenization, also known as microfluidization, is a high energy emulsification method affected by several parameters: pressure and number of homogenization cycles (together determine the energy input), the carrier material, the carrier to core ratio, and the solids content in the emulsion. Microfluization also exhibits an important advantage: the industrial application due to flexibility to control the emulsion droplet size (EDS) and the ability to produce emulsions from a variety of materials [4]. Among the different encapsulation materials, vegetable proteins are trendy, due to their properties and the possibility to be used in pharma, cosmetics and food industries [5]. More specifically, pea proteins present the most interesting properties such as emulsifying easiness, high nutritional value and non-allergenic characteristics [6]. For these reasons, and the wall forming properties pea proteins have, key in microencapsulation processes, they were used in this work. The effect of working pressure (60-150 MPa), composition of the carrier (mixtures of pea protein isolate (PPI) and maltodextrin (MD), (from 50 to 90% of PPI) and carrier to oil ratio (COR) (from 2 to 4) on the emulsion droplet size (EDS) was studied, using the response surface methodology. The number of passes through the homogenization chamber was previously determined and set in 7. The experimental work, revealed that in order to minimize the EDS, moderate pressures (114 MPa), a carrier composed mainly by PPI (64%) and carrier to oil ratios around 3.2 are required. Important interactions between the experimental factors were also observed. In the second stage, the emulsion obtained in the optimal conditions (EDS=189±3nm) was dried using different technologies: spray-drying (Buchi B-290 mini Spray-dryer, inlet temperature 155 ºC, outlet temperature 92-96 ºC and emulsion flow rate of 3 g/min); PGSS-drying (apparatus extensively described by Melgosa et al. [7], being the main working conditionsgas to product ratio (GPR) equal to 30 g/g temperature and pressure in the static mixer of 105 ºC and 10 MPa, respectively) and freeze drying (Labconco Freeze Dry System, 0.15 mbar for, at least, 48 h). All of them were suitable to get dry powders, spray drying provided high encapsulation efficiencies (around 73%) and monomodal powders (around 18 μm), whereas PGSS drying provided lower encapsulation efficiencies (around 52%) but perfect spheres with lower particle size (around 11 μm). Freeze drying yielded powders with almost complete encapsulation efficiencies, and higher stability when stored at 4 ºC, since spray-dried and PGSS-dried powders increased the amount of free oil after two weeks of storage., JCyL and ERDF for financial support of project BU301P18. CampofríoFood Group -SIGMA through project ref. 10/16/BU/0017, funded by ERDF of the EU and JCyL, through ICE

Póster presentado en: 1er Encuentro Ibérico de Fluidos Supercríticos/1º Encontro Ibérico de Fluidos Supercríticos. 2020, 18-19 de febrero, Santiago de Compostela, Brewer’s spent grain (BSG) has been traditionally used only in animal feed despite its high nutritional value, with a protein content of ~20% in dry weight basis [1]. This work is part of a wider project for a complete biomass valorization by using pressurized fluids, namely SC-CO2 to recover the lipophilic fraction and water. In this work we proposed the use of subcritical water at 50 bar and 4 ml/min in a semicontinuous reactor at different temperatures (125 to 185ºC) to extract and hydrolyze BSG proteins. Figure1 shows protein and amino acid values on SubCW extracts accumulated after 240 minutes of extraction. The highest protein content, 15.6 g/100g BSG, dry, was obtained at 185ºC. On the other hand, the highest free amino acid content in extracts, 77 μmol aa/g BSG, dry, was achieved at 160ºC due to amino acid decomposition at this operation conditions. This trend has been also observed in the literature for SCWF of oyster at 150 ºC [2]. Free amino acid formed during BSG protein hydrolysis in SubCW are listed in Table 1 together with the amino acid composition of the BSG protein fraction. Results appears to show a trend related to the hydrophobicity of each group of amino acid since an increase in temperature improves the yield of aliphatic amino acids extraction, while the highest yield for charged amino acids, was reached at 145ºC, and neutral amino acids with a polar side chain, had the highest yield at 160ºC. This agrees with the fact that aliphatic amino acids are stable at higher temperatures. The decrease of water polarity with temperature may favor the affinity for these amino acids due to their hydrophobic character. In addition, small aliphatic amino acids are formed during the decomposition of the other amino acids [3]. BSG offers a great potential as raw material to obtain protein hydrolyzates and amino acids due to its high protein content (~20%) and the elevated rate of EAA. Despite different amino acids has found to be a key parameter on the yield obtained for each amino acid as function of operating temperature. Results obtained in this work suggest that SCWF of BSG is able to recover and hydrolyze BSG proteins., JCyl and ERDF for financial support of project BU301P18. To Hiperbaric, S.A. for financial support of Project BIOLIGNO.

Trabajo presentado en: 1er Encuentro Ibérico de Fluidos Supercríticos/1º Encontro Ibérico de Fluidos Supercríticos. 2020, 18-19 de febrero, Santiago de Compostela, Gelidium sesquipedale is a red alga that provides the best raw material to obtain the highest quality agar in the spanish agar industry [1]. Industrial process generates a residue that is usually discarded; however, this by-product still contains important amounts of different valuable compounds, such as structural carbohydrates, bioactive compounds and proteins. To valorize by-product supplied by Hispanagar company (Burgos), subcritical water extraction (SWE) is a promising green technology since water presents unique properties as solvent [2]. This work is focused on the valorization of the protein fraction to produce free amino acids and to extract and hydrolyze protein into small peptides. A semi-continuous reactor has been used to perform the extraction, and protein fraction hydrolysis was studied at different temperatures (125, 140, 155, 170, 185, 200ºC), times, and solvent flow rate (2 and 6 mL/min). Total protein content in alga residue was 20.11±1.53% in which it was determined free amino acids, among which VAL, LEU, ILE, PHE, LYS and HIS are majority (Table 1), being total essential amino acids found 10.50±0.16%, whereas non-essential amino acids like GLY, PRO and GLU+GLN represent 4.57±0.06% Protein extraction grows with increasing temperature at constant flow, reaching a maximum at 200ºC. Moreover, when flow and temperature are increased, protein recovery shows the highest and fastest extraction because of its less residence time, what makes able to get a marked extraction yield improvement (Fig. 1). Amino acids extraction follows a similar trend than proteins: an increasing extraction up to a maximum at 185ºC when flow rate is 2 ml/min, but lower than 6 ml/min. Greatest extraction was found for ALA, GLY, SER and the mixture of ASN+ASP; also for MET, TYR and GLU+GLN mixture, with lower but remarkable yield (Table 1). At constant flow rate, maximum extraction is reached at 185ºC for all amino acids determined, decreasing at 200ºC. Nevertheless, flow rate increasing makes 200ºC extraction much higher, about all for GLU+GLN mixture, LYS, HIS, TYR, PRO, LEU and MET (Fig. 2). This agrees with other studies carried out with fish protein and amino acids standard [3,4]. Moreover, it has been noted than amino acids extraction presents a similar behaviour according to its properties: basic amino acids extraction remains practically constant, whereas than neutral and acid, hydrophobic and sulfur amino acids extraction increase when temperature raises. SWE is a useful technique to extract bioactive compounds. Parameters as temperature or solvent flow rate have much influence on the protein and amino acids extraction yield. All of this makes SWE an interesting alternative to conventional treatments., JCyL and ERDF for financial support of project BU301P18. To Hiperbaric, S.A. for financial support of Project BIOLIGNO. To JCyL and ESF for the predoctoral contracts of E. Trigueros and P. Alonso-Riaño and

Póster presentado en: 1er Encuentro Ibérico de Fluidos Supercríticos/1º Encontro Ibérico de Fluidos Supercríticos. 2020, 18-19 de febrero, Santiago de Compostela, The biorefinery concept involves the valorization and conversion of biomass into different bioproducts, including energy, materials and chemicals that can replace products derived from fossil carbon sources. The integral valorization of biomass requires the extraction and fractionation of the different constituents thereof, extractables, lipids, proteins and structural components such as cellulose, hemicellulose and lignin. In this work, the use of emerging and clean technologies for the integral valorization of different types of biomass is proposed. First, the use supercritical CO2 (SCCO2) extraction is proposed for recovering the lipid fraction and ultrasonic assisted extraction to recover the hydrophilic fraction, to subsequently perform a fractionation and hydrolysis of the residual biomass using pressurized liquid water. Several types of biomass are being studied, both second and third generation. Among second generation biomass, the brewery spent grain (BSG), which accounts for 85% of the by-products generated in beer processing [1], has been selected as case study of this communication. The extraction of the lipid fraction with SCCO2 has been carried out in a 26.5 mL capacity extractor at different pressures (20-40 MPa) and temperatures (40-80 °C). The lipid fraction obtained has been characterized by determining its lipid profile and antioxidant capacity. The experiments of fractionation and hydrolysis in subcritical water (scW) have been carried out in a semi-continuous reactor, varying the extraction temperature. The different fractions obtained have been characterized by HPLC with two series detectors, UVD and RID, to determine the content in monomeric sugars and oligomers. Protein, free amino acids, total polyphenols and total organic carbon have been also determined. The composition of the BSG according to the biomass characterization protocols of NREL [2] was 21.1 % arabinoxylans, 25.6 % glucanes, 5.1 % soluble lignin, 10.5 % insoluble lignin, 1.2 % ashes, 16.7 % proteins, 5.6 % lipids and 14.4 % extractables. Worth noting the presence of insoluble lignin as well as the high content of arabinoxylans and glucanes, 10% of which were residual starch. The extraction curves obtained when studying the extraction of the lipid fraction of BSG with SCCO2, showed that the extraction rate and the extraction yield increased with increasing temperature and pressure, with the major fatty acid being linoleic acid. Regarding fractionation of the carbohydrate fraction, we have observed that, as temperature increases, hydrolysis increases. Figure 1a shows the sugars yield, including monomeric sugars and oligomers. Degradation of sugars due to the high residence times produces acids (Fig. 1b), hydroxymethylfurfural (HMF) and furfural (Fig. 1c). The treatment of biomass by scW, allows also recovering the entire protein fraction by increasing the temperature up to around 185ºC. In addition, partial hydrolysis of the protein fraction occurs, obtaining as major free aminoacids valine, aspartic acid, alanine and glycine. We can conclude that the fractionation of BSG through emerging and clean technologies allows an integral recovery of BSG, obtaining extracts with high antioxidant capacity. Pressurized water hydrolysis allows the recovery and fractionation of the carbohydrate and protein fraction., JCyL and ERDF for financial support of project BU301P18. To Hiperbaric, S.A. for financial support of Project BIOLIGNO.

Published

Trabajo presentado en: 9ª Reunión de Expertos en Tecnologías de Fluidos Supercríticos (Flucomp), 13 a 15 de junio de 2018, Madrid. Jornada Academia-Empresa (one‐to‐one)

Póster presentado en: 6th International Congress on Green Process Engineering (GPE 2018), 3 a 6 de junio de 2018, Toulouse, JCyL and ERDF for financial support of project BU055U16. MINECO and ERDF for financial support of AEI's contract through project CTQ2015-64396-R.

Trabajo presentado en: 9ª Reunión de Expertos en Tecnologías de Fluidos Supercríticos (Flucomp), 13 a 15 de junio de 2018, Madrid. Jornada Academia-Empresa (one‐to‐one), En las últimas décadas, los ácidos grasos poliinsaturados omega–3 han captado la atención de la industria alimentaria debido a sus propiedades saludables1. La principal fuente de omega–3 es el aceite de pescado, un subproducto pesquero cuya valorización constituye a la vez una oportunidad de negocio y un importante desafío, ya que las exigencias de pureza y baja oxidación en los concentrados de omega–3 son cada vez mayores. Las tecnologías de fluidos supercríticos (FSCs) evitan el uso de disolventes orgánicos tóxicos y perjudiciales para el medio ambiente y utilizan condiciones suaves de temperatura en una atmósfera inerte, lo que retrasa la oxidación de los omega–3. De esta forma, se plantea la posibilidad de obtener concentrados de omega–3 mediante etanolisis de aceite de pescado catalizada por lipasas, utilizando dióxido de carbono supercrítico (CO2-SC) como medio de reacción. En primer lugar, Se han evaluado los efectos de la exposición al CO2-SC sobre el catalizador (Lipozyme RM IM), variando la temperatura (35–70 C), presión (10–25 MPa), tiempo (1–3 h) y ciclos de tratamiento (1–3). La variable que más afectó a la actividad residual (RA) fue la temperatura, con descensos hasta RA = 80 % de la inicial. Presión, tiempo y ciclos de tratamiento no tuvieron un efecto significativo. A continuación, se han obtenido datos experimentales de equilibrio entre fases del sistema de reacción (dióxido de carbono + etanol + aceite de pescado) en el intervalo 50-70 ºC y 10-30 MPa. Dependiendo de las condiciones experimentales, se han observado mezclas bi- o trifásicas constituidas por uno o dos líquidos expandidos y un gas; o bien un sistema con una única fase supercrítica. Por último, se ha realizado un estudio cinético de la reacción de etanolisis de aceite de pescado en CO2-SC, evaluando la influencia de la temperatura (50-80 C), presión (7.5-20 MPa) y relación molar inicial de sustratos (MR) (2:1-38:1, etanol:aceite). MR y presión afectaron positivamente al desarrollo de la reacción, no observándose inhibición por exceso de etanol. El aumento de temperatura aceleró la reacción hasta un máximo (343.15 K) a partir del cual comienza a observarse inactivación térmica., research projects CTQ2012-39131-C02-01 and BU055U16, financed by MINECO and JCyL, respectively, and both co-financed by ERDF

Póster presentado en: 9ª Reunión de Expertos en Tecnologías de Fluidos Supercríticos (Flucomp), 13 a 15 de junio de 2018, Madrid. Jornada Academia-Empresa (one‐to‐one), Este trabajo se centra en la aplicación de la tecnología no térmica HPCD (High Pressure Carbon Dioxide), basada en el empleo del dióxido de carbono a alta presión, generalmente inferior a 50 MPa, para la conservación de alimentos. Se ha estudiado específicamente su aplicación en el proceso de pasteurización de zumos con pH ácido como el de naranja1, manzana2 y tomate. Para ello se han determinado las cinéticas de inactivación de diferentes enzimas como polifenoloxidasa (PPO), responsable del pardeamiento enzimático y pectinmetilesterasa (PME), que junto con poligalacturonasa (PG) juegan un papel importante en la viscosidad de los zumos. El grado de inactivación enzimática encontrado depende tanto de la fuente de la enzima, como de las condiciones de procesado, tiempo, presión y temperatura. Así, en el caso del zumo de manzana se consiguieron actividades residuales en torno a 10 % para la PPO, mientras que para la PME fueron en torno a 65 % en las mismas condiciones de procesado, 60 min, 45ºC y 20 MPa. En este caso, la estructura más compleja de la PME, con tres subunidades frente a una de la PPO, la hace más resistente al tratamiento. Otra de las variables importantes en el tratamiento mediante HPCD es la relación CO2:volumen tratado. Se realizaron estudios con diversas enzimas comerciales (PPO y PME), encontrando que relaciones superiores a 3 g CO2:mL no conllevan un mayor grado de inactivación, pudiendo así optimizar el proceso desde un punto de vista económico. Estudios de espectroscopía de fluorescencia en estas enzimas comerciales tratadas mediante HPCD correlacionaron cambios en la estructura terciaria de la enzima con la pérdida de actividad enzimática de la mismas a diferentes condiciones de presión y temperatura., MINECO y FEDER, proyecto CTQ2015-64396-R y el contrato de AEI. MINECO, contrato FPI (BES-2013-063937) de RM y JCyL y FEDER, contrato de OBR (BU055U16).

Trabajo presentado en: 6th International Congress on Green Process Engineering (GPE 2018), 3 a 6 de junio de 2018, Toulouse, HPCD is a promising technology to inactivate the enzymes responsible for the juice spoilage, such as PPO and PME. In order to understand the mechanism that induces this inactivation, a study using two commercial enzymes (PPO from mushroom and PME from Aspergillus niger) was carried out. The effect of pressure, temperature, exposure time and ratio CO2/enzyme ratio loaded in the reactor were studied. The experimental results (residual activity) were fitted to a kinetic model that served to develop a complete kinetic study: the kinetic constants, activation volume and activation energy were calculated, as well as the pressure and temperature sensitivity parameters (ZP and ZT, respectively). The changes in the tertiary structure of the enzymes after different treatments were analyzed by fluorescence spectroscopy running different tests: intrinsic fluorescence measurement, KI quenching and ANS binding experiments. In the case of PPO, the experimental results revealed that this enzyme inactivation kinetics fitted the two fraction model, which indicates the presence of labile and stable isoenzymes. Exposure time (2 to 15 minutes), temperature (25 to 45°C) and pressure (50 to 200bar) were the studied experimental conditions that led to different physical states for the CO2 (gas, liquid and supercritical). Despite the different experimental combinations of pressure and temperature tried, a similar inactivation pattern was observed: a sudden decrease in activity (more than 75% of the total activity loss was observed within the first 2 minutes) was followed by a slowed decay. At constant temperature, higher inactivation rates were observed the higher the pressure, obtaining an almost complete inactivation at 200bar after 5 minutes regardless the temperature. When temperature was increased, much faster inactivation rates were observed. ZP and ZT were in the range 69-78bar and 27-40°C, respectively. In the case of the commercial PME, the use of supercritical CO2 (pressure 60-180bar, temperature 40-55°C and times up to 75 minutes) increased dramatically the PME inactivation rate, showing that pressure had a limited effect on PME inactivation but temperature had an important effect. The pressure and temperature sensitivity parameters (ZP and ZT) confirmed that trend, being in the range from 276 to 450bar and 8.7°C, respectively. The experimental data fitted the first order model and the inactivation kinetics study of PME was completed with the calculations of the activation energy and volume of activation. The ratio CO2/volume of enzyme (g/mL) loaded in the reactor was found to be critical for both enzymes. It was seen that ratios higher than 3 did not improve the inactivation kinetics, being a waste of CO2 from the economic point of view. Bellow that critical value, the inactivation of the enzyme strongly depended on pressure and temperature. In both cases the structure of the enzyme was dramatically affected after exposure to HPCD, as revealed by the fluorescence spectroscopy analysis that showed significant changes in the tertiary structure of the enzyme, which were compatible with the losses in activity observed., Spanish Governme nt (MINECO) and the European Regional Development Fund (ERDF) for financial support of project CTQ2015-64396-R and A.E. Illera’s contract. To Junta de Castilla y León and ERDF for financial support of project BU055U16 and O.Benito-Román’s Post-doctoral contract

Póster presentado en: 6th International Congress on Green Process Engineering (GPE 2018), 3 a 6 de junio de 2018, Toulouse, MINECO and ERDF for financial support of AEI's contract through project CTQ2015-64396-R To JCyL and ERDF for financial support of project BU055U16 and O.Benito-Román’s Post-doctoral contract To MINECO for RM's pre-doctoral contract (BES-2013-063937).