Uso de procesos convencionales e innovadores para la valorización de los subproductos de la horchata

La “Horchata de chufa" o más comúnmente conocida como “Horchata” es una bebida tradicional, no alcohólica, dulce y de aspecto blanquecino obtenida a partir de tubérculos de chufa (Cyperus esculentus L. var. Sativus Boeck) consumida fundamentalmente en la Comunidad Valenciana durante la época de verano, aunque en los últimos años ha experimentado un crecimiento en su consumo tanto a nivel español como en el resto del mundo. Durante el proceso de elaboración de la “Horchata” se producen una gran cantidad de residuos y subproductos tanto sólidos como líquidos, donde los sólidos pueden representar hasta el 60% del material vegetal. Estos subproductos se han considerado tradicionalmente como desechos industriales sin valor comercial y se han utilizado fundamentalmente como pienso para animales. Los subproductos líquidos destacan por su alto contenido en prebióticos, antioxidantes y como sustituto del agua en productos cárnicos. Por otra parte, los subproductos sólidos pueden ser utilizados para diferentes usos, debido a su alto contenido en fibra dietética, compuestos antioxidantes bioactivos, además de tener buenas propiedades tecnológicas (capacidad de retención de agua, capacidad de emulsión y estabilidad de emulsión). El objetivo de la presente Tesis Doctoral es la recuperación caracterización de diversos nutrientes y compuestos bioactivos obtenidos a partir de subproductos sólidos del proceso de elaboración de la “horchata”. Fundamentalmente, el estudio se ha centrado en la extracción de aceite y evaluación del contenido graso y perfil de ácidos así como vitamina E del mismo. También se ha evaluado el contenido de compuestos antioxidantes bioactivos, capacidad antioxidante total y parámetros fisicoquímicos de los extractos obtenidos. Para obtener los extractos, se utilizaron diferentes técnicas de extracción, tanto métodos convencionales (ej. extracción convencional sólido/líquido, Soxhlet, etc.) como procesos innovadores (ej. fluidos supercríticos (SC-CO₂)), también conocidos como “extracción verde”. Posteriormente, los extractos se analizaron mediante la utilización de diferentes técnicas analíticas, como espectrofotometría y métodos cromatográficos como por ejemplo cromatografía de gases con detector de ionización de llama, cromatografía líquida acoplada a espectrometría de masas en tándem, etc. También se revisó en la literatura disponible el potencial uso de otros procesos innovadores como ultrasonidos, pulsos eléctricos, altas presiones, solas y/o en combinación con enzimas, expresión mecánica asistida por gas, microondas, los cuales también se han mostrado interesantes de cara a la valorización de los subproductos obtenidos durante la elaboración de la “Horchata”. Tras evaluar los resultados obtenidos, se observó cómo los subproductos de la "horchata" presentan un contenido importante de aceite, y compuestos antioxidantes bioactivos, aunque es necesario seleccionar el método de extracción apropiado para conseguir recuperar estos compuestos de la manera adecuada. Se observó que el contenido de nutrientes y compuestos antioxidantes bioactivos difiere de acuerdo al método de extracción utilizado. Por ejemplo, tras utilizar la extracción con fluidos supercríticos, se observó un aumento en la extracción de aceite tras aumentar la presión utilizada, aunque el rendimiento fue inferior al obtenido tras utilizar extracción convencional mediante el método Folch o Soxhlet. Eso sí, en el caso de los fluidos supercríticos se evitó la utilización de disolventes tóxicos. Asimismo, el aceite extraído presentó una mayor estabilidad oxidativa tras aplicar SC-CO2 a 30 y 40 MPa, en comparación con las extracciones realizadas a presiones más bajas o al procedimiento de extracción convencional. La extracción con SC-CO2 fue más eficiente para recuperar compuestos fenólicos lipofílicos con alta capacidad antioxidante en comparación con el procedimiento convencional de extracción (método Folch). Se observó una mayor extracción de los diferentes compuestos individuales, identificados y cuantificados por cromatografía líquida acoplada a espectrometría de masas en tándem cuando se aumentó la presión de tratamiento obteniendo los valores más altos cuando la presión fue de 40 MPa. Por otro lado, también se observó que tras utilizar la extracción convencional sólido/líquido, modificando el tiempo y la temperatura de extracción así como el disolvente utilizado, la eficiencia de extracción de los compuestos fenólicos totales fue mayor al aumentar la temperatura. Asimismo, concentraciones más altas de etanol y tiempos de extracción prolongados también promovieron una mejora en la extracción de estos compuestos. La capacidad antioxidante también aumentó cuando se utilizaron mayores concentraciones de etanol, con un mayor tiempo de extracción y a mayor temperatura. Estos resultados se podrían traducir en beneficios económicos derivados de la valorización de los subproductos frente a su eliminación con el fin de preservar el medio ambiente de residuos procedentes de la industria horchatera de una manera sostenible, reduciendo la huella de dióxido de carbono y proteger la salud del consumidor mediante la prevención o reducción de los impactos globales del uso de los recursos y la mejora de la eficacia de dicho uso."Horchata de chufa" or more commonly known as "Horchata" is a traditional, non-alcoholic, sweet and whitish-looking drink obtained from tubers of tiger nut (Cyperus esculentus L. Sativus Boeck), consumed mainly in the Valencian Community during the summer season, although over the last years it has experienced a growth in its consumption both in Spain and in the rest of the world. During "Horchata" production process, a large amount of wastes and by-products are produced, both solid and liquids, where solid ones can represent up to 60% of the plant material. These by-products have been traditionally considered as industrial waste without commercial value and have been used mainly as animal feed. The liquid by-products stand out for their high content in prebiotics, antioxidants and as a substitute for water in meat products. On the other hand, solid by-products an be used for different applications, due to their high content of dietary fiber, antioxidant bioactive compounds, in addition to having good technological properties (water retention capacity, emulsion capacity and emulsion stability, among others). The aim of the present PhD Thesis is the recovery and characterization of nutrients and antioxidant bioactive compounds obtained from "Horchata" solid by-products. The study has been mainly focused on the extraction of oil and the evaluation of its fatty acid content and profile as well as vitamin E amount. The content of antioxidant bioactive compounds, total antioxidant capacity and physicochemical parameters of the obtained extracts have been also evaluated. To obtain the extracts, different extraction techniques were used, both conventional methods (eg conventional solid/liquid extraction, Soxhlet, etc.) and innovative processes (eg. supercritical fluids (SC-CO₂)), also known as "green extraction". Subsequently, the extracts were analyzed using different analytical techniques, such as spectrophotometry and chromatographic methods such as gas chromatography with flame ionization detector, liquid chromatography coupled to tandem mass spectrometry, etc. The potential use of other innovative processes such as ultrasounds, electric pulses, high pressures, alone and/or in combination with enzymes, gas-assisted mechanical expression, microwaves, which are also promising tools for the valorization of the by-products obtained during the elaboration of the "Horchata". After evaluating the results obtained, it was observed how "Horchata" by-products present an important content of oil, and antioxidant bioactive compounds, although it is necessary to select the appropriate extraction method in order to recover these compounds in the proper way. It was observed that the content of nutrients and antioxidant bioactive compounds differs according to the extraction method used. For example, after using extraction with supercritical fluids, an increase in the oil extraction was observed after increasing the pressure used, although the yield was lower than that obtained after using conventional extraction using the Folch or Soxhlet method. Of course, in the case of supercritical fluids the use of toxic solvents was avoided. Likewise, the extracted oil presented a greater oxidative stability after applying SC-CO2 at 30 and 40 MPa, in comparison with the extractions made at lower pressures or the conventional extraction procedure. Extraction with SC-CO2 was more efficient to recover lipophilic phenolic compounds with high antioxidant capacity compared to the conventional extraction method (Folch method). A greater extraction of the different individual compounds was observed, identified and quantified by liquid chromatography coupled to tandem mass spectrometry, when the treatment pressure was increased, obtaining the highest values when the pressure was 40 MPa. On the other hand, it was also observed that after using the conventional solid/liquid extraction, modifying the extraction time and temperature as well as the solvent used, the extraction efficiency of the total phenolic compounds was higher with increasing temperature. Moreover, higher concentrations of ethanol and longer extraction times also promoted an improvement in the extraction of these compounds. Antioxidant capacity also increased when higher ethanol concentrations were used, with a longer extraction time and higher temperature. These results could translate into economic benefits derived from the valuation of by-products as opposed to their elimination in order to preserve the environment

Influence of temperature, solvent and pH on the selective extraction of phenolic compounds from tiger nuts by-products: Triple-TOF-LC-MS-MS characterization.

The aim of this study was to assess the effect of temperature, solvent (hydroethanolic mixtures) and pH on the recovery of individual phenolic compounds from 'horchata' by-products. These parameters were optimized by response surface methodology and triple-TOF-LC-MS-MS was selected as the analytical tool to identify and quantify the individual compounds. The optimum extraction conditions were 50% ethanol, 35 °C and pH 2.5, which resulted in values of 222.6 mg gallic acid equivalents (GAE)/100 g dry matter and 1948.1 µM trolox equivalent (TE)/g of dry matter for total phenolic content (TPC) and trolox equivalent antioxidant capacity (TEAC), respectively. The extraction of phenolic compounds by the conventional solvent method with agitation was influenced by temperature (p = 0.0073), and more strongly, by the content of ethanol in the extraction solution (p = 0.0007) while the pH did not show a great impact (p = 0.7961). On the other hand, the extraction of phenolic acids was affected by temperature (p = 0.0003) and by ethanol amount (p < 0.0001) but not by the pH values (p = 0.53). In addition, the percentage of ethanol influenced notably the extraction of both 4-vinylphenol (p = 0.0002) and the hydroxycinnamic acids (p = 0.0039). Finally, the main individual phenolic extracted with hydroethanolic mixtures was 4-vinylphenol (303.3 μg/kg DW) followed by spinacetin3-O-glucosyl-(1→6)-glucoside (86.2 μg/kg DW) and sinensetin (77.8 μg/kg DW)

Obtaining antioxidants and natural preservatives from food by-products through fermentation: A review

Industrial food waste has potential for generating income from high-added-value compounds through fermentation. Solid-state fermentation is promising to obtain a high yield of bioactive compounds while requiring less water for the microorganism’s growth. A number of scientific studies evinced an increase in flavonoids or phenolics from fruit or vegetable waste and bioactive peptides from cereal processing residues and whey, a major waste of the dairy industry. Livestock, fish, or shellfish processing by-products (skin, viscera, fish scales, seabass colon, shrimp waste) also has the possibility of generating antioxidant peptides, hydrolysates, or compounds through fermentation. These bioactive compounds (phenolics, flavonoids, or antioxidant peptides) resulting from bacterial or fungal fermentation are also capable of inhibiting the growth of commonly occurring food spoilage fungi and can be used as natural preservatives. Despite the significant release or enhancement of an-tioxidant compounds through by-products fermentation, the surface areas of large-scale bioreactors and flow patterns act as constraints in designing a scale-up process for improved efficiency. An in-process purification method can also be the most significant contributing factor for raising the overall cost. Therefore, future research in modelling scale-up design can contribute towards mitigating the discard of high-added-value generating residues. Therefore, in this review, the current knowledge on the use of fermentation to obtain bioactive compounds from food by-products, emphasizing their use as natural preservatives, was evaluated

Phase I clinical trial in healthy adults of a nasal vaccine candidate containing recombinant hepatitis B surface and core antigens

SummaryBackgroundThe nasal vaccine candidate (NASVAC), comprising hepatitis B virus (HBV) surface (HBsAg) and core antigens (HBcAg), has been shown to be highly immunogenic in animal models.MethodsA phase I double-blinded, placebo-controlled randomized clinical trial was carried out in 19 healthy male adults with no serologic markers of immunity/infection to HBV. This study was aimed at exploring the safety and immunogenic profile of nasal co-administration of both HBV recombinant antigens. The trial was performed according to Good Clinical Practice guidelines. Participants ranged in age from 18 to 45 years and were randomly allocated to receive a mixture of 50μg HBsAg and 50μg HBcAg or 0.9% physiologic saline solution, as a placebo, via nasal spray in a five-dose schedule at 0, 7, 15, 30, and 60 days. A total volume of 0.5ml was administered in two dosages of 125μl per nostril. Adverse events were actively recorded 1h, 6h, 12h, 24h, 48h, 72h, 7 days and 30 days after each dose. Anti-HBs and anti-HBc titers were evaluated using corresponding ELISA kits at days 30 and 90.ResultsThe vaccine candidate was safe and well tolerated. Adverse reactions included sneezing (34.1%), rhinorrhea (12.2%), nasal stuffiness (9.8%), palate itching (9.8%), headache (9.8%), and general malaise (7.3%). These reactions were all self-limiting and mild in intensity. No severe or unexpected events were recorded during the trial. The vaccine elicited anti-HBc seroconversion in 100% of subjects as early as day 30 of the immunization schedule, while a seroprotective anti-HBs titer (≥10IU/l) was at a maximum at day 90 (75%). All subjects in the placebo group remained seronegative during the trial.ConclusionThe HBsAg–HBcAg vaccine candidate was safe, well tolerated and immunogenic in this phase I study in healthy adults. To our knowledge, this is the first demonstration of safety and immunogenicity for a nasal vaccine candidate comprising HBV antigens

gSeaGen: The KM3NeT GENIE-based code for neutrino telescopes

Program summary Program Title: gSeaGen CPC Library link to program files: http://dx.doi.org/10.17632/ymgxvy2br4.1 Licensing provisions: GPLv3 Programming language: C++ External routines/libraries: GENIE [1] and its external dependencies. Linkable to MUSIC [2] and PROPOSAL [3]. Nature of problem: Development of a code to generate detectable events in neutrino telescopes, using modern and maintained neutrino interaction simulation libraries which include the state-of-the-art physics models. The default application is the simulation of neutrino interactions within KM3NeT [4]. Solution method: Neutrino interactions are simulated using GENIE, a modern framework for Monte Carlo event generators. The GENIE framework, used by nearly all modern neutrino experiments, is considered as a reference code within the neutrino community. Additional comments including restrictions and unusual features: The code was tested with GENIE version 2.12.10 and it is linkable with release series 3. Presently valid up to 5 TeV. This limitation is not intrinsic to the code but due to the present GENIE valid energy range. References: [1] C. Andreopoulos at al., Nucl. Instrum. Meth. A614 (2010) 87. [2] P. Antonioli et al., Astropart. Phys. 7 (1997) 357. [3] J. H. Koehne et al., Comput. Phys. Commun. 184 (2013) 2070. [4] S. Adrián-Martínez et al., J. Phys. G: Nucl. Part. Phys. 43 (2016) 084001.The gSeaGen code is a GENIE-based application developed to efficiently generate high statistics samples of events, induced by neutrino interactions, detectable in a neutrino telescope. The gSeaGen code is able to generate events induced by all neutrino flavours, considering topological differences between tracktype and shower-like events. Neutrino interactions are simulated taking into account the density and the composition of the media surrounding the detector. The main features of gSeaGen are presented together with some examples of its application within the KM3NeT project.French National Research Agency (ANR) ANR-15-CE31-0020Centre National de la Recherche Scientifique (CNRS)European Union (EU)Institut Universitaire de France (IUF), FranceIdEx program, FranceUnivEarthS Labex program at Sorbonne Paris Cite ANR-10-LABX-0023 ANR-11-IDEX-000502Paris Ile-de-France Region, FranceShota Rustaveli National Science Foundation of Georgia (SRNSFG), Georgia FR-18-1268German Research Foundation (DFG)Greek Ministry of Development-GSRTIstituto Nazionale di Fisica Nucleare (INFN)Ministry of Education, Universities and Research (MIUR)PRIN 2017 program Italy NAT-NET 2017W4HA7SMinistry of Higher Education, Scientific Research and Professional Training, MoroccoNetherlands Organization for Scientific Research (NWO) Netherlands GovernmentNational Science Centre, Poland 2015/18/E/ST2/00758National Authority for Scientific Research (ANCS), RomaniaMinisterio de Ciencia, Innovacion, Investigacion y Universidades (MCIU): Programa Estatal de Generacion de Conocimiento, Spain (MCIU/FEDER) PGC2018-096663-B-C41 PGC2018-096663-A-C42 PGC2018-096663-BC43 PGC2018-096663-B-C44Severo Ochoa Centre of Excellence and MultiDark Consolider (MCIU), Junta de Andalucia, Spain SOMM17/6104/UGRGeneralitat Valenciana: Grisolia, Spain GRISOLIA/2018/119GenT, Spain CIDEGENT/2018/034La Caixa Foundation LCF/BQ/IN17/11620019EU: MSC program, Spain 71367

Growth And The Growth Hormone-Insulin Like Growth Factor 1 Axis In Children With Chronic Inflammation:Current Evidence, Gaps In Knowledge And Future Directions

Growth failure is frequently encountered in children with chronic inflammatory conditions like juvenile idiopathic arthritis, inflammatory bowel disease and cystic fibrosis. Delayed puberty and attenuated pubertal growth spurt is often seen during adolescence. The underlying inflammatory state mediated by pro-inflammatory cytokines, prolonged use of glucocorticoid and suboptimal nutrition contribute to growth failure and pubertal abnormalities. These factors can impair growth by their effects on the growth hormone-insulin like growth factor axis and also directly at the level of the growth plate via alterations in chondrogenesis and local growth factor signaling. Recent studies on the impact of cytokines and glucocorticoid on the growth plate studies further advanced our understanding of growth failure in chronic disease and provided a biological rationale of growth promotion. Targeting cytokines using biologic therapy may lead to improvement of growth in some of these children but approximately one third continue to grow slowly. There is increasing evidence that the use of relatively high dose recombinant human growth hormone may lead to partial catch up growth in chronic inflammatory conditions, although long term follow-up data is currently limited. In this review, we comprehensively review the growth abnormalities in children with juvenile idiopathic arthritis, inflammatory bowel disease and cystic fibrosis, systemic abnormalities of the growth hormone-insulin like growth factor axis and growth plate perturbations. We also systematically reviewed all the current published studies of recombinant human growth hormone in these conditions and discuss the role of recombinant human insulin like growth factor-1

Ultrasound-Assisted Lactic Acid Fermentation of Bakraei (<i>Citrus reticulata</i> cv. Bakraei) Juice: Physicochemical and Bioactive Properties

In this study, ultrasonication (US) (50 W, 30 kHz, 1–6 min) was used to increase the efficiency of Limosilactobacillus reuteri PTCC 1655 fermentation process (37 °C; 30 h) of Bakraei juice. Total sugars, pH, Brix, organic acids, vitamin C, polyphenols, antioxidant activity, α-amylase inhibition and anti-inflammatory properties were measured during the fermentation period. The results showed that by increasing the ultrasound time up to 5 min, pH, vitamin C, citric acid, and polyphenolic compounds decreased, while lactic acid, antioxidant capacity, α-amylase inhibition and anti-inflammatory properties were increased. When the ultrasound time was increased up to 6 min, compared to the non-ultrasound-treated sample, the efficiency of the fermentation process decreased and promoted a decrease in the microbial population, lactic acid levels, antioxidant activity, α-amylase inhibition, and anti-inflammatory properties of the juices. The initial anti-inflammatory activity (11.3%) of juice reached values of 33.4% and 19.5%, after US treatments of 5 and 6 min, respectively, compared to the non-sonicated juice (21.7%), after 30 h of fermentation. As a result, the use of ultrasound in the controlled fermentation process can increase the efficiency of fermentation process

Chapter 11 - Valorization of waste and by-products from food industries through the use of innovative technologies

Food waste and by-products, which are normally regarded as not being valuable, have great potential for valuable compounds production. Valorization of these materials not only means more nutrients or bioactive compounds, but also addresses environmental issues caused by discarding these “waste” materials. This chapter discusses the nutritive and economic potential of food waste and by-products, as well as the application of innovative technologies (i.e., electrotechnologies, ultrasound, high hydrostatic pressure, nanotechnology, pressurized fluids) in food waste and by-products valorization. Such technologies improve the perspectives regarding waste utilization, paving the ways toward sustainable industrial development, which is one of the basic pillars for public health.Fil: Zhu, Zhenzhou. Universidad Politécnica de Wuhan. Facultad de Ciencias e Ingeniería de los Alimentos; República Popular de China.Fil: Gavahian, Mohsen. Centro de investigación de productos y procesos. Instituto de investigación y desarrollo de la industria alimentaria; República Popular de China.Fil: Barba, Francisco J. Universidad de Valencia. Facultad de Farmacia. Departamento de Medicina Preventiva y Salud Pública, Ciencias de la Alimentación, Toxicología y Medicina Legal; España.Fil: Roselló-Soto, Elena. Universidad de Valencia. Facultad de Farmacia. Departamento de Medicina Preventiva y Salud Pública, Ciencias de la Alimentación, Toxicología y Medicina Legal; España.Fil: Danijela Bursać Kovačević. Universidad de Zagreb. Facultad de Tecnología y Biotecnología de los Alimentos; Croacia.Fil: Putnik, Predrag. Universidad de Zagreb. Facultad de Tecnología y Biotecnología de los Alimentos; Croacia.Fil: Denoya, Gabriela Inés. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Tecnología de Alimentos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina