Milk Proteins, Peptides, and Oligosaccharides: Effects against the 21st Century Disorders

Milk is the most complete food for mammals, as it supplies all the energy and nutrients needed for the proper growth and development of the neonate. Milk is a source of many bioactive components, which not only help meeting the nutritional requirements of the consumers, but also play a relevant role in preventing various disorders. Milk-derived proteins and peptides have the potential to act as coadjuvants in conventional therapies, addressing cardiovascular diseases, metabolic disorders, intestinal health, and chemopreventive properties. In addition to being a source of proteins and peptides, milk contains complex oligosaccharides that possess important functions related to the newborn's development and health. Some of the health benefits attributed to milk oligosaccharides include prebiotic probifidogenic effects, antiadherence of pathogenic bacteria, and immunomodulation. This review focuses on recent findings demonstrating the biological activities of milk peptides, proteins, and oligosaccharides towards the prevention of diseases of the 21st century. Processing challenges hindering large-scale production and commercialization of those bioactive compounds have been also addressed

Lunasin Peptide is a Modulator of the Immune Response in the Human Gastrointestinal Tract

[Introduction]: Lunasin is a soybean bioactive peptide with a variety of beneficial properties against chronic disorders. However, its effect in human primary intestinal cells remains unknown. Hence, this study aims to characterize its ex vivo biological activity in the human intestinal mucosa. [Methods and Results]: Human intestinal biopsies, obtained from healthy controls, are ex vivo conditioned with lunasin both in the presence/absence of lipopolysaccharide (LPS). Peptide maintains its stability during biopsy culture by HPLC-MS/MS analysis. Lunasin is bioactive in the human mucosa, as it induces IL-1β, TNF-α, IL-17A, CCL2, and PGE2/COX-2 gene expression together with an increased expression of tolerogenic IL-10 and TGFβ, while it also downregulates the expression of iNOS and subunit p65 from NF-κB. Indeed, lunasin also abrogates the LPS-induced pro-inflammatory response, downregulating IL-17A, IFNγ, and IL-8 expression, and inducing IL-10 and TGFβ expression. These results are also mirrored in the cell-free culture supernatants at the protein level by Multiplex. Moreover, lunasin further induces a regulatory phenotype and function on human intestinal conventional dendritic cell and macrophage subsets as assessed by flow cytometry. [Conclusions]: We hereby have characterized lunasin as an immunomodulatory peptide with potential capacity to prevent immune and inflammatory-mediated disorders in the human gastrointestinal tract.This work was supported by the Spanish Ministry of Economy (SAF2014-56642-JIN), the Spanish Ministry of Health (PIE13/00041), the Spanish Ministry of Science, Innovation and Universities (AGL2015-66886-R, PID2019-104218RB-I00), and the Community of Madrid (Consejería de Educación, Juventud y Deporte, Programa de Garantía Juvenil 2015 and 2016). S.F.T. is currently funded by the Instituto de Salud Carlos III (Sara Borrell fellowship CD17/00014). L.O.M. is funded by the Community of Madrid (BMD-5800). D.B. is funded by the Spanish Ministry of Science (RYC-2017-21606)

Effect of soybean peptide lunasin on the inflammatory and immune response of healthy human intestinal mucosa

Trabajo presentado al 2nd International Symposium on Bioactive Peptides, celebrado en Valencia (España) del 22 al 24 de mayo de 2019.Peer reviewe

In Vitro and In Vivo Response of Zinc-Containing Mesoporous Bioactive Glasses in a Sheep Animal Model

Zinc-enriched mesoporous bioactive glasses (MBGs) are bioceramics with potential antibacterial and osteogenic properties. However, few assays have been performed to study these properties in animal models.In this study, MBGs enriched with up to 5% ZnO were synthesized, physicochemically characterized, and evaluated for their osteogenic activity both in vitro and in vivo. The ZnO MBGs showed excellent textural properties despite ZnO incorporation. However, the release of Zn2+ ions inhibited the mineralization process when immersed in simulated body fluid. In vitro assays showed significantly highe r values of viability and expression of early markers of celldifferentiation and angiogenesis in a ZnO-content-dependent manner. The next step was to study the osteogenic potential in a sheep bone defect model. Despite their excellent textural properties and cellular response in vitro, the ZnO MBGs were not able to integrate into the bone tissue, which can be explained in terms of inhibition of the mineralization process caused by Zn2+ ions. This work highlights the need to develop nanostructured materials for bone regeneration that can mineralize to interact with bone tissue and induce the processes of implant acceptance, cell colonization by osteogenic cells, and regeneration of lost bone tissue

Injectable mesoporous bioactive nanoparticles regenerate bone tissue under osteoporosis conditions

The osteogenic capability of mesoporous bioactive nanoparticles (MBNPs) in the SiO2–CaO system has been assessed in vivo using an osteoporotic rabbit model. MBNPs have been prepared using a double template method, resulting in spherical nanoparticles with a porous core-shell structure that has a high surface area and the ability to incorporate the anti-osteoporotic drug ipriflavone. In vitro expression of the pro-inflammatory genes NF-κB1, IL-6, TNF-α, P38 and NOS2 in RAW-264.7 macrophages, indicates that these nanoparticles do not show adverse inflammatory effects. An injectable system has been prepared by suspending MBNPs in a hyaluronic acid-based hydrogel, which has been injected intraosseously into cavitary bone defects in osteoporotic rabbits. The histological analyses evidenced that MBNPs promote bone regeneration with a moderate inflammatory response. The incorporation of ipriflavone into these nanoparticles resulted in a higher presence of osteoblasts and enhanced angiogenesis at the defect site, but without showing significant differences in terms of new bone formation

Efectividad de los sistemas de formación multimedia para el aprendizaje de la exploración ecográfica abdominal en la especie canina

Algunas de las principales ventajas que presenta la ecografía como herramienta diagnóstica es que permite la captura de imágenes en tiempo real y que no emite ninguna radiación ionizante perjudicial. Por el contrario, su mayor inconveniente reside en la dificultad de aprender a realizar una ejecución e interpretación correctas. Nuestra hipótesis es que el uso de las herramientas multimedia, tales como las herramientas interactivas para PC, las presentaciones tipo Power pointTM y los libros electrónicos podrían acortar la curva de aprendizaje de la ecografía en los estudiantes de grado de Veterinaria. El objetivo de este trabajo es comparar la eficacia de varias herramientas multimedia frente a un método tradicional, evaluando la asimilación de los principios básicos de la ecografía y el protocolo de exploración abdominal del perro, así como la ejecución de ejercicios prácticos en un simulador ex vivo. En este estudio participaron 70 estudiantes de Veterinaria que se dividieron en cuatro grupos de experiencia similar en ecografía (Grupos A, B, C y D) y 20 veterinarios con más de 4 años de experiencia clínica (Grupos E, expertos en ecografía, y F, inexpertos). Cada grupo de estudiantes llevó a cabo una sesión de entrenamiento sobre ecografía básica abdominal del perro con una herramienta diferente. La sesión de entrenamiento del Grupo A se realizó con un sistema interactivo para PC, el grupo B con una vídeo-presentación en Power pointTM, el grupo C con un libro electrónico específico para iPad® y en el grupo D con un documento impreso. Todas las herramientas presentan contenidos educativos similares. Todos los grupos de estudiantes respondieron a un examen sobre “los principios básicos de ecografía y la exploración ecográfica abdominal en perro” antes (prueba inicial) y después (prueba final) de la sesión de entrenamiento. Por otra parte, los grupos de veterinarios (E y F) no realizaron la sesión de entrenamiento, pero sí la prueba de evaluación teórica. Finalmente todos los participantes realizaron un examen práctico que consistió en ejecutar 14 ejercicios sobre un simulador ex vivo. En la prueba inicial no se observaron diferencias significativas entre los grupos de estudiantes. Por otro lado, después de la sesión de entrenamiento, las pruebas finales mostraron una puntuación mayor en los grupos que emplearon herramientas multimedia. El Grupo C (8,69±1,6 ) presentó una puntuación significativamente mayor en comparación con los grupos A, B y D (A=7,64 ±2,04; B=7,83±1,97 y D=6,92±2,81, p <0,003). En cuanto a la comparación de la prueba final entre los grupos de estudiantes con el grupo E, no se observaron diferencias significativas entre los grupos C y E. Sin embargo, en la prueba práctica realizada sobre el simulador ex vivo, ningún grupo de estudiantes obtuvo resultados similares a los del grupo E. En conclusión, el uso de herramientas multimedia mejora la retención de conocimientos a corto plazo para la asimilación de los principios básicos de la ecografía y el protocolo de exploración abdominal del perro, con respecto el empleo de una herramienta tradicional.The ultrasound main advantages are that it allows real time images capture and it does not involve ionizing radiation. On the other hand, one of the main disadvantages is the difficulty to interpret. We believe that the use of e-books, web-based-learning and Power PointTM video-presentations implemented with multimedia tools could shorten the learning curve of ultrasonography of Veterinary Medicine students. The aim of this work is to assess the effectiveness of the multimedia methods versus a traditional method, determining the assimilation of dog ultrasound scan protocol and the implementation of specific examination exercises on ex vivo ultrasound simulator. This study involved 70 Veterinary students who were divided into four groups of similar experience in ultrasound (groups A, B, C and D) and 20 veterinarians (groups E, expert in ultrasound, and F, inexpert). Each group of students performed a specific training on basic ultrasound and abdominal ultrasound examination in dogs with different tools. The training session was performed in Group A by means of a specific e-book for iPad®, in group B with a video-presentation Power point, in group C with a web-based-learning and in group D with a printed paper document. All tools presented similar educational contents. Student groups responded to a test about “Abdominal Ultrasound examination in dogs” before (pre-test) and after (post-test) the training session. On the other hand, the veterinarians groups did not performed the training sessions but completed the same test that students. Finally every participant executed a practical ultrasound scanning test on an ex vivo simulator. In the pre-test no significant differences were observed between groups. On the other hand, post-test showed a greater score in multimedia than traditional tools. Namely, Group C (8,69±1,63 ) presented a significantly greater score compared with groups A, B and D (A=7,64±2,04; B=7,83±1,97 and D=6,92±2,81, p< 0,003). Regarding the comparisons of student’s post-test with group E, no significant differences were observed between group C and group E. Moreover, no student’s group had similar results to group E in the scanning test. In conclusion, the use of multimedia tools seems to improve to a greater extent the short-term memory retention for learning of basic principles of ultrasound and abdominal examination protocol in dogs

Efectividad de los sistemas de formación multimedia para el aprendizaje de la exploración ecográfica abdominal en la especie canina

Algunas de las principales ventajas que presenta la ecografía como herramienta diagnóstica es que permite la captura de imágenes en tiempo real y que no emite ninguna radiación ionizante perjudicial. Por el contrario, su mayor inconveniente reside en la dificultad de aprender a realizar una ejecución e interpretación correctas. Nuestra hipótesis es que el uso de las herramientas multimedia, tales como las herramientas interactivas para PC, las presentaciones tipo Power pointTM y los libros electrónicos podrían acortar la curva de aprendizaje de la ecografía en los estudiantes de grado de Veterinaria. El objetivo de este trabajo es comparar la eficacia de varias herramientas multimedia frente a un método tradicional, evaluando la asimilación de los principios básicos de la ecografía y el protocolo de exploración abdominal del perro, así como la ejecución de ejercicios prácticos en un simulador ex vivo. En este estudio participaron 70 estudiantes de Veterinaria que se dividieron en cuatro grupos de experiencia similar en ecografía (Grupos A, B, C y D) y 20 veterinarios con más de 4 años de experiencia clínica (Grupos E, expertos en ecografía, y F, inexpertos). Cada grupo de estudiantes llevó a cabo una sesión de entrenamiento sobre ecografía básica abdominal del perro con una herramienta diferente. La sesión de entrenamiento del Grupo A se realizó con un sistema interactivo para PC, el grupo B con una vídeo-presentación en Power pointTM, el grupo C con un libro electrónico específico para iPad® y en el grupo D con un documento impreso. Todas las herramientas presentan contenidos educativos similares. Todos los grupos de estudiantes respondieron a un examen sobre “los principios básicos de ecografía y la exploración ecográfica abdominal en perro” antes (prueba inicial) y después (prueba final) de la sesión de entrenamiento. Por otra parte, los grupos de veterinarios (E y F) no realizaron la sesión de entrenamiento, pero sí la prueba de evaluación teórica. Finalmente todos los participantes realizaron un examen práctico que consistió en ejecutar 14 ejercicios sobre un simulador ex vivo. En la prueba inicial no se observaron diferencias significativas entre los grupos de estudiantes. Por otro lado, después de la sesión de entrenamiento, las pruebas finales mostraron una puntuación mayor en los grupos que emplearon herramientas multimedia. El Grupo C (8,69±1,6 ) presentó una puntuación significativamente mayor en comparación con los grupos A, B y D (A=7,64 ±2,04; B=7,83±1,97 y D=6,92±2,81, p <0,003). En cuanto a la comparación de la prueba final entre los grupos de estudiantes con el grupo E, no se observaron diferencias significativas entre los grupos C y E. Sin embargo, en la prueba práctica realizada sobre el simulador ex vivo, ningún grupo de estudiantes obtuvo resultados similares a los del grupo E. En conclusión, el uso de herramientas multimedia mejora la retención de conocimientos a corto plazo para la asimilación de los principios básicos de la ecografía y el protocolo de exploración abdominal del perro, con respecto el empleo de una herramienta tradicional.The ultrasound main advantages are that it allows real time images capture and it does not involve ionizing radiation. On the other hand, one of the main disadvantages is the difficulty to interpret. We believe that the use of e-books, web-based-learning and Power PointTM video-presentations implemented with multimedia tools could shorten the learning curve of ultrasonography of Veterinary Medicine students. The aim of this work is to assess the effectiveness of the multimedia methods versus a traditional method, determining the assimilation of dog ultrasound scan protocol and the implementation of specific examination exercises on ex vivo ultrasound simulator. This study involved 70 Veterinary students who were divided into four groups of similar experience in ultrasound (groups A, B, C and D) and 20 veterinarians (groups E, expert in ultrasound, and F, inexpert). Each group of students performed a specific training on basic ultrasound and abdominal ultrasound examination in dogs with different tools. The training session was performed in Group A by means of a specific e-book for iPad®, in group B with a video-presentation Power point, in group C with a web-based-learning and in group D with a printed paper document. All tools presented similar educational contents. Student groups responded to a test about “Abdominal Ultrasound examination in dogs” before (pre-test) and after (post-test) the training session. On the other hand, the veterinarians groups did not performed the training sessions but completed the same test that students. Finally every participant executed a practical ultrasound scanning test on an ex vivo simulator. In the pre-test no significant differences were observed between groups. On the other hand, post-test showed a greater score in multimedia than traditional tools. Namely, Group C (8,69±1,63 ) presented a significantly greater score compared with groups A, B and D (A=7,64±2,04; B=7,83±1,97 and D=6,92±2,81, p< 0,003). Regarding the comparisons of student’s post-test with group E, no significant differences were observed between group C and group E. Moreover, no student’s group had similar results to group E in the scanning test. In conclusion, the use of multimedia tools seems to improve to a greater extent the short-term memory retention for learning of basic principles of ultrasound and abdominal examination protocol in dogs

Current state of art after twenty years of the discovery of bioactive peptide lunasin

Non-communicable diseases have become the medical challenge of the 21st century because of their high incidence and mortality rates. Accumulating evidence has suggested that the modulation of diet and other lifestyle habits is the best strategy for the prevention of these diseases. An increasing number of dietary compounds have been found to exert health promoting benefits beyond their nutritional effects. Among them, lunasin is considered one of the most studied bioactive peptides. Since its discovery in soybean twenty years ago, many researchers around the world have focused their studies on demonstrating the chemopreventive and chemotherapeutic activity of lunasin. Moreover, in the last years, promising protective effects of this peptide against hypercholesterolemia, obesity, metabolic syndrome and associated cardiovascular disorders, and inflammatory and immune-regulated diseases have been described. This review summarizes recent remarkable advances on the use of peptide lunasin as a potential functional ingredient to provide health benefits. Moreover, novel aspects related to the influence of lunasin's digestion and bioavailability, the mechanisms of action proposed to explain the underlying biological properties, and the incorporation of this peptide into nutritional supplements are critically discussed.This work has received financial support from Ministry of Economy and Competitiveness (MINECO, Spain) through the project AGL2015-66886-R. SFT is currently funded by the Instituto de Salud Carlos III (Sara Borrell fellowship CD17/00014).Peer reviewe

Gastrointestinal digestion of food proteins under the effects of released bioactive peptides on digestive health

The gastrointestinal tract represents a specialized interface between the organism and the external environment. Because of its direct contact with lumen substances, the modulation of digestive functions by dietary substances is supported by a growing body of evidence. Food‐derived bioactive peptides have demonstrated a plethora of activities in the organism with increasing interest toward their impact over the digestive system and related physiological effects. This review updates the biological effects of food proteins, specifically milk and soybean proteins, associated to gastrointestinal health and highlights the study of digestion products and released peptides, the identification of the active form/s, and the evaluation of the mechanisms of action underlying their relationship with the digestive cells and receptors. The approach toward the modifications that food proteins and peptides undergo during gastrointestinal digestion and their bioavailability is a crucial step for current investigations on the field. The recent literature on the regulation of digestive functions by peptides has been mostly considered in terms of their influence on gastrointestinal motility and signaling, oxidative damage and inflammation, and malignant cellular proliferation. A final section regarding the actual challenges and future perspectives in this scientific topic is critically discussed.This work received financial support from the Ministry of Science and Innovation, Spain (MINECO, Spain) through project AGL2015‐66886‐R. S.F.T. is currently funded by the Instituto de Salud Carlos III (Sara Borrell fellowship CD17/00014).Peer reviewe

Italian legumes: effect of sourdough fermentation on lunasin-like polypeptides

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License.-- Rizzello et al.[Background]: There is an increasing interest toward the use of legumes in food industry, mainly due to the quality of their protein fraction. Many legumes are cultivated and consumed around the world, but few data is available regarding the chemical or technological characteristics, and especially on their suitability to be fermented. Nevertheless, sourdough fermentation with selected lactic acid bacteria has been recognized as the most efficient tool to improve some nutritional and functional properties. This study investigated the presence of lunasin-like polypeptides in nineteen traditional Italian legumes, exploiting the potential of the fermentation with selected lactic acid bacteria to increase the native concentration. An integrated approach based on chemical, immunological and ex vivo (human adenocarcinoma Caco-2 cell cultures) analyses was used to show the physiological potential of the lunasin-like polypeptides.[Results]: Italian legume varieties, belonging to Phaseulus vulgaris, Cicer arietinum, Lathyrus sativus, Lens culinaris and Pisum sativum species, were milled and flours were chemically characterized and subjected to sourdough fermentation with selected Lactobacillus plantarum C48 and Lactobacillus brevis AM7, expressing different peptidase activities. Extracts from legume doughs (unfermented) and sourdoughs were subjected to western blot analysis, using an anti-lunasin primary antibody. Despite the absence of lunasin, different immunoreactive polypeptide bands were found. The number and the intensity of lunasin-like polypeptides increased during sourdough fermentation, as the consequence of the proteolysis of the native proteins carried out by the selected lactic acid bacteria. A marked inhibitory effect on the proliferation of human adenocarcinoma Caco-2 cells was observed using extracts from legume sourdoughs. In particular, sourdoughs from Fagiolo di Lamon, Cece dell’Alta Valle di Misa, and Pisello riccio di Sannicola flours were the most active, showing a decrease of Caco-2 cells viability up to 70 %. The over-expression of Caco-2 filaggrin and involucrin genes was also induced. Nine lunasin-like polypeptides, having similarity to lunasin, were identified.[Conclusions]: The features of the sourdough fermented legume flours suggested the use for the manufacture of novel functional foods and/or pharmaceuticals preparations.Peer reviewe