Safety and Efficacy of a Beverage Containing Lupine Protein Hydrolysates on the Immune, Oxidative and Lipid Status in Healthy Subjects: An Intervention Study (the Lupine-1 Trial)

Scope: We have previously demonstrated the anti-inflammatory and antioxidant properties of in vitro administered Lupinus angustifolius protein hydrolysates (LPHs) on human peripheral blood mononuclear cells (PBMCs). This study aims to evaluate the safety and efficacy of a beverage containing LPHs (LPHb) on the immune, oxidative and metabolic status of healthy subjects. Methods and Results: In this open-label intervention, 33 participants daily ingest a LPHb containing 1 g LPHs for 28 days. Biochemical parameters are assayed in fasting peripheral blood and urine samples before, during (14 days) and after LPHb ingestion. Participants’ health status and the immune and antioxidant responses of PBMCs are also evaluated throughout the trial. The LPHb ingestion is safe and effective in both increasing the anti-/pro-inflammatory response of PBMCs and improving the cellular anti-oxidant capacity. LPHb also reduces the low-density lipoprotein-cholesterol (LDL-C)/high-density lipoprotein-cholesterol (HDL-C) atherogenic index. LPHb effect is particularly beneficial on decreasing not only the LDL-C/HDL-C index but also serum total cholesterol levels in the male cohort that shows the highest baseline levels of well-known cardiovascular risk factors. Conclusion: This is the first study to show the pleiotropic actions of a lupine bioactive peptides-based functional food on key steps of atherosclerosis including inflammation, oxidative stress, and cholesterol metabolism.Ministerio de Economía y Competitividad AGL2012-40247-C02-01, AGL2012-40247- C02-02Junta de Andalucía PC-0111-2016- 0111, CTS160Ministerio de Ciencia e Innovación RD06/0013/0001, RD12/0043/001

Effect of thermal processing on the profile of bioactive compounds and antioxidant capacity of fermented orange juice

Previously, we reported that alcoholic fermentation enhanced flavanones and carotenoids content of orange juice. The aim of this work was to evaluate the influence of pasteurization on the qualitative and quantitative profile of bioactive compounds and the antioxidant capacity of fermented orange juice. Ascorbic acid (203 mg/L), total flavanones (647 mg/L), total carotenoids (7.07 mg/L) and provitamin A (90.06 RAEs/L) values of pasteurized orange beverage were lower than those of fermented juice. Total phenolic remained unchanged (585 mg/L) and was similar to that of original juice. The flavanones naringenin-7-O-glucoside, naringenin-7-O-rutinoside, hesperetin-7-O-rutinoside, hesperetin-7-O-glucoside and isosakuranetin-7-O-rutinoside, and the carotenoids karpoxanthin and isomer, neochrome, lutein, ζ-carotene, zeaxanthin, mutatoxanthin epimers, β-cryptoxanthin and auroxanthin epimers were the major compounds. Pasteurization produced a decrease in antioxidant capacity of fermented juice. However, TEAC (5.45 mM) and ORAC (6353 μM) values of orange beverage were similar to those of original orange juice. The novel orange beverage could be a valuable source of bioactive compounds with antioxidant capacity and exert potential beneficial effects.We are grateful for the support of Junta de Andalucía (Projects P09-AGR4814M, P08-AGR-03477 and Grupo PAI BIO311), and of National Funding Agencies (Projects AGL2010-14850/ALI, AGL2011-23690, CSD007-0063 and CSIC 201170E041). We are also grateful to Fundación Séneca - CARM “Group of Excellence in Research” 04486/GERM/06. The research project grant of B.E.-L. is supported by Junta de Andalucía. A.G.-I., F.F. and S.M. are members of the CORNUCOPIA Network 112RT0460 and D.H.-M. of the IBERCAROT Network 112RT0445 financed by CYTED. We are also grateful to Grupo Hespérides Biotech S.L. for providing the samples.Peer Reviewe

Bioactive Peptides from Lupin (Lupinus angustifolius) Prevent the Early Stages of Atherosclerosis in Western Diet-Fed ApoE-/- Mice

11 Páginas.-- 5 Figuras.-- 2 TablasWe have previously reported the in vitro hypocholesterolemic, anti-inflammatory, and antioxidant effects of Alcalase-generated lupin protein hydrolysate (LPH). Given that lipoprotein deposition, oxidative stress, and inflammation are the main components of atherogenesis, we characterized the LPH composition, in silico identified LPH-peptides with activities related to atherosclerosis, and evaluated the in vivo LPH effects on atherosclerosis risk factors in a mouse model of atherosclerosis. After 15 min of Alcalase hydrolysis, peptides smaller than 8 kDa were obtained, and 259 peptides out of 278 peptides found showed biological activities related to atherosclerosis risk factors. Furthermore, LPH administration for 12 weeks reduced the plasma lipids, as well as the cardiovascular and atherogenic risk indexes. LPH also increased the total antioxidant capacity, decreased endothelial permeability, inflammatory response, and atherogenic markers. Therefore, this study describes for the first time that LPH prevents the early stages of atherosclerosis.Ministerio de Economía y Competitividad, Gobierno de España [AGL2012-40247-C02-01 and AGL2012-40247-C02-02], Consejería de Salud, Junta de Andalucía [PC-0111-2016-0111 and PEMP-0085-2020], and the Programa PAIDI from the Junta de Andalucía [CTS160]. G.S.-S. was supported by Formación Profesorado Universitario grants from the Ministerio de Educación, Cultura y Deporte, Gobierno de España [FPU16/02339]. I.C.-C. was supported by a postdoctoral fellowship from the Consejería de Economía, Conocimiento, Empresas y Universidad, Junta de Andalucía [DOC_00587/2020]. N.Á.-S. was supported by a fellowship from the Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF) [RD12/0043/0012 from the Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación, Gobierno de España]. B.R.-O. was supported by a grant from the Programa de Empleo Juvenil of Ministerio de Empleo y Seguridad Social, Gobierno de España [EJ-086]. A.I.Á.-L. was funded by the Consejería de Salud, Junta de Andalucía [PI-0136-2019]. I.B. was supported by the VI Plan Propio de Investigación y Transferencia of Universidad de Sevilla [VI PPIT-US].Peer reviewe

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

Capacidad de absorción de radicales de oxígeno de los compuestos antioxidantes del vino: Ensayos in vitro e in vivo

El vino es un alimento propio de la dieta mediterránea y recientemente se ha relacionado su consumo moderado y regular con la prevención y disminución del riesgo de padecer enfermedades cardiovasculares, crónicas y degenerativas en la población occidental, sobre todo desde la aparición de la llamada “Paradoja Francesa”. La diferencia fundamental entre el vino y otras bebidas alcohólicas es la presencia de compuestos fenólicos a los que se les atribuye la potencial acción cardiosaludable del vino, ya que presentan actividad antioxidante eficaz como vasodilatadores, antiinflamatorios e incluso anticancerígenos. El organismo humano posee mecanismos para defender sus propias células de las lesiones que les pueden causar los radicales libres de oxígeno, sean estos generados como parte de los procesos metabólicos normales o anormales. Estos mecanismos de defensa incluyen sistemas antioxidantes enzimáticos y no enzimáticos; en determinadas situaciones esta defensa endógena se satura y es necesario un aporte externo de antioxidantes en la dieta para evitar así el desarrollo de las enfermedades que tienen su origen en el daño oxidativo, como las enfermedades cardiovasculares, cáncer, envejecimiento, enfermedades oculares y otros desórdenes coronarios, inmunes y neurodegenerativos. Los compuestos fenólicos presentes en el vino son los responsables de sus propiedades antioxidantes. Se hacen necesarios tanto ensayos in vitro como in vivo en orden a explicar las propiedades que se les atribuyen. La mayoría de los estudios que miden el efecto in vivo del consumo de alimentos se basan en el cambio que experimenta la capacidad antioxidante de diferentes fluidos biológicos (plasma, suero, orina). Junto a la determinación de la composición en nutrientes de un alimento, cada día cobra mayor importancia la evaluación de su potencial antioxidante. En este sentido en los últimos años se han desarrollado y validado varios métodos para determinar la capacidad antioxidante total de alimentos y muestras biológicas. Los ensayos ORAC (Capacidad de Absorbancia de Radicales de Oxígeno), TRAP (Captura Total de Radicales Peroxilos), FRAP (Poder Antioxidante de Reducción del Férrico) y TEAC (Capacidad Antioxidante Equivalente al Trolox) han sido los más utilizados. La producción de vinos que tengan mayor potencial saludable es un reto para la industria enológica. Conocer cuales son los compuestos que contribuyen a incrementar este potencial, y dirigir las prácticas enológicas hacia su consecución es un objetivo prioritario. Este trabajo se propone analizar la actividad antioxidante de diferentes vinos y de los compuestos fenólicos de su composición, así como determinar el efecto que su ingesta produce en el organismo humano para poder recomendar así su consumo moderado a la dieta. OBJETIVOS: 1. Estudiar la capacidad de absorbancia de radicales de oxígeno (método ORAC) de los principales compuestos polifenólicos del vino por separado para analizar la relación estructura química-actividad antioxidante. 2. Determinar si los metabolitos formados en la flora del colon a partir de los compuestos fenólicos del vino presentan actividad antioxidante. En caso afirmativo, comparar los valores obtenidos con los de sus compuestos de origen. 3. Relacionar la capacidad de absorbancia de radicales de oxígeno de distintos tipos de vinos (blancos, generosos y tintos) con su composición fenólica. 4. Evaluar la influencia de distintos procesos tecnológicos de vinificación en la capacidad antioxidante y composición fenólica de los vinos: - Efecto del prensado en mostos blancos. - Efecto del tiempo de maceración en mostos tintos procedentes de distintas variedades de uva (Cabernet-Sauvignon, Tempranillo y Syrah). - Efecto del tipo de clarificante y el sistema de filtrad en mostos blancos y tintos. 5. Estudiar la modificación de la capacidad antioxidante del plasma de voluntarios humanos tras la ingesta de vino tinto: - Evaluar la idoneidad de distintos métodos de medida (ORAC-PE, ORAC-FL y FRAP) y de la matriz biológica usada (suero y plasma). - Determinar la capacidad antioxidante del plasma a diferentes tiempos tras la ingesta del vino. - Analizar por CLAE los compuestos fenólicos o metabolitos que aparecen en el plasma después del consumo del vino. - Estudiar la posible influencia de antioxidantes endógenos (albúmina, bilirrubina y ácido úrico) en el aumento de la capacidad antioxidante del plasma

Effect of alcoholic fermentation on the carotenoid composition and provitamin A content of orange juice

Orange juice is considered a rich source of carotenoids, which are thought to have diverse biological functions. In recent years, a fermentation process has been carried out in fruits resulting in products that provide higher concentrations of bioactive compounds than their original substrates. The aim of this study was to evaluate the effect of a controlled alcoholic fermentation process (15 days) on the carotenoid composition of orange juice. Twenty-two carotenoids were identified in samples. The carotenoid profile was not modified as result of the fermentation. Total carotenoid content and provitamin A value significantly increased from day 0 (5.37 mg/L and 75.32 RAEs/L, respectively) until day 15 (6.65 mg/L and 90.57 RAEs/L, respectively), probably due to a better extractability of the carotenoids from the food matrix as a result of processing. Therefore, the novel beverage produced could provide a rich source of carotenoids and exert healthy effects similar to those of orange juice. © 2014 American Chemical Society.This study was supported by funding from the Ministerio de Ciencia e Innovación (Spanish government, Project AGL2010-14850/ALI) and the Consejerı́a de Economı́a, Innovación, Ciencia y Empleo (Junta de Andalucı́a, Projects P08-AGR-03477 and P09-AGR4814, and Grupo PAI BIO311). The Research Project grant of B.E.-L. is supported by the Junta de Andalucı́a. D.H.-M. is member of the IBERCAROT Network 112RT0445 financed by CYTED.Peer Reviewe

Changes in Antioxidant Endogenous Enzymes (Activity and Gene Expression Levels) after Repeated Red Wine Intake

6 páginas, 7 figuras, 3 tablas.The antioxidant properties of wine have been largely related to the reactive oxygen species (ROS) scavenging ability of phenolic compounds. Polyphenolic compounds are hardly absorbed and quickly transformed into metabolites. Their antioxidant activities just as radical scavenging properties are therefore limited, but it is worth looking to other mechanisms. This study intended to test whether wine consumption affects antioxidant enzyme activity and gene expression. For this purpose, eight subjects drank 300 mL of red wine every day for a week and ate a low phenolic diet (LPD + W) specifically designed to avoid interferences from other polyphenols in the diet. The control period was a week with this diet, and volunteers refrained from drinking wine (LPD). Blood samples were taken at 0, 1, and 7 days. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR) activities were determined in erythrocytes (SOD), plasma (CAT and GR), and blood (GPx). Gene expression was determined in macrophages. Oxidative stress caused by LPD reduced SOD, CAT, and GR activities. After wine consumption, these activities significantly increased (P < 0.05), and this overcame the effect of oxidative stress on enzyme activity. The modulation of CAT activity may be independent of changes in their gene expression, which significantly increased after LPD. However, SOD gene expression increased only during the LPD + W week. Enzyme activities are not all regulated in the same way. The results show that subacute moderate wine ingestion modulated antioxidant enzyme expression and activity, which is important for the prevention of ROS-associated diseases.We thank the Spanish Ministry of Science and Education for financial support (AGL 2007-64622).Peer reviewe

Beta-cryptoxanthin is more bioavailable in humans from fermented orange juice than from orange juice

Trabajo presentado en el XVII Congreso de la Sociedad Española de Nutrición (SEN): Alimantación 5S, celebrado en Barcelona del 27 al 29 de junio de 2018Orange juice (OJ) has one of the highest carotenoid content reported in any fruit‐derived food1. The consumption of OJ is estimated to provide a majority of the intake of β‐cryptoxanthn2. This pigment is a vitamin A precursor and have found to exerts antioxidant and anticancer effects, among other biological activities in the organism3. Food matrix characteristics and food processing techniques influence on the bioaccessibility and absorption of carotenoids, respectively4. In previous studies, our group developed and characterized a novel fermented orange beverage (FOB) obtained by the alcoholic fermentation of OJ and its subsequent heat treatment, finding no differences between FOB and OJ in qualitative or quantitative carotenoid profile5,6. However, it is plausible that the bioavailability of carotenoids may be higher from FOB than from OJ due to food matrix modifications.Peer reviewe

β-Cryptoxanthin is more bioavailable in humans from fermented orange juice than from orange juice

24 Páginas.-- 3 Tablas.-- 1 FiguraCarotenoids, especially β-cryptoxanthin, exert multiple biological activities in the organism. Various processing techniques can improve carotenoid bioavailability in relation to the food matrix. The study objective was to compare the bioavailability of carotenoids from orange juice (OJ) with that from a beverage obtained by alcoholic fermentation of orange juice (FOB). Seven volunteers were recruited for a randomized, controlled, and crossover study. Post-intake plasma carotenoid concentrations were measured by HPLC in the subjects at 0–8 h after their consumption of OJ or FOB. β-Cryptoxanthin and lutein absorption was significantly higher from FOB than from OJ, but no significant difference in zeaxanthin absorption was found. The mean baseline-corrected area under the concentration curve (AUC0-8 h) for β-cryptoxanthin, lutein and zeaxanthin was 24.6-, 1.3- and 4.65-fold larger, respectively, after FOB versus OJ consumption. This fermented orange beverage could be an abundant source of bioavailable carotenoids, and its regular consumption may exert healthy effects.The authors thank the Spanish Government (projects AGL2010-14850/ALI and AGL2014-53195R) and Junta de Andalucía (projects P09-AGR4814M and P08-AGR-03477, and group PAI BIO311) for financial support. DHM is member of CaRed Network funded by MINECO (BIO2015-71703-REDT) and participates in EUROCAROTEN COST Action (CA15136). They are also grateful to Grupo Hespérides Biotech S.L. for supplying the fermented orange beverage.Peer reviewe

Consumption of orange fermented beverage reduces cardiovascular risk factors in healthy mice

The consumption of fruits prevents the risk of cardiovascular diseases. Alcoholic fermentation has been carried out in fruits resulting in products which provide high concentration of bioactive compounds and variable alcohol content. The aim of this study was to assess the potential beneficial effect of an orange beverage obtained by alcoholic fermentation and pasteurization of orange juice on cardiovascular risk biomarkers. For this purpose, four mice groups (n = 8) ingested orange beverage (equivalent volume to 250 mL/day in human), orange juice, alcoholic solution (at the proportional amount of orange beverage) or water during 12 weeks. The equivalent amount to double serving of orange beverage (500 mL/day) was administered to mice in a subsequent intervention, and a control group was also evaluated. Orange beverage consumption increased levels of glutathione and uric acid, improved lipid profile, decreased oxidized LDL and maintained levels of IL-6 and C-reactive protein. Synergistic effects between the bioactive compounds and the alcohol content of orange beverage may occur. The intake of double serving also increased antioxidant enzyme activities, bilirubin content and plasma antioxidant capacity. These results suggest that orange beverage may produce greater protection against cardiovascular risk factors than orange juice in healthy mice.We are grateful for the support of the Junta de Andalucía through Projects P09-AGR4814M and Grupo PAI BIO311. The Research Project grant of BE-L is supported by the Junta de Andalucía. We also thank Grupo Hespérides Biotech S.L. for providing the orange beverage