Vaccination prepartum enhances the beneficial effects of melatonin on the immune response and reduces platelet responsiveness in sheep

BACKGROUND: Melatonin regulates several physiological processes and its powerful action as antioxidant has been widely reported. Melatonin acts modulating the immune system, showing a protective effect on the cardiovascular system and improving vaccine administration as an adjuvant-like agent. Here, we have investigated the role of melatonin as an adjuvant of the Clostridium perfringens vaccine in prepartum sheep and whether melatonin modulates platelet physiology during peripartum. RESULTS: The experiments were carried out in peripartum sheep from a farm located in an area of Mediterranean-type ecosystem. Plasma melatonin levels were determined by ELISA and sheep platelet aggregation was monitored using an aggregometer. Here we demonstrated for the first time that plasma melatonin concentration were higher in pregnant (125 pg/mL) than in non-pregnant sheep (15 pg/mL; P < 0.05). Administration of melatonin prepartum did not significantly modify platelet function but significantly improved the immune response to vaccination against C. perfringens. CONCLUSION: Administration of melatonin as an adjuvant provides a significant improvement in the immune response to vaccine administration prepartum against C. perfringens

Lupinus angustifolius Protein Hydrolysates Reduce Abdominal Adiposity and Ameliorate Metabolic Associated Fatty Liver Disease (MAFLD) in Western Diet Fed-ApoE−/− Mice

Metabolic-associated fatty liver disease (MAFLD) is the most important cause of liver disease worldwide. It is characterized by the accumulation of fat in the liver and is closely associated with abdominal obesity. In addition, oxidative stress and inflammation are significant features involved in MAFLD. Recently, our group demonstrated that lupin protein hydrolysates (LPHs) had lipid lowering, antioxidant, and anti-inflammatory effects. Sixty male mice fed with a Western diet were intragastrically treated with LPHs (or vehicle) for 12 weeks. Liver and adipose tissue lipid accumulation and hepatic inflammatory and oxidant status were evaluated. A significant decrease in steatosis was observed in LPHs-treated mice, which presented a decreased gene expression of CD36 and LDL-R, crucial markers in MAFLD. In addition, LPHs increased the hepatic total antioxidant capacity and reduced the hepatic inflammatory status. Moreover, LPHs-treated mice showed a significant reduction in abdominal adiposity. This is the first study to show that the supplementation with LPHs markedly ameliorates the generation of the steatotic liver caused by the intake of a Western diet and reduces abdominal obesity in ApoE−/− mice. Future clinical trials should shed light on the effects of LPHs on MAFLD.Ministerio de Economía y Competitividad [AGL2012-40247-C02-01 and AGL2012-40247-C02-02]Andalusian Government Ministry of Health [PC-0111-2016-0111]PAIDI Program from the Andalusian Government [CTS160]Spanish Ministerio de Educación, Cultura y Deporte [FPU16/02339] and [FPU13/01210]The National Net RETICEF for Aging Studies (RD12/0043/0012 from the Instituto de Salud Carlos III, Spanish Ministerio de Ciencia e Innovación)European Social Fund and Spanish Ministerio de Empleo y Seguridad Social [EJ-086]Andalusian Government Ministry of Health [PI-0136-2019]VI Program of Inner Initiative for Research and Transfer of University of Seville (VI PPIT-US

Anxiolytic-like effects of Lupinus angustifolious protein hydrolysates in alzheimer model mice

Alzheimer’s disease (AD), which is characterized by a devastating and progressive loss of memory, is the principal neurodegenerative disease in the elderly population worldwide. As a consequence, AD patients present neuropsychiatric symptoms, such as anxiety, causing sleeping difficulty, irritability, agitation, or aggressiveness. Previous studies have demonstrated that a high-fat diet, in addition to exacerbating AD, aggravates anxiety. We have demonstrated that Lupinus angustifolius protein hydrolysates (LPHs) have anti-inflammatory and antioxidant effects, key factors for AD and anxiety. Thus, this study aimed to evaluate the potential effects of LPHs on spatial memory and anxiety of a preclinical model of AD. ApoE−/− mice fed with a western diet were intragastrically treated with LPHs (or a vehicle) for 14 weeks. Spatial memory and anxiety were then assessed through the Morris water maze and elevated plus maze, respectively. The results did not show significant differences in spatial memory between groups. However, a significant increase (p < 0.05) in time in open arms, center time, the number of crossings, and a reduction of anxiety behavior were observed in LPH-treated mice. This is the first study showing that an LPH treatment causes anxiolytic effects, pointing to LPHs as a potential component of future nutritional therapies.Ministerio de Economía y Competitividad AGL2012-40247-C02-01Ministerio de Economía y Competitividad AGL2012-40247-C02-02Ministerio de Salud del Gobierno Andaluz PC-0111-2016-0111Gobierno Andaluz CTS160Ministerio de Educación, Cultura y Deporte FPU16/02339Ministerio de Educación, Cultura y Deporte FPU13/01210Universidad de Sevilla VI PPIT-U

On the toxicity of e-cigarettes consumption: Focus on pathological cellular mechanisms

Tobacco smoking remains without a doubt one of the leading causes of premature death worldwide. In combination with conventional protocols for smoking cessation, e-cigarettes have been proposed as a useful tool to quit smoking. Advertised as almost free of toxic effects, e-cigarettes have rapidly increased their popularity, becoming a sought-after device, especially among young people. Recently some health concerns about e-cigarette consumption are being raised. It is well known that they can release several toxic compounds, some of which are carcinogenic to humans, and emerging results are now outlining the risks related to the onset of respiratory and cardiovascular diseases and even cancer. The present review shows the emerging evidence about the role of technical components of the devices, the e-liquid composition as well as customization by consumers. The primary topics we discuss are the main toxicological aspects associated with e-cigarette consumption, focusing on the molecular pathways involved. Here it will be shown how exposure to e-cigarette aerosol induces stress/mitochondrial toxicity, DNA breaks/fragmentation following the same pathological pathways triggered by tobacco smoke, including the deregulation of molecular signalling axis associated with cancer progression and cell migration. Risk to fertility and pregnancy, as well as cardiovascular risk associated with e-cigarette use, have also been reported.This work was supported by a grant from the Italian Ministry of Education, University and Research. S.G., PhD fellowship grants were awarded from the Italian Ministry of Education, University and Research. L.R. and F.V. postdoctoral fellowship grant was cofounded by D.C., M.P., S.C., and P.R.; I.C-C. was supported by a postdoctoral fellowship from the Andalusian Government Ministry of Economy, Knowledge, Business, and University (DOC_00587/2020).Peer reviewe

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

A Lupinus angustifolius protein hydrolysate exerts hypocholesterolemic effects in Western diet-fed ApoE-/- mice through the modulation of LDLR and PCSK9 pathways

6 Figuras.-- 2 TablasLupin protein hydrolysates (LPHs) are gaining attention in the food and nutraceutical industries due to their several beneficial health effects. Recently, we have shown that LPH treatment reduces liver cholesterol and triglyceride levels in hypercholesterolemic mice. The aim of this study was to elucidate the effects of LPH treatment on the molecular mechanism underlying liver cholesterol metabolism in ApoE-/- mice fed the Western diet. After identifying the composition of the peptide within the LPH mixture and determining its ability to reduce HMGCoAR activity in vitro, its effect on the LDLR and PCSK9 pathways was measured in liver tissue from the same mice. Thus, the LPH reduced the protein levels of HMGCoAR and increased the phosphorylated inactive form of HMGCoAR and the pHMGCoAR/HMGCoAR ratio, which led to the deactivation of de novo cholesterol synthesis. Furthermore, the LPH decreased the protein levels of SREBP2, a key upstream transcription factor involved in the expression of HMGCoAR and LDLR. Consequently, LDLR protein levels decreased in the liver of LPH-treated animals. Interestingly, the LPH also increased the protein levels of pAMPK responsible for HMGCoAR phosphorylation. Furthermore, the LPH controlled the PSCK9 signal pathway by decreasing its transcription factor, the HNF1-α protein. Consequently, lower PSCK9 protein levels were found in the liver of LPH-treated mice. This is the first study elucidating the molecular mechanism at the basis of the hypocholesterolemic effects exerted by the LPH in an in vivo model. All these findings point out LPHs as a future lipid-lowering ingredient to develop new functional foods.This research was funded by the Andalusian Government Ministry of Health (PC-0111-2016-0111), and the PAIDI Program from the Andalusian Government (CTS160). I. C.-C. was supported by the VI Program of Inner Initiative for Research and Transfer of University of Seville (VIPPIT-2020-II.4) and a postdoctoral fellowship from the Andalusian Government Ministry of Economy, Knowledge, Business, and University (DOC_00587/2020). G. S.-S. was supported by a FPU grant from the Spanish Ministerio de Educación, Cultura y Deporte (FPU16/02339), and by an Erasmus+ Mobility Programme. The authors gratefully acknowledge the Carlo Sirtori Foundation (Milan, Italy) for having provided part of the equipment used in this experimentation. We thank all the staff from the Instituto de Biomedicina de Sevilla (IBiS) Animal Facility for their valuable assistance.Peer reviewe

Chemical and biological characterization of the DPP-IV inhibitory activity exerted by lupin (Lupinus angustifolius) peptides: From the bench to the bedside investigation

10 Páginas.-- 5 Figuras.-- 1 TablaDipeptidyl peptidase IV (DPP-IV) is considered a key target for the diabetes treatment, since it is involved in glucose metabolism. Although lupin protein consumption shown hypoglycemic activity, there is no evidence of its effect on DPP-IV activity. This study demonstrates that a lupin protein hydrolysate (LPH), obtained by hydrolysis with Alcalase, exerts anti-diabetic activity by modulating DPP-IV activity. In fact, LPH decreased DPP-IV activity in a cell-free and cell-based system. Contextually, Caco-2 cells were employed to identify LPH peptides that can be intestinally trans-epithelial transported. Notably, 141 different intestinally transported LPH sequences were identified using nano- and ultra-chromatography coupled to mass spectrometry. Hence, it was demonstrated that LPH modulated the glycemic response and the glucose concentration in mice, by inhibiting the DPP-IV. Finally, a beverage containing 1 g of LPH decreased DPP-IV activity and glucose levels in humans.Ministerio de Economía y Competitividad, Gobierno de España [AGL2012-40247-C02-01 and AGL2012-40247-C02-02]. Consejería de Salud [PC-0111-2016-0111], and the PAIDI Program [CTS160] from Junta de Andalucía. I.C.-C. was supported by Ministerio de Educación, Cultura y Deporte, Gobierno de España [FPU13/01210], by the Universidad de Sevilla [VIPPIT-2020-II.4], and by a postdoctoral fellowship from the Consejería de Transformación Económica, Industria, Conocimiento y Universidades, Junta de Andalucía [DOC_00587/2020]. G.S.-S. was supported by Ministerio de Educación, Cultura y Deporte, Gobierno de España [FPU16/02339], and by an Erasmus+ Mobility Programme.Peer reviewe

A Lupin (Lupinus angustifolius) Protein Hydrolysate Exerts Anxiolytic-Like Effects in Western Diet-Fed ApoE−/− Mice

16 Páginas.-- 6 Figuras.-- 3 TablasAnxiety is the most prevalent psychiatric disorder worldwide, causing a substantial economic burden due to the associated healthcare costs. Given that commercial anxiolytic treatments may cause important side effects and have medical restrictions for prescription and high costs, the search for new natural and safer treatments is gaining attention. Since lupin protein hydrolysate (LPH) has been shown to be safe and exert anti-inflammatory and antioxidant effects, key risk factors for the anxiety process and memory impairment, we evaluated in this study the potential effects of LPH on anxiety and spatial memory in a Western diet (WD)-induced anxiety model in ApoE-/- mice. We showed that 20.86% of the 278 identified LPH peptides have biological activity related to anxiolytic/analgesic effects; the principal motifs found were the following: VPL, PGP, YL, and GQ. Moreover, 14 weeks of intragastrical LPH treatment (100 mg/kg) restored the WD-induced anxiety effects, reestablishing the anxiety levels observed in the standard diet (SD)-fed mice since they spent less time in the anxiety zones of the elevated plus maze (EPM). Furthermore, a significant increase in the number of head dips was recorded in LPH-treated mice, which indicates a greater exploration capacity and less fear due to lower levels of anxiety. Interestingly, the LPH group showed similar thigmotaxis, a well-established indicator of animal anxiety and fear, to the SD group, counteracting the WD effect. This is the first study to show that LPH treatment has anxiolytic effects, pointing to LPH as a potential component of future nutritional therapies in patients with anxiety.This research was funded by the Spanish Government, Ministerio de Economía y Competitividad (AGL2012-40247-C02-01, and AGL2012-40247-C02-02), the Andalusian Government Ministry of Health (PC-0111-2016-0111, and PEMP-0085-2020) and the PAIDI Program from the Andalusian Government (CTS160). G.S.-S. was supported by a FPU grant from the Spanish Ministerio de Educación, Cultura y Deporte (FPU16/02339). E.P.-E. and I.B. were supported by the VI Program of Inner Initiative for Research and Transfer of University of Seville (VI PPIT-US). I.C.-C. was supported by the VI Program of Inner Initiative for Research and Transfer of the University of Seville (VIPPIT-2020-II.4) and by a postdoctoral fellowship from the Andalusian Government Ministry of Economy, Knowledge, Business, and University (DOC_00587/2020). N.A.-S. was supported by a fellowship from the National Net RETICEF for Aging Studies (RD12/0043/0012 from the Instituto de Salud Carlos III, Spanish Ministerio de Ciencia e Innovación). A.I.Á.-L. was funded by Andalusian Government Ministry of Health (PI-0136-2019).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