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11 Páginas.-- 5 Figuras.-- 2 Tablas, We 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].

Toxicities according to the criteria by National Cancer Institute are described in this table.

Purpose: We describe the results of a prospective multicenter phase I/II trial evaluating the impact of the use of vitamin D (VitD) from day −5 to +100 on the outcome of patients undergoing allogeneic transplantation (EudraCT: 2010-023279-25; ClinicalTrials.gov: NCT02600988).Experimental Design: A total of 150 patients were included in three consecutive cohorts of 50 patients each group: control group (CG, not receive VitD); low-dose group (LdD, received 1,000 IU VitD daily); and high-dose group (HdD, 5,000 IU VitD daily). We measured levels of VitD, cytokines, and immune subpopulations after transplantation.Results: No significant differences were observed in terms of cumulative incidence of overall and grades 2–4 acute GVHD in terms of relapse, nonrelapse mortality, and overall survival. However, a significantly lower cumulative incidence of both overall and moderate plus severe chronic GVHD (cGVHD) at 1 year was observed in LdD (37.5% and 19.5%, respectively) and HdD (42.4% and 27%, respectively) as compared with CG (67.5% and 44.7%, respectively; P < 0.05). In multivariable analysis, treatment with VitD significantly decreased the risk of both overall (for LdD: HR = 0.31, P = 0.002; for HdD: HR = 0.36, P = 0.006) and moderate plus severe cGVHD (for LdD: HR = 0.22, P = 0.001; for HdD: HR = 0.33, P = 0.01). VitD modified the immune response, decreasing the number of B cells and naïve CD8 T cells, with a lower expression of CD40L.Conclusions: This is the first prospective trial that analyzes the effect of VitD postransplant. We observed a significantly lower incidence of cGVHD among patients receiving VitD. Interestingly, VitD modified the immune response after allo-SCT. Clin Cancer Res; 22(23); 5673–81. ©2016 AACR.

Serum levels of vitamin D. Serum levels of vitamin D. The plasma levels of 25-hydroxi-VitD are summarized in this table

Serum levels of cytokines by flow cytometry. Serum levels of Th1/Th2 cytokines are summarized in this table.

Cumulative incidence of relapse (A) or NRM (B): No significant differences between the different cohorts of patients were observed in terms of cumulative incidence of relapse or NRM (p=0.1 and p= 0.8).

Immune recovery after allo-stem cell transplantation. Mean and standard error of the mean (SEM)of different subpobpopulation of immune system are summarized in this table.

23 Páginas.-- 4 Figuras.-- 4 Tablas, Background: Over the last two decades, the demand for meat-free food has increased for health, environmental, and animal welfare reasons. Thus, research on the value of vegetable compounds as beneficial agents for human health has gained much attention. In particular, great interest has been shown in protein hydrolysation since this food technology facilitates the release of encrypted peptides with biological activities. In this sense, during the last 10 years, the biological activities of Lupinus spp. protein hydrolysates (LPHs) have been extensively studied. Scope and approach: The aim of this review is to address all studies (in silico, in vitro, and in vivo) in which the biological activities (antioxidant, anti-inflammatory, etc.) of LPHs were described, both the whole hydrolysate and the derived peptides. Moreover, the physicochemical characteristics of LPH peptides are evaluated, and challenges and future perspectives for their fast application have been discussed. Key findings and conclusions: LPHs exert many important biological effects in both the in vitro and in vivo systems. The main activities of LPHs described are antioxidant, immunomodulatory, hypotensive, hypoglycemic, and hypolipidemic. These findings point to LPHs as a possible new nutraceutical for human health, capable of preventing or treating some chronic diseases such as diabetes, dyslipidemia, and atherosclerosis. Finally, the development of a new generation of nanonutraceuticals to improve the metabolic stability and bioactivity of LPHs has been discussed., This work 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, PEMP 0085–2020 (co-financed with FEDER funds, call Resolution of 7 July 2021 of the General Secretary for Research, Development and Innovation in Health, which calls for grants to finance research, development and innovation in biomedicine and health sciences in Andalusia by 2021), and the PAIDI Program from the Andalusian Government (CTS160). ICC was supported by a postdoctoral fellowship from Consejería de Transformación Económica, Industria, Conocimiento y Universidades (Junta de Andalucía) (DOC_00587/2020). GSS was supported by a FPU grant from Ministerio de Educación, Cultura y Deporte (Gobierno de España) (FPU16/02339). AIAL was funded by Consejería de Salud (Junta de Andalucía) (PI-0136-2019 and PEMP 0085–2020).

Background: Recently, the study of hydrolysates from food proteins has been increasingly due to their wide range of biological activities. Hydrolysates contain peptides of 2–20 amino acids that are inactive within the sequence of the parent protein, but, once released after proteolytic processes, they exert numerous beneficial health effects. Hemp, the non-drug variety of Cannabis sativa, is known as an important source of bioactive peptides due to the high quality of hempseeds protein (20–25%) and well-balanced amino acid profile. For this reason, during the last decade, numerous investigations have searched to elucidate the beneficial effects on the health of these hempseed protein hydrolysates. Scope and approach: The aim of this review was to collect all the scientific evidence on the demonstrated beneficial effects of hempseed protein hydrolysates (HHs). Key findings and conclusions: HHs have showed to possess antioxidant, immunomodulatory, hypotensive, hypoglycemic, and lipid-lowering capacities in vitro systems. All these effects have pointed out HHs as future ingredient for the development of functional foods or dietary supplements useful for the prevention of chronic diseases such as metabolic syndrome, diabetes or hypertension. However, few studies have evaluated the in vivo effects of HHs. For this reason, further studies carried out in animal models or human are necessary to better exploit the use of HHs for the development of new dietary supplements., I.C.-C. was supported by the VI Program of Inner Initiative for Re- search 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). 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. A.I.A.-L. was funded by Andalusian Government Ministry of Health (PI-0136-2019). E.P.-E. was supported by the VI Pro- gram of Inner Initiative for Research and Transfer of University of Seville.

16 Páginas.-- 6 Figuras.-- 3 Tablas, Anxiety 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).

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).

Anxiety 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

Both immune and neurodegenerative mechanisms underlie multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). MS/EAE are triggered by encephalitogenic immune cells, including Th1 and Th17 cells, whereas T regulatory (Treg) cells are involved in inflammation resolution. Pro-inflammatory macrophages/microglia also play a deleterious role in the disease. Seasonal variations in MS relapses, active lesions, and pro- and anti-inflammatory cytokine levels have been described in MS patients and have been related with both perinatal and adult exposure to sunlight and other environmental factors. However, some data in EAE mice suggest that these variations might be, at least partially, endogenously determined. Thus, our objective was to study the effect of the season of birth and disease induction on the course of EAE, and immune cell infiltration in the central nervous system (CNS) in myelin oligodendrocyte glycoprotein (MOG35–55)-induced EAE in 8 weeks old, female C57BL/6N mice maintained under constant, controlled conditions. EAE severity as well as pathogenic (Th1, Th17, macrophages/microglia) and protective (Treg) subsets was found to vary according to the season of birth or of EAE induction. Summer-born or summer-immunized animals developed a milder disease, which coincided with variations in numbers of T effector/regulatory subsets, and significantly low numbers of macrophages/microglia. These results suggest that endogenous rhythms in immune responses might cause seasonal variations in EAE severity, and, maybe, in the course of MS, and that they might be related to macrophages/microglia., The work reported in the present manuscript was supported by the Department of Health of the Andalusian Government (PI-0209-2010, PI-0485-2014, and PC-0019-2017) and the PAIDI Program of the Andalusian Government (CTS160). NA-S was supported by fellowships from the National Net RETICEF for Aging Studies (Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad; RD06/0013/0001 and RD12/0043/0012; from the Instituto de Salud Carlos III, Spanish Ministry of Science and Innovation) and by a fellowship from the Department of Health of the Andalusian Government (PC-0111-2016-0111), and IC-C was supported by an FPU grant from the Spanish Ministry of Education, Culture and Sport (FPU13/01210).

6 Figuras.-- 2 Tablas, Lupin 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.

Lupin 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

In recent decades, people in the industrialized world have increased the demand for meat-free foods motivated by health, environmental, and animal welfare reasons [...]

La asignatura donde se ha realizado el ciclo de mejora en aula (CIMA) se deno- mina Bioquímica y Biología Molecular, correspondiente al primer curso del grado de Enfermería. Consta de seis créditos, repartidos en 50 horas de clases presen- ciales al grupo completo, impartidas durante 15 semanas (tres o cuatro horas se- manales), más seis horas de seminarios teórico prácticos de dos horas cada uno y 4 horas de prácticas de laboratorio a grupos pequeños. Es una asignatura tron- cal y de formación básica (obligatoria), que se imparte en el primer cuatrimestre del curso. La asignatura consta de un programa docente con 30 temas, agrupados en ocho bloques temáticos. En el curso 2021-2022, imparto la docencia de los 5 pri- meros bloques en dos de los cuatro grupos de la asignatura, con un total de 150 estudiantes entre los dos grupos. El CIMA se ha realizado en los bloques 2, 3 y 4, co- rrespondientes a la bioquímica estructural. Con antelación al abordaje de cada uno de los bloques en la clase presencial, el estudiante recibe el material docente que debe trabajar durante una semana y rellenar un cuestionario sobre el grado de en- tendimiento del material. La clase presencial se utiliza como un entorno de puesta en común de los contenidos estructuradores a través de actividades teórico-prácti- cas. Al final de cada bloque/s se llevan a cabo actividades de evaluación, que eng- lobarán los contenidos clave de todos los contenidos tratados hasta ese momento. Palabras clave: Bioquímica, enfermería, medicina, docencia universitaria, de- sarrollo profesional docente.

High-sensitivity C-reactive protein (hsCRP) and homocysteine (Hcy) are inflammation markers but are also related to cardiovascular diseases, disability, or higher risk of death. Although inflammation is considered to be associated with frailty, data regarding the association between hsCRP or Hcy and frailty are controversial or scarce, especially with respect to their association with prefrailty. Thus, our objective was to study the association of hsCRP and Hcy with prefrailty and frailty in 1,211 Spanish men and women aged 65â 98 years from the Toledo Study for Healthy Aging (TSHA) cohort, classified according to Friedâ s criteria. Hcy was independently associated with frailty (odds ratio [OR] = 1.06; 95% confidence interval [CI]: 1.01â 1.12), whereas hsCRP was independently associated with both prefrailty (OR = 1.03; 95% CI: 1.01â 1.06) and frailty (OR = 1.07; 95% CI: 1.02â 1.12). Furthermore, both markers were positively correlated with the number of Friedâ s criteria that were met and were independently associated with the criteria of exhaustion (Hcy: OR = 1.03, 95% CI: 1.00â 1.06), weakness (hsCRP: OR = 1.03, 95% CI: 1.01â 1.05), and low physical activity (hsCRP: OR = 1.04, 95% CI: 1.02â 1.06). Thus, our results highlight the importance of inflammation in age-related physical decline and, in particular, its association with fatigue, low strength, and decreased physical activity., This study was supported by the Instituto de Salud Carlos III (Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad; RD06/0013/0001, and RD12/0043/0012). Ministerio de Ciencia e Innovación; PI07/90175). NA-S was supported by fellowships from the Instituto de Salud Carlos III (Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad; RD06/0013/0001 and RD12/0043/0012) and by a fellowship from the Consejería de Salud, Junta de Andalucía (PC-0111-2016-0111). IC-C was supported by fellowships from the Ministerio de Educación, Cultura y Deporte (FPU13/01210) and Junta de Andalucía (CTS160).

Objectives To obtain greater knowledge of the extra-pineal sources of melatonin during development, the amount of indolamine and the expression levels of the last two enzymes involved in its biosynthesis, Arylalkylamine N-acetyltransferase (AANAT) and acetylserotonin O-methyltransferase (ASMT), were analyzed in the human thymus from children from three different age groups (from days to years). The melatonin membrane and nuclear receptor expression levels also were studied. Methods Quantitative reverse transcriptase PCR and western blot were performed to investigate the receptor and enzyme expression levels. The results were examined and correlated with the ages of the thymuses. Results We found high levels of indolamine in the thymuses of newborns (younger than 1 month), which decreased during development; thymuses from the months (from 2 to 11 months) and years (from 1 to 12 years) groups showed lower levels. A similar decline was also observed in the mRNA of the AANAT enzyme and the expression levels of melatonin receptors. However, ASMT expression was exactly the opposite, with low levels in the newborn group and higher levels in the years group. Our results show that the thymic synthesis of melatonin occurs very early in childhood. Additionally, this is the first report that is focused on melatonin receptors expression in the human thymus. Conclusion Considering the limited melatonin synthesis performed by the newborn pineal gland, we suggest that the high levels of melatonin found in human thymus in this experimental group arise from synthesis in the tissue itself, which could be contributing to the immune efficiency at the thymic level., Highlights • Melatonin biosynthetic machinery in the human thymus from children changes with age. • Melatonin content in the human thymus is higher in newborns. • The thymus not only synthesizes melatonin but also responds to it. • The melatonin effector system decreases throughout childhood.

15 Páginas.-- 6 Figuras.-- 1 Tabla, 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., 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 the PAIDI Program from the Andalusian Government [CTS160]. G.S.-S. and I.C.-C. were supported by FPU grants from the Spanish Ministerio de Educación, Cultura y Deporte, [FPU16/02339] and [FPU13/01210], respectively. N.Á.-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). B.R.-O. was supported by a grant from European Social Fund and Spanish Ministerio de Empleo y Seguridad Social [EJ-086]. A.I.Á.-L. was funded by the Andalusian Government Ministry of Health [PI-0136-2019]. I.B. was supported by the VI Program of Inner Initiative for Research and Transfer of University of Seville (VI PPIT-US).

10 Páginas.-- 4 Figuras.-- 8 Tablas, 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. Grant Numbers: AGL2012-40247-C02-01, AGL2012-40247-C02-02 Programa PAIDI Junta de Andalucia. Grant Number: CTS160 Ministerio de Educación, Cultura y Deporte. Grant Numbers: FPU13/01210, FPU16/02339 Ministerio de Ciencia e Innovación (Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad; RETICEF). Grant Numbers: RD06/0013/0001, RD12/0043/0012 Universidad de Sevilla. Grant Numbers: V PPIT-US, VI PPIT-US Consejería de Salud y Familias, Junta de Andalucía. Grant Number: PC-0111-2016-0111

Peptides from several plant food proteins not only maintain the nutritional values of the original protein and decrease the environmental impact of animal agriculture, but also exert biological activities with significant health-beneficial effects. Wheat is the most important food grain source in the world. However, negative attention on wheat-based products has arose due to the role of gluten in celiac disease. A controlled enzymatic hydrolysis could reduce the antigenicity of wheat gluten protein hydrolysates (WGPHs). Therefore, the aims of the present study were to evaluate the effects of the in vitro administration of Alcalase-generated WGPHs on the immunological and antioxidant responses of human peripheral blood mononuclear cells (PBMCs) from 39 healthy subjects. WGPH treatment reduced cell proliferation and the production of the Type 1 T helper (Th1) and Th17 pro-inflammatory cytokines IFN-γ and IL-17, respectively. WPGHs also improved the cellular anti-inflammatory microenvironment, increasing Th2/Th1 and Th2/Th17 balances. Additionally, WGPHs improved global antioxidant capacity, increased levels of the reduced form of glutathione and reduced nitric oxide production. These findings, not previously reported, highlight the beneficial capacity of these vegetable protein hydrolysates, which might represent an effective alternative in functional food generation., This work was supported 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 the PAIDI Program from the Andalusian Government (CTS160). I.C.-C. was supported by Ministerio de Economía y Competitividad (AGL2012-40247-C02-02) and by a FPU grant from the Spanish Ministerio de Educación, Cultura y Deporte (FPU13/01210). N.Á.-S. was supported by Instituto de Salud Carlos III, Ministerio de Economía y Competitividad (RD06/0013/0001 and RD12/0043/0012). G.S.-S. was supported by a FPU grant from the Spanish Ministerio de Educación, Cultura y Deporte (FPU16/02339). I.B. was supported by the VI Program of Inner Initiative for Research and Transfer of University of Seville (VI PPIT-US).

20 Páginas.-- 2 Figuras.-- 1 Tabla, The effect of GPETAFLR, a peptide isolated from Lupinus angustifolius L. protein hydrolysate (LPH), on osteoclastogenesis was investigated. Human osteoclasts generated from monocytes were used to analyse the effects of GPETAFLR (50–100 µg/mL) on osteoclastogenesis using TRAP reaction, RT-qPCR, and ELISA procedures. LPS enhanced TRAP activity and the expression of osteoclast marker genes (TRAP, OSCAR, RANK, and CATHK) while downregulated the expression of OPG gene in human monocyte-derived osteoclasts. These effects were reduced with GPETAFLR. Moreover, LPS increased the release of osteoclastogenic cytokines (TNF-α, IL-1β, and IL-6) meanwhile GPETAFLR increased the release of anti-osteoclastogenic cytokines (IL-4 and IL-10) in the medium of human monocyte-derived osteoclasts. For the first time, we show that plant peptides from lupine protein hydrolysates have anti-osteoclastogenic activity. These exciting findings open opportunities for developing nutritional strategies with Lupinus angustifolius L. as dietary source of plant proteins, notably GPETAFLR, to prevent development and progression of osteoclast-related diseases., This work was supported by grant AGL2012-40247-C02-01 from the Spanish Ministry of Economy and Competitiveness. Authors thank Cell Biology Unit of Instituto de la Grasa for its assistance during the fulfillment of this work. SMP acknowledges financial support from “V Own Research Plan” (University of Seville).

Background Homocysteine (Hcy) high levels are associated with fractures, bone resorption and an early onset of osteoporosis in elderly persons; a relationship between Hcy and bone formation has also been suggested but is still controversial. Frailty, an independent predictor of fractures and decreased bone mineral density is associated with altered bone metabolism in women. However, no previous works have studied the relationship among frailty, Hcy levels and bone turnover. Methods We studied the association among Hcy, osteoporosis and N-terminal propeptide of type I procollagen (PINP), C-terminal telopeptide of type I collagen (β-CTX), parathyroid hormone (PTH), calcium and 25-hydroxyvitamin D (25(OH)D) in 631 Spanish women between the ages of 65–78 from the Toledo Study for Healthy Aging (TSHA) cohort, who were classified as highly functional (robust subjects) or non-robust (pre-frail or frail subjects) according to Fried's criteria. Results Hcy was independently associated with β-CTX in the entire population (B = 0.22; 95% CI, 0.09–0.34; p = 0.001) and in the non-robust group (B = 0.24; 95% CI, 0.09–0.39; p = 0.002). Hcy was also associated with PINP in the entire and non-robust populations, but the association was lost after including the levels of β-CTX, but not the other bone biomarkers, in the multivariate analysis. This suggests that the controversial relationship between Hcy and bone formation might be explained, at least to a certain extent, by the confounding effects of β-CTX. Conclusions This work highlights the important implication of frailty status in the association between Hcy and increased bone turnover in older women.

Cerebral vasospasm, one of the main complications of subarachnoid hemorrhage (SAH), is characterized by arterial constriction and mainly occurs from day 4 until the second week after the event. Urotensin-II (U-II) has been described as the most potent vasoconstrictor peptide in mammals. An analysis is made of the serum U-II concentrations and mRNA expression levels of U-II, urotensin related peptide (URP) and urotensin receptor (UT) genes in an experimental murine model of SAH.An experimental study was carried out.Experimental operating room of the Biomedicine Institute of Seville (IBiS), Virgen del Rocío University Hospital (Seville, Spain).96 Wistar rats: 74 SAH and 22 sham intervention animals.Day 1: blood sampling, followed by the percutaneous injection of 100μl saline (sham) or blood (SAH) into the subarachnoid space. Day 5: blood sampling, followed by sacrifice of the animals.Weight, early mortality, serum U-II levels, mRNA values for U-II, URP and UT.Serum U-II levels increased in the SAH group from day 1 (0.62pg/mL [IQR 0.36-1.08]) to day 5 (0.74pg/mL [IQR 0.39-1.43]) (p0.05), though not in the sham group (0.56pg/mL [IQR 0.06-0.83] day 1; 0.37pg/mL [IQR 0.23-0.62] day 5; p=0.959). Between-group differences were found on day 5 (p0.05). The ROC analysis showed that the day 5 serum U-II levels (AUC=0.691), URP mRNA (AUC=0.706) and UT mRNA (AUC=0.713) could discriminate between sham and SAH rats. The normal serum U-II concentration range in rats was 0.56pg/mL (IQR 0.06-0.83).The urotensinergic system is upregulated on day 5 in an experimental model of SAH.

La prevalencia de la esclerosis múltiple (EM) en los países asiáticos se considera que es más baja que en los países occidentales, las poblaciones asiáticas tienen un 80% menos de riesgo de EM que las caucásicas. No se conocen bien las cifras de incidencia, prevalencia y su relación con otros países de su entorno y su relación con factores étnicos, ambientales y socioeconómicos. Hemos realizado una revisión bibliográfica exhaustiva de los datos epidemiológicos existentes en China y países limítrofes, para estudiar la frecuencia de la enfermedad, centrándonos en la prevalencia, sus cambios evolutivos a lo largo del tiempo y su relación con factores de género, ambientales, alimenticios y socioculturales. La prevalencia en China oscila en cifras que van desde 0,88 en 1986 a 5,2 en 2013 por 100.000 habitantes con una tendencia a aumentar que no es estadísticamente significativa (p = 0,08), mientras que en Japón este incremento es muy significativo, con cifras que oscilan entre 8,1 y 18,6 (p < 0,001). La prevalencia en los países donde predomina la raza caucásica son mucho más elevadas y aumentan con el tiempo, llegando a 115 por 100.000 habitantes en 2015 (r2 = 0,79, p = 0,0001). En conclusión, la prevalencia de la EM en China parece está aumentando en los últimos años, aunque la raza amarilla (chinos y japoneses, entre otros) tienen menor riesgo de padecerla que el resto de las poblaciones. La latitud no parece ser un factor muy determinante en Asia para presentar un mayor riesgo de padecer EM. The prevalence of multiple sclerosis (MS) in Asian countries is thought to be lower than in Western countries, with Asian populations presenting 80% less risk of MS than white populations. Incidence and prevalence rates in Asian countries are therefore not well defined and their association with rates in neighboring countries, as well as with ethnic, environmental, and socioeconomic factors, are not well understood. We performed a comprehensive literature review of epidemiological data from China and neighbouring countries to study the frequency of the disease, focusing on prevalence, and the progression over time and the influence of sex-related, environmental, dietary, and sociocul- tural factors. Prevalence rates in China range between 0.88 cases/100,000 population in 1986 and 5.2 cases/100,000 population in 2013, with a non-significant upwards trend (p = .08). The increase observed in Japan, where figures ranged between 8.1 and 18.6 cases/100,000 population was highly significant (p < .001). Prevalence rates in countries with predominantly white populations are considerably higher and have increased over time, reaching 115 cases/100,000 population in 2015 (r2 = 0.79, p < .0001). In conclusion, the prevalence of MS in China appears to have risen in recent years, although Asian populations (including Chinese and Japanese populations, among others) appear to pre- sent less risk than other populations. Within Asia, geographical latitude appears not to be a determining factor for developing MS.

CD39, an ectonucleotidase that hydrolyses pro-inflammatory ATP, is a marker of highly active and suppressive T regulatory cells (Tregs). Although CD39 has a role in Treg suppression and might be important in the control of neuroinflammation in relapsing-remitting multiple sclerosis (RR-MS), to date, there are contradictory reports concerning the Tregs expression of CD39 in RR-MS patients. Thus, our objectives were to assess the activity and expression of CD39, especially in Tregs from peripheral blood mononuclear cells (PBMCs) of relapsing RR-MS patients compared with control subjects and to evaluate the association of CD39+ Tregs with disability and the odds of RR-MS. The activity and expression of CD39 and the CD39+ Treg frequency were measured in PBMCs from 55 relapsing RR-MS patients (19 untreated and 36 receiving immunomodulatory treatment) and 55 age- and sex-paired controls. Moreover, the association between CD39+ Tregs and RR-MS was assessed by multivariate logistic regression. CD39 activity and the frequency of CD39-expressing Tregs were elevated in relapsing RR-MS patients. Moreover, CD39+ Tregs were significantly correlated with the EDSS score and were independently associated with the odds of RR-MS. Our results highlight the relevance of CD39+ Treg subset in the clinical outcomes of RR-MS., This work was supported by the Andalusian Government Ministry of Health (PI-0209-2010 PI-0485-2014, PC-0019-2017) and the PAIDI Program from the Andalusian Government (CTS160). NA-S was supported by a fellowship from the National Net RETICEF for Aging Studies (Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad; RD12/0043/0012 from the Instituto de Salud Carlos III, Spanish Ministerio de Ciencia e Innovación). IC-C was supported by an FPU grant from the Spanish Ministerio de Educación, Cultura y Deporte (FPU13/01210).

[Background] Seasonal influenza virus infection is a significant cause of morbimortality in the elderly. However, there is poor vaccine efficacy in this population due to immunosenescence. We aimed to explore several homeostatic parameters in the elderly that could impact influenza vaccine responsiveness., [Methods] Subjects (> 60 years old) who were vaccinated against influenza virus were included, and the vaccine response was measured by a haemagglutination inhibition (HAI) test. At baseline, peripheral CD4 and CD8 T-cells were phenotypically characterized. Thymic function and the levels of different inflammation-related biomarkers, including Lipopolysaccharide Binding Protein (LBP) and anti-cytomegalovirus (CMV) IgG antibodies, were also measured., [Results] Influenza vaccine non-responders showed a tendency of higher frequency of regulatory T-cells (Tregs) before vaccination than responders (1.49 [1.08–1.85] vs. 1.12 [0.94–1.63], respectively, p = 0.061), as well as higher expression of the proliferation marker Ki67 in Tregs and different CD4 and CD8 T-cell maturational subsets. The levels of inflammation-related biomarkers correlated with the frequencies of different proliferating T-cell subsets and with thymic function (e.g., thymic function with D-dimers, r = − 0.442, p = 0.001)., [Conclusions] Age-related homeostatic dysregulation involving the proliferation of CD4 and CD8 T-cell subsets, including Tregs, was related to a limited responsiveness to influenza vaccination and a higher inflammatory status in a cohort of elderly people., This work was supported by grants from the Fondo de Investigación Sanitaria (FIS; PI18/01216), which is co-funded by Fondos Europeos para el Desarrollo Regional (FEDER) and the Junta de Andalucía, Consejería de Economía, Innovación, Ciencia y Empleo (Proyecto de Investigación de Excelencia; CTS2593). The Spanish AIDS Research Network of Excellence also supported this study (RD16/0025/0019). YM.P was supported by the Consejería de Salud y Bienestar Social of Junta de Andalucía through the “Nicolás Monardes” programme (C-0013-2017).

Systemic lupus erythematosus (SLE) is an autoimmune disorder characterized by the production of antinuclear autoantibodies. In addition, the involvement of CD4+ T-helper (Th) cells in SLE has become increasingly evident. Although the role of melatonin has been tested in some experimental models of lupus with inconclusive results, there are no studies evaluating the melatonin effect on cells from patients with SLE. Therefore, the aim of this study was to analyse the role of in vitro administered melatonin in the immune response of peripheral leukocytes from treated patients with SLE (n = 20) and age- and sex-matched healthy controls. Melatonin was tested for its effect on the production of key Th1, Th2, Th9, Th17 and innate cytokines. The frequency of T regulatory (Treg) cells and the expression of FOXP3 and BAFF were also explored. Our results are the first to show that melatonin decreased the production of IL-5 and to describe the novel role of melatonin in IL-9 production by human circulating cells. Additionally, we highlighted a two-faceted melatonin effect. Although it acted as a prototypical anti-inflammatory compound, reducing exacerbated Th1 and innate responses in PHA-stimulated cells from healthy subjects, it caused the opposite actions in immune-depressed cells from patients with SLE. Melatonin also increased the number of Treg cells expressing FOXP3 and offset BAFF overexpression in SLE patient cells. These findings open a new field of research in lupus that could lead to the use of melatonin as treatment or cotreatment for SLE.

Multiple sclerosis (MS) is a neuroinflammatory disease of the central nervous system in which the immune system plays a central role. In particular, effector populations such as T helper (Th) 1, Th9, Th17, and Th22 cells are involved in disease development, whereas T regulatory cells (Tregs) are associated with the resolution of the disease. Melatonin levels are impaired in patients with MS, and exogenous melatonin ameliorates the disease in MS animal models by modulating the Th1/Th17/Treg responses and also improves quality of life and several symptoms in patients with MS. However, no study has examined melatonin's effect on T cells from relapsing-remitting MS (RR-MS) patients. Therefore, the objectives of the present study were to evaluate the effects of the in vitro administration of melatonin to peripheral blood mononuclear cells (PBMCs) from 64 RR-MS patients and 64 sex- and age-matched healthy subjects on Th1, Th9, Th17, Th22, and Treg responses and to analyze the expression of the melatonin effector/receptor system in these cells. Melatonin decreased Th1 and Th22 responses in patients, whereas it did not affect the Th17 and Treg subsets. Melatonin also promoted skewing toward a more protective cytokine microenvironment, as shown by an increased anti-inflammatory/Th1 ratio. Furthermore, for the first time, we describe the overexpression of the melatonin effector/receptor system in PBMCs from patients with MS; this alteration might be relevant to the disease because acetylserotonin O-methyltransferase expression significantly correlates with disease progression and T effector/regulatory responses in patients. Therefore, our data suggest that melatonin may be an effective treatment for MS.

5 Figuras.-- 2 Tablas, Although cell-free systems and immortalized cell lines have been used to demonstrate the potential health benefits of lupine proteins and peptides, no study has examined the effects of lupine protein hydrolysates (LPHs) on the immune and oxidative responses of non-immortalized human cells. Therefore, the aims of this study were to evaluate the effects of the in vitro administration of LPHs from Lupinus angustifolius on the immunological and oxidative statuses of human peripheral blood mononuclear cells (PBMCs) from 53 healthy donors. LPHs reduced PBMCs proliferation and the levels of Th1, Th9 and Th17 pro-inflammatory cytokines without being cytotoxic. LPHs also skewed the pro-/anti-inflammatory balance towards a Th2 protective response. Additionally, LPHs increased superoxide dismutase and catalase activities, and the total antioxidant capacity (TAC). This study is the first to show that LPHs reduce T cell inflammatory responses and improve the anti-inflammatory/pro-inflammatory cytokine balance and the TAC by PBMCs. Thus, LPHs may represent an effective option for developing nutritional strategies to prevent pathologies with underlying inflammation and oxidative stress., This work was supported by the Spanish Government, Ministerio de Economía y Competitividad [AGL2012-40247-C02-01, AGL2012-40247-C02-02]; the Andalusian Government, Consejería de Salud [PC-0111-2016-0111] and the PAIDI Program from the [CTS160]. IC-C was supported by a FPU grant from the Ministerio de Educación, Cultura y Deporte [FPU13/01210].

S100B is a calcium-binding protein released into the blood from astroglial cells due to brain injury. Some authors have described a correlation between S100B serum concentration and severity of brain damage. There is not much information about the accuracy of urinary S100B for predicting outcome after severe traumatic brain injury (TBI). 55 patients with severe TBI were included in the study. Blood and urine samples were drawn to determine S100B levels on admission and on the subsequent 24, 48, 72 and 96 h. S100B concentrations (serum and urine) were significantly higher in patients who were dead a month after the accident compared to survivors. ROC-analysis showed that S100B at 24h post-severe TBI is a useful tool for predicting mortality (serum: AUC 0.958, urine: AUC 0.778). The best cut-offs for S100B were 0.461 μg/L and 0.025 μg/L (serum and urine respectively), with a sensitivity of 90% for both measurements and a specificity of 88.4% (serum) and 62.8% (urine). We can state that the determination of S100B levels both in urine and serum acts as a sensitive and an effective biomarker for the early prediction of mortality after severe TBI.

Melatonin modulates a wide array of physiological events with pleiotropic effects on the immune system. While the relevance of specific melatonin membrane receptors has been well established for several biological functions, retinoic acid-related orphan receptor alpha (RORα) has been suggested as a mediator of nuclear melatonin signalling by results obtained from pharmacological approaches. However, a melatonin-mediated downstream effect cannot be ruled out, and further evidence is needed to support a direct interaction between melatonin and RORα. Here, we show that RORα is mainly located in human Jurkat T-cell nucleus, and it is co-immunoprecipitated with melatonin. Moreover, immunocytochemistry studies confirmed the co-localization of melatonin and RORα. Melatonin promoted a time-dependent decrease in nuclear RORα levels, suggesting a role in the RORα transcriptional activity. Interestingly, RORα acts as a molecular switch implicated in the mutually exclusive generation of Th17 and Treg cells, both involved in the harm/protection balance of immune conditions such as autoimmunity or acute transplant rejection. Therefore, the identification of melatonin as a natural modulator of RORα gives it a tremendous therapeutic potential for a variety of clinical disorders.

Breakdown in immunological self tolerance, leading to autoimmune diseases such as multiple sclerosis, might arise from immune recognition of self proteins that have undergone heightened posttranslational modification under pathophysiological conditions. A posttranslational modification of particular interest is the deimination of Arg to citrulline, catalyzed by peptidylarginyl deiminase (PAD) enzymes. As a CD4+ T cell-driven model of multiple sclerosis, we used experimental autoimmune encephalomyelitis (EAE) induced with the immunodominant 35–55 peptide of myelin oligodendrocyte glycoprotein (pMOG) in C57BL/6 mice to test whether citrullination of a T cell epitope can contribute to disease etiopathology. Immunization with an altered peptide ligand (APL) of pMOG with an Arg→citrulline conversion at a TCR contact (residue 41) led to the activation of two populations of APL-responsive T cells that either did, or did not cross-react with the native pMOG peptide. This APL could induce EAE. However, this reflected the activation of T cells that cross-reacted with the native pMOG epitope, because prior tolerization of these T cells using pMOG prevented APL-induced EAE. Using a passive transfer model, we found that T cells that responded specifically to the citrullinated form of pMOG were neither necessary, nor sufficient to initiate the EAE lesion. Nevertheless, these cells could provoke exacerbation of pathology if transferred into mice with ongoing EAE. The PAD2 and PAD4 enzymes were markedly upregulated in the inflamed CNS. Therefore, once inflammation is established, citrullination of target autoantigens can allow an expanded repertoire of T cells to contribute to CNS pathology.

Purpose: We describe the results of a prospective multicenter phase I/II trial evaluating the impact of the use of vitamin D (VitD) from day −5 to +100 on the outcome of patients undergoing allogeneic transplantation (EudraCT: 2010-023279-25; ClinicalTrials.gov: NCT02600988). Experimental Design: A total of 150 patients were included in three consecutive cohorts of 50 patients each group: control group (CG, not receive VitD); low-dose group (LdD, received 1,000 IU VitD daily); and high-dose group (HdD, 5,000 IU VitD daily). We measured levels of VitD, cytokines, and immune subpopulations after transplantation. Results: No significant differences were observed in terms of cumulative incidence of overall and grades 2–4 acute GVHD in terms of relapse, nonrelapse mortality, and overall survival. However, a significantly lower cumulative incidence of both overall and moderate plus severe chronic GVHD (cGVHD) at 1 year was observed in LdD (37.5% and 19.5%, respectively) and HdD (42.4% and 27%, respectively) as compared with CG (67.5% and 44.7%, respectively; P < 0.05). In multivariable analysis, treatment with VitD significantly decreased the risk of both overall (for LdD: HR = 0.31, P = 0.002; for HdD: HR = 0.36, P = 0.006) and moderate plus severe cGVHD (for LdD: HR = 0.22, P = 0.001; for HdD: HR = 0.33, P = 0.01). VitD modified the immune response, decreasing the number of B cells and naïve CD8 T cells, with a lower expression of CD40L. Conclusions: This is the first prospective trial that analyzes the effect of VitD postransplant. We observed a significantly lower incidence of cGVHD among patients receiving VitD. Interestingly, VitD modified the immune response after allo-SCT. Clin Cancer Res; 22(23); 5673–81. ©2016 AACR.

Melatonin is an indoleamine widely distributed in the evolution that shows a great functional versatility, playing an important role as a transmitter of photoperiodic information and exhibiting antioxidant, oncostatic, anti-aging and immunomodulatory properties. In vertebrates, this molecule is produced by the pineal gland and other extrapineal sites. The present study was carried out to investigate the presence of melatonin in thymus and the possibility of an endogenous melatonin synthesis in this organ, in which T cells are matured. In this work, we demonstrate in humans and rats that thymus contains melatonin, expresses the mRNAs encoding N-acetyltransferase and hydroxyindol-O-methyltransferase, the two key enzymes of the melatonin synthesis, and has this biosynthetic machinery activated. In addition, rat thymocytes cultured for 24 h exhibited high levels of melatonin. The results presented here suggest that human and rat thymuses are able to synthesize melatonin, which could have intracrine, autocrine and paracrine functions.

Experimental autoimmune encephalomyelitis (EAE), the experimental model for multiple sclerosis (MS), is triggered by myelin-specific Th1 and Th17 cells. The immunomodulatory activities of melatonin have been shown to be beneficial under several conditions in which the immune system is exacerbated. Here, we sought to elucidate the basis of the melatonin protective effect on EAE by characterizing the T effector/regulatory responses, particularly those of the memory cell subsets. Melatonin was tested for its effect on Th1, Th17 and T regulatory (Treg) cells in the lymph nodes and CNS of immunodominant peptide of myelin oligodendrocyte glycoprotein (pMOG)-immunized and EAE mice, respectively. The capacity of melatonin to ameliorate EAE as well as modifying both T cell response and effector/regulatory balance was surveyed. T cell memory subsets and CD44, a key activation marker involved in the EAE pathogenesis, were also examined. Melatonin protected from EAE by decreasing peripheral and central Th1/Th17 responses and enhancing both the Treg frequency and IL-10 synthesis in the CNS. Melatonin reduced the T effector memory population and its pro-inflammatory response and regulated CD44 expression, which was decreased in T effector cells and increased in Tregs. The alterations in the T cell subpopulations were associated with a reduced mononuclear infiltration (CD4 and CD11b cells) of the melatonin-treated mice CNS. For the first time, we report that melatonin protects against EAE by controlling peripheral and central T effector/regulatory responses, effects that might be partially mediated by CD44. This immunomodulatory effect on EAE suggests that melatonin may represent an effective treatment option for MS.

To assess whether oxidative damage in some tissues was related to their melatonin concentration, endogenous melatonin levels and the age-linked protein and lipid damage in spleen, thymus and liver in 5-month-old SAM P8 mice were examined. The results show that high levels of melatonin in spleen and thymus correlate with lower protein and lipid damage. The liver, which had much lower melatonin concentrations than the other two tissues, had much higher levels of oxidatively damaged protein, as measured by carbonyl values. These results add new evidence concerning the protective role of endogenous melatonin as an antioxidant agent, and suggest that a treatment with this molecule might help to reduce age-associated functional deficits in many organs, including those of the immune system.

To gain insight into the relationship between thymus and pineal gland during rat development, the melatonin content as well as the activity and expression of the two key enzymes for melatonin biosynthesis, i.e. N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT), were studied in the thymus at fetal and postnatal stages. Moreover, melatonin-membrane receptor (MT1) expression was also analyzed. We found both the expression and activity of thymic NAT and HIOMT at 18 days of fetal life. Additionally, there is production of melatonin in the thymus as well as MT1 expression at this fetal age. These results show values higher in day-time than at night-time. The pineal gland begins to produce significant levels of melatonin around postnatal day 16, and this synthesis shows a circadian rhythm with high values during the dark period; therefore the nocturnal serum melatonin may inhibit thymic melatonin production. To document this, we report an increased melatonin content of the thymus in pinealectomized rats compared with sham-pinealectomized. In conclusion, these results show, for the first time, the presence of the biosynthetic machinery of melatonin and melatonin production in developing rat thymus and that the pineal gland may regulate this process.