Estudio de antiinflamatorios naturales para el diseño de alimentos de uso específico para la salud

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Química Física Aplicada. Fecha de lectura: 16-12-2013Trabajo realizado en el Instituto de Investigación en Ciencias de la AlimentaciónEn la presente Memoria se recoge el trabajo realizado y los resultados más relevantes obtenidos durante la investigación llevada a cabo para optar al título de Doctor. El objetivo ha sido contribuir al campo del desarrollo de productos de uso específico para la salud, estudiando las propiedades de extractos naturales obtenidos mediante tecnologías avanzadas de aislamiento y concentración. Se han obtenido extractos de diversas especies de plantas labiadas, empleando las tecnologías de fluidos supercríticos y líquidos presurizados, y se ha determinado su actividad antiinflamatoria in vitro. Para ello, se han usado dos modelos de inflamación, uno donde los macrófagos humanos se activan con lipopolisacárido bacteriano y otro con lipoproteínas de baja densidad oxidadas. Los extractos procedentes de todas las especies presentaron actividad antiinflamatoria, ya que han reducido tanto la secreción como la expresión génica de las citoquinas proinflamatorias TNF-α, IL-1β e IL-6. Sin embargo, de todos los extractos ensayados, el que ha presentado una mayor actividad antiinflamatoria ha sido el extracto supercrítico de romero, relacionándose esta actividad con su contenido en ácido carnósico y carnosol principalmente, además de con la presencia de 1,8-cineol y alcanfor. La incorporación de un 5% de etanol durante el proceso de extracción supercrítica de romero, ha incrementado el contenido en ácido carnósico y carnosol del extracto, lo que se traduce en una mayor actividad antiinflamatoria del mismo. La fracción biodisponible de este extracto de romero, determinada in vitro mediante células Caco-2, presenta un 25% del ácido carnósico y un 3% del carnosol presentes en el extracto de partida, tras 12 horas de ensayo. Asimismo, esta fracción biodisponible también posee una importante actividad antiinflamatoria, que no se puede atribuir exclusivamente al ácido carnósico y al carnosol. Para evitar posibles degradaciones del extracto de romero durante el proceso de digestión y con el fin de aumentar su biodisponibilidad, se ha encapsulado en micelas de caseínas. Los resultados han indicado que esta encapsulación protege al ácido carnósico presente en el extracto de su oxidación durante el proceso de digestión y aumenta su biodisponibilidad tras los ensayos de absorción, ya que solo son necesarias 4 horas para detectar ácido carnósico en la fracción biodisponible. Por todo ello, ha quedado demostrado que el extracto de romero supercrítico puede emplearse como un ingrediente funcional, incorporándolo a formulaciones biodisponibles idóneas para obtener alimentos de uso específico para la salud, donde el extracto de romero aporte su elevada actividad antiinflamatoria.This report describes the scientific methodology and the most relevant results obtained during the experimental training period required to obtain the PhD degree. The aim of the work was to investigate the potential development of specific products with beneficial health properties by studying the biological properties of natural extracts obtained using advanced technologies for isolation and concentration of particular compounds. Specific plant extracts from different Lamiaceae sp. species were obtained using supercritical fluid technologies and pressurized solvents. In order to study their in vitro anti-inflammatory activities, two inflammation models were used: in one of them human macrophages activated with bacterial lipopolysaccharides were utilized, the other used oxidized low density lipoproteins. The extracts from all the tested species showed antiinflammatory activity because they reduced both secretion and gene expression of the proinflammatory cytokines TNF-α, IL-1β and IL-6. However, the supercritical extract obtained from rosemary showed the highest anti-inflammatory activity mainly due to its carnosic acid and carnosol content and the presence of 1,8-cineole and camphor. Addition of 5% (v/v) ethanol as co-solvent during the supercritical extraction process, increased carnosic acid and carnosol levels in the obtained rosemary extract resulting in a higher anti-inflammatory activity than extracts obtained without ethanol. The bioavailable fraction of the rosemary extract obtained with ethanol, determined by in vitro Caco-2 transport assays, contained 25% carnosic acid and 3% carnosol of the initially applied extract after 12 h incubation. This bioavailable fraction also showed an important anti-inflammatory activity that might be not exclusively attributed to carnosic acid and carnosol. In order to avoid the extract degradation during digestion and to increase its bioavailability, the rosemary extract was encapsulated in casein micelles. Results indicated that this encapsulation process protected the carnosic acid of the extract from oxidation during the digestion process. Absorption tests confirmed that encapsulation also increased its bioavailability because after only 4 h carnosic acid was detected in the bioavailable fraction. Therefore, it can be concluded that the supercritical rosemary extract can be used as a functional ingredient to be incorporated into bioavailable formulations suitable to design foodstuffs with specific beneficial health properties because of its high antiinflammatory activities

Supercritical sage extracts as anti-inflammatory food ingredients

This is the author’s version of a work that was accepted for publication in Industrial Crops and Products. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Industrial Crops and Products, [54, (2014)] DOI: 10.1016/j.indcrop.2014.01.021The anti-inflammatory capacity of supercritical CO2extracts (S1 and S2) obtained from sage (Salvia offi-cinalis) was evaluated using THP-1 human macrophages activated with human ox-LDL, a specific in vitromodel to determine the anti-inflammatory effect of the extracts in an atherosclerotic environment. Theexpression of pro-inflammatory cytokines, with an important role in the atherogenic process, such asTNF- , IL-1 and IL-6, in presence of different extracts concentrations was evaluated. Results showedthat 30 g/mL of both supercritical extracts (S1 and S2) markedly suppressed the ox-LDL induced pro-duction of TNF- , IL-1 and IL-6, as well as their mRNA expression. Data showed that S1 presented ahigher anti-inflammatory activity than S2.A characterization by GC–MS of sage extracts identified 16 compounds, mainly camphor, borneol and1,8-cineole. These three compounds represented a 62.4% of S1 and a 48.1% of S2. Camphor, borneol and1,8-cineole presented an important anti-inflammatory activity in the proposed model, with a decreasein the release and gene expression of TNF- , IL-1 and IL-6 and an increase in IL-10 expression. Theseresults explained the higher activity found in S1.This study suggested that supercritical sage extracts could be used as food ingredients in the develop-ment of anti-inflammatory/anti-atherogenic products.Financial support from Spanish Ministry of Science and Innovation (CICYT) (Project: IPT-300000-2010-034, Ingredientes Saludables Mediterráneos Innovadores (INNSAMED) And Comunidad Autónoma de Madrid (ALIBIRD, project S-505/AGR-0153). E.A. thanks Spanish Ministry of Education for a FPU grant

Plan de formación docente de jóvenes investigadores pre- y postdoctorales del Departamento de Farmacología y Toxicología.

Los objetivos que se han alcanzado son los siguientes: 1. Teniendo en cuenta los resultados obtenidos durante el curso 2018-19 (además de los resultados obtenidos en el curso 2017-18) los participantes han alcanzado una formación por encima de lo esperado en Farmacología habiendo asistido a una media de 57% del curso en su primer año de participación (cuando lo estipulado en el Plan de Formacion Docente es del 30%). Además, han superado un 42,3% de la materia entre su primer y segundo año con una calificación media de 8,3. 2. Los jóvenes investigadores han realizado una media de 10 horas de prácticas docentes contabilizando aquellas dedicadas a la asistencia a prácticas como oyentes, el ensayo de las prácticas con tutores y la impartición misma de las sesiones de prácticas. El número de horas está muy limitado por el bajo número de horas prácticas en las asignaturas de Farmacología del Dpto. y el elevado número de jóvenes investigadores incorporados al Dpto. 3. Con todo lo anterior, los jóvenes investigadores han alcanzado la formación en competencias docentes y las horas realizadas han sido acreditadas a las respectivas autoridades de sus becas/contratos. Su participación en la docencia práctica les permitirá en el futuro solicitar un certificado de actividades docentes emitido por las autoridades académicas de la Facultad de Medicina que avalaran su experiencia docente en solicitudes de acreditación a las diferentes figuras de profesor ante la ANECA. Además, el Dpto. de Farmacología y Toxicología ha emitido informes detallados de Aptitud Docente en Farmacología reflejando su participación en el Plan de Formación Docente del Dpto. que podrán ser consideradas en solicitudes a puestos docentes en el futuro

Immigrant IBD Patients in Spain Are Younger, Have More Extraintestinal Manifestations and Use More Biologics Than Native Patients

BackgroundPrevious studies comparing immigrant ethnic groups and native patients with IBD have yielded clinical and phenotypic differences. To date, no study has focused on the immigrant IBD population in Spain. MethodsProspective, observational, multicenter study comparing cohorts of IBD patients from ENEIDA-registry who were born outside Spain with a cohort of native patients. ResultsWe included 13,524 patients (1,864 immigrant and 11,660 native). The immigrants were younger (45 +/- 12 vs. 54 +/- 16 years, p < 0.001), had been diagnosed younger (31 +/- 12 vs. 36 +/- 15 years, p < 0.001), and had a shorter disease duration (14 +/- 7 vs. 18 +/- 8 years, p < 0.001) than native patients. Family history of IBD (9 vs. 14%, p < 0.001) and smoking (30 vs. 40%, p < 0.001) were more frequent among native patients. The most prevalent ethnic groups among immigrants were Caucasian (41.5%), followed by Latin American (30.8%), Arab (18.3%), and Asian (6.7%). Extraintestinal manifestations, mainly musculoskeletal affections, were more frequent in immigrants (19 vs. 11%, p < 0.001). Use of biologics, mainly anti-TNF, was greater in immigrants (36 vs. 29%, p < 0.001). The risk of having extraintestinal manifestations [OR: 2.23 (1.92-2.58, p < 0.001)] and using biologics [OR: 1.13 (1.0-1.26, p = 0.042)] was independently associated with immigrant status in the multivariate analyses. ConclusionsCompared with native-born patients, first-generation-immigrant IBD patients in Spain were younger at disease onset and showed an increased risk of having extraintestinal manifestations and using biologics. Our study suggests a featured phenotype of immigrant IBD patients in Spain, and constitutes a new landmark in the epidemiological characterization of immigrant IBD populations in Southern Europe

Effectiveness and Safety of the Sequential Use of a Second and Third Anti-TNF Agent in Patients With Inflammatory Bowel Disease: Results From the Eneida Registry

Background: The effectiveness of the switch to another anti-tumor necrosis factor (anti-TNF) agent is not known. The aim of this study was to analyze the effectiveness and safety of treatment with a second and third anti-TNF drug after intolerance to or failure of a previous anti-TNF agent in inflammatory bowel disease (IBD) patients. Methods: We included patients diagnosed with IBD from the ENEIDA registry who received another anti-TNF after intolerance to or failure of a prior anti-TNF agent. Results: A total of 1122 patients were included. In the short term, remission was achieved in 55% of the patients with the second anti-TNF. The incidence of loss of response was 19% per patient-year with the second anti-TNF. Combination therapy (hazard ratio [HR], 2.4; 95% confidence interval [CI], 1.8-3; P < 0.0001) and ulcerative colitis vs Crohn's disease (HR, 1.6; 95% CI, 1.1-2.1; P = 0.005) were associated with a higher probability of loss of response. Fifteen percent of the patients had adverse events, and 10% had to discontinue the second anti-TNF. Of the 71 patients who received a third anti-TNF, 55% achieved remission. The incidence of loss of response was 22% per patient-year with a third anti-TNF. Adverse events occurred in 7 patients (11%), but only 1 stopped the drug. Conclusions: Approximately half of the patients who received a second anti-TNF achieved remission; nevertheless, a significant proportion of them subsequently lost response. Combination therapy and type of IBD were associated with loss of response. Remission was achieved in almost 50% of patients who received a third anti-TNF; nevertheless, a significant proportion of them subsequently lost response

Risk Factors for COVID-19 in Inflammatory Bowel Disease: A National, ENEIDA-Based Case–Control Study (COVID-19-EII)

(1) Scant information is available concerning the characteristics that may favour the acquisition of COVID-19 in patients with inflammatory bowel disease (IBD). Therefore, the aim of this study was to assess these differences between infected and noninfected patients with IBD. (2) This nationwide case-control study evaluated patients with inflammatory bowel disease with COVID-19 (cases) and without COVID-19 (controls) during the period March-July 2020 included in the ENEIDA of GETECCU. (3) A total of 496 cases and 964 controls from 73 Spanish centres were included. No differences were found in the basal characteristics between cases and controls. Cases had higher comorbidity Charlson scores (24% vs. 19%; p = 0.02) and occupational risk (28% vs. 10.5%; p < 0.0001) more frequently than did controls. Lockdown was the only protective measure against COVID-19 (50% vs. 70%; p < 0.0001). No differences were found in the use of systemic steroids, immunosuppressants or biologics between cases and controls. Cases were more often treated with 5-aminosalicylates (42% vs. 34%; p = 0.003). Having a moderate Charlson score (OR: 2.7; 95%CI: 1.3-5.9), occupational risk (OR: 2.9; 95%CI: 1.8-4.4) and the use of 5-aminosalicylates (OR: 1.7; 95%CI: 1.2-2.5) were factors for COVID-19. The strict lockdown was the only protective factor (OR: 0.1; 95%CI: 0.09-0.2). (4) Comorbidities and occupational exposure are the most relevant factors for COVID-19 in patients with IBD. The risk of COVID-19 seems not to be increased by immunosuppressants or biologics, with a potential effect of 5-aminosalicylates, which should be investigated further and interpreted with caution

Correction : Chaparro et al. Incidence, Clinical Characteristics and Management of Inflammatory Bowel Disease in Spain: Large-Scale Epidemiological Study. J. Clin. Med. 2021, 10, 2885

The authors wish to make the following corrections to this paper [...]

Incidence, Clinical Characteristics and Management of Inflammatory Bowel Disease in Spain : Large-Scale Epidemiological Study

(1) Aims: To assess the incidence of inflammatory bowel disease (IBD) in Spain, to describe the main epidemiological and clinical characteristics at diagnosis and the evolution of the disease, and to explore the use of drug treatments. (2) Methods: Prospective, population-based nationwide registry. Adult patients diagnosed with IBD-Crohn's disease (CD), ulcerative colitis (UC) or IBD unclassified (IBD-U)-during 2017 in Spain were included and were followed-up for 1 year. (3) Results: We identified 3611 incident cases of IBD diagnosed during 2017 in 108 hospitals covering over 22 million inhabitants. The overall incidence (cases/100,000 person-years) was 16 for IBD, 7.5 for CD, 8 for UC, and 0.5 for IBD-U; 53% of patients were male and median age was 43 years (interquartile range = 31-56 years). During a median 12-month follow-up, 34% of patients were treated with systemic steroids, 25% with immunomodulators, 15% with biologics and 5.6% underwent surgery. The percentage of patients under these treatments was significantly higher in CD than UC and IBD-U. Use of systemic steroids and biologics was significantly higher in hospitals with high resources. In total, 28% of patients were hospitalized (35% CD and 22% UC patients, p < 0.01). (4) Conclusion: The incidence of IBD in Spain is rather high and similar to that reported in Northern Europe. IBD patients require substantial therapeutic resources, which are greater in CD and in hospitals with high resources, and much higher than previously reported. One third of patients are hospitalized in the first year after diagnosis and a relevant proportion undergo surgery

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality