Bioaccesibilidad y efectos biológicos de esteroles vegetales y sus óxidos en una bebida láctea enriquecida en esteroles vegetales

El colesterol ha sido asociado directamente como factor clave en las enfermedades de riesgo cardiovascular. El efecto de los esteroles vegetales (EV) en la reducción del colesterol (10-15%) es ampliamente conocido, si bien, la ingesta necesaria (2-3 g/día) para este efecto debe ser superior a la dietética. Por ello, los EV se utilizan en el enriquecimiento de alimentos, siendo la margarina el primer alimento enriquecido. Este tipo de alimentos es contrario a las recomendaciones dietéticas actuales orientadas hacia un menor consumo de grasa, por ello actualmente se han enriquecido otros alimentos con mejor perfil nutricional (leche, zumo con leche, yogur…). Los EV pueden oxidarse dando lugar a productos de oxidación (POPs). Los estudios en los que se evalúan los efectos biológicos relativos a los POPs son escasos y, en muchos casos, contradictorios. Debido a la similitud estructural, se atribuyen los efectos causados por los productos de oxidación del colesterol (COPs) a los POPs, los cuales se han relacionado con una serie de patologías relacionadas con la modulación del metabolismo lipídico y la disfunción celular. El objetivo de la presente tesis doctoral es el estudio de los EV y POPs en bebidas a base de zumo de frutas y/o leche enriquecidas en EV, mediante la selección de un ingrediente fuente de EV, el estudio de estabilidad de los EV y la bioaccesibilidad (BA) de los EV y POPs presentes en dichas bebidas utilizadas en el marco de un estudio de intervención clínica en humanos. Paralelamente se han evaluado los efectos biológicos de 7-cetoderivados de estigmasterol y colesterol en cultivos celulares del epitelio intestinal (Caco-2) como primera barrera fisiológica para la absorción de estos compuestos. El estudio de los efectos biológicos incluye la determinación de la citotoxicidad causada por los 7-cetoesteroles (evaluando la actividad lisosomal, la funcionalidad e integridad mitocondrial y el contenido de ácidos nucleicos), el potencial inflamatorio (mediante la expresión relativa (ARNm) de biomarcadores (NFκB, TNFα, receptor IL-1β) la cuantificación de citoquinas (TNFα, IL-1β, IL-8, IL-10) y su efecto sobre la modulación del metabolismo del colesterol (monitorización de transportadores (NPC1L1, ABCG5/8) y enzimas (HMGCoA, ACAT)). Se optimiza (energías de colisión) la identificación de POPs por cromatografía de gases acoplada a espectrometría de masas (GC-MS) para la posterior detección de estos compuestos en las bebidas utilizadas en la presente tesis doctoral. De los dos ingredientes ensayados (tall oil y aceites vegetales), se selecciona el tall oil. Este ingrediente permite una mejor cuantificación (disminución de la formación de emulsiones y mejor reproducibilidad en ambas matrices (zumo y zumo con leche)) de los EV en las bebidas a las que se ha adicionado. El almacenamiento (24ºC, 4 meses) de las bebidas enriquecidas no produce cambios en el contenido de EV, si bien, existe un incremento significativo del contenido de POPs durante dicho almacenamiento. Los valores de BA indican diferencias en la solubilidad de los EV dependiendo de la longitud de la cadena alquílica, siendo el campesterol el más bioaccesible. En el caso de los POPs, la BA es superior que en el caso de los EV y, de entre ellos, el sitostanotriol es el más bioaccesible. Los resultados relativos a los efectos biológicos causados por los 7-cetoesteroles en células Caco-2 señalan, principalmente, el compartimento mitocondrial como principal diana de su citotoxicidad, siendo el efecto causado por el 7-cetocolesterol superior al del 7-cetoestigmasterol. La presencia conjunta de ambos 7-cetoderivados, indica una interacción negativa, mostrando una reducción del efecto tóxico del derivado del colesterol. No obstante, la expresión de biomarcadores (NFκB, TNFα, receptor IL-1β) y la cuantificación de citoquinas (TNFα, IL-8, IL-10) pone de manifiesto el mayor potencial inflamatorio del 7-cetoestigmasterol. La monitorización de la expresión relativa de transportadores (NPC1L1, ABCG5/8) y enzimas (HMGCoA, ACAT) relacionados con el metabolismo del colesterol, indican que el poder reductor y el ATP están directamente relacionados con el metabolismo de los 7-cetoesteroles, sin embargo, la influencia del calcio intracelular parece ser determinante en las células incubadas con 7-cetoestigmasterol. El estudio preliminar sobre el potencial efecto citotóxico de los 7-cetoderivados sobre células HepG2 indica una mayor susceptibilidad de esta línea celular versus las células Caco-2, hecho de especial relevancia dado que el metabolismo de los compuestos lipídicos tiene lugar en el hígado. En general, los efectos observados indican que pequeños cambios en la estructura química de los productos de oxidación de los esteroles condicionan el efecto fisiológico determinado in vitro.Cholesterol has been associated with diseases related to cardiovascular risk. The cholesterol-lowering effect (10-15%) of plant sterols (PS) is widely known, but the intake required (2-3 g/day) to achieve this effect is higher than the dietetic intake. Plant sterols are therefore used in food enrichment – margarine being the first enriched food of this kind. However, foods of this type are contrary to current dietary recommendations which indicate a lower fat intake. As a result, other foods with a better nutritional profile have been enriched with PS (milk, juice with milk, yogurt, etc.). Plant sterols can be oxidized, forming oxidation products (POPs). Studies on the biological effects of POPs are scarce and sometimes contradictory. Due to their structural similarity, the effects caused by cholesterol oxidation products (COPs) are attributed to POPs, which have been implicated in a number of pathologies related to the modulation of lipid metabolism and cellular dysfunction. The objective of this thesis is the study of PS and POPs in fruit and/or milk-enriched beverages, by selecting an ingredient source of PS, investigating the stability of PS and the bioavailability (BA) of the PS and POPs present in these beverages and used as part of a clinical intervention study in humans. Moreover, the biological effects of 7-ketoderivatives of stigmasterol and cholesterol have been evaluated in intestinal epithelial cell cultures (Caco-2) as a first physiological barrier for the absorption of these compounds. The evaluation of biological effects included the determination of cytotoxicity caused by 7-ketosterols (evaluating lysosomal activity, mitochondrial function and integrity, and nucleic acids content), their inflammatory potential (via the relative expression (mRNA) of biomarkers (NFκB, TNFα, IL-1β receptor) and the quantification of cytokines (TNFα, IL-1β, IL-8, IL-10)), and their effect upon the modulation of cholesterol metabolism (monitoring protein transporters (NPC1L1, ABCG5/8) and enzymes (HMGCoA, ACAT)). The identification of POPs by gas chromatography coupled with mass spectrometry (GC-MS) has been optimized (collision energies) for the subsequent detection of these compounds in the beverages used in this thesis. After evaluating two ingredients for PS enrichment, tall oil has been chosen. This ingredient allows better PS quantification (decreasing formation of emulsions and increasing reproducibility in both matrixes (juice and juice with milk)) in the enriched beverages. The storage (24°C, 4 months) of fortified beverages induces no changes in PS content, though there is a significant increase in POPs content. BA values indicate differences in the solubility of PS depending on the length of the alkyl chain - campesterol being the most bioavailable molecule. In the case of POPs, BA is higher than for PS - the most bioavailable molecule in this case being sitostanotriol. The results of the biological effects caused by 7-ketosterols in Caco-2 cells show the main subcellular target to be the mitochondrial compartment - the effect caused by 7-ketocholesterol being greater than that of 7-ketostigmasterol. The combined presence of both 7-ketoderivatives indicates a negative interaction through reduction of the toxic effect induced by the cholesterol derivative. However, the biomarkers expression (NFκB, TNFα, receptor IL-1β) and the cytokines quantification (TNFα, IL-8, IL-10) attributes the greatest inflammatory potential to 7-ketostigmasterol. The relative expression of transporters (NPC1L1, ABCG5/8) and enzymes (HMGCoA, ACAT) related to cholesterol homeostasis indicates that the reducing power and ATP are directly related to the metabolism of 7-ketosterols, but the influence of intracellular calcium appears to be determinant in cells incubated with 7-ketostigmasterol. The preliminary study on the potential cytotoxic effect of 7-ketoderivatives upon HepG2 cells indicates a greater susceptibility of this cell line versus Caco-2 cells. This is of particular relevance, because lipid metabolism takes place in the liver. In general, the observed effects indicate that small changes in the chemical structure of the sterol oxidation products condition the physiological effects observed in vitro

Carteles Mondo

[ES] Este Trabajo Final de Grado nace del interés sobre los medios de reproducción gráfica y más concretamente sobre aquellos que posibilitaron la aparición del cartel de cine, así como de la influencia del arte en su propia creación configurando nuestra memoria colectiva. Es por ello necesario plantear una propuesta práctica donde utilizando ciertos recursos plásticos (algunos propios de este momento y otros propios del conocimiento adquirido) se pueda reproducir e interpretar de nuevo aquellos carteles que supusieron una aportación importante al mundo del arte, del cine y/o del diseño, en busca de nuevas formas de expresión y comunicación más acordes a la actualidad y por qué no decirlo, a una propuesta artística de carácter personal, más alternativa si cabe, que ayude a rememorar y revisar los planteamientos que los originaron.[EN] This Final Degree Project arises from the interest in the means of graphic reproduction and more specifically in those that made possible the appearance of the cinema poster, as well as the influence of art in its own creation, configuring our collective memory. It is therefore necessary to propose a practical proposal where using certain plastic resources (some of them from this moment and others from acquired knowledge) can be reproduced and interpreted again those posters that supposed an important contribution to the world of art, cinema and / or of design, in search of new forms of expression and communication more in tune with current events and why not say so, to an artistic proposal of a personal nature, more alternative if possible, that helps to recall and revise the ideas that originated them.Alemany Dominguis, L. (2017). Carteles Mondo. Universitat Politècnica de València. http://hdl.handle.net/10251/165139TFG

Advances in testing for human papillomavirus-mediated head and neck cancer

PURPOSE OF REVIEW: New evidence has recently emerged regarding the utility and benefits of dual p16 INKa (p16) and Human papillomavirus (HPV) status testing when determining the diagnosis and prognosis of patients with oropharyngeal cancer.RECENT FINDINGS: HPV RNA polymerase chain reaction (PCR) is the most accurate diagnostic test. The other assays (HPV DNA PCR, HPV DNA/RNA in-situ hybridization (ISH) and p16) applied to formalin fixed tumour tissue have varying but high sensitivities and specificities. Dual p16 and HPV testing identifies discordant (p16+/HPV- or p16-/HPV+) results in 9.2% of cases, who have significantly poorer prognoses than p16+/HPV+, particularly in smokers. The proportion of discordant cases varies by region, and appears to be highest in regions with lowest attributable (p16+/HPV+) fractions. Dual testing improves prognostication for oropharyngeal cancer cases by identifying discordant cases and improving the prognostic power of the Tumour Node Metastasis (TNM) classification, especially in regions with high discordant rates.SUMMARY: Dual testing is essential when considering patients for clinical trials of treatment de-escalation, and may be important when counselling patients on prognosis, especially in regions with high discordant rates and in smokers.</p

The food energy/protein ratio regulates the rat urea cycle but not total nitrogen losses.

Nitrogen balance studies have shown that a portion of the N ingested but not excreted is not accounted for. We compared several diets (standard, high-fat, high-protein, and self-selected cafeteria) to determine how diet-dependent energy sources affect nitrogen handling, i.e., the liver urea cycle. Diet components and rat homogenates were used for nitrogen, lipid, and energy nalyses. Plasma urea and individual amino acids, as well as liver urea cycle enzyme activities, were determined. Despite ample differences in N intake, circulating amino acids remained practically unchanged in contrast to marked changes in plasma urea. The finding of significant correlations between circulating urea and arginine-succinate synthase and lyase activities supported their regulatory role of urea synthesis, the main N excretion pathway. The cycle operation also correlated with the food protein/energy ratio, in contraposition to total nitrogen losses and estimated balance essentially independent of dietary energy load. The different regulation mechanisms observed have potentially important nutritional consequences, hinting at nitrogen disposal mechanisms able to eliminate excess nitrogen under conditions of high availability of both energy and proteins. Their operation reduces urea synthesis to allow for a safe (albeit unknown) mechanism of N/energy excess accommodation

Unconnected body accrual of dietary lipid and protein in rats fed diets with different lipid and protein content

Scope: Eating large amounts of fat is usually associated with fat accumulation. However, different types of diets (not only lipids) elicit different metabolic responses. Methods and results: Male and female rats (10 week-old) are distributed in four groups and fed for 1 month a standard diet (SD), or this diet enriched with either lipid (high-fat diet, HF) or protein (high-protein diet, HP), or a cafeteria diet (CAF). Both HF and CAF diets share the percentage of energy from lipids (40%) but these are different. Protein-derived energy in the HP diet is also 40%. Feeding SD, HF, and HP diets does not result in differences in energy intake, energy expenditure, total body weight, or lipid content. However, the CAF-fed groups show increases in these parameters, which are more marked in the male rats. The CAF diet increases the mass of adipose tissue while the HF diet does not. Conclusion: Different diets produce substantial changes in the fate of ingested nutrient energy. Dietary lipids are not essential for sustaining an increase in body lipid (or adipose tissue) content. Body protein accrual is unrelated to dietary lipids and overall energy intake. Both protein and lipid accrual are more efficient in male rats

Cervical cancer screening programmes and age-specific coverage estimates for 202 countries and territories worldwide: a review and synthetic analysis

Q1Q1Background: Cervical cancer screening coverage is a key monitoring indicator of the WHO cervical cancer elimination plan. We present global, regional, and national cervical screening coverage estimates against the backdrop of the 70% coverage target set by WHO. Methods: In this review and synthetic analysis, we searched scientific literature, government websites, and official documentation to identify official national recommendations and coverage data for cervical cancer screening for the 194 WHO member states and eight associated countries and territories published from database inception until Oct 30, 2020, supplemented with a formal WHO country consultation from Nov 27, 2020, to Feb 12, 2021. We extracted data on the year of introduction of recommendations, the existence of individual invitation to participate, financing of screening tests, primary screening and triage tests used, recommended ages and screening intervals, use of selfsampling, and use of screen-and-treat approaches. We also collected coverage data, either administrative or surveybased, as disaggregated as possible by age and for any available screening interval. According to data completeness and representativeness, different statistical models were developed to produce national age-specific coverages by screening interval, which were transformed into single-age datapoints. Missing data were imputed. Estimates were applied to the 2019 population and aggregated by region and income level. Findings: We identified recommendations for cervical screening in 139 (69%) of 202 countries and territories. Cytology was the primary screening test in 109 (78%) of 139 countries. 48 (35%) of 139 countries recommended primary HPV-based screening. Visual inspection with acetic acid was the most recommended test in resource-limited settings. Estimated worldwide coverage in women aged 30–49 years in 2019 was 15% in the previous year, 28% in the previous 3 years, and 32% in the previous 5 years, and 36% ever in lifetime. An estimated 1·6 billion (67%) of 2·3 billion women aged 20–70 years, including 662 million (64%) of 1·0 billion women aged 30–49 years, had never been screened for cervical cancer. 133 million (84%) of 158 million women aged 30–49 years living in high-income countries had been screened ever in lifetime, compared with 194 million (48%) of 404 million women in upper-middle-income countries, 34 million (9%) of 397 million women in lower-middle-income countries, and 8 million (11%) of 74 million in low-income countries. Interpretation: Two in three women aged 30–49 years have never been screened for cervical cancer. Roll-out of screening is very low in low-income and middle-income countries, where the burden of disease is highest. The priority of the WHO elimination campaign should be to increase both screening coverage and treatment of detected lesions; however, expanding the efforts of surveillance systems in both coverage and quality control are major challenges to achieving the WHO elimination target. Funding: Instituto de Salud Carlos III, European Regional Development Fund, Secretariat for Universities and Research of the Department of Business and Knowledge of the Government of Catalonia, and Horizon 2020.https://orcid.org/0000-0001-7187-9946Revista Internacional - IndexadaA1N

Might Oral Human Papillomavirus (HPV) Infection in Healthy Individuals Explain Differences in HPV-Attributable Fractions in Oropharyngeal Cancer? A Systematic Review and Meta-analysis

Background. Differences in oral human papillomavirus (HPV) prevalence and contrasts in HPV-attributable fractions (AFs) in oropharyngeal cancer (OPC) have not been evaluated in depth. Methods. A systematic review was performed to identify studies in which at least 50 healthy individuals were tested for oral HPV infection. Information on sex, age, tobacco/alcohol consumption, sex practices, specimen collection, HPV detection, and population type was extracted. Prevalences were pooled using random-effects models for meta-analyses of binomial data. Correlations were assessed by the Spearman test. Results. Forty-eight reports comprising 28 544 individuals fulfilled inclusion criteria. Global oral HPV prevalence was 4.9%. Estimates were highest in Europe, although regional differences were not statistically significant. HPV16 prevalence was 1.0% globally, and regional differences became statistically significant. A lifetime history of >6 sex partners showed a higher risk of oral HPV infection. The age-specific HPV distribution revealed a prevalence of >= 5% over 40 years of age and a lower prevalence at younger ages. There was no association between oral HPV prevalence and HPV-AFs or age-standardized rates (ASRs) of OPC, genital HPV in healthy women, or tobacco use. Conclusions. Differences in HPV-AFs or ASRs of OPC cannot be explained by differences in the prevalence of oral HPV infection across healthy populations. Consistent research on determinants of oral HPV prevalence, acquisition, clearance, and persistence is warranted

Potential impact of a nine-valent vaccine in human papillomavirus related cervical disease

Background: Information on human papillomavirus (HPV) type distribution is necessary to evaluate the potential impact of current and future HPV vaccines. We estimated the relative contribution (RC) to invasive cervical cancer (ICC) and precancerous cervical lesions of the nine HPV types (HPV 6/11/16/18/31/33/45/52/58) included in an HPV vaccine currently under development. Methods: Estimations on ICC were based on an international study of 8,977 HPV positive cases and estimations on precancerous cervical lesions were extracted from a published meta-analysis including 115,789 HPV positive women. Globocan 2008 and 2010 World Population Prospects were used to estimate current and future projections of new ICC cases. Results: RC of the 9 HPV types in ICC was 89.4%, with 18.5% of cases positive for HPV 31/33/45/52/58. Regional variations were observed. RCs varied by histology, ranging between 89.1% in squamous cell carcinomas (SCC) and 95.5% in adenocarcinomas (ADC). HPV 16/18/45 were detected in 94.2% of ADC. RC of the 9 types altogether decreased with age (trend test p < 0.0001), driven by the decrease in older ages of HPV 16/18/45. In contrast, the RC of HPV 31/33/52/58 increased with age. Due to population growth alone, projected estimates of ICC cases attributable to the 9 types are expected to rise from 493,770 new cases in 2012 to 560,887 new cases in 2025. The RCs of individual high risk HPV types varied by cytological and histological grades of HPV-positive precancerous cervical lesions, and there was an under representation of HPV 18 and 45 compared to ICC. Conclusions: The addition of HPV 31/33/45/52/58 to HPV types included in current vaccines could prevent almost 90% of ICC cases worldwide. If the nine-valent vaccine achieves the same degree of efficacy than previous vaccines, world incidence rates could be substantially reduced

HPV distribution in cervical cancer in Portugal. A retrospective study from 1928 to 2005

Objectives: To determine human papillomavirus (HPV) types in invasive cervical cancer in Portugal. Methods: Cases diagnosed at the Institute Portugues de Oncologia de Lisboa de Francisco Gentil from the year 1928 to 2005 were selected for HPV DNA detection and genotyping using SPF10/DEIA/LiPA25 system. Results: Of the 1214 samples that were considered appropriate for HPV detection, 714 (58.8%; 95% CI: 56.0-61.6%) were positive for HPV DNA. This detection rate varied being lower in the first 3 decades (31.3%; 50.1%; 46.5%) and higher in the last decades (77.4-95.1%). This difference was due probably to the fixative used in the first three decades. The five most common types identified among HPV positive cases were HPV16 (58.2%), HPV18 (9.2%), HPV33 (6.2%), HPV45 (4.7%) and HPV31 (4.4%). Multiple infections were detected in 2.8% of the cases. HPV16 and 18 accounted for 67.4% of infections. There were no statistically significant changes of these types over the studied period. An increase at patient's age at diagnosis was observed in the last decades (p < 0.001). Conclusion: HPV16 and 18 accounts for almost 70% of cervical cancers in all 9 decades studied and support data that effective vaccination against these 2 types will reduce the cervical burden in Portuguese women

Estimation of the overall burden of cancers, precancerous lesions, and genital warts attributable to 9-valent HPV vaccine types in women and men in Europe

Background: In addition to cervical cancer, human papillomavirus (HPV) is responsible for a significant proportion of cancers and precancerous lesions of the vulva, vagina, anus, penis, head and neck, as well as genital warts. We estimated the annual number of new cases of these diseases attributable to 9-valent HPV vaccine types in women and men in Europe. Methods: The annual number of new cancers of the cervix, vulva, vagina, anus, penis, and selected head and neck sites in the population of the European Medicines Agency territory was estimated based on age-specific incidence rates extracted from Cancer Incidence in 5 Continents, Volume X and Eurostat population data for 2015. The annual number of new cancers attributable to 9-valent HPV vaccine types was estimated by applying the HPV attributable fraction from reference publications based on a large European multicenter study. For non-cervical cancers, HPV attributable fractions were based on oncogenically-active HPV infections only (i.e., detection of HPV DNA and either mRNA and/or p16 positivity). For precancerous lesions of the cervix, vulva, vagina, and anus, and for genital warts, previously published estimations were updated for the 2015 population. Results: The annual number of new cancers attributable to 9-valent HPV vaccine types was estimated at 47,992 (95% bound: 39,785-58,511). Cervical cancer showed the highest burden (31,130 cases), followed by head and neck cancer (6,786 cases), anal cancer (6,137 cases), vulvar cancer (1,466 cases), vaginal cancer (1,360 cases), and penile cancer (1,113 cases). About 81% were estimated to occur in women and 19% in men. The annual number of new precancerous lesions (CIN2+, VIN2/3, VaIN2/3, and AIN2/3) and genital warts attributable to 9-valent HPV vaccine types was estimated at 232,103 to 442,347 and 680,344 to 844,391, respectively. Conclusions: The burden of cancers associated with 9-valent HPV vaccine types in Europe is substantial in both sexes. Head and neck cancers constitute a heavy burden, particularly in men. Overall, about 90% of HPV-related cancers, 80% of precancerous lesions, and 90% of genital warts are expected to be attributable to 9-valent HPV vaccine types each year, demonstrating the important preventive potential of the 9-valent HPV vaccine in Europe