Evaluación de la calidad ovocitaria y del potencial de desarrollo de ovocitos humanos inmaduros vitrificados antes o después de su maduración in vitro

En la actualidad una de las estrategias empleadas para la preservación de la fertilidad, es la obtención de ovocitos PI de ciclos naturales con el fin de madurarlos in vitro (MIV) y posteriormente vitrificarlos hasta su transferencia. En este trabajo se propone invertir el orden de las dos técnicas empleadas, y valorar si mejoran las tasas de supervivencia (TSu), maduración (TM), activación partenogenota (TA) y desarrollo hasta blastocisto (TD). También, se pretende examinar si la configuración de la placa metafásica del ovocito MII madurado in vitro, se ve afectada por realizar su vitrificación en estadio inmaduro (PI) o maduro (MII). Además, describir los cambios en la ultraestructura ovocitaria tras desvitrificar PI y MIV hasta MII, en comparación con los PI MIV sin preservación. Para ello se incluyeron 1113 ovocitos procedentes de ciclos estimulados para FIV/ICSI, rechazando 28 ovocitos que presentaban vacuolas o signos de degeneración en su citoplasma. El resto de ovocitos fueron aleatoriamente distribuidos en los dos grupos de estudio [G1 (497): VITRI + MIV y G2 (543): MIV + VITRI] y en un grupo control [G0 (45): MIV]. Todo ello se llevó a cabo en dos fases, en la primera se empleó como medio de MIV un medio de cultivo para gametos, mientras que en la segunda fase se utilizó un medio de cultivo embrionario hasta blastocisto, suplementado con SSS y hMG. Todos los medios y componentes utilizados en el medio de MIV estaban comercializados, siendo altamente reproducibles. En la primera fase se establecieron las condiciones de trabajo para la segunda fase, optimizando el protocolo de vitrificación, activación partenogenota y desarrollo embrionario hasta blastocisto. En la segunda fase se incluyó un grupo control y se realizó técnicas de: inmunocitoquímica (TCN) al G1 y G2 y microscopía electrónica de transmisión al G1 y G0. Cada técnica se realizó de forma individualizada para poder mantener la trazabilidad ovocitaria y comparar las tasas generadas de cada uno de los procedimientos con las 17 variables clínicas y de laboratorio recogidas. Se obtuvieron TA, TD y TCN comparables en los dos grupos de estudio. Sin embargo, la TSu y TM fue mejor en el G1 vs G2. El patrón ultraestructural de ovocitos humanos MIV fue similar entre los ovocitos vitrificados en PI y no vitrificados, salvo las microvellosidades y gránulos corticales que modificaron su número, distribución y electrodensidad (G1 vs G0). Estos resultados muestran que: (i) la vitrificación es una técnica útil para la criopreservación tanto de ovocitos maduros (MII) como inmaduros (PI), (ii) el medio de cultivo embrionario hasta blastocisto suplementado adecuadamente puede ser una alternativa válida para sustituir a los medios de MIV comerciales, (iii) la vitrificación puede tener un efecto activador del reinicio de la meiosis, pero se desconoce cuál es el mecanismo de acción, (iv) la configuración de la placa metafísica no presentó diferencias significativas al comparar los ovocitos MIV previa o posteriormente a su vitrificación (G1 vs G2), (v) el TEM reveló que la vitrificación determinaba un patrón alterado de microvellosidades y gránulos corticales, y (vi) el estudio individual de cada una de las variables clínicas y de laboratorio nos permitió observar: la influencia de los días de estímulo en la TSu, el protocolo de supresión hipofisiaria en la TA y la fecundación/estradiol en la TDE. Por todo ello, este trabajo de tesis valida la estrategia de vitrificar ovocitos PI y posteriormente MIV.Nowadays, one of the strategies used to preserve fertility is the obtaining of PI oocytes from natural cycles so as to mature them in vitro (IVM) and then vitrify them until their transference. This work intends to reverse the order of the two techniques used and assess if the rates of survival (SuR), maturation (MR), parthenogenetic activation (AR) and development until blastocyst (DR) improve. In addition, it aims to study whether the metaphase plate configuration of the MII oocytes matured in vitro is affected by the use of vitrification at its immature (PI) or mature (MII) stage. Also, this work pretends to describe changes in the oocyte ultrastrucure after devitrification of PI and IVM to MII, compared to PI IVM without preservation. For this purpose, 1113 oocytes from stimulated cycles of IVF / ICSI were used, rejecting 28 oocytes that showed vacuoles or sings of degeneration in the cytoplasm. The oocytes were randomly distributed in the two groups of study [G1 (497): VITRI + IVM and G2 (543): IVM + VITRI] and one control group [G0 (45): IVM]. All this was carried out in two phases; in the first one, a gametes culture media was used as the media for IVM, while in the second phase, embryonic culture media supplemented with SSS and hMG was used until the blastocyst stage. All media and components used in the IVM media are commercialized, being highly reproducible. In the first phase, the working conditions for the second phase were established, optimizing the vitrification protocol, the parthenogenetic activation and the embryonic development until blastocyst stage. In the second phase, a control group was included and the following techniques were carried out: immunocytochemistry (TCN) to G1 and G2 groups and transmission electron microscopy to G1 and G0 groups. Each technique was performed individually in order to maintain the oocyte traceability and to compare the generated rates of each of the procedures with the 17 clinic and lab variables collected. AR, DR and CNR comparable in the two study groups were obtained. However, the SuR and RT was better in the G1 vs G2. The ultrastructural pattern of human IVM oocytes was similar between the oocytes vitrified at PI stage and no vitrified oocytes, except from microvilli and cortical granules that changed their number, distribution and electrodensity (G1 vs G0). These results show that: (i) vitrification is a useful technique for the cryopreservation of both mature (MII) and immature (PI) oocytes, (ii) the embryonic culture media adequately supplemented until blastocyst stage may be a valid alternative to replace IVM commercial media, (iii) vitrification can have and activating effect on the restart of meiosis, but the mechanism of action is still unknown, (iv) the configuration of the metaphase plate did not show significant differences when comparing IVM oocytes before or after its vitrification (G1 vs G2), (v) TEM revealed that vitrification determined an altered pattern of microvilli and cortical granules, and (vi) the individual study of each of the clinic and lab variables allowed us to observe: the influence of the stimulus days in the SuR, the protocol of hypophyseal suppression in the AR and the fertilization/estradiol in the DET. Therefore, this thesis validates the strategy of vitrification of PI oocytes and its further IVM.1 yea

Estudio de utilización de isomaltulosa en el desarrollo de productos untables de fresa de bajo indice glicémico

Cada vez es más habitual la demanda de alimentos que no provoquen ciertos efectos indeseables relacionados, por ejemplo con el consumo de azúcar, como pueden ser el desarrollo de caries o la diabetes. Junto a esto, el hecho de que la población sepa apreciar los continuos avances en la mejora de la calidad organoléptica y su contenido nutricional, lleva a la industria agroalimentaria al desarrollo de nuevos productos que satisfagan todas las expectativas del consumidor. La existencia en el mercado de nuevos carbohidratos funcionales que pueden sustituir la sacarosa permite desarrollar y/o reformular productos que cumplan dichas expectativas. La deshidratación osmótica tradicional o por vía húmeda (DOH) es una técnica ampliamente extendida en el procesado de frutas y verduras debido a la elevada calidad de los productos obtenidos. Sin embargo, existen una serie de desventajas relacionadas con el manejo de grandes volúmenes de las disoluciones, que deberían ser tenidas en cuenta. La deshidratación osmótica por vía seca (DOS) podría ser una alternativa, ya que el volumen de la disolución generada es considerablemente menor y además, ésta es más rica en compuestos aromáticos e hidrosolubles (vitaminas y minerales) que provienen de la propia fruta. El trabajo realizado en la siguiente tesis docotral ha consistido en la optimización del proceso de formulación de un producto untable de fresa mediante deshidratación osmótica sustituyendo además, parcial o totalmente, la sacarosa por otros azúcares más saludables. Para ello se estudió la influencia de diferentes variables de proceso (tipo de azúcar, tipo de deshidratación osmótica, % de pectina, % de ácido cítrico) sobre la cinética de transferencia de materia y, sobre diferentes parámetros relacionados con la calidad funcionalidad (antocianinas y actividad antioxidante) y sensorial (color, textura, reología y perfil aromático) en los untables de fresa formulados. Los resultados obtenidos permiten afirmar que la cinética de transferencia de materia es mayor cuando la concentración del medio envolvente es variable, siendo ligeramente superior en los procesos de deshidratación por vía húmeda que en los de vía seca.Peinado Pardo, I. (2011). Estudio de utilización de isomaltulosa en el desarrollo de productos untables de fresa de bajo indice glicémico [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/1167

INFLUENCIA DE LAS CONDICIONES DE DESHIDRATACIÓN EN LOS NIVELES DE COMPUESTOS CON ACTIVIDAD ANTIOXIDANTE DEL TOMATE CHERRY

El objetivo de este estudió fue analizar el efecto de las variables del proceso de deshidratación sobre los compuestos con actividad antioxidante del tomate cherryPeinado Pardo, I. (2007). INFLUENCIA DE LAS CONDICIONES DE DESHIDRATACIÓN EN LOS NIVELES DE COMPUESTOS CON ACTIVIDAD ANTIOXIDANTE DEL TOMATE CHERRY. http://hdl.handle.net/10251/1243

La cooperación al desarrollo como parte de la estrucuta económica del capitalismo global

En este artículo se examina la relación entre la Cooperación al Desarrollo y el desarrollo capitalista, prestando especial atención a los cambios en ambos en las últimas décadas. Se argumenta que la Cooperación al Desarrollo forma parte de la propia estructura económica mundial diseñada por el capitalismo en sus diversas fases (inter-nacional y después global), por lo que su funcionalidad está sujeta a la propia lógica estructural capitalista. Se examinan, así, las estrechas relaciones entre los modelos de cooperación y las lógicas de reproducción del Sistema Mundial, tanto en su estructura como en su superestructura. Se distinguen una Cooperación Pro-sistema y una Cooperación Antisistema, y se extraen algunas primeras conclusiones sobre los cambios a introducir para conseguir una cooperación verdaderamente transformadora

Lipids digestibility and polyphenols release under in vitro digestion of dark, milk and white chocolate

[EN] This study evaluated the influence of intestinal conditions on lipolysis and polyphenols release and bioaccessibility in dark, milk and white chocolates. Chocolates were in vitro digested under different intestinal conditions of pH (6 and 7), bile concentration (1 and 10 mM) and pancreatic concentrations (1000-3000 LU/g Fat). The lipolysis varied from 300 to 500 mg FFA/g fat in dark chocolate and ranged between 600 and 1000 mg FFA/g fat in both, milk and white. Polyphenols release in dark chocolate (upto 12 mg GA/g), seems to be related to the absence of dairy compounds. Finally, no effect of intestinal pH or biliary concentration was found on the lipolysis in digested dark and milk chocolates. The oral pancreatic supplementation, however, was crucial to lipolysis and polyphenols release in all chocolates, even if no differences were found on these parameters from 2000 LU/g fat.Authors of this paper, on behalf of MyCyFAPP consortium, acknowledge the European Union and the Horizon 2020 Research and Innovation Framework Programme for funding the above-mentioned project under grant agreement number 643806. The authors would like to thank the Secretaria de Educacion Superior, Ciencia, Tecnologia e Innovacion (Ecuador) for the PhD scholarship given to Carolina Alicia Paz Yepez.Paz-Yépez, C.; Peinado, I.; Heredia Gutiérrez, AB.; Andrés Grau, AM. (2019). Lipids digestibility and polyphenols release under in vitro digestion of dark, milk and white chocolate. Journal of Functional Foods. 52:196-203. https://doi.org/10.1016/j.jff.2018.10.028S1962035

Dietary acrylamide: What happens during digestion

[EN] Acrylamide is a well-known potentially carcinogen compound formed during thermal processing as an intermediate of Maillard reactions. Three objectives were addressed: the impact of gastric digestion on acrylamide content of French Fries, chips, chicken nuggets, onions rings, breakfast cereals, biscuits, crackers, instant coffee and coffee substitute; the acrylamide content evolution during gastrointestinal digestion of French fries and chips; and the effectiveness of blanching and air-frying on acrylamide mitigation after gastrointestinal digestion. A significant increase (p-value < 0.05) in acrylamide content was observed for most of the products after gastric digestion (maximum registered for sweet biscuits, from 30 +/- 8 to 150 +/- 48 mu g/kg). However, at the end of the intestinal stage, acrylamide values were statistically similar (p-value = 0.132) for French fries and lower than the initial values (before digestion) in potato chips (p-value = 0.027). Finally, the low acrylamide content found in blanched and air-fried samples, remained still lower than for deep fried samples even after gastrointestinal digestion. (C) 2017 Elsevier Ltd. All rights reserved.The authors would like to thank the Universitat Politecnica de Valencia for the PhD scholarship given to Mariola Sansano Tomas.Sansano Tomás, M.; Heredia Gutiérrez, AB.; Peinado Pardo, I.; Andrés Grau, AM. (2017). Dietary acrylamide: What happens during digestion. Food Chemistry. 237:58-64. doi:10.1016/j.foodchem.2017.05.104S586423

Effect of cooking methods and intestinal conditions on lipolysis, proteolysis and xanthophylls bioaccessibility of eggs

[EN] Digestibility of macro and micronutrients depends on the ingested food as well as on gastrointestinal conditions, being those suboptimal in exocrine pancreatic insufficiency (EPI) patients. Under this scenario, oral enzyme supplementation improves enzymatic hydrolysis of nutrients. In this study, a static in vitro model was used to assess lipids and protein digestibility as well as lutein and zeaxanthin bioaccessibility of eggs cooked differently and submitted to different intestinal conditions. Boiled, poached and omelette eggs were digested under different intestinal conditions of pH (6 or 7), bile concentration (1 or 10 mM) and doses of the enzyme supplement (1000-4000 LU/g fat). Results showed that poaching resulted in higher digestibility of lipids and proteins, compared to boiling or omelette preparations, under gastrointestinal conditions of EPI (pH 6, bile 1 mM). Concerning xanthophylls bioaccessibility, boiling and poaching led to higher bioaccessibility of lutein and zeaxanthin than omelette under EPI conditions.Authors of this paper, on behalf of MyCyFAPP consortium, acknowledge the European Union and the Horizon 2020 Research and Innovation Framework Programme for funding the above-mentioned project under grant agreement number 643806. The authors would like to thank the Conselleria de Educacio i Investigacio de la Generalitat Valenciana for the PhD scholarship given to Andrea Asensio Grau.Asensio-Grau, A.; Peinado Pardo, I.; Heredia Gutiérrez, AB.; Andrés Grau, AM. (2018). Effect of cooking methods and intestinal conditions on lipolysis, proteolysis and xanthophylls bioaccessibility of eggs. Journal of Functional Foods. 46:579-586. https://doi.org/10.1016/j.jff.2018.05.025S5795864

Could secondary flows make possible the cross-strait transport of passive floating organisms in the Strait of Gibraltar?

The Gibraltar Strait suffers an unprecedented invasion of the alien alga Rugulopteryx okamurae of North Pacific origin. Seemingly, algae first settled in the south shore around year 2015, probably following commercial exchanges with French ports, but there is no certainty that algae first colonized the south shore and then spread to the north one. The opposite could well have happened. Whatever the case, it spread with amazing rapidity over the whole area. Human-mediated vectors (algae attached to ship hulls or fishing nets) can be behind the spread from the shore initially settled to the opposite one. But secondary cross-strait flows within frictional Ekman boundary layers associated to the large along-strait velocity typical of this region could also have propitiated the connection without human intervention. Historical currentmeter profiles collected in the Strait show an intermediate layer of north-going cross-strait velocity near the interface of the mean baroclinic exchange, and an overlying surface layer of southward velocity, whose lower part overlaps the interfacial zone. The first one would facilitate south-to-north transport of algal fragments (or any other neutrally buoyant material) able to settle near the interface depth, while the second one would do the opposite. Cross-strait currents at this depth are of few cm/s, which implies crossing times of several days in low-illuminated conditions. Living organisms must be able to overcome these demanding conditions of darkness and maintain good photosynthetic activity after such period for a successful colonization. Rugulopteryx okamurae can do it.Universidad de Málaga. Campus de Excelencia Internacional del Mar CEIMAR

Influence of particle size and intestinal conditions on in vitro lipid and protein digestibility of walnuts and peanuts

Authors of this paper, on behalf of MyCyFAPP consortium, acknowledge the European Union and the Horizon 2020 Research and Innovation Framework Programme for funding the above-mentioned project under grant agreement number 643806. The authors would like to thank the Secretaria de Educacion Superior, Ciencia, Tecnologia e Innovation (Ecuador) for the PhD scholarship given to Carolina Alicia Paz Yepez.Paz-Yépez, CA.; Peinado Pardo, I.; Heredia Gutiérrez, AB.; Andrés Grau, AM. (2019). Influence of particle size and intestinal conditions on in vitro lipid and protein digestibility of walnuts and peanuts. Food Research International. 119:951-959. https://doi.org/10.1016/j.foodres.2018.11.014S95195911

Efficacy of bortezomib in sarcomas with high levels of MAP17 (PDZK1IP1

Sarcomas are malignant tumors accounting for a high percentage of cancer morbidity and mortality in children and young adults. Surgery and radiation therapy are the accepted treatments for most sarcomas; however, patients with metastatic disease are treated with systemic chemotherapy. Many tumors display marginal levels of chemoresponsiveness, and new treatment approaches are needed. MAP17 is a small non-glycosylated membrane protein overexpressed in carcinomas. The levels of MAP17 could be used as a prognostic marker to predict the response to bortezomib in hematological malignancies and in breast tumors. Therefore, we analyzed the expression of this oncogene in sarcomas and its relationship with clinico-pathological features, as well as tested whether it can be used as a new biomarker to predict the therapeutic response to bortezomib and new therapies for sarcomas. We found that the levels of MAP17 were related to clinical features and poor survival in a cohort of 69 patients with different sarcoma types, not being restricted to any special subtype of tumor. MAP17 expression is associated with poor overall survival (p<0.001) and worse disease-free survival (p=0.002). Cell lines with high levels of MAP17 show a better response to bortezomib in vitro. Furthermore, patient-derived xenografts (PDX) with high levels of MAP17 respond to bortezomib in vivo. Our results showed that this response is due to the lower levels of NFκB and autophagy activation. Therefore, we suggest that MAP17 is a new biomarker to predict the efficacy of bortezomib as a new therapy for sarcomas.España, Consejeria de Ciencia e innovación CTS-6844 and CTS-1848Andalucia, Consejería de Salud PI-0029-2013Andalucia, Consejería de Salud PI-0096-2014Andalucia, Consejería de Salud PI-0306-201